Nabil Tahri

Oxidative Stress Markers in Intestinal Mucosa of Tunisian Inflammatory Bowel Disease Patients

Background/Aims: Inflammatory bowel diseases (IBDs), Crohns disease (CrD) and ulcerative colitis (UC), are chronic gastrointestinal inflammatory disorders. The precise etiology of IBD remains unclear, and it is thought that interactions among various factors, including, genetic factors, the host immune system and environmental factors, cause disruption of intestinal homeostasis, leading to dysregulated inflammatory responses of the gut. As inflammation is intimately related to formation of reactive intermediates, including, reactive oxygen species, oxidative stress has been proposed as a mechanism underlying the pathophysiology of IBD. The purpose of this study is to examine the lipid peroxidation, protein oxidation and anti-oxidative profile in Tunisian IBD. Materials and Methods: Malondialdehyde (MDA), conjugated dienes (CD), protein thiol levels, as well as the catalase (CAT) activity were evaluated in intestinal biopsies of 17 patients affected by IBD (12 CrD and 5 UC) and 12 healthy control individuals. Results: Oxidative stress was confirmed in these two types of disease biopsies as compared to controls. MDA and CD levels were significantly increased in both UC and CrD patients’ biopsies as compared to controls’ biopsies (P < 0.001). CAT activity was similar in UC and CrD biopsies’ and was not significantly increased in IBD patients’ biopsies compared with controls’ biopsies (P > 0.05). Anon-significant decrease in thiol (SH) level was observed in both UC and CrD patients’ biopsies compared with controls’ biopsies (P > 0.05). Conclusion: Increased levels of MDA and CD in IBD patients’ biopsies underline the implication of oxidative stress in the physiopathology of IBD.

Expression of COX-2 and E-cadherin in Tunisian patients with colorectal adenocarcinoma.

Cyclo-oxygenase 2 (COX-2) and E-cadherin are promising biomarkers for cancer diagnosis and therapy. The aim of this study was to examine the expression of these two proteins in primary colorectal adenocarcinomas and to investigate their association with clinicopathological characteristics including survival of patients. Immunostaining of E-cadherin and COX-2 was assessed in 70 primary colorectal adenocarcinomas from Tunisian patients. Membranous E-cadherin immunostaining and cytoplasmic COX-2 expression were observed in 74.3% and 68.6% of cases respectively. A significant association was found between COX-2 expression and age at diagnosis (P=0.02), and vessel invasion (P=0.037). The expression of E-cadherin correlated with age at diagnosis (P=0.01), and tumor size (P=0.02). In addition, by multivariate analysis, we revealed a significant association with 1-year disease free survival and a tendency with distant metastasis (P=0.017 and P=0.065 respectively). On the other hand, tumors exhibiting COX-2+/E-cadherin-profile were larger (P=0.006), and in an advanced stage (P=0.001). Survival analysis showed that COX-2 over-expression confers a reduced overall survival rate (Plog rank=0.036) and is an independent factor predictive for prognosis.

Polymorphisms in the IL2RA and IL2RB Genes in Inflammatory Bowel Disease Risk

Associations with different autoimmune diseases of polymorphisms in genes encoding the IL2RA and IL2RB subunits (located in 10p15 and 22q13, respectively), were identified through genome-wide studies. Polymorphisms in these two genes were studied in 107 inflammatory bowel disease (IBD) patients (39 Crohns disease [CD] and 68 ulcerative colitis [UC]) and in 162 ethnically healthy controls from Tunisia (Sfax). Two of the 15 IL2RA single-nucleotide polymorphisms (SNPs) genotyped (rs4749924 and rs706778) were significantly associated with UC (pcorr=0.018 and 0.048, respectively), but no evidence of association with CD was observed. The IL2RA GTCT haplotype was also more frequent in UC patients compared to controls (2.6% vs. 0%; p=0.002). One of the 6 IL2RB SNPs genotyped (rs743776) was significantly associated with CD (pcorr= 0.039), but no evidence of association with UC was observed. No significant association between IL2RB haplotypes was observed among investigated groups. Our study identified markers in the IL2RA and IL2RB genes that are significantly associated with UC and CD, respectively. Our results supporting IL2RA and IL2RB as promising candidate genes for IBD and suggesting a potential role of IL2R in the pathogenesis of IBD, likely involves regulatory T cells.

Inflammatory Bowel Disease: Susceptibility and Disease Heterogeneity Revealed by Human Leukocyte Antigen Genotyping

