Arthur Hoffman

High definition colonoscopy combined with i-Scan is superior in the detection of colorectal neoplasias compared with standard video colonoscopy: a prospective randomized controlled trial

INTRODUCTION Colonoscopy is the accepted gold standard for the detection of colorectal cancer. The aim of the current study was to prospectively compare high definition plus (HD+) colonoscopy with I-Scan functionality (electronic staining) vs. standard video colonoscopy. The primary endpoint was the detection of patients having colon cancer or at least one adenoma. METHODS A total of 220 patients due to undergo screening colonoscopy, postpolypectomy surveillance or with a positive occult blood test were randomized in a 1 : 1 ratio to undergo HD+ colonoscopy in conjunction with I-Scan surface enhancement (90i series, Pentax, Tokyo, Japan) or standard video colonoscopy (EC-3870FZK, Pentax). Detected colorectal lesions were judged according to type, location, and size. Lesions were characterized in the HD+ group by using further I-Scan functionality (p- and v-modes) to analyze pattern and vessel architecture. Histology was predicted and biopsies or resections were performed on all identified lesions. RESULTS HD+ colonoscopy with I-Scan functionality detected significantly more patients with colorectal neoplasia (38 %) compared with standard resolution endoscopy (13 %) (200 patients finally analyzed; 100 per arm). Significantly more neoplastic (adenomatous and cancerous) lesions and more flat adenomas could be detected using high definition endoscopy with surface enhancement. Final histology could be predicted with high accuracy (98.6 %) within the HD+ group. CONCLUSIONS HD+ colonoscopy with I-Scan is superior to standard video colonoscopy in detecting patients with colorectal neoplasia based on this prospective, randomized, controlled trial.

Antibodies Against Tumor Necrosis Factor (TNF) Induce T-Cell Apoptosis in Patients With Inflammatory Bowel Diseases via TNF Receptor 2 and Intestinal CD14+ Macrophages

BACKGROUND & AIMS The anti-tumor necrosis factor (TNF) antibodies infliximab, adalimumab, and certolizumab pegol have proven clinical efficacy in Crohns disease. Here, we assessed the effects of anti-TNF antibodies on apoptosis in inflammatory bowel disease (IBD). METHODS CD14(+) macrophages and CD4(+) T cells were isolated from peripheral blood and lamina propria mononuclear cells from patients with IBD and control patients. Cell surface markers and apoptosis were assessed by immunohistology and fluorescence-activated cell sorting techniques. RESULTS Lamina propria CD14(+) macrophages showed significantly more frequent and higher membrane-bound TNF (mTNF) expression than CD4(+) T cells in IBD, whereas mTNF-dependent signaling proteins such as TNF receptor (TNFR) 2, TNFR-associated factor (TRAF) 2, and nuclear factor κB were induced in IBD mucosal CD4(+) T cells. Most anti-TNF antibodies did not induce T-cell apoptosis in purified peripheral or mucosal CD4(+) T cells. However, in contrast to etanercept, administration of all clinically effective anti-TNF antibodies resulted in a significant induction of T-cell apoptosis in IBD when lamina propria CD4(+) T cells expressing TNFR2(+) were cocultured with mTNF(+) CD14(+) intestinal macrophages. In contrast, no effects in control patients were noted. T-cell apoptosis in IBD occurred in vivo after treatment with adalimumab and infliximab, was critically dependent on TNFR2 signaling, and could be prevented via interleukin-6 signal transduction. Blockade of interleukin-6R signaling augmented anti-TNF-induced T-cell apoptosis in IBD. CONCLUSIONS Clinically effective anti-TNF antibodies are able to induce T-cell apoptosis in IBD only when mucosal TNFR2(+) T cells are cocultured with mTNF-expressing CD14(+) macrophages. The finding that anti-TNF antibodies induce apoptosis indirectly by targeting the mTNF/TNFR2 pathway may have important implications for the development of new therapeutic strategies in IBD.

