Validation of podocalyxin-like protein as a biomarker of poor prognosis in colorectal cancer

ABSTRACT: BACKGROUND: Podocalyxin-like 1 (PODXL) is a cell-adhesion glycoprotein and stem cell marker that has been associated with an aggressive tumour phenotype and adverse outcome in several cancer types. We recently demonstrated that overexpression of PODXL is an independent factor of poor prognosis in colorectal cancer (CRC). The aim of this study was to validate these results in two additional independent patient cohorts and to examine the correlation between PODXL mRNA and protein levels in a subset of tumours. METHOD: PODXL protein expression was analyzed by immunohistochemistry in tissue microarrays with tumour samples from a consecutive, retrospective cohort of 270 CRC patients (cohort 1) and a prospective cohort of 337 CRC patients (cohort 2). The expression of PODXL mRNA was measured by real-time quantitative PCR in a subgroup of 62 patients from cohort 2. Spearman's Rho and Chi-Square tests were used for analysis of correlations between PODXL expression and clinicopathological parameters. Kaplan Meier analysis and Cox proportional hazards modelling were applied to assess the relationship between PODXL expression and time to recurrence (TTR), disease free survival (DFS) and overall survival (OS). RESULTS: High PODXL protein expression was significantly associated with unfavourable clinicopathological characteristics in both cohorts. In cohort 1, high PODXL expression was associated with a significantly shorter 5-year OS in both univariable (HR=2.28; 95% CI 1.43-3.63, p=0.001) and multivariable analysis (HR=2.07; 95% CI 1.25-3.43, p=0.005). In cohort 2, high PODXL expression was associated with a shorter TTR (HR=2.93; 95% CI 1.26-6.82, p=0.013) and DFS (HR=2.44; 95% CI 1.32-4.54, p=0.005), remaining significant in multivariable analysis, HR=2.50; 95% CI 1.05-5.96, p=0.038 for TTR and HR=2.11; 95% CI 1.13-3.94, p=0.019 for DFS. No significant correlation could be found between mRNA levels and protein expression of PODXL and there was no association between mRNA levels and clinicopathological parameters or survival. CONCLUSIONS: Here, we have validated the previously demonstrated association between immunohistochemical expression of PODXL and poor prognosis in CRC in two additional independent patient cohorts. The results further underline the potential utility of PODXL as a biomarker for more precise prognostication and treatment stratification of CRC patients.     

Correlation of optical coherence tomography with clinical and histopathological findings in experimental autoimmune uveoretinitis

Optical coherence tomography (OCT) is becoming the state-of-the-art method for the non-invasive imaging of a variety of ocular diseases. The aim of this study was to assess the application of OCT for the in vivo monitoring and follow-up of pathological changes during experimental autoimmune uveoretinitis (EAU) in rats. Initially we established OCT imaging in healthy brown Norway rats and correlated it with retinal histology. Subsequently, we induced EAU and imaged animals by OCT throughout the pre-peak, peak, and post-peak phases of the disease. The sensitivity of OCT imaging was determined by comparison with clinical EAU and histopathology scores obtained ex vivo at several time points throughout the disease course. Our data demonstrate that OCT imaging of the healthy rat retina closely correlates with histological observations and allows the clear visualization of all retinal layers. After induction of EAU, the first pathological changes could be detected by OCT at day (d) 8 post-immunization (p.i.) which corresponded to the time point of clinical disease onset. An increase in retinal thickness (RT) was detected from d10 p.i. onwards which peaked at d16 p.i. and decreased again to near control levels by d20 p.i. We introduce a novel semi-quantitative OCT scoring which correlates with histopathological findings and complements the clinical scores. Therefore, we conclude that OCT is an easily accessible, non-invasive tool for detection and follow-up of histopathological changes during EAU in rats. Indeed, significant differences in RT between different stages of EAU suggest that this OCT parameter is a sensitive marker for distinguishing disease phases in vivo.     

Effect of food preservatives on in situ biofilm formation

The aim of this double-blind, controlled crossover study was to evaluate the influence of food preservatives on in situ dental biofilm growth. Twenty-four volunteers wore appliances with six specimens each of bovine enamel to build up intra-oral biofilms. During three test cycles, the subjects had to put one half of the appliance twice a day in one of the assigned active solutions (0.1% benzoate, BA; 0.1% sorbate, SA or 0.2% chlorhexidine, CHX) and the other into NaCl. After 5 days, the developed biofilms were stained with two fluorescent dyes to visualise vital (green) and dead bacteria (red). Biofilms were scanned by confocal laser scanning microscopy and biofilm thickness (BT) and bacterial vitality (BV%) were calculated. After a washout period of 7 days, a new test cycle was started. The use of SA, BA and CHX resulted in a significantly reduced BT and BV compared to NaCl (p < 0.001). Differences between SA and BA were not significant (p > 0.05) for both parameters, while CHX showed significantly lower values. Both preservatives showed antibacterial and plaque-inhibiting properties, but not to the extent of CHX. The biofilm model enabled the examination of undisturbed oral biofilm formation influenced by antibacterial components under clinical conditions.     

