Search

Upalne bolesti crijeva kronične su bolesti nepoznate etiologije i patogeneze u kojih postoji doprinos genetskih čimbenika pojavi bolesti. MDR1/ABCB1 gen zanimljiv je kandidat gen za upalne bolesti crijeva iz nekoliko razloga. Prvo, knock out miševi za mdr1a gen razvijaju kolitis nalik ulceroznom kolitisu. Nadalje, smješten je na 7 kromosomu u regiji povezanoj s pojavom upalnih bolesti crijeva. Konačno, postoje visoke razine ekspresije Pgp-a, proteinskog produkta MDR1 gena koji funkcionira kao o ATP-u ovisna membranska transportna pumpa, na apikalnim površinama crijevnih epitelnih stanica. Nakon inicijalnih istraživanja koja su uputila na povezanost polimorfizama MDR1 gena i upalnih bolesti crijeva, kasnija istraživanja ponudila su oprečne rezultate te je uloga MDR1 u patogenezi upalnih bolesti crijeva nejasna. Dodatno zanimanje za MDR1 pobudila je i činjenica da su glukokortikoidi supstrat Pgp-a, proteinskog produkta MDR1. Spoznaja da pojedini polimorfizmi MDR1 gena utječu na ekspresiju proteina otvorila je mogućnost da različite razine ekspresije mogu utjecati na uspjeh terapije glukokortikoidima. Međutim, rezultati dosadašnjih istraživanja također su proturječni. Stoga su ciljevi ovog istraživanja bili dokazati povezanost između polimorfizama MDR1 gena i upalnih bolesti crijeva te pronaći povezanost između prisutnosti određenog genotipa i pojedinih kliničkih karakteristika bolesti. Nadalje, cilj istraživanja bilo je dokazati povezanost polimorfizama MDR1 gena i učinka terapije glukokortikoidima s krajnjim ciljem identificiranja rizičnih faktora na razini alela odnosno genotipova za terapijski neuspjeh. U istraživanje smo uključili ukupno 308 bolesnika s upalnim bolestima crijeva, 199 s Crohnovom bolesti i 109 s ulceroznim kolitisom, te 120 zdravih kontrola. U svih bolesnika učinjena je genotipizacija za G2677T/A i C3435T polimorfizma metodom RT-PCR. U bolesnika s upalnim bolestima crijeva učinjen je studiozan pregled dosadašnje medicinske dokumentacije te su bolesnici fenotipizirani prema Montrealskoj klasifikaciji. Također, učinjen je i pregled dosadašnjeg načina liječenja posebno u pogledu uzimanja glukokortikoida u tijeku bolesti te su bolesnici grupirani u skupinu s dobrim odgovorom, ovisne ili refraktorne na glukokortikoide. Dokazali smo povezanost polimorfizama MDR1 gena i upalnih bolesti crijeva na razini alela, genotipova i haplotipova dva lokusa. Na temelju naših rezultata povezanost polimorfizama MDR1 gena i upalnih bolesti crijeva je skromna. Također smo dokazali povezanost ispitivanih polimorfizama i fenotipskih karakteristika u bolesnika s Crohnovom bolesti. Nadalje, dokazali smo utjecaj ovih polimorfizama na uspjeh terapije glukokortikoidima. Opaženi utjecaj na ishod terapije je skroman te je nešto izraženiji u bolesnika s ulceroznim kolitisom. Zaključno, polimorfizmi MDR1 gena igraju ulogu u patogenezi upalnih bolesti crijeva. Izuzev učinka na predispoziciju na pojavu bolesti modificiraju i tijek Crohnove bolesti. Također, MDR1 gen ima skroman utjecaj na uspjeh liječenja glukokortikoidima posebno u bolesnika s ulceroznim kolitisom., Inflammatory bowel diseases are chronic diseases of unknown etiology and pathogenesis in which genetic factors contribute to development of disease. MDR1/ABCB1 is an interesting candidate gene in inflammatory bowel disease for several reasons. First, mdr1a knock out mice develop colitis that resembles ulcerative colitis. Second, it is located in the region of chromosome 7 associated with inflammatory bowel disease. Finally, high levels of its protein product Pgp, functioning as an ATP-dependant membrane efflux pump, are found on apical surfaces of intestinal epithelial cells. Initial reports confirmed association of polymorphisms of MDR1 and IBD. However, results of subsequent studies were contradictory making the role of MDR1 gene in IBD pathogenesis and susceptibility unclear. Additional interest in these gene arose from the fact that glucocorticoids are know substrates of Pgp. The fact that certain polymorphisms influence expression levels of Pgp leads to the possibility that different expression levels might influence outcome of glucocorticoid therapy. However, results of current studies are contradictory. Thus, the aims of this research were to investigate the association of MDR1polymorphisms and inflammatory bowel disease and to investigate the association of these polymorphisms with certain clinical characteristics. We also aimed to investigate the association of MDR1 polymorphisms with outcome of glucocorticoid therapy with an attempt to identify aleles or genotypes risk conffering for therapy failure. A total of 310 inflammatory bowel disease patients, 199 Crohn's disease and 109 ulcerative colitis patients, and 120 healthy controls were included in the study. All subjects were genotyped for G2677T/A i C3435T polymorphism using RT-PCR. In IBD patients review of medical records was performed and patients were meticulously phenotyped according to the Montreal classification. Additionaly, data regarding exposure to glucocorticoids and therapy outcome were recorded and patients were categorised as having good response, dependant or refractory to glucocorticoids. We report association of MDR1 gene polymorphisms and inflammatory bowel disease on the level of alleles, genotypes and two locus haplotypes. Based on our findings, the observed association of MDR1 polymorphisms and IBD is modest. We also report an association of investigated polymorphisms and phenotypic characteristics of Crohn's disease patients. Furthermore, we found an association with outcome of glucocorticoid therapy. The observed effect is also modest but is somewhat more prononced in ulcerative colitis patients. In conclusion, MDR1 gene polymorphisms are associated with inflammatory bowel disease. Apart from their effect on disease susceptibility they also exhibit a disease-modifying effect in Crohn's disease. Furthermore, MDR1 gene has a modest impact glucocorticoid therapy outcome that is more pronounced in ulcerative colitis.

