Francisca Rius-Diaz

Clinical applicability of in vivo reflectance confocal microscopy for the diagnosis of actinic keratoses.

BACKGROUND In vivo reflectance confocal microscopy (RCM) has been used for evaluation of the morphologic features of nonmelanoma skin cancer. The application of RCM for diagnosis of basal cell carcinoma has been reported; however, the evaluation of actinic keratoses (AKs) has only been the subject of preliminary studies. STUDY GOAL The goal of this study was to evaluate the applicability of RCM in the diagnosis of AK in correlation with routine histology. MATERIALS AND METHODS Forty-four Caucasians with a minimum of one AK participated in this study. Evaluation consisted of clinical examination, RCM, and routine histology, including a total of 46 AKs in the final analysis. Ten normal skin sites served as controls. RCM features of AK included parakeratosis, architectural disarray, and keratinocyte pleomorphism. Following blinded evaluations, sensitivity/specificity, kappa analysis, and Spearmans correlation were performed on all parameters. RESULTS Sensitivity/specificity values of RCM features ranged from 80% to 98.6%. The presence of architectural disarray and cellular pleomorphism appeared to be the best predictor of AK. CONCLUSION In summary, RCM may be a promising technology for the noninvasive detection of AK and as adjunct tool to clinical diagnosis and monitoring. However, the preliminary nature of this study warrants further investigations.

Peritumoral clefting in basal cell carcinoma: correlation of in vivo reflectance confocal microscopy and routine histology.

Background: Histopathologically, basal cell carcinoma (BCC) is characterized by basaloid tumor nodules of varying size showing peripheral palisading of cells and nuclei, and separation from surrounding stroma by optically empty appearing clefts. These are usually regarded as an artifact, occurring during routine tissue processing. Recently, reflectance confocal microscopy (RCM) has been applied for noninvasive, in vivo evaluation of BCC. Besides other features, small areas of low refractility separating tumor islands from the surrounding tissue can be observed in vivo, suggesting that the presence of amorphous material like mucin might be the causal factor for these clefts.

Imaging mitochondrial dynamics in human skin reveals depth-dependent hypoxia and malignant potential for diagnosis

In vivo, label-free evaluation of mitochondrial function in human skin epithelia noninvasively differentiates healthy from cancerous tissues. Mitochondria expose tumors’ misbehavior Mitochondria, the powerhouses inside cells, change their shape and function according to the needs of the cell. Such mitochondrial dynamics go awry in a variety of human diseases, and if these abnormalities are detected early, they can be useful for diagnosis and fast treatment. Now, Pouli et al. show that such signs of mitochondrial dysfunction can be spotted in living human skin with two-photon–excited fluorescence by monitoring the mitochondrial metabolic coenzyme NADH. The authors detected rapid changes in mitochondrial organization in response to oxygen deprivation and successfully distinguished healthy skin from two common skin cancers—basal cell carcinoma and melanoma. Active changes in mitochondrial structure and organization facilitate cellular homeostasis. Because aberrant mitochondrial dynamics are implicated in a variety of human diseases, their assessment is potentially useful for diagnosis, therapy, and disease monitoring. Because current techniques for evaluating mitochondrial morphology are invasive or necessitate mitochondria-specific dyes, their clinical translation is limited. We report that mitochondrial dynamics can be monitored in vivo, within intact human skin by relying entirely on endogenous two-photon–excited fluorescence from the reduced metabolic coenzyme nicotinamide adenine dinucleotide (NADH). We established the sensitivity of this approach with in vivo, fast temporal studies of arterial occlusion-reperfusion, which revealed acute changes in the mitochondrial metabolism and dynamics of the lower human epidermal layers. In vitro hypoxic-reperfusion studies validated that the in vivo outcomes were a result of NADH fluorescence changes. To demonstrate the diagnostic potential of this approach, we evaluated healthy and cancerous human skin epithelia. Healthy tissues displayed consistent, depth-dependent morphological and mitochondrial organization patterns that varied with histological stratification and intraepithelial mitochondrial protein expression. In contrast, these consistent patterns were absent in cancerous skin lesions. We exploited these differences to successfully differentiate healthy from cancerous tissues using a predictive classification approach. Collectively, these results demonstrate that our label-free, automated, near real-time assessments of mitochondrial organization—relying solely on endogenous contrast—could be useful for accurate, noninvasive in vivo diagnosis.

Prognostic value of serum angiogenic activity in colorectal cancer patients

Angiogenesis, resulting from an imbalance between angiogenic activator factors and inhibitors, is required for tumour growth and metastasis. The determination of the circulating concentration of all angiogenic factors (activators and inhibitors) is not feasible at present. We have evaluated diagnostic and prognostic values of the measurement of serum angiogenic activity in colorectal carcinoma (CRC) patients. Serum proliferative activity (PA) on human umbilical vein endothelial cells (HUVEC) in vitro, and serum vascular endothelial growth factor (VEGF) levels were determined by ELISA in 53 patients with primary CRC, 16 subjects with non‐neoplastic gastrointestinal disease (SC) and 34 healthy individuals. Data were compared with clinical outcome of the patients. Although serum from CRC patients significantly increased the PA of HUVEC, compared to culture control (HUVEC in medium + 10% foetal bovine serum (FBS); P < 0.001); our results indicate that serum PA in CRC patients was similar to that of SC or healthy individuals. There was no correlation between serum PA and circulating VEGF concentrations. Surgery produced a decrease of PA at 8 hrs after tumour resection in CRC patients compared to pre‐surgery values (186 ± 47 versus 213 ± 41, P < 0.001). However, an increase in serum VEGF values was observed after surgery (280 [176–450] versus 251 [160–357] pg/ml, P = 0.004). Patients with lower PA values after surgery showed a worse outcome that those with higher PA values. Therefore, this study does not support a diagnostic value for serum angiogenic activity measured by proliferative activity on HUVEC but suggests it could have a prognostic value in CRC patients.

