ADOA3R as a therapeutic target in experimental colitis: proof by validated high-density oligonucleotide microarray analysis

Adenosine A3 receptors (ADOA3Rs) are emerging as novel purinergic targets for treatment of inflammatory diseases. Our goal was to assess the protective effect of the ADOA3R agonist N(6)-(3-iodobenzyl)-adenosine-5-N-methyluronamide (IB-MECA) on gene dysregulation and injury in a rat chronic model of 2,4,6-trinitrobenzene sulfonic acid (TNBS)--induced colitis. It was necessary to develop and validate a microarray technique for testing the protective effects of purine-based drugs in experimental inflammatory bowel disease. High-density oligonucleotide microarray analysis of gene dysregulation was assessed in colons from normal, TNBS-treated (7 days), and oral IB-MECA-treated rats (1.5 mg/kg b.i.d.) using a rat RNU34 neural GeneChip of 724 genes and SYBR green polymerase chain reaction. Analysis included clinical evaluation, weight loss assessment, and electron paramagnetic resonance imaging/spin-trap monitoring of free radicals. Remarkable colitis-induced gene dysregulation occurs in the most exceptional cluster of 5.4% of the gene pool, revealing 2 modes of colitis-related dysregulation. Downregulation occurs in membrane transporter, mitogen-activated protein (MAP) kinase, and channel genes. Upregulation occurs in chemokine, cytokine/inflammatory, stress, growth factor, intracellular signaling, receptor, heat shock protein, retinoid metabolism, neural, remodeling, and redox-sensitive genes. Oral IB-MECA prevented dysregulation in 92% of these genes, histopathology, gut injury, and weight loss. IB-MECA or adenosine suppressed elevated free radicals in ex vivo inflamed gut. Oral IB-MECA blocked the colitis-induced upregulation (90% of genes tested (33 of 37 genes). We conclude that our validated high-density oligonucleotide microarray analysis is a powerful technique for molecular gene dysregulation studies to assess the beneficial effects of purine-based or other drugs in experimental colitis. ADOA3R is new potential therapeutic target for inflammatory bowel disease.     

The transfer of skills from cognitive and physical training to activities of daily living: a randomised controlled study

Ageing is associated with the deterioration of all cognitive functions, including attention, memory and psychomotor speed. It has not yet been clearly confirmed whether the effects of cognitive and physical interventions can improve activities of daily living (ADL). This study compared the effectiveness of cognitive and physical training on cognitive functions and the transfer to ADL. Eighty older people with mild cognitive impairment (mean age 67.07 ± 4.3 years) were randomly divided into an experimental group (n = 40) and a control group (n = 40). Data were collected in an outpatient psychiatric clinic in a randomised controlled trial. Primary outcome measures included the following: cognitive functions were evaluated using the mini mental state examination, the AVLT—Auditory verbal learning test, the Stroop test, the TMT—trail making test, the DRT—disjunctive reaction time and the NHPT—nine hole peg test. Secondary outcome measure was the Bristol activities of daily living scale. The experimental group underwent a CogniPlus and physical training; consisting of 20 training sessions over 10 weeks. Both groups went through 30 min of daily physical training for 10 weeks. After the training, significant differences in favour of the experimental group were found in almost all the tests. In memory (AVLT) (p ≤ 0.0001, effect size (ES) η ²  = 0.218. In reduction of the response time on attention tasks (Stroop tasks) (p ≤ 0.006, ES = 0.092–0.115). In lower error rates in all tests: Stroop tasks, DRT, TMT, NHPT (p ≤ 0.02–0.001, ES = 0.062–0.176). In ADL (p ≤ 0.0001, ES = 0.176). The combined cognitive and physical training had better efficacy for most cognitive functions and for ADL when compared with the physical training only.     

Clopidogrel IBS Patients Have Higher Incidence of Gastrointestinal Symptoms Influenced by Age and Gender

Background

Clopidogrel is an irreversible antagonist of P2Y₁₂ receptors (P2Y₁₂Rs) used as an antiplatelet drug to reduce risk of thrombosis. P2Y₁₂Rs are expressed in gastrointestinal (GI) tract where they might regulate GI function.

Aim

To evaluate if blockade of P2Y₁₂Rs by clopidogrel is associated with higher incidence of GI symptoms in patients with irritable bowel syndrome (IBS).

Methods

A retrospective analysis of our institutional database was conducted for a 13-year period. IBS patients were identified, and their demographics, GI symptoms and clopidogrel therapy were collected. Logistic regression models were used to characterize symptoms in clopidogrel versus no-clopidogrel IBS-groups, adjusting for Age and Sex differences. An additional study characterized the P2Y₁₂R distribution in human gut.

Results

The search identified 7217 IBS patients (6761 no-clopidogrel/456 clopidogrel). There were a higher proportion of patients with GI symptoms on clopidogrel (68%) compared to controls (60%, p = 0.0011) that were Females (70 vs. 60%, p = 0.0003) not Males (61 vs. 60%; p = 0.8312). In Females, clopidogrel was associated with higher incidence of GI symptoms (Age adjusted; p < 0.0001) for pain, constipation, gastroparesis (p ≤ 0.0001) and psychogenic pain (p = 0.0006). Age or Sex (adjusted models) influenced one or more GI symptoms (i.e., pain, p < 0.0001; constipation, p < 0.0001/p = 0.008; diarrhea, flatulence, p = 0.01). P2Y₁₂R immunoreactivity was abundant in human ENS; glial-to-neuron ratio of P2Y₁₂Rs expressed in Females ? Males.

Conclusions

Irreversible blockade of P2Y₁₂R by clopidogrel is associated with higher incidence of GI symptoms in Female IBS patients, although Age or Sex alone contributes to symptomatology. Prospective studies can determine clinical implications of P2Y₁₂Rs in IBS.

