Clinical evaluation of Bladder CARE, a new epigenetic test for bladder cancer detection in urine samples.

BACKGROUND: Bladder cancer (BC) is the 5th most common cancer in the USA. Non-muscle invasive bladder cancer represents about 70% of all cases and has generally a favorable outcome. However, recurrence rates as high as 60 to 70% and progression rates of 10 to 20% necessitate intensive surveillance with cystoscopy. The invasiveness and high cost of cystoscopy poses significant burden on BC patients as well as on the healthcare system. In this study we test the feasibility of a simple, sensitive, and non-invasive detection of BC using Bladder CARE test in urine samples. RESULTS: Urine from 136 healthy and 77 BC subjects was collected using the at-home Bladder CARE Urine Collection Kit and analyzed with Bladder CARE test. The test measures the methylation level of three BC-specific biomarkers and two internal controls using methylation-sensitive restriction enzymes coupled with qPCR. Bladder CARE showed an overall sensitivity of 93.5%, a specificity of 92.6%, and a PPV and NPV of 87.8% and 96.2%, respectively. Bladder CARE has an LOD as low as 0.046%, which equates to detecting 1 cancer cell for every 2,200 cells analyzed. We also provided evidence that bisulfite-free methods to assess DNA methylation, like Bladder CARE, are advantageous compared to conventional methods that rely on bisulfite conversion of the DNA. CONCLUSION: Highly sensitive detection of BC in urine samples is possible using Bladder CARE. The low LOD of the test and the measurement of epigenetic biomarkers make Bladder CARE a good candidate for the early detection of BC and possibly for the routine screening and surveillance of BC patients. Bladder CARE and the at-home urine sample collection system have the potential to (1) reduce unnecessary invasive testing for BC (2) reduce the burden of surveillance on patients and on the healthcare system, (3) improve the detection of early stage BC, and (4) allow physicians to streamline the monitoring of patients.

Baseline corticosterone is associated with parental care in virgin Japanese quail (Coturnix japonica).

Glucocorticoids (GCs) are thought to impact reproductive success, and ultimately fitness. In this study we focus specifically on the relationship between GCs and parental care. Captive bred Japanese quail (Coturnix japonica) do not show spontaneous parental care behavior, however this behavior can be induced through a sensitization procedure. We investigated how the GC status of Japanese quail relates to parental care in animals of both sexes exposed or not to a chick sensitization treatment. To assess GC status, we obtained baseline plasma and feather cort samples, and used the HPA-axis function test to assess stress responsiveness by examining the response to a standardized stressor as well as negative feedback efficacy through dexamethasone injection. Next, birds were either exposed to chicks overnight in a small enclosure (sensitization treatment) or were enclosed but not exposed to chicks (control). The following morning, adult behaviors were filmed in the presence of a fresh set of chicks for 20 min. A final serum GC sample was obtained to assess if exposure to novel chicks was perceived as stressful. In control animals, baseline GCs were associated with increased total parental care duration and decreased latency to first parental care event. Interestingly, the opposite relationship was found in the sensitization group. Finally, exposure to novel chicks was not associated with an increase in corticosterone in either group. Overall it appears that baseline GCs are correlated with parental care in captive bred Japanese quail, and that the relationship changes direction depending on whether or not sensitization has occurred.