Consistency and distribution of reflectance confocal microscopy features for diagnosis of cutaneous T cell lymphoma

Reflectance confocal microscopy (RCM) represents a noninvasive imaging technique that has previously been used for characterization of mycosis fungoides (MF) in a pilot study. We aimed to test the applicability of RCM for diagnosis and differential diagnosis of MF in a clinical study. A total of 39 test sites of 15 patients with a biopsy-proven diagnosis of either MF, parapsoriasis, Sézary syndrome, or lymphomatoid papulosis were analyzed for presence and absence of RCM features of MF. Cochran and Chi(2) analysis were applied to test the concordance between investigators and the distribution of RCM features, respectively. For selected parameters, the Cochran analysis showed good concordance between investigators. Inter-observer reproducibility was highest for junctional atypical lymphocytes, architectural disarray, and spongiosis. Similarly, Chi(2) analysis demonstrated that selected features were present at particularly high frequency in individual skin diseases, with values ranging from 73% to 100% of all examined cases.

The Effect of Peripapillary Detachment on Retinal Nerve Fiber Layer Measurement by Spectral Domain Optical Coherence Tomography in High Myopia

Purpose: The aim of this study was to investigate the repercussions of peripapillary detachment on retinal nerve fiber layer (RNFL) measurements in patients with highly myopic eyes. Methods: A total of 244 highly myopic eyes underwent a complete ophthalmologic examination that included optical coherence tomography (OCT) to analyze the peripapillary retina and RNFL thickness. Based on the OCT findings, patients were grouped as follows: group A: eyes with a peripapillary intrachoroidal cavitation (PIC); group B: eyes with a peripapillary neurosensory retinal detachment (PNRD), and group C: eyes without a peripapillary detachment. Results: The OCT scans identified a peripapillary detachment in 42 eyes (17.21%). Out of these 42 eyes, 22 showed PIC (52.38%; group A) and 20 had a PNRD (47.62%; group B). The average overall RNFL thickness in groups A, B and C was 74.11 ± 10.88, 88.26 ± 25.72 and 72.75 ± 16.24 μm, respectively (ANOVA test, p = 0.00). Conclusion: Eyes with a PNRD had a significantly greater average RFNL thickness than those without peripapillary detachment in pathologic myopia due to a misidentification of the outer profile of the RFNL. This fact makes the interpretation of RNFL thickness in highly myopic eyes more challenging.

Analysis of peripapillary choroidal thickness in non-arteritic anterior ischaemic optic neuropathy

Purpose To analyse peripapillary choroidal thickness (PCT) in non-arteritic ischaemic optic neuropathy (NAION). Methods 28 patients diagnosed with NAION (37 affected and 19 unaffected eyes) and 38 disease-free control individuals (38 eyes) were analysed using enhanced-depth imaging of spectral-domain optical coherence tomography. A vertical and a horizontal raster scan centred on the optic nerve were obtained per eye. PCT was measured at the superior, inferior, nasal and temporal quadrants from the posterior edge of the retinal pigment epithelium to the choroid–sclera junction at 500u2005μm intervals up to 2000u2005μm away from the optic nerve. Statistical analysis was used to compare average PCT and to correlate PCT with other ocular and systemic parameters. Results Mean PCT in NAION eyes and control group was 148.18±42.68u2005μm and 182.90±59.81u2005μm, respectively (p=0.005). Except for inferior PCT (p=0.158), superior, nasal and temporal PCT were significantly thinner in the NAION eyes than in the control group (p=0.006, 0.002 and 0.046). Thinner PCT, adjusted for refractive error, was associated with the diagnosis of NAION (p=0.048). Similarly, unaffected contralateral eyes showed a significant thinner PCT compared with the control group (p=0.024). Diagnosis of NAION was negatively associated with PCT in NAION eyes (p=0.008; OR 0.98; 95% CI 0.97 to 0.99) and in their contralateral unaffected eyes (p=0.043; OR 0.98; 95% CI 0.97 to 0.99). Conclusions Eyes affected by NAION and contralateral unaffected eyes showed significantly thinner PCT compared with disease-free control eyes after adjusting for refractive error.

Grosor coroideo macular en la neuropatía óptica isquémica anterior no arterítica

OBJECTIVEnTo analyse macular choroidal thickness (MCT) in non-arteritic ischaemic optic neuropathy (NAION).nnnMATERIALS AND METHODSnAn analysis was made on 22 patients diagnosed with NAION (22 eyes) and 42 healthy controls (42 eyes) using enhanced-depth imaging of spectral-domain optical coherence tomography. A horizontal raster scan centred on the fovea was obtained per eye 3 months after the onset of NAION. Three measurements of MCT were obtained from the posterior edge of the retinal pigment epithelium to the choroid-sclera junction at 500μm intervals. Statistical analysis was used to compare the mean MCT and to correlate MCT with other ocular and systemic parameters.nnnRESULTSnExcept for refractive error (P=.01), there were no statistically significant differences between both groups in axial length (P=.53), age (P=.88) and other epidemiological and ocular parameters. Mean MCT in NAION eyes and control group was 236.21±63.29μm and 269.13±52.28, respectively. Mean MCT was significantly thinner in NAION eyes than in healthy eyes (P=.03). Thinner MCT, adjusted for refractive error, was associated with the diagnosis of NAION (P=.04).nnnCONCLUSIONSnEyes affected by NAION showed significantly thinner MCT compared with healthy control eyes after adjusting for refractive error.