Recognition and characterization of small colonic neoplasia with high-definition colonoscopy using i-Scan is as precise as chromoendoscopy

BACKGROUND The EPKi system (Pentax, Japan) enables resolution above HDTV. Aim of the study was to test the efficacy of HD+ alone and with the new post-processing digital filter i-Scan or chromoendoscopy (Methylene blue 0.1%) in screening for colorectal cancer. We focused on lesions less than 5 mm as a surrogate marker for the optical possibilities of the EPKi system. METHODS The last 30 cm of the colon in a screening population were inspected with HD+ alone, in combination with i-Scan (2:1 randomisation) and subsequently with chromoendoscopy. All lesions were characterized and targeted biopsies were performed. RESULTS i-Scan augmented in 69 patients the identification of lesions from 176 to 335 (p<0.001) and chromoendoscopy to 646 (p<0.001). The additional lesions were mainly flat (type IIb, 74%), which were only recognized using i-Scan or chromoendoscopy. The amount of neoplasias was not significantly different (HD+: 5, i-Scan: 11, Chromoendoscopy: 11), but all could correctly be predicted using i-Scan or chromoendoscopy. CONCLUSIONS HD+ colonoscopy with and without i-Scan unmask a plethora of small lesions but chromoendoscopy can even advance the number. However, i-Scan was able to predict neoplasia as precisely as chromoendoscopy and might shortly replace chromoendoscopy as a more time efficient tool.

In vivo diagnosis of collagenous colitis by confocal endomicroscopy

Collagenous colitis is a form of microscopic colitis which has recently been recognised as an entity of its own, characterised by chronic watery diarrhoea of unknown aetiology. The diagnosis of collagenous colitis relies on histopathological examination of biopsy specimens from colorectal mucosa, which is usually of normal macroscopic appearance. The typical histological feature is diffuse thickening of the subepithelial collagen layer beneath the basement membrane and an unspecific chronic inflammatory infiltrate of the lamina propria.1–3 Recently, a confocal laser endomicroscope has been developed that is integrated into the distal tip of a conventional video endoscope. This confocal laser microscope (EC-3870CIFK; Pentax, Tokyo, Japan) was designed to enable subsurface imaging of living tissue during ongoing endoscopy and allows confocal microscopy in addition to standard video endoscopy. Images are generated by intravenously administered fluorescein sodium as the fluorescent contrast agent and an argon ion laser integrated into the system that generates an excitation wavelength of 488 nm.4,5 For the first time, we used …

The transcription factor IFN regulatory factor–4 controls experimental colitis in mice via T cell–derived IL-6

The proinflammatory cytokine IL-6 seems to have an important role in the intestinal inflammation that characterizes inflammatory bowel diseases (IBDs) such as Crohn disease and ulcerative colitis. However, little is known about the molecular mechanisms regulating IL-6 production in IBD. Here, we assessed the role of the transcriptional regulator IFN regulatory factor-4 (IRF4) in this process. Patients with either Crohn disease or ulcerative colitis exhibited increased IRF4 expression in lamina propria CD3+ T cells as compared with control patients. Consistent with IRF4 having a regulatory function in T cells, in a mouse model of IBD whereby colitis is induced in RAG-deficient mice by transplantation with CD4+CD45RB(hi) T cells, adoptive transfer of wild-type but not IRF4-deficient T cells resulted in severe colitis. Furthermore, IRF4-deficient mice were protected from T cell-dependent chronic intestinal inflammation in trinitrobenzene sulfonic acid- and oxazolone-induced colitis. In addition, IRF4-deficient mice with induced colitis had reduced mucosal IL-6 production, and IRF4 was required for IL-6 production by mucosal CD90+ T cells, which it protected from apoptosis. Finally, the protective effect of IRF4 deficiency could be abrogated by systemic administration of either recombinant IL-6 or a combination of soluble IL-6 receptor (sIL-6R) plus IL-6 (hyper-IL-6). Taken together, our data identify IRF4 as a key regulator of mucosal IL-6 production in T cell-dependent experimental colitis and suggest that IRF4 might provide a therapeutic target for IBDs.

Confocal laser endomicroscopy is a new imaging modality for recognition of intramucosal bacteria in inflammatory bowel disease in vivo