Precurrence risk of birth defects in Hawaii

The aim of the investigation was to describe the risk of selected types of birth defects among older siblings of infants and fetuses with specific birth defects. Using data from a population-based birth defects registry in Hawaii for deliveries during 1986-2000, the precurrence risk (risk among older siblings) for any major birth defect, birth defects of the same organ system, and same specific birth defect were calculated for nine major organ systems and 54 specific birth defects. The precurrence risk of any major birth defect (3.5%) was substantially less than the risk of any major birth defect among the entire population (4.7%). The precurrence risk of a birth defect of the same organ system was significantly higher than the reference rate for six (67%) of the major organ systems and eight (15%) of the specific birth defects. The precurrence risk of the same birth defect was substantially elevated for 15 (28%) of the specific birth defects. Precurrence risk of any major birth defect was not elevated, but tended to be lower than expected. However, for several categories, precurrence risk of birth defects of the same organ system or the same specific birth defect was substantially elevated.     

Behavior, knowledge and perception of risks about sexually transmitted diseases in a group of people over 50 years old

The goal was to analyze the behavior, knowledge and risk perception about sexually transmitted diseases / AIDS (STD/AIDS) in people over 50 years old. 165 public servants at a State Secretary in Mato Grosso, Brazil were interviewed. The chi-square test and logistic regression tests were used for the analysis of risk perception and the behavior and knowledge variables about STD/AIDS. Most of the respondents were female (60.6%), 63.2 had a fixed partner, 72.4% had sexual relations in the past six months, and only 13.3% always wore condoms, with 21.5% being male and 8% female. Perception of risk was associated with non-use of condoms in their last sexual relation (p<0.001) and answering that anyone may acquire a STD/AIDS (p=0.039). The challenge of public policies is to increase actions, directing them to promote the health of the adult and elderly populations, especially regarding sexuality and vulnerability to STD/AIDS.     

Transgenic expression of AQP1 in the fiber cells of AQP0 knockout mouse: Effects on lens transparency

Mutations and knockout of aquaporin 0 (AQP0) result in dominant lens cataract. To date, several functions have been proposed for AQP0; however, two functions, water permeability and cell-to-cell adhesion have been supported by several investigators and only water channel function has been readily authenticated by in vitro and ex vivo studies. Lens shifts protein expression from the more efficient AQP1 in the equatorial epithelial cells to the less efficient water channel, AQP0, in the differentiating secondary fiber cells; perhaps, AQP0 performs a distinctive function. If AQP0 has only water permeability function, can the more efficient water channel AQP1 transgenically expressed in the fiber cells compensate and restore lens transparency in the AQP0 knockout (AQP0^−/−) mouse? To investigate, we generated a transgenic wild-type mouse line expressing AQP1 in the fiber cells using αA-crystallin promoter. These transgenic mice (TgAQP1^+/+) showed increase in fiber cell membrane water permeability without any morphological, anatomical or physiological defects compared to the wild type indicating that the main purpose of the shift in expression from AQP1 to AQP0 may not be to lessen the membrane water permeability. Further, we transgenically expressed AQP1 in the lens fiber cells of AQP0 knockout mouse (TgAQP1^+/+/AQP0^−/−) to determine whether AQP1 could restore AQP0 water channel function and regain lens transparency. Fiber cells of these mice showed 2.6 times more water permeability than the wild type. Transgene AQP1 reduced the severity of lens cataract and prevented dramatic acceleration of cataractogenesis. However, lens fiber cells showed deformities and lack of compact cellular architecture. Loss of lens transparency due to the absence of AQP0 was not completely restored indicating an additional function for AQP0. In vitro studies showed that AQP0 is capable of cell-to-cell adhesion while AQP1 is not. To our knowledge, this is the first report which uses an animal model to demonstrate that AQP0 may have an additional function, possibly cell-to-cell adhesion.     

Computer-assisted detection of pulmonary embolism: evaluation of pulmonary CT angiograms performed in an on-call setting

Purpose

The purpose of the study was to assess the stand-alone performance of computer-assisted detection (CAD) for evaluation of pulmonary CT angiograms (CTPA) performed in an on-call setting.

Methods

In this institutional review board-approved study, we retrospectively included 292 consecutive CTPA performed during night shifts and weekends over a period of 16 months. Original reports were compared with a dedicated CAD system for pulmonary emboli (PE). A reference standard for the presence of PE was established using independent evaluation by two readers and consultation of a third experienced radiologist in discordant cases.

Results

Original reports had described 225 negative studies and 67 positive studies for PE. CAD found PE in seven patients originally reported as negative but identified by independent evaluation: emboli were located in segmental (n?=?2) and subsegmental arteries (n?=?5). The negative predictive value (NPV) of the CAD algorithm was 92% (44/48). On average there were 4.7 false positives (FP) per examination (median 2, range 0–42). In 72% of studies ≤5 FP were found, 13% of studies had ≥10 FP.

Conclusion

CAD identified small emboli originally missed under clinical conditions and found 93% of the isolated subsegmental emboli. On average there were 4.7 FP per examination.