Plasmid- and strain-specific factors drive variation in ESBL-plasmid spread in vitro and in vivo

Horizontal gene transfer, mediated by conjugative plasmids, is a major driver of the global rise of antibiotic resistance. However, the relative contributions of factors that underlie the spread of plasmids and their roles in conjugation in vivo are unclear. To address this, we investigated the spread of clinical Extended Spectrum Beta-Lactamase (ESBL)-producing plasmids in the absence of antibiotics in vitro and in the mouse intestine. We hypothesised that plasmid properties would be the primary determinants of plasmid spread and that bacterial strain identity would also contribute. We found clinical Escherichia coli strains natively associated with ESBL-plasmids conjugated to three distinct E. coli strains and one Salmonella enterica serovar Typhimurium strain. Final transconjugant frequencies varied across plasmid, donor, and recipient combinations, with qualitative consistency when comparing transfer in vitro and in vivo in mice. In both environments, transconjugant frequencies for these natural strains and plasmids covaried with the presence/absence of transfer genes on ESBL-plasmids and were affected by plasmid incompatibility. By moving ESBL-plasmids out of their native hosts, we showed that donor and recipient strains also modulated transconjugant frequencies. This suggests that plasmid spread in the complex gut environment of animals and humans can be predicted based on in vitro testing and genetic data

Estimating plasmid conjugation rates: A new computational tool and a critical comparison of methods

Plasmids are important vectors for the spread of genes among diverse populations of bacteria. However, there is no standard method to determine the rate at which they spread horizontally via conjugation. Here, we compare commonly used methods on simulated and experimental data, and show that the resulting conjugation rate estimates often depend strongly on the time of measurement, the initial population densities, or the initial ratio of donor to recipient populations. Differences in growth rate, e.g. induced by sub-lethal antibiotic concentrations or temperature, can also significantly bias conjugation rate estimates. We derive a new ‘end-point’ measure to estimate conjugation rates, which extends the well-known Simonsen method to include the effects of differences in population growth and conjugation rates from donors and transconjugants. We further derive analytical expressions for the parameter range in which these approximations remain valid. We present an easy to use R package and web interface which implement both new and previously existing methods to estimate conjugation rates. The result is a set of tools and guidelines for accurate and comparable measurement of plasmid conjugation rates

Host-specific plasmid evolution explains the variable spread of clinical antibiotic-resistance plasmids

Antibiotic resistance encoded on plasmids is a pressing global health problem. Predicting which plasmids spread in the long term remains very challenging, even though some key parameters influencing plasmid stability have been identified, such as plasmid growth costs and horizontal transfer rates. Here, we show these parameters evolve in a strain-specific way among clinical plasmids and bacteria, and this occurs rapidly enough to alter the relative likelihoods of different bacterium-plasmid combinations spreading. We used experiments with Escherichia coli and antibiotic-resistance plasmids isolated from patients, paired with a mathematical model, to track long-term plasmid stability (beyond antibiotic exposure). Explaining variable stability across six bacterium-plasmid combinations required accounting for evolutionary changes in plasmid stability traits, whereas initial variation of these parameters was a relatively poor predictor of long-term outcomes. Evolutionary trajectories were specific to particular bacterium-plasmid combinations, as evidenced by genome sequencing and genetic manipulation. This revealed epistatic (here, strain-dependent) effects of key genetic changes affecting horizontal plasmid transfer. Several genetic changes involved mobile elements and pathogenicity islands. Rapid strain-specific evolution can thus outweigh ancestral phenotypes as a predictor of plasmid stability. Accounting for strain-specific plasmid evolution in natural populations could improve our ability to anticipate and manage successful bacterium-plasmid combinations.ISSN:0027-8424ISSN:1091-649

Fair enough? Decreased equity of dyadic coping across the transition to parenthood associated with depression of first-time parents