This study aimed to investigate the association between HLA DR/DQ and inflammatory bowel diseases (IBD) in Tunisian patients and to determine the relationship between HLA DR/DQ alleles with the clinical disease patterns. DNA typing of human leukocyte antigen (HLA) genes was performed in 70 ulcerative colitis (UC) patients, 40 Crohns disease (CD) patients, and 123 healthy controls (HC) using a polymerase chain reaction sequence specific primer technique. Data were analyzed using Cochran-Mantel-Haenszel test and binary logistic regression. Compared with HC, IBD patients showed an increased frequency of the homozygous DRB1*07 genotype. This positive association was maintained when UC and CD were separately compared to HC. In UC patients, DQB1*03:02 was predictive of colonic extension whereas DRB1*13 and DQB1*03:01 were associated limited disease localization (left-sided colitis and proctitis). The DRB1*15 allele increased in patients with extraintestinal manifestations. In CD, female patients showed an increased frequency of DRB1*13, DRB1*15, and DQB1*06 alleles and DRB1*13-DQB1*06 haplotype, whereas a significant increase of DRB1*07, DQB1*02 alleles, and DRB1*07-DQB1*02 haplotype was noted in male patients. These results show a significant association of the homozygous HLA-DRB1*07 genotype with UC and CD and of several HLA DR/DQ alleles and haplotypes with the clinical phenotypes of these diseases in Tunisian patients. Because of limited statistical power, our study findings are subject to further investigation.

The CREM gene is involved in genetic predisposition to inflammatory bowel disease in the Tunisian population

The identification of susceptibility genes for inflammatory bowel disease (IBD) is key to understanding pathogenic mechanisms. Recently, the results of genetic association studies have highlighted many loci that are shared among several autoimmune diseases. We aimed to study the genetic epidemiology of polymorphisms in specific genes previously associated with other autoimmune diseases, namely the CREM, STAT4, STAT5a, Stat5b, and IRF5 genes. Twelve polymorphisms in the CREM, STAT4, STAT5a, Stat5b, and IRF5 genes were genotyped in a cohort of 107 IBD patients (39 Crohns disease [CD] and 68 ulcerative colitis [UC]) and 162 controls from southern Tunisia. One CREM single nucleotide polymorphism (SNP) displayed evidence for genetic association with IBD (p=8.7×10(-4), odds ratio [OR]=2.84 [1.58; 5.09]). One STAT4 SNP (p=0.026; OR=1.65 [1.06; 2.58]) exhibited a marginal association with UC but not with CD. No significant association was observed with the SNPs in STAT5a, IRF5, and STAT5b. These results suggest that common variants of the CREM gene are involved in the genetic component conferring general susceptibility to IBD, whereas STAT4 appears to be more specifically associated with UC. This work provides motivation for studies aiming to replicate these findings in larger populations.

Association of ZAP70 and PTPN6, but Not BANK1 or CLEC2D, with inflammatory bowel disease in the Tunisian population.

Inflammatory bowel diseases (IBDs), consisting of ulcerative colitis (UC) and Crohns disease (CD), are complex disorders with multiple genes contributing to disease pathogenesis. We aimed to identify the associations of genetic variations in the ZAP70, PTPN6, BANK1, and CLEC2D genes encoding for intracellular signaling molecules with IBDs. One hundred seven patients (39 CD and 68 UC) with IBD and 162 healthy control subjects from the Southern Tunisia were recruited. We genotyped 4 single-nucleotide polymorphisms (SNPs) in ZAP70 (rs1020396, rs11686881, rs13420683, and rs17695937), 2 SNPs in PTPN6 (rs7310161 and rs759052), 3 SNPs in BANK1 (rs10516487, rs17266594, and rs3733197), and 1 SNP in CLEC2D (rs3764021). ZAP70 displayed a strong genetic association with CD for rs13420683 [allele C, p=0.003, P(corr)=0.006, odds ratio (OR)=2.25 (1.32; 3.85); genotype CC, p=0.016, P(corr)=0.048, OR=2.57 (1.22; 5.41)]. However, in UC, a weak association with PTPN6 was observed [TT (p=0.01; P(corr)=0.03; OR=2.11 (1.18; 3.76)]. No significant association in the BANK1 and CLEC2D genes was observed. These results suggest the involvement of the ZAP70 and PTPN6 genes in the genetic component conferring a general susceptibility to CD and UC, respectively. This work provides motivation for studies aiming to replicate these findings in larger populations.

Contribution of immunofluorescence to identification and characterization of antineutrophil cytoplasmic antibodies in inflammatory bowel disease

We evaluated the combined use of different fixatives for the identification of atypical perinuclear anti-neutrophil cytoplasmic antibodies (p-ANCA) in patients with inflammatory bowel diseases (IBD) by indirect immunofluorescence (IIF). Sera from 59 ulcerative colitis (UC) and 37 Crohn’s disease (CD) patients, and from 64 healthy controls were studied. The IIF on ethanol-, formalin-, and methanol-fixed neutrophils was used for the detection of ANCA. Enzyme linked immunosorbant assay (ELISA) was performed to identify the antigens recognized by ANCA. ANCAs were present in 35 of 59 (59.3%) UC patients and in 10 of 37 (27.02%) CD patients. Atypical p-ANCA positivity was strongly associated with UC disease (44.1% in UC vs. 8.1% in CD; p = 0.0002). The combined application of different fixatives contributed to make easy the differentiation between typical p-ANCA and atypical p-ANCA. Atypical p-ANCA determination appears to be a useful parameter for the distinction between UC and CD.

