Resistant mutants of Mycobacterium tuberculosis selected in vitro do not reflect the in vivo mechanism of isoniazid resistance

The high prevalence of isoniazid-resistant Mycobacterium tuberculosis is often explained by a high mutation rate for this trait, although detailed information to support this theory is absent. We studied the development of isoniazid resistance in vitro, making use of a laboratory strain of M. tuberculosis. Spontaneous isoniazid-resistant mutants were characterized by molecular methods allowing identification of the most commonly encountered resistance-conferring mutations. Additionally, we determined the in vitro mutation rates for isoniazid and rifampicin resistance, and characterized the genome of a triple-resistant strain. Results confirm that the in vitro mutation rate for isoniazid resistance (3.2 x 10(-7) mutations/cell division) is much higher than the rate for rifampicin resistance (9.8 x 10(-9) mutations/cell division). However, in the majority of the in vitro mutants katG was partially or completely deleted and neither of the two most common in vivo mutations, katG-S315T or inhA-C(-)15T, were found in 120 isogenic mutants. This implies that clinically prevalent resistance mutations were present in <0.8% of isoniazid-resistant strains selected in vitro (95% CI 0%-2.5%). The triple-resistant strain had acquired isoniazid resistance via a 49 kbp deletion, which included katG. Apart from previously identified resistance-conferring mutations, three additional point mutations were acquired during sequential selection steps. These outcomes demonstrate that the in vivo mechanism of isoniazid resistance is not reflected by in vitro experiments. We therefore conclude that the high in vitro mutation rate for isoniazid resistance is not a satisfactory explanation for the fact that isoniazid monoresistance is significantly more widespread than monoresistance to rifampici

Blinatumomab versus historical standard therapy in pediatric patients with relapsed/refractory Ph-negative B-cell precursor acute lymphoblastic leukemia

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Neuropsychosocial profiles of current and future adolescent alcohol misusers

A comprehensive account of the causes of alcohol misuse must accommodate individual differences in biology, psychology and environment, and must disentangle cause and effect. Animal models1 can demonstrate the effects of neurotoxic substances; however, they provide limited insight into the psycho-social and higher cognitive factors involved in the initiation of substance use and progression to misuse. One can search for pre-existing risk factors by testing for endophenotypic biomarkers2 in non-using relatives; however, these relatives may have personality or neural resilience factors that protect them from developing dependence3. A longitudinal study has potential to identify predictors of adolescent substance misuse, particularly if it can incorporate a wide range of potential causal factors, both proximal and distal, and their influence on numerous social, psychological and biological mechanisms4. Here we apply machine learning to a wide range of data from a large sample of adolescents (n = 692) to generate models of current and future adolescent alcohol misuse that incorporate brain structure and function, individual personality and cognitive differences, environmental factors (including gestational cigarette and alcohol exposure), life experiences, and candidate genes. These models were accurate and generalized to novel data, and point to life experiences, neurobiological differences and personality as important antecedents of binge drinking. By identifying the vulnerability factors underlying individual differences in alcohol misuse, these models shed light on the aetiology of alcohol misuse and suggest targets for prevention

Salivary and gingival CXCL8 correlation with periodontal status, periodontal pathogens, and smoking

Objectives Neutrophil granulocytes have been proposed to play a major role in the mediation of periodontitis-associated tissue destruction. Their recruitment and activation are regulated by the chemokine CXCL8. This study aimed to delineate the dependency of CXCL8 expression in gingival crevicular fluid (GCF) and saliva on periodontal status, bacterial infection, and smoking, in patients with periodontitis. Methods The study cohort comprised 279 subjects with untreated periodontitis. Probing pocket depth (PPD), gingival recession, bleeding on probing (BOP), plaque index, and bone loss were evaluated. CXCL8 was determined in saliva and GCF using flow cytometry. Results Considering the entire study sample, CXCL8 levels were correlated with the mean PPD (ρ = 0.131; p = 0.029), severity of periodontitis (ρ = 0.121; p = 0.043), BOP (ρ = 0.204; p = 0.001), and smoking (ρ = −0.219; p 3mm. Conclusion The current study revealed a strong dependency of CXCL8 expression in GCF on the severity and activity of periodontal disease. Smoking causes a significant reduction in CXCL8 expression in saliva and GCF

Tree species identity drives soil organic carbon storage more than species mixing in major two-species mixtures (pine, oak, beech) in Europe

International audienceMixed forests are usually associated with higher aboveground carbon storage compared to the corresponding monocultures but information on the impact of tree species mixing on soil organic carbon (SOC) is still limited. Yet, maximizing SOC storage is crucial for ecosystem C sequestration and many other ecosystem services. This study used a triplet approach (ie. two-species mixed stand and respective pure stands at the same site) to assess the impact of tree species identity and mixing on SOC storage in eight pine-oak, eight pine-beech and five beech-oak triplets in Europe. We sampled the forest floor (FF) and 0–40 cm in the mineral soil per 10 cm interval. For each triplet type, we fitted basal area (BA) proportion of one component species (for species identity) and a BA-based plot-level True Shannon Diversity index (for species mixing) as explanatory variables for SOC stocks in linear mixed effects models, which included stone content and plot BA as covariates, and site as a random intercept. Considering the total soil depth (FF + 0–40 cm), species identity effect on SOC stocks was only significant for pine-beech and pine-oak triplets but explained more variability in SOC stocks than species mixing across triplet types. Species mixing effect was not significant for any triplet type in the total soil depth. While species identity consistently drove SOC storage in the topsoil layers across triplet types, species mixing explained more variability in SOC stocks in the deeper soil layers except for pine-oak triplets. The results showed that species identity is a stronger driver of SOC storage than species mixing. While tree species identity effect was strongly related to a conifers vs broadleaves signature, the drivers behind mixing effects remained elusive. The results suggest that targeted selection of tree species could better enhance SOC storage in European forests than a mere increase in species richness

