The incidence of esophageal adenocarcinoma has increased significantly in the western world over the last 20 yr. Most cases arise in a background of chronic gastroesophageal reflux, and specialized intestinal metaplasia in Barrett's esophagus is frequently an antecedent phenotype or evident in association with adenocarcinoma. The molecular events that characterize the pathway from inflammation to metaplasia to dysplasia and adenocarcinoma are poorly understood. To examine the expression of the proinflammatory cytokines IL-8 and IL-1β along the esophagitis, metaplasia, dysplasia, and adenocarcinoma pathway, and to correlate this with histological changes and expression of the transcription factor NF-κB. Fresh biopsy specimens were collected from patients with reflux esophagitis (n= 15), Barrett's esophagus (n = 35), Barrett's adjacent to adenocarcinoma (n = 8), and esophageal adenocarcinoma (n = 35). IL-8 and IL-1β expression were measured using enzyme-linked immunosorbent assay. NF-κB expression was measured by electrophoretic mobility shift assay. Elevated expression of NF-κB was found in 2 (13%) out of 15 patients with reflux esophagitis, 21 (60%) out of 35 patients with Barrett's esophagus, and 28 (80%) out of 35 patients with esophageal adenocarcinoma. All 5 patients with Barrett's esophagus and high-grade dysplasia showed elevated expression of NF-κB. IL-8 and IL-1β were significantly increased in esophagitis, Barrett's, and adenocarcinoma compared with squamous epithelium, and in adenocarcinoma compared with all other groups. There was a stepwise increase in the expression of IL-8, IL-1β, and NF-κB from normal through Barrett's epithelium to adenocarcinoma in eight cases of esophageal adenocarcinoma. The levels of both IL-8 and IL-1β in adenocarcinoma patients correlated with stage of disease. Patients with adenocarcinoma who were NF-κB positive had significantly higher levels of both IL-8 (p= 0.04) and IL-1β (p= 0.03) compared to adenocarcinoma patients who were NF-κB negative. The proinflammatory cytokines IL-8 and IL-1β are elevated in esophagitis and Barrett's epithelium, and markedly elevated in adenocarcinoma. NF-κB activation is infrequent in esophagitis, but is increased in Barrett's epithelium and adenocarcinoma. The association of NF-κB activation with cytokine upregulation was only evident in patients with adenocarcinoma. These patterns may play an important role in Barrett's inflammation and tumourigenesis, and inhibition of the NF-κB/proinflammatory cytokine pathway may be an important target for future chemoprevention strategies.

To examine the expression of the transcription factor nuclear factor kappa B (NF-kappaB) in Barrett's epithelium and adenocarcinoma and the impact of NF-kappaB expression on tumor stage and response to neoadjuvant chemotherapy and radiation therapy. Progression of Barrett's esophagus to adenocarcinoma is associated with a wide range of cellular and molecular abnormalities. Nuclear factor-kappa B (NF-kappaB) regulates several genes involved in inflammatory, immune and apoptotic responses, but its role in esophageal inflammation and tumorigenesis has not been reported. Mobility shift assay was used to measure NF-kappaB activity in nuclear extracts of fresh-frozen biopsies from tumor and uninvolved tissues (n = 30) and esophageal cell lines OE33, SKGT-4, and OE21. RelA expression was assessed by immunohistochemical staining (n = 97). The NF-kappaB/RelA and IkappaB protein expressions were also examined by Western blotting. NF-kappaB was not expressed in normal esophageal squamous epithelium, in contrast to increased expression in 40% of patients with Barrett's epithelium. Sixty-one percent of resected tumors (n = 97) displayed NF-kappaB immunoreactivity, and 87.5% of the NF-kappaB-positive tumors were Stage IIb and III compared with only 12.5% of patients with Stage I and IIa disease (P < 0.05). The expression of NF-kappaB inversely correlated with major or complete pathologic responses to neoadjuvant chemotherapy and radiation therapy, with 15/20 (75%) responders in the NF-kappaB-negative group compared with 7/38 (18%) in the NF-kappaB-positive group (P < 0.00001). Moreover, incubation of esophageal cell lines OE33, SKGT-4, and OE21 with deoxycholic acid or low pH induced NF-kappaB expression. Bile acids and low pH induce NF-kappaB expression in esophageal cell lines. NF-kappaB activation is common in esophageal adenocarcinoma. In patients with Barrett's epithelium and an associated esophageal adenocarcinoma, there is a progressive expression of NF-kappaB through Barrett's tumorigenesis. The absence of NF-kappaB expression in esophageal adenocarcinoma correlates with response to neoadjuvant chemoradiotherapy and may be of value in predicting response to neoadjuvant therapy.