     

Association of F11 polymorphism rs2289252 with deep vein thrombosis and related phenotypes in population of Latvia

Introduction

Deep vein thrombosis (DVT) has a strong inherited predisposition that is partly explained by the strong genetic risk factors such as mutations in factor V, prothrombin, antithrombin III, protein C and S genes. Only recently the first GWAS have been performed on DVT resulting in discovery of novel genetic variants, however, the information on the common polymorphisms predisposing to the risk of DVT is still scarce.

Materials and Methods

Here we selected six SNPs (rs5361 in SELE, rs2066865 in FGG, rs2227589 in SERPINC1, rs1613662 in GP6, rs13146272 in CYP4V2, rs2289252 in F11) reported to be associated with venous thrombosis conditions and studied the association of these common variants in selected case (n = 177) and control (n = 235) groups from population of Latvia. Genotyping was performed using TaqMan hybridization probe SNP genotyping assay.

Results

Patients with DVT had a significantly higher frequency of F11 rs2289252 polymorphism (p = 0.001; OR [95%CI] = 1.61 [1.20-2.14]). When stratified by recurrence of DVT the tendency was observed that the same SNP had higher OR value in group of DVT patients with repeated episodes of DVT compared to patients with single DVT episode (p = 0.009; OR [95%CI] = 2.27[1.22-4.21] and p = 0.009; OR [95%CI] = 1.52[1.11-2.08] respectively), but due to limited group of cases this finding should be replicated.

Conclusion

We conclude that F11 gene variant rs2289252 contribute to inherited forms of DVT incidence and correlation of other analysed SNPs should be explored in populations with greater sample size and associated with various thrombosis related traits.     

Determinants of daily smoking in Estonia, Latvia, Lithuania, and Finland in 1994-2002

AIMS: To investigate time trends in the smoking prevalence and the sociodemographic and psychosocial background of smoking in the Baltic countries in comparison with Finland during 1994-2002. METHODS: Differences in daily smoking according to age, education, urbanization, and psychological distress in the Baltic countries and Finland were studied using postal surveys in 1994, 1996, 1998, 2000, and 2002 among adults (20-64 years old) in Estonia (n = 6,271), Latvia (n = 6,106), Lithuania (n = 9,824), and Finland (n = 15,764). RESULTS: In 1994, 1996, 1998, 2000, and 2002 the prevalence of smoking in Estonia, Latvia, Lithuania, and Finland was 47%, 54%, 46%, and 29% among men, and 21%, 19%, 11%, and 19% among women, respectively. Smoking increased among Lithuanian women from 6% in 1994 to 13% in 2002, but decreased among Estonian men and women. Smoking was generally more common among younger individuals, the less educated, and people with distress in all four countries. The odds ratios for smoking for those with low education compared with those with high education were 2.18 (1.69-2.81), 3.32 (2.55-4.31), 2.20 (1.79-2.70) and 2.80 (2.40-3.27) in men, and 1.90 (1.42-2.52). 3.09 (2.28-4.18), 0.86 (0.59-1.26), and 3.00 (2.53-3.55) in women, in Estonia, Latvia, Lithuania, and Finland, respectively. There were indications of increasing educational differences in Latvian men. Smoking was less common among rural women in all countries except Estonia. CONCLUSIONS: Estonia, Latvia, and Finland show characteristics of the "mature" phase of a smoking epidemic, and smoking may not increase in these countries. In Lithuanian women smoking may increase. Smoking may be increasingly unequally distributed in the future in all the studied countries.     

Effects of probiotics on the composition of the intestinal microbiota following antibiotic therapy

The effects of probiotic supplementation on the intestinal re-growth microbiota following antibiotic therapy were studied in a double-blind placebo-controlled study. In the placebo group, numbers of facultative anaerobes and enterobacteria increased significantly, and at day 35 the numbers were significantly higher in the placebo group than in the active group; in the active group, the numbers of bacteroides increased significantly. Although the numbers of enterococci in both groups did not change, in the placebo group the number of patients harbouring antibiotic-resistant enterococci post therapy increased significantly. There was no change in the incidence rate of antibiotic resistance among the patients in the probiotic group.     

Trends in the magnitude of educational inequalities in health in Estonia, Latvia, Lithuania and Finland during 1994-2004

BACKGROUND: Socioeconomic inequalities in health in the Baltic countries are possibly increasing due to concomitant pressures. This study compared time trends from 1994 to 2004 in the pattern and magnitude of educational inequalities in health in Estonia, Latvia, Lithuania and Finland. METHODS: The data were gathered from cross-sectional postal surveys of the Finbalt project, conducted every second year since 1994 on adult populations (aged 20-64 years) in Estonia (n=9049), Latvia (n=7685), Lithuania (n=11,634) and Finland (n=18,821). Three self-reported health indicators were used: (i) less than good perceived health, (ii) diagnosed diseases, and (iii) symptoms. RESULTS: The existing educational inequalities in health in three Baltic countries and Finland remained generally stable over time from 1994 to 2004. Also, the overall prevalence of all three health indicators was generally stable, but in the Baltic countries improvement in perceived health was mainly found among the better-educated men and women. Diagnosed diseases increased in the Baltic countries, except Lithuania, where diseases decreased among the better-educated women. Symptoms increased among the better-educated Estonian and Finnish women. CONCLUSIONS: The period from 1994 to 2004 of relative stabilization since the worst conditions of the social transition has not been followed by notable changes in self-reported health, and this appears to be the situation across all educational groups in the Baltic countries. While health inequalities did not markedly change, substantial inequalities do remain, and there were indications of favourable developments mainly among the better-educated respondents. The factors contributing towards increasing health inequalities may only be visible in the future.