Prevalence of Helicobacter Pylori Genotypes and Their Association with Upper Gastrointestinal Diseases: A Cross Sectional Study in Southern Iran

Background: Investigating the prevalence of vacuolating cytotoxin (vacA), cytotoxin associated gene A (cagA), glm M genotypes, and subtypes of vacA of Helicobacter pylori (H. pylori) isolate in Jahrom, Southern Iran.Methods: DNA extracted from H. pylori samples retrieved from gastric biopsy isolated from 113 dyspeptic patients with positive rapid urease test (RUT). Genotyping was done by polymerase chain reaction (PCR) technique, using primers for vacA (s1a, s1b, s1c, s1, s2, m2, and m1), cagA, and glmM. Endoscopy was done for all the patients to screen gastrointestinal (GI) pathologies. Results: GlmM was detected in 100% of the cases. VacA subtypes s1am2, s2m2, s1am1, s1b, and s1c were detected in 27.9%, 25.6%, 22.1%, 3.5% and 2.4% of the isolates, respectively, while cagA was detected in 60.5% of the isolates. VacA alleles m1, s1, and s2 were detected in 54%, 50%, and 44% of isolates, respectively. Also, 60.5% of the isolates were cagA-vacA-positive. A significant correlation was observed between vacAs1bm1 and gastroesophageal reflux disease (GERD), and glmM positive isolates had normal esophagus. The presence of vacAs1bm1 and vacAs1bm2 has a significant association with gastric erythema. The presence of cagA showed a significant association with normal esophagus and hiatal hernia.Conclusion: In our research, the number of glmM and cagA positive isolates is higher among other genotypes, and cagA is correlated with hiatal hernia, and normal esophageal finding is correlated with glmM genotype. There was no association between the age or sex of the patients and bacterial genotype.

Identification and Validation of Esophageal Squamous Cell Carcinoma Targets for Fluorescence Molecular Endoscopy

Dysplasia and intramucosal esophageal squamous cell carcinoma (ESCC) frequently go unnoticed with white-light endoscopy and, therefore, progress to invasive tumors. If suitable targets are available, fluorescence molecular endoscopy might be promising to improve early detection. Microarray expression data of patient-derived normal esophagus (n = 120) and ESCC samples (n = 118) were analyzed by functional genomic mRNA (FGmRNA) profiling to predict target upregulation on protein levels. The predicted top 60 upregulated genes were prioritized based on literature and immunohistochemistry (IHC) validation to select the most promising targets for fluorescent imaging. By IHC, GLUT1 showed significantly higher expression in ESCC tissue (30 patients) compared to the normal esophagus adjacent to the tumor (27 patients) (p < 0.001). Ex vivo imaging of GLUT1 with the 2-DG 800CW tracer showed that the mean fluorescence intensity in ESCC (n = 17) and high-grade dysplasia (HGD, n = 13) is higher (p < 0.05) compared to that in low-grade dysplasia (LGD) (n = 7) and to the normal esophagus adjacent to the tumor (n = 5). The sensitivity and specificity of 2-DG 800CW to detect HGD and ESCC is 80% and 83%, respectively (ROC = 0.85). We identified and validated GLUT1 as a promising molecular imaging target and demonstrated that fluorescent imaging after topical application of 2-DG 800CW can differentiate HGD and ESCC from LGD and normal esophagus.

Endoscopic resection combined with the Cryoballoon focal ablation system in the porcine normal esophagus: a preclinical study

Background The Cryoballoon focal ablation system (CbFAS) for dysplastic Barrett’s esophagus is simple, time-saving and has high therapeutic efficacy. This study aimed to evaluate the technical feasibility and tissue damage with combination therapy of endoscopic resection (ER) and CbFAS in porcine models. Methods Three pigs (A, B, and C) were included, and all ER procedures were performed by endoscopic mucosal resection using the Cap method (EMR). Combination therapy for each pig was performed as follows: (a) CbFAS was performed for a post-EMR mucosal defect for Pig A; (b) CbFAS for post-EMR scar for Pig B, and (c) EMR for post-CbFAS scar for Pig C. All pigs were euthanized at 32 days after the initial procedure, and the tissue damage was evaluated. Results All endoscopic procedures were followed as scheduled. None of the subjects experienced anorexia, rapid weight loss, bleeding, and perforation during the observation period. They were euthanized at 32 days after the initial endoscopic procedure. On histological assessment, there was little difference between the tissue that was treated with CbFAS alone and that treated with CbFAS in combination with ER. Conclusion Combination therapy with ER and CbFAS can be technically feasible, and its outcome was not significantly different from CbFAS alone in terms of tissue damage.

