Reliable identification of cardiac liability in drug discovery using automated patch clamp: Benchmarking best practices and calibration standards for improved proarrhythmic assessment.

INTRODUCTION: Screening compounds for activity on the hERG channel using patch clamp is a crucial part of safety testing. Automated patch clamp (APC) is becoming widely accepted as an alternative to manual patch clamp in order to increase throughput whilst maintaining data quality. In order to standardize APC experiments, we have investigated the effects on IC50 values under different conditions using several devices across multiple sites. METHODS: APC instruments SyncroPatch 384i, SyncroPatch 384PE and Patchliner, were used to record hERG expressed in HEK or CHO cells. Up to 27 CiPA compounds were used to investigate effects of voltage protocol, incubation time, labware and time between compound preparation and experiment on IC50 values. RESULTS: All IC50 values of 21 compounds recorded on the SyncroPatch 384PE correlated well with IC50 values from the literature (Kramer et al., 2013) regardless of voltage protocol or labware, when compounds were used immediately after preparation, but potency of astemizole decreased if prepared in Teflon or polypropylene (PP) compound plates 2-3 h prior to experiments. Slow acting compounds such as dofetilide, astemizole, and terfenadine required extended incubation times of at least 6 min to reach steady state and therefore, stable IC50 values. DISCUSSION: Assessing the influence of different experimental conditions on hERG assay reliability, we conclude that either the step-ramp protocol recommended by CiPA or a standard 2-s step-pulse protocol can be used to record hERG; a minimum incubation time of 5 min should be used and although glass, Teflon, PP or polystyrene (PS) compound plates can be used for experiments, caution should be taken if using Teflon, PS or PP vessels as some adsorption can occur if experiments are not performed immediately after preparation. Our recommendations are not limited to the APC devices described in this report, but could also be extended to other APC devices.

Socioeconomic position and mortality among patients with prostate cancer: influence of mediating factors.

INTRODUCTION: Men with low socioeconomic position experience higher mortality after a prostate cancer diagnosis compared to men with a higher socioeconomic position, however, the specific mediators of this association are unclear. We therefore evaluated the influence of potential mediators on the association between socioeconomic position, and prostate cancer-specific and all-cause death in prostate cancer patients. MATERIALS AND METHODS: We conducted a cohort study of prostate cancer patients in the Danish Diet, Cancer and Health study. All patients completed questionnaires and anthropometric measurements at enrollment. Information on educational level, income, comorbidity and vital status was obtained by linkage to Danish nationwide registries. Clinical data and anthropometric measures were collected from medical records at diagnosis. Cox proportional hazard models were used to compute hazard ratios (HR) for all-cause and prostate cancer-specific death according to socioeconomic position and potential mediators. RESULTS: We included 953 prostate cancer patients identified among 27 179 male participants in the Diet, Cancer and Health study who were followed for a median of 6.5 years (interquartile range 6.4-11.2 years). Patients with low socioeconomic position were more often overweight or obese at baseline. Low socioeconomic position was associated with increased prostate cancer-specific and all-cause death. The increased mortality could largely be explained by tumor aggressiveness, comorbidity, treatment and metabolic indicators, except for patients in the lowest income group. DISCUSSION: Our study confirmed the a priori assumption that socioeconomic position is associated with increased mortality after prostate cancer. The increased mortality could largely be explained by lifestyle and clinical parameters.

Socioeconomic position and breast reconstruction in Danish women.