Background and objectives Interaction of bacteria with the immune system within the intestinal mucosa plays a key role in the pathogenesis of inflammatory bowel disease (IBD). The aim of the current study was to develop a fluorescein-aided confocal laser endomicroscopy (CLE) method to visualise intramucosal enteric bacteria in vivo and to determine the involved mucosal area in the colon and ileum in patients with ulcerative colitis (UC) and Crohns disease (CD). Methods Initially, E coli strains expressing enhanced green fluorescent protein (pEGFP) were endomicroscopically imaged in mice. In addition, ex vivo and in vivo imaging of fluorescent human enteric bacteria was performed to specify the distinct endomicroscopic appearance of enteral bacteria. Targeted mucosal biopsies towards endomicroscopic identifiable intramucosal bacteria and negative mucosal areas were prospectively obtained during colonoscopy and correlated with bench-top fluorescence microscopy (FISH) to prove the endomicroscopic visualisation of intramucosal bacteria. Finally, a retrospective analysis as well as a prospective study was performed in patients with UC and CD to confirm the presence and distribution of intramucosal bacteria within the gut. Results Confocal endomicroscopy was able to identify intramucosal pEGFP E coli in mice and strains of enteric microflora in the mucosa of humans. Using FISH as the gold standard, evaluation of 21 patients showed that CLE had a sensitivity of 89% and specificity of 100% to identify intramucosal bacteria. In a retrospective study, 113 patients with CD and UC had intramucosal bacteria significantly more often than 50 control patients (66% vs 60% vs 14%, p<0.001). This result was confirmed in a prospective study in which 10 patients with CD and 10 with UC had a significantly wider distribution of involvement with intramucosal bacteria in the colon and terminal ileum compared with 10 controls (85.2% vs 75.9% vs 16.8%, p<0.0001). Conclusions CLE is a new tool that can image intramucosal bacteria in vivo in patients with IBD. Intramucosal bacteria are found more frequently and with a wider distribution in patients with IBD than in patients with a normal intestine.

Simultaneous confocal laser endomicroscopy and chromoendoscopy with topical cresyl violet

BACKGROUND Confocal laser endomicroscopy (CLE) has been shown to reliably predict histology during ongoing endoscopy. To unmask lesions for CLE, chromoendoscopy has been mandated. Usually fluorescein then serves as a contrast agent for CLE, but it does not allow direct nuclear visualization, must be injected, leads to a transient skin discoloration, and may have allergic side effects. OBJECTIVE To establish a single topical dye, cresyl violet (CV), for simultaneous chromoendoscopy and in vivo CLE of the lower GI tract. DESIGN Animal preclinical study, prospective clinical trial. SETTING Mainz University Clinic (tertiary care center). PATIENTS, METHODS, AND INTERVENTIONS: To establish the staining characteristics and optimal concentration of CV, the ileum and colon of 7 BL6 mice were stained with CV (0.1%-2%), and in vivo confocal imaging was performed with FIVE1. In a subsequent clinical trial, 67 sites in 36 patients were topically stained with CV 0.13%, and subsurface serial images were generated at different depths with an endomicroscope. MAIN OUTCOME MEASUREMENTS Prediction of histology according to the Mainz confocal classification and nuclear visualization with topical CV. RESULTS Endomicroscopy with topical CV yielded (sub-)cellular details of normal mucosa, and regenerative and neoplastic changes at variable imaging depths in high resolution comparable to those with intravenous fluorescein. By cytoplasmic enrichment of CV, nuclear morphology could be negatively visualized. Reliable differentiation of nonneoplastic versus neoplastic changes during ongoing endoscopy and a high interobserver agreement based on the microscopic images generated in vivo could be achieved. LIMITATIONS Single-center study, nonrandomized, limited number of patients. CONCLUSIONS CV can be applied topically and allows simultaneous chromoendoscopy and endomicroscopy with accurate prediction of histology with visualization of nuclear morphology. It may therefore be a single-agent alternative to chromoendoscopy and fluorescein in endomicroscopy.

Near-infrared confocal imaging during mini-laparoscopy: a novel rigid endomicroscope with increased imaging plane depth.

BACKGROUND & AIMS Histopathology is the gold standard in the diagnosis of liver diseases but may be complicated by sampling error and bleeding. Confocal laser endomicroscopy (CLE) is a novel imaging modality providing in vivo histology. Previous studies using CLE for liver microscopy suffered from limited imaging depth using blue laser light and fluorescein. The aim of the current study was to evaluate a novel near-infrared (NIR) light probe with indocyanine green (ICG) contrast for in vivo confocal imaging of the human liver during mini-laparoscopy. METHODS The hand-held rigid laparoscopy probe (diameter 6.3mm) used a 780 nm diode laser. Subsurface images at different depths with < 1 microm lateral resolution were generated in real time by gently placing the sterile probe onto the liver in 22 patients under conscious analgo-sedation. Targeted biopsy was performed for histopathological correlation. RESULTS Maximum imaging depth was >350 microm. Typical aspects of normal liver architecture and liver diseases, such as nuclei, sinusoids, fibrous tissue, fatty inclusions, and bile ducts but not inflammation could be visualized at high resolution. The presence of steatosis and fibrosis was predicted correctly in 81% and 90%, respectively. No liver damage or severe adverse events occurred. CONCLUSIONS For the first time, ICG-augmented confocal mini-laparoscopy with a novel NIR probe allowed in vivo microscopy of human liver diseases, and with deeper imaging compared to the so far available blue laser systems. Such an increase of imaging depth could potentially also be used for submucosal imaging of the GI tract.

