Substantial Intestinal Microbiota Differences Between Patients With Ulcerative Colitis From Ghana and Denmark:Ulcerative Colitis in Denmark vs Ghana

BACKGROUND: Ulcerative colitis (UC) is a relapsing nontransmural inflammatory disease that is restricted to the colon and is characterized by flare-ups of bloody diarrhea. In this study, we aimed to investigate intestinal bacterial diversity in healthy controls and patients with UC with and without active disease, from Ghana and Denmark. METHODS: The study included 18 UC patients (9 with active and 9 with inactive disease) and 18 healthy controls from Ghana. In addition 16 UC patients from Denmark (8 UC with active and 8 UC with inactive disease) and 19 healthy controls from Denmark. Microbiota diversity analysis relied on sequencing of ribosomal small subunit genes. Purified genomic DNA was submitted to PCR using a primer set targeting prokaryotes and eukaryotes. The purified DNA was sequenced on the Illumina MiSeq system in a 2 × 250 bp set up (Illumina, San Diego, CA, USA). Blinded analysis of the taxonomy table was performed using BioNumerics-7.5 (Applied Maths NV, Sint-Martens-Latem, Belgium). RESULTS: When analyzing the taxonomy data for prokaryotes, cluster and principal component analysis shows Danish healthy controls clustered together, but separate from healthy controls from Ghana, which also clustered together. The Shannon diversity index (SDI) for prokaryotes shows significant differences between Danish healthy controls and patients in comparison with the corresponding groups from Ghana (p = 0.0056). Significant increased abundance of Escherichia coli was detected in healthy controls from Ghana in comparison with healthy controls from Denmark. The SDI of the prokaryotes ranges between 0 and 3.1 in the Ghana study groups, while in the Danish study groups it ranges between 1.4 and 3.2, the difference is however not significant (p = 0.138). Our data show a significant increased abundance of eukaryotes species in the healthy control group from Ghana and Denmark in comparison with patient groups from Ghana and Denmark. CONCLUSION: Overall, healthy controls and patients with UC from Denmark have increased diversity of prokaryotes. Healthy controls from Denmark and Ghana have increased abundance of eukaryotes in comparison with UC patient groups from Denmark and Ghana

Factors associated with gastro-duodenal disease in patients undergoing upper GI endoscopy at the Korle-Bu Teaching Hospital, Accra, Ghana.

Background: There is a high prevalence of gastro-duodenal disease in sub-Saharan Africa. Peptic ulcer disease in dyspeptic patients, 24.5%, was comparable to prevalence of gastro-duodenal disease among symptomatic individuals in developed countries (12 \u2013 25%). Limited data exists regarding its associated risk factors despite accumulating evidence indicating that gastroduodenal disease is common in Ghana. Objectives: This study investigates risk factors associated with gastro-duodenal disease at the Korle-Bu Teaching Hospital, Accra, Ghana. Methods: This study utilized a cross-sectional design to consecutively recruit patients referred with upper gastro-intestinal symptoms for endoscopy. The study questionnaire was administered to study participants. Helicobacter pylori infection was confirmed by rapid-urease examination at endoscopy. Results: Of 242 patients sampled; 64 had duodenal ulcer, 66 gastric ulcer, 27gastric cancer and 64 non-ulcer dyspepsia. Nineteen (19) had duodenal and gastric ulcer while 2 had gastric ulcer and cancer. A third (32.6%) of patients had history of NSAIDuse. H. pylori was associated with gastric ulcer (p=0.033) and duodenal ulcer (p=0.001). There was an increased prevalence of duodenal ulcer in H. pylori-infected patients taking NSAIDs, P=0.003. Conclusion: H. pylori was a major risk factor for peptic ulcer disease. However, NSAID-related gastro-duodenal injury has been shown to be common in H. pylori infected patients. It highlights the need for awareness of the adverse gastro-intestinal effects in a H. pylori endemic area