Supplemental material, sj-docx-1-tag-10.1177_17562848231174290 for Barriers in inflammatory bowel disease care in Central and Eastern Europe: a region-specific analysis by Michal Prokopič, Georgiana Gilca-Blanariux, Peter Lietava, Anca Trifan, Anna Pietrzak, Agata Ladic, Marko Brinar, Svetlana Turcan, Tamás Molnár, Peter Bánovčin and Milan Lukáš in Therapeutic Advances in Gastroenterology

Mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) is a rare disorder commonly diagnosed in later disease stages when it prominently manifests as malnutrition. We report on a female patient diagnosed with MNGIE at the age of 36. She was severely malnourished due to loss of resorptive surface after several surgical procedures, gastrointestinal dysmotility, and small intestinal bacterial overgrowth. Therefore, early and aggressive total parenteral nutrition was introduced. Although no reports have shown that nutritional support can modify the clinical outcome, this case suggests that adequate nutritional support, particularly parenteral nutrition, supervised by an experienced nutritional team, may prolong the lifespan of patients with MNGIE.

SAŽETAK Hipofosfatemija kao nuspojava parenteralne primjene željeza dobro je opisana u dosadašnjoj literaturi, no jasni protokoli ili preporuke o prevenciji, pravodobnom prepoznavanju i liječenju nisu dostupni. Čimbenici rizika koji dovode do pojave hipofosfatemije kod bolesnika s upalnim bolestima crijeva (IBD) nisu jasno utvrđeni te dodatno otežavaju razumijevanje nastanka hipofosfatemije i dugoročnih komplikacija, a time i potrebne korake u adekvatnom pristupu prevenciji i liječenju. Prikazom serije kliničkih slučajeva četiriju IBD bolesnika želimo pokazati heterogenost kliničke prezentacije hipofosfatemije, utjecaja nutritivnih, upalnih i drugih čimbenika te moguće modalitete liječenja. Također, želimo istaknuti koja je skupina IBD bolesnika izložena dodatnom riziku razvoja ove nuspojave s ciljem pravodobnog dijagnosticiranja i početka liječenja., Hypophosphatemia as a side effect of parenteral iron administration has been well described in the recent literature. However clear protocols or recommendations for prevention, timely recognition and treatment are not available. Different data on risk factors in patients with inflammatory bowel disease (IBD) according to the published papers further complicate the understanding of hypophosphatemia and long-term complications, and thus the necessary steps in an adequate approach to prevention and treatment. By presenting a series of clinical cases of four IBD patients, we want to show the heterogeneity of the clinical presentation of hypophosphatemia, the influence of nutritional, inflammatory and other factors and also possible treatment modalities. Furthermore, we want to point out which group of IBD patients is at additional risk of developing hypophoshatemia, with the aim of timely diagnosis and treatment initiation.