Endoscopic Resection Combined with the Cryoballoon Focal Ablation system in the Porcine Normal Esophagus: A Preclinical Study

Background: The Cryoballoon focal ablation system (CbFAS) for dysplastic Barrett’s esophagus is simple, time-saving and has high therapeutic efficacy. This study aimed to evaluate the technical feasibility and tissue damage with combination therapy of endoscopic resection (ER) and CbFAS in porcine models.Methods: Three pigs (A, B, and C) were included, and all ER procedures were performed by endoscopic mucosal resection using the Cap method (EMR). Combination therapy for each pig was performed as follows: (a) CbFAS was performed for a post-EMR mucosal defect for Pig A; (b) CbFAS for post-EMR scar for Pig B, and (c) EMR for post-CbFAS scar for Pig C. All pigs were euthanized at 32 days after the initial procedure, and the tissue damage was evaluated.Results: All endoscopic procedures were followed as scheduled. None of the subjects experienced anorexia, rapid weight loss, bleeding, and perforation during the observation period. They were euthanized at 32 days after the initial endoscopic procedure. On histological assessment, there was little difference between the tissue that was treated with CbFAS alone and that treated with CbFAS in combination with ER.Conclusion: Combination therapy with ER and CbFAS can be technically feasible, and its outcome was not significantly different from CbFAS alone in terms of tissue damage.

Differential Expression of Desmocollin2, Desmoglein2, and Plakophilin2 in the Progression of Esophagitis to Esophageal Adenocarcinoma

BackgroundDesmosomes play a key role in intercellular adhesive, but also contribute to tumorigenesis. This study aimed to examine the differential expression of desmocollin2 (DSC2), desmoglein2 (DSG2), and plakophilin2 (PKP2) in the progression of reflux esophagitis to esophageal adenocarcinoma (EAC) in the rat model of reflux disease established by esophagogastroduodenal anastomosis (EGDA).MethodsEGDA was performed on rats to induce gastroesophageal reflux leading to the development of EAC. All rats were randomly divided into four groups: group A rats received EGDA only (n=27); group B rats received EGDA and iron supplementation (n=28); group C received pseudo surgery only (n=20); group D received pseudo surgery and iron supplementation (n=20). Animals were randomly selected from each group and euthanized at 8 weeks and 32 weeks following EGDA. Esophageal tissues were harvested and divided into 4 types (normal esophagus, esophagitis, dysplasia, and EAC). On these tissue types, immunohistochemistry was performed to characterize the localization and distribution of DSC2, DSG2, and PKP2, while qRT-PCR and western blot were performed to detect the expression of DSC2, DSG2, and PKP2 at the gene and protein levels.ResultsAt 8 weeks after surgery, 80% of rats in group A and 100% in group B had esophagitis. At 32 weeks, 29.41% and 17.65% of rats in group A developed dysplasia and EAC, respectively, while in group B, dysplasia and EAC accounted for 44.44% and 38.89%, respectively. The expression of DSC2, DSG2, and PKP2 at both the gene and protein levels increased progressively from esophagitis to dysplasia, and EAC. Of note, all of the three genes were significantly upregulated in EAC tissues compared with tissues of esophagitis.ConclusionDSC2, DSG2, and PKP2 may play an important role in the progression of esophagitis to EAC. Their expression levels may therefore be utilized as molecular biomarkers for early diagnosis and targeted therapy for EAC.

Prevalence of Helicobacter Pylori Genotypes and Their Association With Upper Gastrointestinal Diseases: a Cross Sectional Study in Southern Iran

Background: Investigating the prevalence of vacuolating cytotoxin (vacA), cytotoxin associated gene A (cagA), glm M genotypes, and subtypes of vacA of Helicobacter pylori (H. pylori) isolate in Jahrom, Southern Iran. DNA extracted from H. pylori samples retrieved from gastric biopsy isolated from 113 dyspeptic patients with positive rapid urease test (RUT). Genotyping was done by polymerase chain reaction (PCR) technique, using primers for vacA (s1a, s1b, s1c, s1, s2, m2, and m1), cagA, and glmM. Endoscopy was done for all the patients to screen upper gastrointestinal (GI) disorders. Results: GlmM was detected in 100% of the cases. VacA subtypes s1am2, s2m2, s1a, s1b, and s1c were detected in 27.9%, 25.6%, 50%, 3.5% and 2.4% of the isolates, respectively, while cagA was detected in 60.5% of the isolates. VacA alleles m1, s1, and s2 were detected in 54%, 50%, and 44% of isolates respectively. Also, 60.5% of the isolates were cagA-vacA-positive. A significant correlation was observed between vacAs1bm1 and gastroesophageal reflux disease (GERD) and glmM and normal esophagus. The presence of vacAs1bm1 and vacAs1bm2 has a significant association with gastric erythema. The presence of cagA showed a significant association with normal esophagus and hiatal hernia.Conclusions: In our research, the number of glmM and cagA positive isolates is higher among other genotypes and cagA is correlated with hiatal hernia, and normal esophageal finding is correlated with glmM genotype. There was no association between age or sex of the patients and bacterial genotype.