Deoxycholic acid (DCA) has been implicated in colorectal carcinogenesis in humans with effects on proliferation and apoptosis, mediated at least in part by activation of transcription factors nuclear factor kappa B (NF‐κB), activator protein 1 (AP‐1) and protein kinase C (PKC) enzymes. Ursodeoxycholic acid (UDCA) is reported to reduce the frequency of colonic carcinogenesis in ulcerative colitis patients. Hence, we postulated that it might differ from DCA in its regulation of these transcription factors. The aim of the study was to determine effects of DCA and UDCA on NF‐κB and AP‐1 activation and explore its relationship to PKC. Human colonic tumour cell lines HCT116 were treated with DCA, UDCA, alone or pretreated with UDCA followed by DCA or IL‐1β. In other experiments, cells were pretreated with PKC inhibitors and then stimulated with DCA and IL‐1β or PMA. Gel shift assays were performed on nuclear extracts of the cells for NF‐κB and AP‐1 analysis. Western blot analyses and immunofluorescence were performed for Rel A (p65) and IκB‐α levels on the treated cells. DCA increased NF‐κB and AP‐1 DNA binding. UDCA did not increase DNA binding of NF‐κB and AP‐1 and UDCA pretreatment inhibited DCA‐induced NF‐κB and AP‐1 DNA binding. PKC inhibitors blocked DCA‐induced NF‐κB and AP‐1 activation. These results were validated by Western blot analysis for RelA and IκB‐α. In conclusion, UDCA did not induce NF‐κB and AP‐1 DNA binding but also blocked DCA‐induced NF‐κB and AP‐1 activation. These findings suggest a possible mechanistic role for UDCA in blocking pathways thought to be involved in colon carcinogenesis.

Caffeic acid phenethyl ester (CAPE), an active component of propolis from honeybee hives (honeybee resin), has anti‐inflammatory, anti‐carcinogenic and anti‐bacterial properties. This study was designed to investigate the anti‐inflammatory effects of CAPE onHelicobacter pylori‐induced NF‐κB and AP‐1 in the gastric epithelial cell line AGS. Electrophoretic mobility shift assay was used to measure NF‐κB‐ and AP‐1‐DNA binding activity. Western blotting was used to detect IκB‐αand COX‐2 expression in AGS cells cocultured withH. pylori. The antiproliferative effect of CAPE was measured by MTT assay. Our results showed that caffeic phenethyl ester inhibitsH. pylori‐induced NF‐κB and AP‐1 DNA‐binding activity in a dose (0.1–25 μg ml⁻¹∼0.35–88 μM) and time‐ (15–240 min) dependent manner in AGS cells. Maximum inhibition by CAPE was observed at concentrations of 25 μg ml⁻¹(∼88 μM) CAPE preventedH. pylori‐ and cytokine‐induced degradation of IκB‐αprotein. Pretreatment of AGS cells with CAPE also blocked cytokine‐ and mitogen‐induced NF‐κB and AP‐1 expression. Furthermore, CAPE suppressedH. pylori‐induced cell proliferation and production of the cytokines TNF‐αand IL‐8. In addition, CAPE blockedH. pylori‐induced COX‐2 expression. The inhibition of such transcription by CAPE could result in suppression of many genes duringH. pylori‐induced inflammation, and also provide new insights into the anti‐cancer and anti‐inflammatory properties of CAPE. British Journal of Pharmacology(2005)146, 1139–1147. doi:10.1038/sj.bjp.0706421

Adverse Drug Reactions (ADRs) are scantly reported with poor contribution by healthcare professionals worldwide and in particular in developing countries. The aim of this study was to assess the knowledge and awareness of adverse drug reactions (ADRs) reporting and pharmacovigilance system among healthcare professionals in Al-Madinah Al-Munawwarah hospitals, Kingdom of Saudi Arabia. A questionnaire was designed addressing; awareness of ADRs, knowledge of pharmacovigilance system, availability of ADRs reporting system, patient counseling about ADRs and documentation of ADRs. The questionnaire was distributed to randomly selected healthcare professionals (n = 585) such as physicians, pharmacists, nurses and pharmacists' technicians of hospitals. Completed questionnaires were collected and data were analyzed. Data were expressed in number as well as percentage. Of the 585 questionnaires circulated, a total of 384 healthcare professionals responded. Healthcare professional categories involved in the study were 148 physicians, 37 pharmacists, 158 nurses and 41 pharmacist technicians. The percent of the respondents who accepted to enroll in the study was 65.64%. Most of the respondents were unable to correctly define the pharmacovigilance term, but they were aware of ADRs. The awareness among healthcare professionals of the national pharmacovigilance system was 39.6%. Pharmacists had a good knowledge of pharmacovigilance and ADRs terminology and showed a more positive attitude to report ADRs. A greater number of the healthcare professionals were aware of ADRs reporting, but practically it is not implemented in hospitals. Most hospitals had follow-up documentation systems, but did not include ADRs reporting. There was no distinct pharmacovigilance system in place. Our study has demonstrated a lack of knowledge and awareness of pharmacovigilance and ADRs reporting among healthcare professionals in hospitals. The poor knowledge of ADRs reporting emphasized the urgent need to implement the appropriate strategies to improve the awareness of pharmacovigilance practices and ADRs reporting in Al-Madinah Al-Munawwarah hospitals.