Impact of DNA ligase IV on the fidelity of end joining in human cells.

A DNA ligase IV (LIG4)-null human pre-B cell line and human cell lines with hypomorphic mutations in LIG4 are significantly impaired in the frequency and fidelity of end joining using an in vivo plasmid assay. Analysis of the null line demonstrates the existence of an error-prone DNA ligase IV-independent rejoining mechanism in mammalian cells. Analysis of lines with hypomorphic mutations demonstrates that residual DNA ligase IV activity, which is sufficient to promote efficient end joining, nevertheless can result in decreased fidelity of rejoining. Thus, DNA ligase IV is an important factor influencing the fidelity of end joining in vivo. The LIG4-defective cell lines also showed impaired end joining in an in vitro assay using cell-free extracts. Elevated degradation of the terminal nucleotide was observed in a LIG4-defective line, and addition of the DNA ligase IV-XRCC4 complex restored end protection. End protection by DNA ligase IV was not dependent upon ligation. Finally, using purified proteins, we demonstrate that DNA ligase IV-XRCC4 is able to protect DNA ends from degradation by T7 exonuclease. Thus, the ability of DNA ligase IV-XRCC4 to protect DNA ends may contribute to the ability of DNA ligase IV to promote accurate rejoining in vivo.

Job stress and burnout in the care staff of Leros PIKPA Asylum 25 years after the first Deinstitutionalisation and Rehabilitation Greek Project.

Aims and method To identify correlates between burnout and job stress of care staff at Leros PIKPA Asylum. Forty-nine asylum employees were assessed by Maslach's Burnout Inventory, a sociodemographic questionnaire, the Perceived Stress Scale and the Job Content Questionnaire. Results Emotional exhaustion is related negatively to social support (P = 0.010, r = -0.362). Lack of job achievements is related positively to overall job responsibility (P = 0.040) and negatively to lack of job satisfaction (r = -0.430). Depersonalisation was negatively associated with support from superiors (P = 0.036). Employees with high levels of perceived stress reported higher levels of fatigue (P = 0.050). Positive associations of perceived stress with depression (P = 0.011) and sleep problems (P<0.001) were also detected. Positive correlation was found between monthly salary and lack of sense of personal achievement (P = 0.020). Clinical implications It is necessary to address these issues through staff education and stress management.