UNLABELLED: Few studies have been conducted on the socioeconomic position of women undergoing breast reconstruction, and none have been conducted in the Danish population. We investigated the association between educational level and breast reconstruction in a nationwide cohort of Danish women with breast cancer. MATERIAL AND METHODS: From nationwide registers, 13 379 women aged 30-80 years who had been treated by mastectomy for breast cancer in Denmark in 1999-2006 were identified and followed up through November 2009. Multivariate logistic regression models were used to investigate the simultaneous influence of educational level on the likelihood of having immediate or delayed (up to three years after mastectomy) breast reconstruction, with adjustment for age, breast cancer characteristics, comorbidity, socioeconomic variables and availability of plastic surgery services at each woman's affiliated hospital. RESULTS: The odds ratios (ORs) for both immediate and delayed breast reconstruction increased significantly with level of education. Being affiliated to a hospital with a plastic surgery department increased the likelihood of both immediate (adjusted OR, 4.02; 95% confidence interval [CI], 2.81-5.75) and delayed breast reconstruction (adjusted OR, 1.41; 95% CI, 1.26-1.66). There was no association between education and breast reconstruction among 30-44 year old women, regardless of type of breast reconstruction; however, medium or higher education was significantly associated with a fourfold increase in the OR for immediate breast reconstruction in women aged 45-59 years and a more than twofold increase in the OR for delayed breast reconstruction in women aged 60-80 years compared to women with short education. CONCLUSION: Increasing education was associated with increasing odds for having immediate or delayed breast reconstruction, but only in the older age groups. The offer of breast reconstruction appears to be unequally distributed in Denmark, and living in an area where the hospital has a plastic surgery department significantly increased the odds for having breast reconstruction.

Social inequality and incidence of and survival from male genital cancer in a population-based study in Denmark, 1994-2003.

We investigated the effects of socioeconomic, demographic and health-related indicators on the incidence of and survival from prostate and testicular cancer diagnosed 1994-2003 with follow-up through 2006 in Denmark using information from nationwide registers. The analyses were based on data on 8279 men with prostate cancer and 1770 with testicular cancer in a cohort of 3.22 million persons born between 1925 and 1973 and aged >or=30 years. We found that men with higher education and the highest disposable income had the highest incidence of prostate cancer. The 1-year and 5-year relative survival after prostate cancer were best amongst men of the highest socioeconomic position. We found no substantial social gradients in the incidence of or survival from testicular cancer.

The Danish Registry for Plastic Surgery of the Breast: establishment of a nationwide registry for prospective follow-up, quality assessment, and investigation of breast surgery.

Although numerous epidemiologic studies have examined the long-term safety of silicone breast implants during the past decade, there is a relative lack of surveillance data on short-term health effects and complications following cosmetic surgery of the breast. The Danish Registry for Plastic Surgery of the Breast, established in May of 1999, provides plastic surgeons with a nationwide system for the collection of preoperative, perioperative, and postoperative data on women undergoing breast implantation, breast reduction, or mastopexy. The purpose of the Registry is to examine short-term and, eventually, long-term local complications and possible health effects, and to contribute to an ongoing evaluation of surgical results and surveillance of the products. Furthermore, the Registry will allow the identification of new areas for research into cosmetic and reconstructive breast surgery. Women accepting registration in the Danish Registry for Plastic Surgery of the Breast complete a self-administered questionnaire focusing on medical history and demographic and behavioral factors. Preoperative blood samples are drawn for storage. Surgical data, postoperative results, and complications are registered following surgery and at postoperative visits. Currently, registration has been initiated at 24 private and public clinics, representing more than 80 percent of the plastic surgery clinics in Denmark. As of November of 2001, a total of 1472 women with breast implants and 560 women with breast reduction were included in the Registry. These figures are expected to increase annually by 1000 women undergoing breast implantation and 500 women undergoing breast reduction or mastopexy. The authors present their experience of establishing the first nationwide comprehensive clinical-epidemiologic database and biological bank for cosmetic and reconstructive surgery procedures.

Characteristics of women with cosmetic breast implants compared with women with other types of cosmetic surgery and population-based controls in Denmark.

Herein the authors describe characteristics of women with breast implants compared with women with other types of cosmetic surgery as well as population controls. All women who acquired breast implants from 1977 to 1997 were identified from the files of two private plastic surgery clinics in Denmark. Patient characteristics were obtained through a self-administered questionnaire. The magnitude of differences between patient and control groups was estimated using odds ratios and 95% confidence intervals. Women with breast implants had a significantly lower body mass index and reported a two-fold greater incidence of current smoking compared with women from the general population and compared with women with other cosmetic surgery. Women with implants reported a greater number of full-term pregnancies and were less likely than controls to have had their first birth at age 30 years or older. Women with implants were not more likely than women in either control group to report a history of diseases, including connective tissue diseases, cancer, or depression before their implant surgery. Women with cosmetic breast implants differ from women with other forms of cosmetic surgery and from general population controls with respect to characteristics that may importantly influence health outcomes and that need to be addressed in future breast implant studies.