Separation of mutation avoidance and antirecombination functions in an Escherichia coli mutS mutant

DNA mismatch repair in Escherichia coli has been shown to be involved in two distinct processes: mutation avoidance, which removes potential mutations arising as replication errors, and antirecombination which prevents recombination between related, but not identical (homeologous), DNA sequences. We show that cells with the mutSΔ800 mutation (which removes the C-terminal 53 amino acids of MutS) on a multicopy plasmid are proficient for mutation avoidance. In interspecies genetic crosses, however, recipients with the mutSΔ800 mutation show increased recombination by up to 280-fold relative to mutS(+). The MutSΔ800 protein binds to O(6)-methylguanine mismatches but not to intrastrand platinated GG cross-links, explaining why dam bacteria with the mutSΔ800 mutation are resistant to cisplatin, but not MNNG, toxicity. The results indicate that the C-terminal end of MutS is necessary for antirecombination and cisplatin sensitization, but less significant for mutation avoidance. The inability of MutSΔ800 to form tetramers may indicate that these are the active form of MutS

Homologous recombination prevents methylation-induced toxicity in Escherichia coli

Methylating agents such as N-methyl-N\u27-nitro-N-nitrosoguanidine (MNNG) and methyl methane sulfonate (MMS) produce a wide variety of N- and O-methylated bases in DNA, some of which can block replication fork progression. Homologous recombination is a mechanism by which chromosome replication can proceed despite the presence of lesions. The two major recombination pathways, RecBCD and RecFOR, which repair double-strand breaks (DSBs) and single-strand gaps respectively, are needed to protect against toxicity with the RecBCD system being more important. We find that recombination-deficient cell lines, such as recBCD recF, and ruvC recG, are as sensitive to the cytotoxic effects of MMS and MNNG as the most base excision repair (BER)-deficient (alkA tag) isogenic mutant strain. Recombination and BER-deficient double mutants (alkA tag recBCD) were more sensitive to MNNG and MMS than the single mutants suggesting that homologous recombination and BER play essential independent roles. Cells deleted for the polA (DNA polymerase I) or priA (primosome) genes are as sensitive to MMS and MNNG as alkA tag bacteria. Our results suggest that the mechanism of cytotoxicity by alkylating agents includes the necessity for homologous recombination to repair DSBs and single-strand gaps produced by DNA replication at blocking lesions or single-strand nicks resulting from AP-endonuclease action

Ecological strategy for soil contaminated with mercury

Aims The paper presents results from plot experiments aimed at the development of an ecological strategy for soil contaminated with mercury. Meadow grass (Poa pratensis) was tested on mercury contaminated soil in a former chlor-alkali plant (CAP) in southern Poland for its phytoremediation potential. Methods The stabilisation potential of the plants was investigated on plots without additives and after the addition of granular sulphur. Biomass production, uptake and distribution of mercury by plants, as well as leachates and rhizosphere microorganisms were investigated, along with the growth and vitality of plants during one growing season. Results The analysed plants grew easily on mercury contaminated soil, accumulating lower amounts of mercury, especially in the roots, from soil with additive of granular sulphur (0.5 % w/w) and sustained a rich microbial population in the rhizosphere. After amendment application the reduction of Hg evaporation was observed. Conclusions The obtained results demonstrate the potential of using Poa pratensis and sulphur for remediation of mercury contaminated soil and reduction of the Hg evaporation from soil. In the presented study, methods of Hg reduction on “hot spots” were proposed, with a special focus on environmental protection. This approach provides a simple remediation tool for large areas heavily contaminated with mercury

Rubella seroprevalence among primary and pre- primary school pupils at Moi's Bridge location, Uasin Gishu District, Kenya

<p>Abstract</p> <p>Background</p> <p>Rubella is an infectious and generally mild childhood viral disease. The disease is of public health importance because infection acquired during early pregnancy often results in foetal abnormalities that are classified as congenital rubella syndrome (CRS). The burden of rubella infection in most developing countries in not well documented because of limited epidemiological data. However, availability of an effective vaccine has made it necessary to have all the countries with no routine vaccination schedule to evaluate the burden of disease in order to make informed decisions on rubella vaccination and strategy. To address this gap we conducted a study to determine age-specific rubella seroprevalence rates and related risk factors among primary and pre-primary school children in Uasin Gishu district, Moi's Bridge location of Kenya.</p> <p>Methods</p> <p>Subjects of the study were 498 pupils from seven primary schools aged 4–20 years. Questionnaire surveys with blood sampling were conducted between January to July 2005. Samples were tested for rubella specific IgG antibody using ELISA test kit (Enzygnost^®Behring, Germany).</p> <p>Results</p> <p>Overall, rubella seropositivity rate was 80% and it increased with age from 59% (among ages 4–6 years) to 94% (ages 14–20 years). Multivariate logistic regression analysis model, showed that age of child and ownership of a television set which is a proxy measure of socio-economic status of family were significantly associated with rubella seropositivity. The odds of rubella seropositivity in a child older than 13 years was more than that in children younger than 7 years (OR = 3.8 95% CI 2.56–5.78). The odds of rubella seropositivity in a child whose family did not own a television set was 3 times higher than that of child whose family owned a set (OR 3.06, 95% CI 1.17–7.97).</p> <p>Conclusion</p> <p>The study provides important and highly useful information on rubella age specific seroprevalence rates in Kenya. Advancing age was found to be associated with increased risk of rubella. Low socio-economic factors suggest an increased risk of infection in certain categories of society, and control measures need to target this. Overall, the findings can also be used by policy makers to model introduction of routine rubella vaccination in the country and also other developing countries facing similar challenges. More than half of the children got infected in pre-primary and efforts to control rubella should target pre-school children. These data provides pre-vaccination information that can be used to guide immunization strategy as well as to determine success of an immunization programme.</p

