Expansion of Human Limbal Epithelial Stem/Progenitor Cells Using Different Human Sera: A Multivariate Statistical Analysis

Transplantation of human cultured limbal epithelial stem/progenitor cells (LESCs) has demonstrated to restore the integrity and functionality of the corneal surface in about 76% of patients with limbal stem cell deficiency. However, there are different protocols for the expansion of LESCs, and many of them use xenogeneic products, being a risk for the patients’ health. We compared the culture of limbal explants on the denuded amniotic membrane in the culture medium—supplemental hormone epithelial medium (SHEM)—supplemented with FBS or two differently produced human sera. Cell morphology, cell size, cell growth rate, and the expression level of differentiation and putative stem cell markers were examined. Several bioactive molecules were quantified in the human sera. In a novel approach, we performed a multivariate statistical analysis of data to investigate the culture factors, such as differently expressed molecules of human sera that specifically influence the cell phenotype. Our results showed that limbal cells cultured with human sera grew faster and contained similar amounts of small-sized cells, higher expression of the protein p63α, and lower of cytokeratin K12 than FBS cultures, thus, maintaining the stem/progenitor phenotype of LESCs. Furthermore, the multivariate analysis provided much data to better understand the obtaining of different cell phenotypes as a consequence of the use of different culture methodologies or different culture components.Research study supported by grants from Mutua Madrileña Foundation (FMM11/02), Basque Foundation for Health Research and Innovation BIOEF (BIO14/TP/002), National Eye Institute (R01EY021797), California Institute for Regenerative Medicine (TR-TR2-01768 and CLIN1-08686) and an unrestricted grant from Research to Prevent Blindness to the Department of Ophthalmology, University of California, Los Angeles. R.H.-M. was supported by fellowships from the University of the Basque Country UPV/EHU and the Jesus de Gangoiti Barrera foundation

Bladder cancer index: cross-cultural adaptation into Spanish and psychometric evaluation

BACKGROUND: The Bladder Cancer Index (BCI) is so far the only instrument applicable across all bladder cancer patients, independent of tumor infiltration or treatment applied. We developed a Spanish version of the BCI, and assessed its acceptability and metric properties. METHODS: For the adaptation into Spanish we used the forward and back-translation method, expert panels, and cognitive debriefing patient interviews. For the assessment of metric properties we used data from 197 bladder cancer patients from a multi-center prospective study. The Spanish BCI and the SF-36 Health Survey were self-administered before and 12 months after treatment. Reliability was estimated by Cronbach's alpha. Construct validity was assessed through the multi-trait multi-method matrix. The magnitude of change was quantified by effect sizes to assess responsiveness. RESULTS: Reliability coefficients ranged 0.75-0.97. The validity analysis confirmed moderate associations between the BCI function and bother subscales for urinary (r = 0.61) and bowel (r = 0.53) domains; conceptual independence among all BCI domains (r ≤ 0.3); and low correlation coefficients with the SF-36 scores, ranging 0.14-0.48. Among patients reporting global improvement at follow-up, pre-post treatment changes were statistically significant for the urinary domain and urinary bother subscale, with effect sizes of 0.38 and 0.53. CONCLUSIONS: The Spanish BCI is well accepted, reliable, valid, responsive, and similar in performance compared to the original instrument. These findings support its use, both in Spanish and international studies, as a valuable and comprehensive tool for assessing quality of life across a wide range of bladder cancer patients

Factores de riesgo y mortalidad de los derrames pleurales que precisan de una toracocentesis diagnóstica