Background Patients with IBD are at higher risk for non- alcoholic fatty liver disease (NAFLD) comparing to general population. Complex pathogenesis of NAFLD in IBD may be related to disease-specific risk factors such as chronic inflammation, steroid exposure, drug induced hepatotoxicity, malnutrition and alteration of gut microbiota, which is emerging as a major factor in the pathogenesis of NAFLD. The goal of the study was to investigate factors associated with NADLF and advanced liver fibrosis (ALF) in patients with CD and UC. Methods This is a retrospective study on IBD patients without extraintestinal manifestations and known liver disease. NAFLD was defined as Hepatic Steatosis Index (HSI) ≥ 36, and ALF was defined as FIB-4 ≥ 2.67. Predictors of NAFLD development were analysed using Kaplan–Meier and Cox regression analyses. Results In this retrospective study, we have included 225 IBD patients ; 72.4% (n = 163) patients with CD and 27.6% (n = 62) patients with UC (median age 41.2 yr, 53.7% males) which were observed for a median of 4.6 years. There were 63.1% (n = 142) patients with normal BMI, 27.6% (n = 62) overweight and 9.3% (n = 21) obese patients. Obese patients had the highest HIS score 43.9 ± 5.9, following with overweight 37.8 ± 5.7 and normal BMI 30 ± 4.3 kg/m2, p < 0.001. During the follow-up obese and overweight patients had higher risk of developing NAFLD comparing to patients with normal BMI (obese HR = 11.1 95% CI 4.3–28.3 and overweight HR = 5.55 95% CI 3.4–9.1, Logrank test p < 0.001) (Figure 1). Regarding FIB-4 score there, was no difference among different BMI categories (p = 0.192), and there was no difference in ALF development in the follow-up period (Logrank test p = 0.91). In Cox proportional-hazards regression significant predictors for NAFLD development were dyslipidaemia HR=2.11, 95% CI 1.2– 3.7, overweight HR=6 95% CI 3.6–10, and obesity HR=13.4, 95% CI 7– 35. graphic Conclusion NAFLD is frequent comorbidity in patients with CD and UC, which can lead to development of advanced liver fibrosis. Our results show that patients with IBD have a high risk of NAFLD development, whereas the increased risk for ALF was not observed. Overweight and obese patients and those with dyslipidemia should be closer monitored due to significantly higher risk of NAFLD. This study points out the complexity disease-specific risk factors and importance of better stratifying IBD patients at risk of NAFLD and advanced liver fibrosis.

ObjectiveThe Epi-IBD cohort is a prospective population-based inception cohort of unselected patients with inflammatory bowel disease from 29 European centres covering a background population of almost 10 million people. The aim of this study was to assess the 5-year outcome and disease course of patients with Crohn’s disease (CD).DesignPatients were followed up prospectively from the time of diagnosis, including collection of their clinical data, demographics, disease activity, medical therapy, surgery, cancers and deaths. Associations between outcomes and multiple covariates were analysed by Cox regression analysis.ResultsIn total, 488 patients were included in the study. During follow-up, 107 (22%) patients received surgery, while 176 (36%) patients were hospitalised because of CD. A total of 49 (14%) patients diagnosed with non-stricturing, non-penetrating disease progressed to either stricturing and/or penetrating disease. These rates did not differ between patients from Western and Eastern Europe. However, significant geographic differences were noted regarding treatment: more patients in Western Europe received biological therapy (33%) and immunomodulators (66%) than did those in Eastern Europe (14% and 54%, respectively, PConclusionDespite patients being treated early and frequently with immunomodulators and biological therapy in Western Europe, 5-year outcomes including surgery and phenotype progression in this cohort were comparable across Western and Eastern Europe. Differences in treatment strategies between Western and Eastern European centres did not affect the disease course. Treatment with immunomodulators reduced the risk of surgery and hospitalisation.

