Analysis of DNA processing reactions in bacterial conjugation by using suicide oligonucleotides

Protein TrwC is the conjugative relaxase responsible for DNA processing in plasmid R388 bacterial conjugation. TrwC has two catalytic tyrosines, Y18 and Y26, both able to carry out cleavage reactions using unmodified oligonucleotide substrates. Suicide substrates containing a 30-Sphosphorothiolate linkage at the cleavage site displaced TrwC reaction towards covalent adducts and thereby enabled intermediate steps in relaxase reactions to be investigated. Two distinct covalent TrwC–oligonucleotide complexes could be separated from noncovalently bound protein by SDS–PAGE. As observed by mass spectrometry, one complex contained a single, cleaved oligonucleotide bound to Y18, whereas the other contained two cleaved oligonucleotides, bound to Y18 and Y26. Analysis of the cleavage reaction using suicide substrates and Y18F or Y26F mutants showed that efficient Y26 cleavage only occurs after Y18 cleavage. Strand-transfer reactions carried out with the isolated Y18–DNA complex allowed the assignment of specific roles to each tyrosine. Thus, only Y18 was used for initiation. Y26 was specifically used in the second transesterification that leads to strand transfer, thus catalyzing the termination reaction that occurs in the recipient cell.

Area factors and suicide: 5-year follow-up of the Northern Ireland population

Background: Suicide rates vary markedly between areas but it is unclear whether this is due to differences in population composition or to contextual factors operating at an area level.
Aims: To determine if area factors are independently related to suicide risk after adjustment for individual and family characteristics.
Method: A 5-year record linkage study was conducted of 1 116 748 non-institutionalised individuals aged 16-74 years, enumerated at the 2001 Northern Ireland census.
Results: The cohort experienced 566 suicides during follow-up. Suicide risks were lowest for women and for those who were married or cohabiting. Indicators of individual and household disadvantage and economic and health status at the time of the census were also strongly related to risk of suicide. The higher rates of suicide in the more deprived and socially fragmented areas disappeared after adjustment for individual and household factors. There was no significant relationship between population density and risk of suicide.
Conclusions: Differences in rates of suicide between areas are predominantly due to population characteristics rather than to area-level factors, which suggests that policies targeted at area-level factors are unlikely to significantly influence suicides rates.

Notes on the History of Social Work: An Examination of the Transition from Philanthropy to Professional Social Work in Ireland

The objective of the article is to examine the way in which social work in Ireland evolved from practices of philanthropy in the late 19th century to a distinct professional strategy in the present. Results: The results of archival research show that philanthropy in Ireland was provided almost exclusively by religious organizations and was constructed within a discourse of sectarianism and rivalry between the two main denominations, Catholic and Protestant, up to the 1960s. It is only in the past 30 years that social work has become firmly established as a secular strategy. Conclusions: It is concluded that although social work is now clearly distinct from voluntary and religious-based social work practices, some of its present principles and practices remain continuous with its historical antecedents.