Resumen: Introducción: El derrame pleural (DP) maligno se asocia a un mal pronóstico, pero la mortalidad de los pacientes con derrames no malignos no ha sido suficientemente estudiada. Nuestro objetivo fue describir la evolución clínica y explorar los factores asociados con la mortalidad por todas las causas a 1, 5 y 10 años de los pacientes que desarrollan un DP. Métodos: Estudio observacional retrospectivo de los pacientes intervenidos mediante toracocentesis diagnóstica durante el decenio 2008-2017 en un servicio de neumología. Se evaluaron variables demográficas, bioquímicas, anatomopatológicas y evolutivas. La etiología de los derrames se determinó mediante criterios estandarizados. Resultados: Se estudió a 358 pacientes con líquido pleural analizado; el 69,2% de los cuales eran varones con una edad media de 68,9 años (DE 15,1). Predominaron los derrames malignos (29,4%), paraneumónicos (19,8%) y secundarios a insuficiencia cardiaca (18,9%). Los pacientes con derrame maligno y por insuficiencia cardiaca tuvieron tasas de mortalidad a un año del 60,0% y 30,8%, respectivamente y del 85% y 64,7% a 5 años. El sexo masculino (hazard ratio [HR] 1,46; IC 95%: 1,03-2,07), la citología positiva para malignidad (HR 1,66; IC 95%: 1,03-2,68) y la recidiva del derrame (HR 1,61; IC 95%: 1,17-2,21) se asociaron a un peor pronóstico y a mortalidad a 5 años. Conclusiones: Los pacientes intervenidos mediante toracentesis por derrame tienen una alta mortalidad a corto y largo plazo. En nuestros pacientes hospitalizados con DP, los factores asociados con mayor mortalidad a 1 y 5 años fueron la edad, el sexo masculino, la recidiva del DP y la coexistencia de neoplasia maligna. Abstract: Introduction: Occurrence of malignant pleural effusion (PE) is known to be associated with a poor prognosis, but the mortality of patients with non-malignant effusions has not been sufficiently studied. Our objective was to describe the clinical course and explore risk factors associated with all-cause mortality at 1, 5 and 10 years in patients who develop a PE. Methods: Retrospective observational study of patients undergoing diagnostic thoracentesis during the decade 2008-2017 in a pulmonology service. Demographic, biochemical, pathological and evolutionary variables were evaluated. The etiology of the effusions was determined using standardized criteria. Results: Pleural fluid samples from 358 patients with a mean age of 68.9 years (SD 15.1 years), 69.2% males, were analyzed. Malignant (29.4%), parapneumonic (19.8%) and secondary to heart failure (18.9%) effusions predominated. Patients with malignant and heart failure related PE had 1-year mortality rates of 60.0% and 30.8%, respectively, and 85% and 64.7% at 5 years. Male gender (hazard ratio [HR] 1.46; 95% CI: 1.03-2.07), positive cytology for malignancy (HR 1.66; 95% CI: 1.03-2.68) and effusion recurrence (HR 1.61; 95% CI: 1.17-2.21) were associated with a worse prognosis and 5-year mortality. Conclusions: Patients undergoing thoracentesis for effusion have a high short and long-term mortality. In our series of hospitalized patients with PE, the factors associated with higher mortality at 1 and 5 years were age, male sex, recurrence of PE, and coexistence of malignancy

Bladder cancer index : Cross-cultural adaptation into Spanish and psychometric evaluation

Background: The Bladder Cancer Index (BCI) is so far the only instrument applicable across all bladder cancer patients, independent of tumor infiltration or treatment applied. We developed a Spanish version of the BCI, and assessed its acceptability and metric properties. Methods: For the adaptation into Spanish we used the forward and back-translation method, expert panels, and cognitive debriefing patient interviews. For the assessment of metric properties we used data from 197 bladder cancer patients from a multi-center prospective study. The Spanish BCI and the SF-36 Health Survey were self-administered before and 12 months after treatment. Reliability was estimated by Cronbach's alpha. Construct validity was assessed through the multi-trait multi-method matrix. The magnitude of change was quantified by effect sizes to assess responsiveness. Results: Reliability coefficients ranged 0.75-0.97. The validity analysis confirmed moderate associations between the BCI function and bother subscales for urinary (r = 0.61) and bowel (r = 0.53) domains; conceptual independence among all BCI domains (r ≤ 0.3); and low correlation coefficients with the SF-36 scores, ranging 0.14-0.48. Among patients reporting global improvement at follow-up, pre-post treatment changes were statistically significant for the urinary domain and urinary bother subscale, with effect sizes of 0.38 and 0.53. Conclusions: The Spanish BCI is well accepted, reliable, valid, responsive, and similar in performance compared to the original instrument. These findings support its use, both in Spanish and international studies, as a valuable and comprehensive tool for assessing quality of life across a wide range of bladder cancer patients