Background: Inflammatory bowel disease (IBD) and its two most common disease subtypes ; Crohn's disease (CD) and ulcerative colitis (UC) are suggested to be related to gut dysbiosis caused by the persistent imbalance of the microbiota. Advances in the next-generation sequencing enable culture-independent analysis of the complex microbial communities and provide insight into gastrointestinal tract and its ecological landscape. The aim of this study was to determine differences in mucosa-associated microbiota composition in adult, newly diagnosed and treatment-naïve IBD patients and control patients with irritable bowel syndrome (IBS). Methods: Mucosal tissue samples were collected from both groups of patients at 6 distinct positions along the gut, as part of diagnostic colonoscopy examination prior to the initiation of treatment, followed by snap-freezing. DNA was extracted using MasterPure DNA purification kit (Epicentre). The composition of gut microbiota from mucosa was determined by amplification and sequencing of bacterial 16S rRNA gene using Illumina MiSeq according to manufacturer- recommended protocols. Raw sequencing files were processed using QIIME 1 pipeline and Operational Taxonomic Units (OTUs) were assigned using the vsearch algorithm and PyNast alignment against the GreenGenes database (version 13_8, May 2013). Subsequent processing and analysis was done using ALDEx2 R (Gloor and Reid. Can. J. Microbiol. 703 ; 2016). Results: The microbiota composition of mucosa sampled at 6 positions (from terminal ileum to rectum) was analyzed at the family level. Contrary to our expectations, no significant position- dependent differences in taxa abundance in individual patients were determined. The major enterotypes (Bacteroidetes- and Prevotella- dominating) were observed among individuals. Samples from inflamed areas of the gut displayed altered abundance of a limited number of families (decrease of Verrucomicrobiaceae and Rikenellaceae, increase of Pasterurellaceae), albeit with modest effect sizes. Alpha-diversity for individuals diagnosed with UC was decreased. Differences in microbiota abundance were most pronounced between two IBD phenotypes – UC and CD with UC having increased abundance of Neisseriaceae, Pasterurellaceae, Peptococcaceae, Peptostreptococcaceae, Turicibacteriaceae, Veillonelaceae, S24-7 and shifts in several of the low abundance families. Decrease in Verrucomicrobiaceae was found in CD. Conclusion: Preliminary results indicate distinct individual gut mucosa bacterial profiles, independent of the sampling position. Microbiota differences between disease phenotypes were found to be more profound than between inflammation status of the colon mucosa.

Aim To analyze the distribution of SLC6A4 gene polymorphisms in Crohn’s disease (CD) patients and their association with the disease. Methods We evaluated the presence/absence of promoter (5- HTTLPR, rs25531) and intron 2 (STin2 VNTR) polymorphic variants of SLC6A4 gene in a retrospective case-control study including 192 CD patients and 157 healthy controls (HC). Genotyping was performed by polymerase chain reaction. The association of polymorphisms with CD and its clinical subtypes was analyzed using χ2 and Fisher exact test, binary logistic regression, and haplotype analysis. Results CD patients and healthy controls had similar sex (88 [45.8%] vs 84 [53.5%] women, respectively ; P = 0.154) and age (41.3 ± 12.8 years vs 41.7 ± 8.8 years, respectively, P = 0.091) distribution. Significant differences were observed in the STin2 genotype and allele distribution between CD patients and healthy controls (P = 0.003 and P = 0.002, respectively) and between the corresponding female subgroups (P = 0.004 and P = 0.007, respectively), with a significant negative association of biallelic ss (STin2.9 and Stin2.10) STin2 genotype with CD (P = 0.013, age- and sexadjusted odds ratio [OR] 0.5, 95% confidence interval [CI] 0.29- 0.86 ; women: P = 0.006, age-adjusted OR 0.32, 95% CI 0.14-0.72) and a significantly higher S- STin2.12 (5-HTTLPR/ rs25531: S-STin2: STin2.12) haplotype distribution in CD patients (P = 0.004, OR 1.62, 95% CI 1.16-2.26). There was no significant association between 5-HTTLRP and rs25531 genotype or allele frequencies and CD and between any SLC6A4 polymorphic loci with clinical CD subtypes. Conclusion STin2 VNTR polymorphism of SLC6A4 gene may contribute to CD pathogenesis.