The transition to parenthood (TTP) is a stressful life event for most couples. Therefore, the way both partners jointly cope with stress (i.e., dyadic coping) is important for the prevention of individual adjustment problems (e.g., depression). For dyadic coping to be effective in reducing depressive symptoms, efforts of both partners should be equal. However, many couples experience a decrease of equity in task division within the domestic sphere across the TTP. The current study investigates the equity of a specific skill within the 'relationship sphere', because similarly to a decreased equity in household and childcare, a decreased equity of dyadic coping is likely to be associated with poorer individual adjustment. We collected longitudinal self-report data on dyadic coping and depressive symptoms from 104 mixed-gender first-time parents (n = 208 individuals) from pregnancy until 40 weeks postpartum. We created an equity score for men and women that measured their perceived difference between received and provided dyadic coping. On average, women reported providing more and receiving less dyadic coping than men. While both genders agreed on this distribution, men did perceive a higher equity of dyadic coping than women. Furthermore, the decrease of equity perceived by women across TTP was not visible in men. In line with our assumptions based on the equity theory, perceived equity of dyadic coping was associated with depressive symptoms in a curvilinear manner: Decreases in women's perceived equity in either direction (over- or underbenefit) were associated with more depressive symptoms in women and their male partners. This association was found above and beyond the beneficial effect of dyadic coping itself. This implies that not only how well partners support each other in times of stress, but also how equal both partners' efforts are, is important for their individual adjustment across TTP

The Zurich 3-step concept for the management of behavioral sleep disorders in children: a before-and-after study

OBJECTIVES Several strategies have been found to be effective for the treatment of childhood behavioral sleep disorders. One which has yet to be evaluated is the Zurich 3-step concept, which combines basic notions of the two-process model of sleep regulation (introducing a regular rhythm and adjusting bedtime to sleep need) with behavioral strategies. This uncontrolled before-and-after study describes our concept and its step-wise approach, assesses changes in sleep-wake variables and behavior problems, and also examines associations between changes in sleep-wake variables and behavior problems. METHODS A total of 79 children with sleep problems (age range 6-47 months, 42% females) were included. Sleep problems were assessed by the Infant Sleep Questionnaire, sleep-wake variables by diary and actigraphy, and behavior problems of children ≥ 18 months by the Child Behavior Checklist. RESULTS A significant decrease in nocturnal wake duration (Cohen's d = -0.34) and a significant increase in the duration of the longest continuous nocturnal sleep period (Cohen's d = 0.19) were found from before to after intervention (on average 2.7 months, SD 1.5). The variability for sleep onset and end time decreased, and actigraphically measured circadian rest-activity cycle measures improved. Parent-reported internalizing and total behavior problems also decreased (Cohen's d = 0.66). CONCLUSIONS The findings of both objective and subjective assessment techniques suggest that the Zurich 3-step concept is effective. Thus, the intervention concept may be useful in clinical practice with sleep-disordered children

Estimating plasmid conjugation rates: A new computational tool and a critical comparison of methods

Plasmids are important vectors for the spread of genes among diverse populations of bacteria. However, there is no standard method to determine the rate at which they spread horizontally via conjugation. Here, we compare commonly used methods on simulated and experimental data, and show that the resulting conjugation rate estimates often depend strongly on the time of measurement, the initial population densities, or the initial ratio of donor to recipient populations. Differences in growth rate, e.g. induced by sub-lethal antibiotic concentrations or temperature, can also significantly bias conjugation rate estimates. We derive a new ‘end-point’ measure to estimate conjugation rates, which extends the well-known Simonsen method to include the effects of differences in population growth and conjugation rates from donors and transconjugants. We further derive analytical expressions for the parameter range in which these approximations remain valid. We present an easy to use R package and web interface which implement both new and previously existing methods to estimate conjugation rates. The result is a set of tools and guidelines for accurate and comparable measurement of plasmid conjugation rates

Fair enough? Decreased equity of dyadic coping across the transition to parenthood associated with depression of first-time parents