Possible gastrointestinal disorders for athletes during Ramadan: an overview

Abstract Ramadan is considered to be the month of the stomach break. It has been reported that Ramadan has some health benefits. In most Muslims countries, there is a huge modification in the diet during this month. These changes could induce some gastrointestinal (GI) disorders (e.g. diarrhea, cramps, fullness, nausea). The aims of the present overview were to present some challenges that could be observed for athletes during the training sessions or competition and present some practical recommendations to avoid GI disorders during Ramadan. Based on previous studies, we could advance that the prevalence of GI disorders will be more pronounced when athletes travel for international competitions during Ramadan. Besides, GI disorders are more frequent for athletes when there is a huge modification in the training load. Dehydration observed during Ramadan is one of the factors that may induce GI disorders. The latter could be exacerbated by sweat loss during training sessions. Carbohydrate is frequently used by athletes to improve performance and is associated with some GI disorders. Therefore, during Ramadan, coaches and athletes should be advised to maintain the same diet and a good hydration as before the fasting month, and avoid air travel when preparing for competitions.

Effect of Ramadan fasting on feelings, dietary intake, rating of perceived exertion and repeated high intensity short-term maximal performance

ABSTRACT The present study was designed to investigate the effect of Ramadan fasting on feelings, dietary intake, rating of perceived exertion (RPE) and repeated high-intensity short-term maximal performance. Thirteen physically active men (age: 21.2 ± 2.9 years, height: 175.6 ± 5.6 cm, body-mass: 72.4 ± 8.6 kg) performed a 5-m shuttle run test (6 × 30-s + 35-s of recovery in-between) during five experimental periods: fifteen days before Ramadan (BR), the first ten days of Ramadan (FR), the last ten days of Ramadan (ER), ten days after Ramadan (AR10) and 20 days after Ramadan (AR20). The study was carried out in Tunisia during the 2016 Ramadan month. During the 5-m shuttle run test, higher distance (HD), total distance (TD) and fatigue index (FI) were recorded. RPE was determined after a 5-min warm-up and after each repetition of the 5-m shuttle run test (the mean RPE score during the test was calculated). Moreover, a feelings scale (FS) was used after the warm-up and after the end of the 5-m shuttle run test. During the five experimental periods, dietary intake was assessed. The results showed that HD, TD and FI during the 5-m shuttle run test were not affected by Ramadan observance (p > 0.05). Likewise, FS scores recorded after the warm-up and the 5-m shuttle run test were not affected by Ramadan fasting (p > 0.05). However, mean RPE scores during the 5-m shuttle run test were significantly lower at ER (4.06 UA), AR10 (3.86 UA) and AR20 (3.71 UA) in comparison to BR (4.51 UA) (p < 0.05). The results showed also that Ramadan fasting has no adverse effect on energy intake, protein (g and %), fat (g and %) and carbohydrate (g). However, the fractional contribution of carbohydrate was significantly higher AR10 than FR (53.1% vs. 45.8%) and ER (53.1% vs. 46.5%) and AR20 than FR (5.92% vs. 45.8%) (p < 0.05). In conclusion, Ramadan fasting has no adverse effect on feelings, dietary intake, and short-term maximal performance. However, the RPE during repeated high intensity short-term maximal exercise was reduced AR20 in comparison to ER. Abbreviations: AR: After Ramadan; AR10: Ten days after Ramadan; AR20: Twenty days after Ramadan; BR: Fifteen days before Ramadan; ER: Last ten days of Ramadan; FI: Fatigue index; FR: First ten days of Ramadan; FS: Feelings scale; HD: Higher distance; PSQI: The Pittsburgh Sleep Quality Index; RPE: Rating of Perceived Exertion Scale; TD: Total distance

HLA-A*31:01 and carbamazepine-induced DRESS syndrom in a sample of North African population

PURPOSE Drug rash with eosinophilia and systemic symptoms (DRESS) is a serious adverse drug reaction. Carbamazepine is the most common causes of this syndrome. The HLA-A*31:01 allele has been shown to be strongly correlated with carbamazepine-induced DRESS syndrome in European, Japanese, Han Chinese and other asian population but not in African populations. So, our purpose is to study there is a correlation between HLA-A*31:01 and carbamazepine-induced DRESS syndrome in africain population? METHODS HLA class I (A and B) typing was performed on 7 subjects with carbamazepine-DRESS syndrome and 25 tolerants controls subjects. DNA typing HLA class I (A) alleles was checked by the polymerase chain reaction amplification Sequence Specific Oligonucleotide Probes (SSO) (reverse-SSO assay). High resolution HLA DNA Kit based on the Luminex technology (One Lambda®) was used according to the manufacturers protocol. RESULTS The HLA-A*31:01 allele, which has a prevalence of 1% in Tunisian population, was significantly associated with DRESS syndrome. It was detected in 57.14% of cases (4/7) and only 4% of controls subjects (1/25). Thus, the carrier frequency of HLA-A*31:01 allele in the cases group was also significantly higher than in the controls group (57, 14% vs 4% P = 0,004). Odds ratio is estimated 32 (OR = 32 [2.6; 389.2]) CONCLUSION: Similarly to other ethnicities, the presence of the HLA-A*31:01 allele was associated with carbamazepine-DRESS syndrome in a sample of North African population. Future study must be conducted on a larger sample in order to confirm these results.