Recent advancements in next-generation sequencing techniques have enabled culture-independent analysis of the gut microbiota, revealing that an altered balance of the gut microbiota constituents (rather than specific pathogens) is involved in the pathogenesis of Crohn’s disease (CD) - one of the principal subsets of inflammatory bowel disease (IBD). Thus far, there were only a handful studies with naïve CD patients on pediatric subjects, and many studies on adult CD patients in late disease stages or post treatment. The aim of our study was to determine differences in microbiota composition in adult, newly diagnosed and treatment-naïve CD and controls with irritable bowel syndrome (IBS) patients. Methods: Mucosal sample (terminal ileum) was collected from both groups of patients as part of diagnostic colonoscopy examination prior to the initiation of treatment and snap-frozen. DNA was extracted using MasterPure DNA purification kit (Epicentre). The composition of gut microbiota from mucosa was determined by amplification and sequencing of bacterial 16S rRNA gene using Illumina MiSeq according to manufacturer-recommended protocols. Raw sequencing files were processed using QIIME pipeline and Operational Taxonomic Units (OTUs) were assigned using the vsearch algorithm and PyNast alignment against the GreenGenes database (version 13_8, May 2013). Subsequent processing and analysis was done using ALDeX2 R package by utilizing multivariate generalized linear model. Results: The difference in abundance of several bacterial taxa was observed between CD and IBS groups. The analyses of effect sizes down to the genus level indicated that taxa belonging to Clostridia, Coriobacteriia and Actinobacteria classes were overrepresented in CD, while Betaproteobacteria were underrepresented, with respect to the IBS group. Conclusions: Preliminary results on limited patient cohort demonstrated differences in mucosa-associated microbiota populations between adult, newly diagnosed, treatment-naïve CD and IBS patients.

Novija istraživanja na široj populaciji upućuju na općeprisutni manjak vitamina D koji je neovisan od izvanjskih čimbenika (npr. mjesto stanovanja, izloženost suncu). Hipovitaminoza vitamina D nastupa vrlo često u stanjima poremećena rada crijeva, kao što je sindrom iritabilnoga crijeva (IBS). Svrha ovoga rada bila je odrediti status vitamina D u serumu 39 bolesnika s dijagnozom IBS- a. Dobiveni rezultati upućuju na manjak vitamina D u serumu većine ispitanika (72%), a uočeno je da ispitanici s višom koncentracijom serumskoga vitamina D pokazuju blaže simptome bolesti. Analizom njihovih demografskih obilježja i navika (prehrana i pušenje) uočeni su zanimljivi trendovi u varijacijama vitamina D, koji pokazuju da ispitanici s BMI

Sažetak. Strano tijelo u probavnom sustavu pripada rjeđim uzrocima crijevne opstrukcije. Većina stranih tijela spontano će proći cijelim probavnim traktom. Endoskopska intervencija bit će potrebna u 10 – 20% bolesnika, a kirurški zahvat u manje od 1% njih. Osobe povišenog rizika od zastoja stranog tijela jesu one s prethodnim kirurškim zahvatom u abdomenu, hernijom, tumorom, Crohnovom bolesti ili prirođenim malformacijama crijeva. U radu smo prikazali bolesnicu s Crohnovom bolesti i stenozama terminalnog ileuma s prestenotičkom dilatacijom, koja je hospitalizirana zbog opstruktivnih smetnja uzrokovanih stranim tijelom u području terminalnog ileuma. Strano tijelo uklonjeno operativnim putem bila je koštica breskve zaglavljena u ileocekalnoj valvuli. Postoperativni tijek protekao je bez komplikacija. Osim najčešćih uzroka opstrukcije tankog crijeva, potrebno je imati na umu i rijetke uzroke opstrukcije i uzeti ih u obzir pri diferencijalnodijagnostičkom razmišljanju, osobito u bolesnika s Crohnovom bolesti, kako bi se pravodobno interveniralo i uklonila mogućnost komplikacija poput perforacije, krvarenja ili razvoja fistula. Summary. Foreign body in digestive tract is a rare cause of intestinal obstruction. Most foreign bodies pass digestive tract spontaneously. Endoscopic intervention will be needed in approximately 10-20%, and surgical intervention in less than 1% of cases. People with increased risk of foreign body obstruction are those with previous abdominal operation, hernia, tumor, Crohn’s disease or congenital intestinal malformations. In this paper we have presented a patient with Crohn’s disease and stenosis of terminal ileum with prestenotic dilatation hospitalized due to bowel obstruction caused by foreign body in terminal ileum. Surgicaly removed foreign body was peach pit stuck in the ileocaecal valve. Except of the most common causes of small bowel obstruction, it is necessary to bear in mind the rare causes of obstruction and consider them in differential diagnosis, especially in patients with Crohn’s disease, due to timely intervention and lowering the possibility of developing complications such as perforation, bleeding or fistula formation.