High-definition endoscopy with i-Scan and Lugol's solution for more precise detection of mucosal breaks in patients with reflux symptoms.

BACKGROUND AND STUDY AIMS Patients with gastroesophageal reflux disease are subdivided into non-erosive (NERD) and erosive reflux disease (ERD). The newly available EPKi processor enables high-definition resolution above HDTV standard (HD+). The aim of the study was to test the efficacy of HD+ esophagogastroduodenoscopy alone and in conjunction with i-Scan (newly developed postprocessing digital filter) and chromoendoscopy (Lugols solution) for differentiation of reflux patients. METHODS The distal esophagus of patients with heartburn was inspected with three imaging modalities. HD+ was followed by i-Scan and 15-mL Lugols solution (1.5 %). The esophagus was evaluated for mucosal breaks (Los Angeles Classification [LA]). Small visible changes were also characterized, and targeted biopsies were performed. End points of the study were the presence and grade of esophagitis and the number of circumscribed changes. RESULTS A total of 50 patients were included (female 29; mean age 54.7 years). HD+ identified nine patients with mucosal breaks (LA 7A; 2C), i-Scan was able to detect 12 patients (LA 8A; 2B; 2C; 0D) ( P = n. s.) and chromoendoscopy identified 25 patients (LA 16A; 7B; 1C, 1D) ( P < 0.01). Furthermore, a higher grade of esophagitis was recognized by using i-Scan and Lugols solution in 19 patients. The number of circumscribed lesions could be increased from 21 (HD+) to 58 (i-Scan) ( P < 0.01), and up to 85 after Lugol spraying ( P < 0.01). CONCLUSIONS Lugols solution in conjunction with HD+ endoscopy significantly improves the identification of patients with esophagitis and reduces misclassification. The i-Scan filter and chromoendoscopy help to identify reflux-associated lesions.

IRF4 regulates IL-17A promoter activity and controls RORγt-dependent Th17 colitis in vivo.

Background: The transcription factor IRF4 is involved in several T‐cell‐dependent chronic inflammatory diseases. To elucidate the mechanisms for pathological cytokine production in colitis, we addressed the role of the IRF transcription factors in human inflammatory bowel disease (IBD) and experimental colitis. Methods: IRF levels and cytokine production in IBD patients were studied as well as the effects of IRF4 deficiency in experimental colitis. Results: In contrast to IRF1, IRF5, and IRF8, IRF4 expression in IBD was augmented in the presence of active inflammation. Furthermore, IRF4 levels significantly correlated with IL‐6 and IL‐17 mRNA expression and to a lesser extent with IL‐22 mRNA expression in IBD. To further explore the role of IRF4 under in vivo conditions, we studied IRF4‐deficient and wildtype mice in experimental colitis. In contrast to DSS colitis, IRF4 deficiency was protective in T‐cell‐dependent transfer colitis associated with reduced ROR&agr;/&ggr;t levels and impaired IL‐6, IL‐17a, and IL‐22 production, suggesting that IRF4 acts as a master regulator of mucosal Th17 cell differentiation. Subsequent mechanistic studies using database analysis, chromatin immunoprecipitation, and electrophoretic mobility shift assays identified a novel IRF4 binding site in the IL‐17 gene promoter. Overexpression of IRF4 using retroviral infection induced IL‐17 production and IL‐17 together with IL‐6 induced ROR&ggr;t expression. Conclusions: IRF4 can directly bind to the IL‐17 promotor and induces mucosal ROR&ggr;t levels and IL‐17 gene expression thereby controlling Th17‐dependent colitis. Targeting of this molecular mechanism may lead to novel therapeutic approaches in human IBD. (Inflamm Bowel Dis 2011;)