JmjC-KDMs KDM3A and KDM6B modulate radioresistance under hypoxic conditions in esophageal squamous cell carcinoma

Esophageal squamous cell carcinoma (ESCC), the most frequent esophageal cancer (EC) subtype, entails dismal prognosis. Hypoxia, a common feature of advanced ESCC, is involved in resistance to radiotherapy (RT). RT response in hypoxia might be modulated through epigenetic mechanisms, constituting novel targets to improve patient outcome. Post-translational methylation in histone can be partially modulated by histone lysine demethylases (KDMs), which specifically removes methyl groups in certain lysine residues. KDMs deregulation was associated with tumor aggressiveness and therapy failure. Thus, we sought to unveil the role of Jumonji C domain histone lysine demethylases (JmjC-KDMs) in ESCC radioresistance acquisition. The effectiveness of RT upon ESCC cells under hypoxic conditions was assessed by colony formation assay. KDM3A/KDM6B expression, and respective H3K9me2 and H3K27me3 target marks, were evaluated by RT-qPCR, Western blot, and immunofluorescence. Effect of JmjC-KDM inhibitor IOX1, as well as KDM3A knockdown, in in vitro functional cell behavior and RT response was assessed in ESCC under hypoxic conditions. In vivo effect of combined IOX1 and ionizing radiation treatment was evaluated in ESCC cells using CAM assay. KDM3A, KDM6B, HIF-1α, and CAIX immunoexpression was assessed in primary ESCC and normal esophagus. Herein, we found that hypoxia promoted ESCC radioresistance through increased KDM3A/KDM6B expression, enhancing cell survival and migration and decreasing DNA damage and apoptosis, in vitro. Exposure to IOX1 reverted these features, increasing ESCC radiosensitivity and decreasing ESCC microtumors size, in vivo. KDM3A was upregulated in ESCC tissues compared to the normal esophagus, associating and colocalizing with hypoxic markers (HIF-1α and CAIX). Therefore, KDM3A upregulation in ESCC cell lines and primary tumors associated with hypoxia, playing a critical role in EC aggressiveness and radioresistance. KDM3A targeting, concomitant with conventional RT, constitutes a promising strategy to improve ESCC patients’ survival.

Clinical features and time trends associated with an endoscopically normal esophagus in active eosinophilic esophagitis

Background A proportion of patients with active eosinophilic esophagitis (EoE) have a normal-appearing esophagus on esophagogastroduodenoscopy (EGD). We aimed to determine the associations between the baseline clinical features and the endoscopically normal esophagus in EoE, as well as time trends in reporting. Methods In this retrospective study of active EoE cases from 2002 – 2018, patients with and without esophageal endoscopic abnormalities were compared. Multivariable logistic regression identified the independent predictors of a normal EGD. The proportion of patients with a normal EGD was determined per year, and before and after the introduction of the first EoE guidelines and the EoE Endoscopic Reference Score (EREFS). Results Of 878 EoE patients, 101 (11.5 %) had an endoscopically normal esophagus; they were younger (8.3 vs. 25.4 years), had shorter median symptom duration before diagnosis (2.8 vs. 5.0 years), were less likely to have dysphagia (40 % vs. 76 %) or food impaction (8 % vs. 33 %), and more likely to have abdominal pain (37 % vs. 19 %) (P < 0.01 for all). On multivariable logistic regression, independent predictors of a normal esophagus were younger age (odds ratio [OR] 0.96, 95 % confidence interval [CI] 0.94 – 0.98), abdominal pain (OR 2.03, 95 %CI 1.13 – 3.67), and lack of dysphagia (OR 0.49, 95 %CI 0.26 – 0.93). The proportion of patients with a normal esophagus decreased from 21 % before the first EoE guidelines to 7 % (P < 0.01) after introduction of the EREFS. Conclusions An endoscopically normal esophagus is seen in ~10 % of active EoE patients and should not preclude biopsies; younger age, abdominal pain, and lack of dysphagia are independent predictors. The proportion of normal EGDs decreased over time, suggesting improved recognition of endoscopic findings.

A case of formic acid poisoning: prompting for a different line of management

Formic acid is an easily available substance and can thus be consumed voluntarily or accidentally. Here is a case who presented to our center 5 days post accidental ingestion of formic acid. He had developed acute renal failure requiring hemodialysis. Post admission he had altered sensorium probably alcohol withdrawal and later aspiration pneumonia and ARDS. He developed a late onset GI bleed on day 8 of ingestion with significant hemoglobin drop. An emergency endoscopy was done which showed a normal esophagus but extensive corrosive damage and active bleeding from gastric area. He continued to have drop in hemoglobin in-spite of aggressive volume and blood product transfusions. He was taken up for an emergency subtotal gastrectomy and viable tissue was demarcated with intraoperative endoscopy and a subtotal gastrectomy was performed. He stabilized initially but worsened again 2 days later with worsening lactic acidosis and succumbed to his illness. The fact that esophagus was completely spared with extensive involvement of stomach could possibly indicate a need for review of initial management of corrosive/organic acid poisoning.