Helicobacter pylori diversity and gastro-duodenal disease in Western Africa

Helicobacter pylori (Hp) is causally linked with gastric pre-malignant lesions, gastric cancer (GCA) and peptic ulcer disease (PUD). In Africa, PUD is prevalent while GCA has reportedly low incidence. Less is known about pre-malignant lesions. Hp genes cagA and vacA are established virulence factors and have been associated with increased gastroduodenal disease (GDD) risk. Further, diversity in the Carboxyl-terminus of cagA can influence GCA risk. Virulence studies have evolved from candidate-based to more robust genome wide association studies (GWAS). Here, a clinical and histopathological assessment of GDD in West Africa was performed, as well as an evaluation of associated Hp genetic variation. In 136 dyspeptic patients from Ghana, pre-malignant lesions had low prevalence, 14.7%, most of which were located distally. vacA and cagA Hp strains were identified by PCR in 69.2% and 74.8% of 159 dyspeptic Ghanaian patients respectively; cagA-(internal duplication region) being more prevalent than cagA-(hydrophilic region). Duodenal ulcer was associated with cagA-(hydrophilic region) and vacAs1m1. Multi-locus sequence typing (MLST) of seven housekeeping genes have previously identified multiple distinct H. pylori populations or lineages such as hpAfrica1 and hpEurope. An agnostic analysis of Hp virulence employed a GWAS using 193 controls from the European-African admixed population of Cape Verde (CV) and 101 cases from reference databases. African controls and cases clustered within hpAfrica1, while European controls and cases clustered within hpEurope. Genes significant for GCA or PUD were established virulence factors cagPAI, vacA and babA2. Prominent among these were lipopolysaccharide and peptidoglycan biosynthesis proteins. Phylogenetic analyses showed a cluster of closely related CV lineages that diverged from European cases. This CV_outgroup was notable for absence of major virulence genes. Although several novel and established variants have been linked to GDD, population structure in bacteria can lead to false positives in linkage dysequilibrium (LD) with genuine risk variants. Hence, validation studies are required to assist in risk stratification of infected individuals for further management. Ultimately, GDD risk is multi-factorial. Therefore, studies ought to evaluate host, bacterial and environmental factors in an integrated design.</p

Helicobacter Pylori Variants with ABC-Type Tyrosine Phosphorylation Motif in Gastric Biopsies of Ghanaian Patients

Background. Helicobacter pylori pathogenicity and disease severity are determined by the tyrosine phosphorylation motifs of CagA protein. This study is aimed at detecting the presence of H. pylori and identifying the CagA tyrosine phosphorylation motifs in Ghanaian patients. Material and Methods. A total of 94 archival genomic DNA samples from gastric biopsies were used for the study, and H. pylori was detected by amplifying the 16S rRNA gene. The 3′-end variable region of the cagA gene was amplified, and the entire 3′-end was sequenced and translated into amino acids. Results. H. pylori was detected in 53.2% (50/94) of the samples, and all the detected bacteria harboured the cagA gene. Two variants of the bacteria were identified based on the size of the amplified cagA gene: 207 bp and 285 bp. The 207 bp and 285 bp variants accounted for 74% and 22%, respectively, and 4% showed both fragments. Translated amino acid sequence of the cagA gene showed EPIYA-A, EPIYA-B, and EPIYA-C (ABC type) motifs, indicating the Western variant. The CagA protein C-terminal showed insertion of amino acids in the sequence flanking the EPIYA-A motif at the N-terminal and a complete deletion of the EPIYA-CC and EPIYA-CCC motifs together with the flanking sequences. Conclusions. H. pylori identified were Western variant (ABC type) with unique amino acid insertions, suggesting unique variants in Ghanaian patients. Further investigation is however required to understand the role of the molecular diversity of the variant in gastric disease outcome

Helicobacter pylori cagA and vacA genes in dyspeptic Ghanaian patients

BACKGROUND: Helicobacter pylori infection is prevalent in Ghana. The development of gastro-duodenal disease is dependent on virulence of the infecting strain, host susceptibility and environmental factors. Helicobacter pylori cagA and vacA strains induce more inflammation, ulceration and oncogenesis. Here, for the first time we present data on H. pylori cagA and vacA genes and their association with gastro-duodenal disease in Ghana. A total of 159 patients with dyspepsia at Korle-Bu Teaching Hospital, Accra, were investigated for H. pylori with urease-CLO, of which 113 (71.1%) were positive. Genomic DNA was extracted from antral biopsies using QIAGEN DNeasy kit. Detection of H. pylori vacA and cagA genes were determined by PCR as previously described. RESULTS: In total, 110 (69.2%) vacAs1, 71 (44.7%) vacAm1, 35 (22.0%) vacAm2, 77 (48.4%) cagA-(hydrophilic region) and 109 (68.6%) cagA-(internal duplication region) were detected. In multivariate analysis, duodenal ulcer was more likely than other diagnoses to have detectable cagA-(hydrophilic region) (OR 3.1 CI 1.2-7.9) or vacAs1m1 (OR 6.5 CI 1.2-34.0). CONCLUSIONS: Majority of biopsies were colonized with H. pylori harboring both cagA and vacA. H. pylori cagA-(internal duplication region) was more prevalent than cagA-(hydrophilic region). Duodenal ulcer was more likely than other diagnoses to have detectable cagA-(hydrophilic region) or vacAs1m1