In vitro simulation of the first technique for non-invasive measurement of volumetric ophthalmic artery blood flow in humans.

BACKGROUND AND OBJECTIVE: To evaluate the accuracy, reproducibility, and variability of volumetric flow measurements taken by color Doppler imaging ultrasound, using an in vitro "phantom" model to simulate the ophthalmic artery. MATERIALS AND METHODS: An agar flow phantom with two wall-less lumens was constructed to simulate the ophthalmic artery. Velocity and volumetric flow measurements were taken for various flow rates and ultrasound probe positions. The measurements were analyzed for accuracy, reproducibility, and variability. RESULTS: Velocity measurements were more accurate than flow measurements (8 of 24 vs 3 of 24 accurate trials). The average coefficient of variation for volumetric blood flow was 11.4% (n = 120). Volumetric flow significantly correlated with velocity (R(2) = 0.408, n = 600, P < .001). The highest correlation was achieved using the large lumen with the probe held at 75 degrees , offset to the flow (R(2) = 0.862, n = 75). CONCLUSION: Based on an in vitro model, non-invasive color Doppler imaging recordings of volumetric flow measurements in the ophthalmic artery significantly correlated with velocity and higher correlations were found using the larger lumens, although the data showed a lack of high accuracy in measurements of flow and velocity.

Interobserver repeatability of heidelberg retinal flowmetry using pixel-by-pixel analysis.

PURPOSE: To determine the interobserver reproducibility of Heidelberg retinal flowmeter (HRF) blood flow measurements using independently selected study areas for pixel-by-pixel analysis. PATIENTS AND METHODS: Blood flow measurements were performed on 257 scans from 15 patients, 14 of whom had glaucoma or ocular hypertension. HRF was used to record capillary perfusion in a 2560x640 mum area of the supratemporal peripapillary region and pixel-by-pixel analysis was performed from an area adjacent to the optic disc with a minimum of 1600 pixels. Each observer independently selected the area for analysis. The percentage of pixels with <1 arbitrary unit of flow (no flow) and 10, 25, 50, 75, and 90th percentiles of flow values was calculated. Interobserver variability was assessed by estimating the intraclass correlation coefficient (ICC) and its 95% confidence interval. Bland-Altman plots of the difference between the 2 physicians versus the average of the 2 physicians for each outcome were created. RESULTS: ICC was 0.79 (range: 0.74 to 0.83) for mean flow values. For 0, 10, 25, 50, 75, and 90th percentiles of flow, the ICC was 0.67 (0.60 to 0.73), 0.74 (0.68 to 0.79), 0.82 (0.78 to 0.86), 0.85 (0.82 to 0.88), 0.85 (0.81 to 0.88), and 0.77 (0.72 to 0.82), respectively. Zero flow pixels had a nonsignificant mean difference between observers (P=0.542), whereas the remainder of the flow values demonstrated significant mean differences. CONCLUSIONS: This study demonstrates that independent observers can review high-quality HRF scans and may produce different absolute values while retaining strong consistency of agreement when independently selecting areas for analysis using the pixel-by-pixel method.

The circadian variations in systemic blood pressure, ocular perfusion pressure, and ocular blood flow: risk factors for glaucoma?

Intraocular pressure, a major risk factor for glaucoma, is known to vary throughout the day, yet glaucoma continues to progress in some patients despite it being well controlled. It is important to understand how other glaucomatous risk factors are affected by circadian variations. The purpose of this review is to analyze the literature concerning circadian variations in systemic blood pressure, ocular perfusion pressure, and ocular blood flow and to identify consensus findings regarding their impact on glaucoma. This review suggests that nonphysiologic nocturnal blood pressure dipping and wider circadian fluctuations in ocular perfusion pressure are linked with the development and progression of glaucoma. No consensus concerning circadian variations in ocular blood flow exists in the current literature, and future investigations of nocturnal changes in blood flow and glaucoma progression are required.

Enhanced TNF-alpha-induced apoptosis in Fanconi anemia type C-deficient cells is dependent on apoptosis signal-regulating kinase 1.

Fanconi anemia (FA) is a chromosomal instability disorder characterized by progressive bone marrow failure. Experimental evidence suggests that enhanced oxidant and myelosuppressive cytokine-mediated apoptosis of hematopoietic stem and progenitor cells contributes to the pathogenesis of marrow failure in FA. However, the molecular mechanisms responsible for the apoptotic phenotype in hematopoietic cells are incompletely understood. Recent data in Fancc-/- murine embryonic fibroblasts (MEFs) implicate increased oxidant-induced apoptotic signaling through the redox-dependent protein, apoptosis signal-regulating kinase 1 (Ask1). Here, we examined whether altered Ask1 signaling participated in the proapoptotic phenotype of primary Fancc-/- MEFs and hematopoietic progenitors treated with the myelosuppressive cytokine tumor necrosis factor-alpha (TNF-alpha). Our data indicate that TNF-alpha induces hyperactivation of Ask1 and the downstream effector p38 in Fancc-/- MEFs. In addition,Ask1 inactivation in Fancc-/- MEFs and hematopoietic progenitors restored survival to wild-type (WT) levels in the presence of TNF-alpha. Furthermore, targeting the Ask1 pathway by using either antioxidants or a p38 inhibitor protected Fancc-/- MEFs and c-kit+ cells from TNF-alpha-induced apoptosis. Collectively, these data argue that the predisposition of Fancc-/- hematopoietic progenitors to apoptosis is mediated in part through altered redox regulation and Ask1 hyperactivation.

Cooperative interactions between flagellin and SopE2 in the epithelial interleukin-8 response to Salmonella enterica serovar typhimurium infection.

Flagellin is an important stimulus for epithelial interleukin-8 (IL-8) secretion because of its ability to activate Toll-like receptor 5 (TLR5). SopE2, a Salmonella guanine nucleotide exchange factor (GEF), is also involved in intestinal inflammation. To clarify the proinflammatory mechanisms of these proteins, we examined their effects on IL-8 secretion and intracellular signaling in T84 epithelial cells. A Salmonella strain lacking SopE2 (and its homolog SopE) induced lower levels of IL-8 than the wild type and exhibited reduced activation of mitogen-activated protein kinases (MAPKs). Overexpression of wild-type SopE2 in this strain restored MAPK activation and augmented IL-8 production, whereas a mutant lacking GEF activity failed to increase IL-8 expression. Additional effects on signaling were demonstrated in transient transfection experiments, in which SopE2 enhanced the ability of TRAF6, a signal transducer downstream of TLR5, to activate the NF-kappaB transcription factor in 293 cells. Flagellin was also found to be required for IL-8 induction in T84 cells. In its absence, the ability of SopE2 overexpression to increase IL-8 secretion was impaired. Part of this impairment was related to the decreased motility of the flagellin-deficient strain, but lack of flagellin also affected translocation of SopE2 into the infected cells. Our results indicate that flagellin and SopE2 interact functionally at multiple levels to increase IL-8 secretion by epithelial cells-flagellin facilitating the translocation of SopE2, and SopE2 enhancing signaling pathways activated by flagellin. These observations offer a mechanistic explanation for the involvement of these proteins in the pathogenesis of Salmonella-induced gastroenteritis.