The transition to parenthood (TTP) is a stressful life event for most couples. Therefore, the way both partners jointly cope with stress (i.e., dyadic coping) is important for the prevention of individual adjustment problems (e.g., depression). For dyadic coping to be effective in reducing depressive symptoms, efforts of both partners should be equal. However, many couples experience a decrease of equity in task division within the domestic sphere across the TTP. The current study investigates the equity of a specific skill within the ‘relationship sphere’, because similarly to a decreased equity in household and childcare, a decreased equity of dyadic coping is likely to be associated with poorer individual adjustment. We collected longitudinal self-report data on dyadic coping and depressive symptoms from 104 mixed-gender first-time parents (n = 208 individuals) from pregnancy until 40 weeks postpartum. We created an equity score for men and women that measured their perceived difference between received and provided dyadic coping. On average, women reported providing more and receiving less dyadic coping than men. While both genders agreed on this distribution, men did perceive a higher equity of dyadic coping than women. Furthermore, the decrease of equity perceived by women across TTP was not visible in men. In line with our assumptions based on the equity theory, perceived equity of dyadic coping was associated with depressive symptoms in a curvilinear manner: Decreases in women’s perceived equity in either direction (over- or underbenefit) were associated with more depressive symptoms in women and their male partners. This association was found above and beyond the beneficial effect of dyadic coping itself. This implies that not only how well partners support each other in times of stress, but also how equal both partners’ efforts are, is important for their individual adjustment across TTP

sST2 translation is regulated by FGF2 via an hnRNP A1-mediated IRES-dependent mechanism

Translation is an energy-intensive process and tightly regulated. Generally, translation is initiated in a cap-dependent manner. Under stress conditions, typically found within the tumor microenvironment in association with e.g. nutrient deprivation or hypoxia, cap-dependent translation decreases, and alternative modes of translation initiation become more important. Specifically, internal ribosome entry sites (IRES) facilitate translation of specific mRNAs under otherwise translation-inhibitory conditions. This mechanism is controlled by IRES trans-acting factors (ITAF), i.e. by RNA-binding proteins, which interact with and determine the activity of selected IRESs. We aimed at characterizing the translational regulation of the IL-33 decoy receptor sST2, which was enhanced by fibroblast growth factor 2 (FGF2). We identified and verified an IRES within the 5'UTR of sST2. Furthermore, we found that MEK/ERK signaling contributes to FGF2-induced, sST2-IRES activation and translation. Determination of the sST2-5'UTR structure by in-line probing followed by deletion analyses identified 23 nucleotides within the sST2-5'UTR to be required for optimal IRES activity. Finally, we show that the RNA-binding protein heterogeneous ribonucleoprotein A1 (hnRNP A1) binds to the sST2-5'UTR, acts as an ITAF, and thus controls the activity of the sST2-IRES and consequently sST2 translation. Specifically, FGF2 enhances nuclear-cytoplasmic translocation of hnRNP A1, which requires intact MEK/ERK activity. In summary, we provide evidence that the sST2-5'UTR contains an IRES element, which is activated by a MEK/ERK-dependent increase in cytoplasmic localization of hnRNP A1 in response to FGF2, enhancing the translation of sST2

Estimating plasmid conjugation rates : A new computational tool and a critical comparison of methods

Plasmids are important vectors for the spread of genes among diverse populations of bacteria. However, there is no standard method to determine the rate at which they spread horizontally via conjugation. Here, we compare commonly used methods on simulated and experimental data, and show that the resulting conjugation rate estimates often depend strongly on the time of measurement, the initial population densities, or the initial ratio of donor to recipient populations. Differences in growth rate, e.g. induced by sub-lethal antibiotic concentrations or temperature, can also significantly bias conjugation rate estimates. We derive a new ‘end-point’ measure to estimate conjugation rates, which extends the well-known Simonsen method to include the effects of differences in population growth and conjugation rates from donors and transconjugants. We further derive analytical expressions for the parameter range in which these approximations remain valid. We present an easy to use R package and web interface which implement both new and previously existing methods to estimate conjugation rates. The result is a set of tools and guidelines for accurate and comparable measurement of plasmid conjugation rates