Prospective observational study of conjunctival scarring after phacoemulsification.

PURPOSE: To examine whether anterior segment-optical coherence tomography (AS-OCT) can detect conjunctival scarring after transscleral phacoemulsification and whether temporal transscleral phacoemulsification causes scarring in the superior conjunctiva. METHODS: Transscleral phacoemulsification was performed in the superior conjunctiva (superior incision group) or the temporal conjunctiva (temporal incision group). Anterior segment-optical coherence tomography (AS-OCT) images of the superior conjunctiva were obtained before and after surgery. We quantified the thickness of the conjunctiva and preservation rates of the borderlines among the subconjunctival layers. The relationship between the AS-OCT images and histology was evaluated in rabbit eyes with phacoemulsification. RESULTS: Each group comprised 25 patients. At ≤1 month after surgery, the superior conjunctiva was significantly thicker in the superior incision group than the temporal incision group (1 day, 7 days, and 1 month after surgery; p < 0.0001, p < 0.0001, and p < 0.001 respectively); however, there were no significant differences between the two groups at ≥2 months. The borderline of the conjunctival stroma/Tenon's capsule and the borderline of the Tenon's capsule/sclera were preserved significantly better in the temporal incision group after surgery (p < 0.0001). The cell densities in the conjunctiva of the rabbit temporal incision group were unchanged after surgery, whereas the rabbit superior incision group had significantly more neutrophils (p = 0.0001) and myofibroblasts (p < 0.0001) in the superior conjunctiva than the temporal incision group. CONCLUSIONS: Anterior segment-optical coherence tomography (AS-OCT) images can detect conjunctival scarring after transscleral phacoemulsification. The layer structures in the superior conjunctiva are unaffected by temporal transscleral incision.

Double-Filtration Plasmapheresis plus Interferon-ß for HCV-1b Patients with Non-Sustained Virological Response to Previous Combination Therapy.

BACKGROUND AND AIMS: Double-filtration plasmapheresis (DFPP) together with interferon (IFN) administration produces a substantial reduction in the viral load during the early stages of treatment. METHODS: Based on their responses to previous pegylated IFN and ribavirin (PEG-IFN/RBV) therapy, 20 patients were divided into null virological response (NVR; n = 12) and relapse (n = 8) groups. DFPP was used in combination with IFN-ß/RBV with subsequent administration of PEG-IFN-α2a/RBV therapy (DFPP + IFN-ß/RBV then PEG-IFN/RBV). Early viral dynamics was assessed, focusing especially on complete early virological response (cEVR) associated with sustained virological response. Additionally, the interleukin 28B gene, the IFN/RBV resistance-determining region, the IFN sensitivity-determining region and the core regions were analyzed. RESULTS: Rapid virological response was achieved in 0% (0/12) of NVR and in 75% (6/8) of relapse patients, with a significant difference between the two groups (p = 0.001). Similarly, cEVR was achieved in 8% (1/12) of NVR and in 88% (7/8) of relapse patients, with a significant difference between the two groups (p = 0.037). By multivariate logistic regression analysis, interleukin-28B major was a significant determiner of cEVR (odds ratio = 24.19, p = 0.037). CONCLUSION: DFPP + IFN-ß/RBV then PEG-IFN/RBV therapy is indicated more for relapse than for NVR patients.

Establishment of framework of the cortical area is influenced by Otx1.

Subcortical projection from layer 5 neurons is the major cortical output. A transcription factor, Otx1, which is expressed in the layer 5 subcortical projection neurons in the visual cortex, was reported to be responsible for the establishment of visual area-specific layer 5 subcortical projections by inducing the sensorimotor cortex to adopt a visual cortex identity. However, we here demonstrate that the area of corticospinal neurons shifted caudo-medially in the Otx1-null mice of which cortex is 9 tenths in size compared with that of the wild-type littermates, while the whole visual cortex did not convert to the sensorimotor cortex in the absence of Otx1. This suggests that Otx1 is not crucial for the development of visual cortex identity but for the determination of the proportion of cortical areas to the whole neocortex.