Alkylation damage causes MMR-dependent chromosomal instability in vertebrate embryos

SN1-type alkylating agents, like N-methyl-N-nitrosourea (MNU) and N-ethyl-N-nitrosourea (ENU), are potent mutagens. Exposure to alkylating agents gives rise to O6-alkylguanine, a modified base that is recognized by DNA mismatch repair (MMR) proteins but is not repairable, resulting in replication fork stalling and cell death. We used a somatic mutation detection assay to study the in vivo effects of alkylation damage on lethality and mutation frequency in developing zebrafish embryos. Consistent with the damage-sensing role of the MMR system, mutant embryos lacking the MMR enzyme MSH6 displayed lower lethality than wild-type embryos after exposure to ENU and MNU. In line with this, alkylation-induced somatic mutation frequencies were found to be higher in wild-type embryos than in the msh6 loss-of-function mutants. These mutations were found to be chromosomal aberrations that may be caused by chromosomal breaks that arise from stalled replication forks. As these chromosomal breaks arise at replication, they are not expected to be repaired by non-homologous end joining. Indeed, Ku70 loss-of-function mutants were found to be equally sensitive to ENU as wild-type embryos. Taken together, our results suggest that in vivo alkylation damage results in chromosomal instability and cell death due to aberrantly processed MMR-induced stalled replication forks

"Don't try to teach me, I got nothing to learn": Management students' perceptions of business ethics teaching

[EN] Interest is growing towards including business ethics in university curricula, aiming at improving ethical behaviour of future managers. Extant literature has investigated the impact of ethics education on different ethics-related students' cognitive and/or behavioural outcomes, considering variables related to training programmes and students' demographic aspects. Accordingly, we aim at assessing students' understanding of business ethics issues, by focusing on the differences in students' perceptions depending on gender, age, work experience, and ethics courses taken. Testing our hypotheses on a sample of 307 management students at a Polish university, and controlling for social desirability bias, we obtained mixed and partially surprising results. We found significant differences in students' understanding of business ethics depending on their gender and age (female and older students showed more ethical inclinations), but not depending on having taken ethics courses-actually perceptions of such courses worsened after taking them. Besides, work experience was not a significant variable. Moreover, course exposure intensiveness (i.e., number of ethics courses completed), and time passed since completion of the latest course, did not confirm hypothesized effects on most of the dependent (sub)variables. These findings stimulate further questions and challenges for future research (e.g., around course design and methodology, and social/cultural/contextual issues).Tormo-Carbó, G.; Oltra, V.; Klimkiewicz, K.; Seguí-Mas, E. (2019). "Don't try to teach me, I got nothing to learn": Management students' perceptions of business ethics teaching. 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RecA maintains the integrity of chloroplast DNA molecules in Arabidopsis

Although our understanding of mechanisms of DNA repair in bacteria and eukaryotic nuclei continues to improve, almost nothing is known about the DNA repair process in plant organelles, especially chloroplasts. Since the RecA protein functions in DNA repair for bacteria, an analogous function may exist for chloroplasts. The effects on chloroplast DNA (cpDNA) structure of two nuclear-encoded, chloroplast-targeted homologues of RecA in Arabidopsis were examined. A homozygous T-DNA insertion mutation in one of these genes (cpRecA) resulted in altered structural forms of cpDNA molecules and a reduced amount of cpDNA, while a similar mutation in the other gene (DRT100) had no effect. Double mutants exhibited a similar phenotype to cprecA single mutants. The cprecA mutants also exhibited an increased amount of single-stranded cpDNA, consistent with impaired RecA function. After four generations, the cprecA mutant plants showed signs of reduced chloroplast function: variegation and necrosis. Double-stranded breaks in cpDNA of wild-type plants caused by ciprofloxacin (an inhibitor of Escherichia coli gyrase, a type II topoisomerase) led to an alteration of cpDNA structure that was similar to that seen in cprecA mutants. It is concluded that the process by which damaged DNA is repaired in bacteria has been retained in their endosymbiotic descendent, the chloroplast