The intestine represents an interface where host tissues come in contact with microbiota in a balanced state of homeostasis. Mounting knowledge on gut microbiota led to many important findings associating the composition of bacterial taxa in the human gastrointestinal tract with many human disorders including the Inflammatory Bowel Disease (IBD). Ulcerative Colitis (UC) and Crohn's Disease (CD) as the most prevalent forms of IBD are characterized by chronic relapsing inflammation affecting the intestinal mucosa and although the etiology of both diseases is unknown, there is increasing evidence that intestinal microbial dysbiosis has a role in the pathogenesis. One of the main objectives of our study is to investigate the contribution of the faecal and intestinal microbiota composition to the disease specific phenotype. Faecal samples are being collected from adult, newly diagnosed, treatment-naïve IBD patients and controls with Irritable Bowel Syndrome (IBS) using OMNIgene.Gut collection system. The composition of gut microbiota from faeces was determined by amplification and sequencing of bacterial 16S rRNA gene using Illumina MiSeq. Raw sequencing files were processed using QIIME pipeline and Operational Taxonomic Units (OTUs) were assigned using the vsearch algorithm and PyNast alignment against the GreenGenes database (version 13_8, May 2013). Relative abundance proportions were analysed using two-way ANOVA with Bonferroni multiple comparisons test (significant if p

Inflammatory bowel disease (IBD) is a chronic gastrointestinal disorder increasing in incidence and severely influencing affected individuals. Although the factors associated with the disease risk include immune status, gut microbiome and several environmental causes including food intake, the current scientific knowledge of specific triggers and diagnostic markers to improve interventional approaches is still scarce. Additional basic and clinical research involving specific groups of patients and obtaining multilevel data are essential for the management of the disease. Investigation of mutual correlation between gut microbiota, inflammatory, endocrine and nutritional status provides integrative framework for better understanding the IBD pathophysiology. MINUTE for IBD project’s scope is to perform the best possible systematic analysis of relevant elements and factors (gut microbiota composition, inflammatory/endocrine markers, nutritional status) contributing to the development and progress of IBD on newly diagnosed, therapy naïve IBD patients and individuals not fulfilling IBD diagnosis criteria as a control. Through an integrated translational approach, two skilled groups of experts (academic and clinical) are focused on accomplishing the main scope of the project. Analysis of microbiota composition (by 16S rRNA gene sequencing) in stools and in mucosa, quantification of relevant immune and endocrine biomarkers in blood samples, determination of overall nutritional status in study participants and integration of the obtained results and setting-up better guiding principles when deciding on the therapy and nutritional intervention in IBD patients

Inflammatory bowel disease (IBD) is a chronic gastrointestinal disorder with increased incidence. The onset of the disease is mainly associated with the immune status, gut microbiota and food intake. The current scientific knowledge of specific triggers and diagnostic markers is limited ; therefore, new data on gut microbiota, inflammatory, endocrine and nutritional status are required to better understand the IBD pathophysiology and ultimately support stratification of patients and initiation of appropriate therapy. In the translational project (Assessment of Microbiota, Inflammatory Markers, Nutritional and Endocrinological Status in IBD Patients ; Acronym Minute for IBD) we are exploring host-gut microbiota interactions in order to define novel strategies for the management of IBD. Stool, intestine biopsies and blood samples from 40 newly diagnosed IBD patients and 20 non-IBD control individuals are collected and their nutritional status recorded. This work has been fully supported by the Croatian Science Foundation under the project number [5467]. To characterize the composition of the microbiota in collected samples next-generation sequencing (MiSeq, Illumina) is used. As DNA isolation from complex biological samples is a major step in obtaining high quality DNA, as the first step, the utility of commercially available systems (OMNIgene.GUT stool collection system, and DNA extraction kits: MP Biomedicals, QIAGEN, MO BIO) was assessed in healthy volunteers. It has been demonstrated that DNA yield and quality varied between DNA extraction kits (MP Biomedicals>QIAGEN>MO BIO). Although donor-specific bacterial diversity was maintained irrespective of the collection, storage or extraction method used, kit-based trends were observed displaying different levels of abundance among the most prevalent families, notably Bacteroidaceae and Lachnospiraceae. These preliminary results showed that commercially available systems OMNIgene.GUT and MP Biomedicals are convenient and reliable tools for the assessment of human faecal microbiota composition. This clinical research study deals with the cutting-edge, hot topics (microbiota composition) and uses state-of-the art sensitive methodologies (biomarkers determination). It will provide valuable insights that will be applied for better IBD patients’ stratification into specific therapeutic and nutritional interventions.