Relationship of BRAF mutation, morphology, and apoptosis in early colorectal cancer.

BACKGROUND AND AIMS: Many investigators have reported flat and depressed lesions as a new type of precursor of colorectal cancer. In our previous study, we determined that mutations in the BRAF gene may contribute to colorectal carcinogenesis by inhibiting apoptosis. However, the relationship among BRAF mutations, morphology and apoptosis in early colorectal cancer has not been clear. Therefore, gene alternation, morphology, and apoptosis in early colorectal cancer were investigated. MATERIALS AND METHODS: Forty-five flat and depressed early colorectal cancer samples and 43 polypoid early colorectal cancer samples were analyzed. Mutations in the BRAF gene and the K-ras gene were examined by direct sequence analysis, and proliferative activity and induction of apoptosis were evaluated using immunohistochemical examination. RESULTS FINDINGS: BRAF mutations were found in 5 (11.1%) of 45 flat and depressed early colorectal cancer samples. No BRAF alteration was found in polypoid early colorectal cancer samples. Mutations in the K-ras gene were detected in 13 (30.2%) of 43 polypoid early colorectal cancer samples. The rate of submucosal invasion of the samples with BRAF mutations was significantly higher than that of the samples with K-ras mutations (p<0.05). INTERPRETATION/CONCLUSIONS: BRAF and K-ras mutations were independent factors that influenced morphology in early colorectal cancer. In this study, the relationship between BRAF mutation and apoptosis is not so clear, but BRAF mutations and inhibition in apoptosis may play an important role in the developmental process of flat and depressed early colorectal cancer.

Real-time in vivo virtual histology of colorectal lesions when using the endocytoscopy system.

BACKGROUND: The histological findings of GI lesions are based on light-microscopic examination of H&E-stained thin-slice specimens. Recently, a concept of optical biopsy has been advocated. A study of the observation of colorectal lesions using endocytoscopy to obtain real-time histological images in vivo during endoscopy was performed. DESIGN: Prospective study. AIM: To evaluate the usefulness of optical biopsy of colorectal lesions with the endocytoscopy (E-C) system. PATIENTS: The subjects were 113 consecutive patients who underwent a complete colonic examination, from April 2003 to March 2004, performed by a single colonoscopist. SETTING: Digestive Disease Center of Showa University Northern Yokohama Hospital. RESULTS: With E-C, it was possible to observe lesions at the cellular level and evaluate cellular atypia in addition to structural atypia in vivo. The correlation was statistically significant between the endocytoscopic diagnosis and the histological diagnosis. LIMITATIONS: The endocytoscope had to be touched to the target colonic glands. CONCLUSIONS: It was possible to distinguish neoplastic from non-neoplastic lesions, and also possible to distinguish invasive cancer from adenoma. "Ultra-high" magnifying endoscopy, the E-C system, provides real-time histological images in vivo, which correspond well with those of H&E-stained microscopic images.

BRAF mutation associated with dysregulation of apoptosis in human colorectal neoplasms.

To understand the role of BRAF dysfunction in the carcinogenesis and progression/development of colorectal tumors, the authors investigated genetic alterations in the BRAF gene in human colorectal neoplasms as well as the effects of an RAS inhibitor in BRAF-mutant cells. Seven colon cancer cell lines and 116 colorectal tumors (34 adenomas and 82 adenocarcinomas) were analyzed. Genetic alterations in the BRAF and K-ras genes were examined using polymerase chain reaction-single strand conformation polymorphism and direct sequencing analyses. The growth-inhibitory and apoptosis-inducing effects of the FTI-277 RAS inhibitor in colon cancer cell lines were analyzed as well. An immunohistochemical study was also performed to investigate the correlations between the clinicopathologic parameters involved in the Ki-67 labeling index and the number of apoptotic bodies in tumor cells. FTI-277 did not suppress the proliferation of BRAF-mutant cells (WiDr and TCO), but remarkably inhibited the growth of K-ras mutant cells (LoVo). Interestingly, LoVo cells underwent apoptosis by FTI-277 in a dose-dependent manner, whereas WiDr cells were resistant to this agent. In tumor samples, BRAF mutations were found in 1 (3.0%) of 33 adenomas and 6 (7.2%) of 83 adenocarcinomas. No tumor exhibited mutations in both the BRAF and K-ras genes. Neither BRAF nor K-ras mutations correlated with the Ki-67 labeling index immunohistochemically. However, the number of apoptotic bodies was significantly decreased in the BRAF-mutant tumors. Mutation in the BRAF gene may contribute to colorectal carcinogenesis by upregulating the antiapoptotic role of the RAS/RAF/MEK/ERK pathway.

Relationship between gastric ulcer and Helicobacter pylori VacA detected in gastric juice using bead-ELISA method.

BACKGROUND: VacA is an important pathogenetic factor produced by Helicobacter pylori. VacA has often been detected in supernatants of liquid cultures or lysates of whole bacterial cells. However, no studies have ever tried to assay VacA produced in the human stomach. We applied a very sensitive and simple method, bead-ELISA, to detect VacA in gastric juice. MATERIALS AND METHODS: Forty-eight H. pylori-positive patients (16 nonulcer dyspepsia, 16 gastric ulcer, and 16 duodenal ulcer) and four H. pylori-negative nonulcer dyspepsia patients had endoscopy performed and gastric juice were aspirated. Polystyrene beads coated with the antibody to VacA, were used in this bead-ELISA method. The nucleotide sequences of vacA in the signal and middle regions were investigated. RESULTS: Of the 48 samples that were positive for H. pylori, 21 [43.8%] were found to be VacA positive in gastric juice. The average and maximum concentrations of detected VacA in gastric juice were 143.2 +/- 216.5 and 840 pg/ml, respectively. The average density of VacA from gastric ulcer patients (227.5 +/- 276.7 pg/ml) was higher than that found in nonulcer dyspepsia (51.8 +/- 39.8 pg/ml) and duodenal ulcer (49.2 +/- 21.5 pg/ml) patients. There was no relationship between VacA in gastric juice and vacA genotype. CONCLUSIONS: VacA in gastric juice could be directly detected by bead-ELISA. In this study, the diversity of disease outcome was associated with not the quality but the quantity of VacA. Therefore, not only the quality but also the quantity of VacA is important etiological factors in the pathogenesis of mucosal damage.

Effect of Helicobacter pylori-induced cyclooxygenase-2 on gastric epithelial cell kinetics: implication for gastric carcinogenesis.

BACKGROUND: Cyclooxygenase (COX)-2 induced by Helicobacter pylori is thought to enhance gastric carcinogenesis by affecting the maintenance of epithelial homeostasis. MATERIALS AND METHODS: Gastric biopsies from 160 subjects, 97 with nonulcer dyspepsia (47 H. pylori negative, 50 H. pylori positive) and 63 with gastric cancer were examined immunohistochemically for COX-2 expression, cell proliferation and apoptotic indices. RESULTS: COX-2 expression in corpus was significantly higher in H. pylori positive than in negative non-ulcer dyspepsia (NUD) (p <.05). Regardless of site, gastric cancer subjects had higher COX-2 expression in both antrum and corpus compared with H. pylori negative and positive NUD (p <.005). Proliferation was higher in cancer and H. pylori positive than in negative NUD (p <.0001). Moreover, cancer had enhanced proliferation than H. pylori positive NUD in corpus greater (p =.0454) and antrum lesser (p =.0215) curvatures. Apoptosis was higher in H. pylori positive than in negative NUD (p <.05). However, both had a higher index than the cancer subjects (p <.0001). Apoptosis : proliferation ratio was higher in corpus of H. pylori negative than in positive NUD in greater (p =.0122) and lesser (p =.0009) curvatures. However, both had a higher A:P ratio than cancer cases (p =.0001). A negative correlation between COX-2 expression and A:P ratio was found in corpus greater (r = -.176, p =.0437) and lesser (r = -.188, p =.0312) curvatures. CONCLUSION: The expression of COX-2 is associated with disruption in gastric epithelial kinetics and hence may play a role in gastric carcinogenesis.