Methodologies for collection and storage of human faecal samples as well as DNA extraction govern the quality of DNA obtained and present crucial steps in obtaining representative models of bacterial community structure after 16S rRNA gene sequencing. Collection of faeces in the clinical settings necessitates the need for uniform, reliable, robust and patient compliant procedures. In this study we compared two different approaches for faeces collection and storage and three commercially available DNA extraction kits. Faeces from 6 healthy donors were collected fresh and in OMNIgene.GUT system, then stored for 14 days at -20°C and at room temperature, respectively. DNA yield and quality varied between DNA extraction kits in the order MP>QIAGEN>MoBio with OMNIgene.GUT preserving integrity of the obtained DNA. Donor-specific bacterial diversity was maintained irrespective of the collection, storage or extraction method used. However, kit-based trends were observed displaying different levels of abundance among the most prevalent families, notably Bacteroidaceae and Lachnospiraceae. Although OMNIgene.GUT faeces collection system slightly decreased the richness of detected bacteria, the change was not statistically significant. Overall, OMNIgene.GUT was proven as convenient and reliable human faecal collection, transport and storage system for human faecal microbiota analyses.

S obzirom na činjenicu da TNF ima važnu ulogu u patogenezi upalnih bolesti crijeva, u liječenju ulceroznog kolitisa upotrebljavaju se anti-TNF lijekovi. Iako su ovi lijekovi značajno unaprijedili liječenje bolesnika, imunogeničnost ograničava njihove mogućnosti u kliničkoj primjeni. Infliksimab i adalimumab su učinkoviti u postizanju i održavanju remisije u ambulantnih bolesnika s umjereno teškim i teškim ulceroznim kolitisom. Biološki lijekovi mogu biti i lijek izbora za liječenje bolesnika s refraktornim proktitisom i refraktornim pouchitis-om. U hospitaliziranih bolesnika s teškim akutnim kolitisom refraktornih na intravensku primjenu kortikosteroida može se primijeniti infliksimab kao druga linija terapije, ali je adekvatna dugotrajna terapija održavanja s anti-TNF lijekom potrebna za dugoročni učinak. S obzirom na poznate rizike i nuspojave anti-TNF lijekova nužna su buduća istraživanja, osobito o problemu konkomitantne terapije tiopurinskim lijekovima.

Objective: Serotonin (5-HT) is an important factor in gut function, playing key roles in intestinal peristalsis, secretion, vasodilatation and sensory signalling. The serotonin-selective reuptake transporter protein (SERT) terminates the action of 5-HT. Human SERT is encoded by a single gene on chromosome 17q11 ; 2 important polymorphic sites in the SERT gene are: variable number tandem repeats in the gene’s second intron (SERTin2), and an insertion/deletion in the promoter region (SERTPR). Consistent with the effects of 5-HT in the gut, SERT polymorphisms could potentially be involved in the development of different CD phenotypes. The aim of this study was to evaluate the relationship between SERT polymorphisms in CD patients vs. controls. Methods: A total of 193 CD patients (phenotyped in 3 group according to Vienna classification) and 217 control were subjected to genotyping. DNA of all subjects was analysed by polymerase chain reaction (PCR-RT). The association of genetic polymorphic variant SERTPR/rs 25531 and SERTin2 polymorphic loci with the CD patients vs. controls was tested using program SPSS 13.0, UNPHASED ver. 3.0. 10. A test for Hardy – Weinberg equilibrium using Markov chain method implemented in Arlequin ver. 3.0. Chi squared test was used for comparisons of the allele and genotype frequencies among groups ; log likelihood ratio tests were done to compare distributions of the estimated haplotypes among groups. Results: Genotype frequencies of the SERTPR/rs25531 LL, LS and SS in the CD patients (and controls) were 58 (74), 97 (96) and 37 (47), respectively and of the SERTin2 ll, ls and ss genotypes were 76 (77), 91 (92) and 25 (48), respectively. No significant deviations from the expected Hardy– Weinberg proportions were observed in the sample. Pair-wise comparisons of the allele, genotypand haplotype frequencies between CD patients and controls revealed statistical differences for SERT in2 loci ; ss genotype. Conclusion: Polymorphisms of SERT gene could be associated with development of different phenotypes of CD.