The distribution of carbon stocks between tree woody biomass and soil differs between Scots pine and broadleaved species (beech, oak) in European forests

While the impacts of forest management options on carbon (C) storage are well documented, the way they affect C distribution among ecosystem components remains poorly investigated. Yet, partitioning of total forest C stocks, particularly between aboveground woody biomass and the soil, greatly impacts the stability of C stocks against disturbances in forest ecosystems. This study assessed the impact of species composition and stand density on C storage in aboveground woody biomass (stem + branches), coarse roots, and soil, and their partitioning in pure and mixed forests in Europe. We used 21 triplets (5 beech-oak, 8 pine-beech, 8 pine-oak mixed stands, and their respective monocultures at the same sites) in seven European countries. We computed biomass C stocks from total stand inventories and species-specific allometric equations, and soil organic C data down to 40 cm depth. On average, the broadleaved species stored more C in aboveground woody biomass than soil, while C storage in pine was equally distributed between both components. Stand density had a strong effect on C storage in tree woody biomass but not in the soil. After controlling for stand basal area, the mixed stands had, on average, similar total C stocks (in aboveground woody biomass + coarse roots + soil) to the most performing monocultures. Although species composition and stand density affect total C stocks and its partitioning between aboveground woody biomass and soil, a large part of variability in soil C storage was unrelated to stand characteristics

Climatic water availability modifies tree functional diversity effects on soil organic carbon storage in European forests

Forest stand and environmental factors influence soil organic carbon (SOC) storage, but little is known about their relative impacts in different soil layers. Moreover, how environmental factors modulate the impact of stand factors, particularly species mixing, on SOC storage, is largely unexplored. In this study, conducted in 21 forest triplets (two monocultures of different species and their mixture on the same site) distributed in Europe, we tested the hypothesis that stand factors (functional identity and diversity) have stronger effects on topsoil (FF + 0–10 cm) C storage than environmental factors (climatic water availability, clay + silt content, oxalate-extractable Al–Alox) but that the opposite occurs in the subsoil (10–40 cm). We also tested the hypothesis that functional diversity improves SOC storage under high climatic water availability, clay + silt contents, and Alox. We characterized functional identity as the basal area proportion of broadleaved species (beech and/or oak), and functional diversity as the product of broadleaved and conifer (pine) proportions. The results show that functional identity was the main driver of topsoil C storage, while climatic water availability had the largest control on subsoil C storage. Functional diversity decreased topsoil C storage under increasing climatic water availability, but the opposite was observed in the subsoil. Functional diversity effects on topsoil C increased with increasing clay + silt content, while its effects on subsoil C were negative at increasing Alox content. This suggests that functional diversity effect on SOC storage changes along gradients in environmental factors and the direction of effects depends on soil depth

Decitabine and vorinostat with FLAG chemotherapy in pediatric relapsed/refractory AML: Report from the therapeutic advances in childhood leukemia and lymphoma (TACL) consortium

Survival outcomes for relapsed/refractory pediatric acute myeloid leukemia (R/R AML) remain dismal. Epigenetic changes can result in gene expression alterations which are thought to contribute to both leukemogenesis and chemotherapy resistance. We report results from a phase I trial with a dose expansion cohort investigating decitabine and vorinostat in combination with fludarabine, cytarabine, and G-CSF (FLAG) in pediatric patients with R/R AML [NCT02412475]. Thirty-seven patients enrolled with a median age at enrollment of 8.4 (range, 1-20) years. There were no dose limiting toxicities among the enrolled patients, including two patients with Down syndrome. The recommended phase 2 dose of decitabine in combination with vorinostat and FLAG was 10 mg/m2 . The expanded cohort design allowed for an efficacy evaluation and the overall response rate among 35 evaluable patients was 54% (16 complete response (CR) and 3 complete response with incomplete hematologic recovery (CRi)). Ninety percent of responders achieved minimal residual disease (MRD) negativity (&lt;0.1%) by centralized flow cytometry and 84% (n&nbsp;=&nbsp;16) successfully proceeded to hematopoietic stem cell transplant. Two-year overall survival was 75.6% [95%CI: 47.3%, 90.1%] for MRD-negative patients vs. 17.9% [95%CI: 4.4%, 38.8%] for those with residual disease (p &lt; .001). Twelve subjects (34%) had known epigenetic alterations with 8 (67%) achieving a CR, 7 (88%) of whom were MRD negative. Correlative pharmacodynamics demonstrated the biologic activity of decitabine and vorinostat and identified specific gene enrichment signatures in nonresponding patients. Overall, this therapy was well-tolerated, biologically active, and effective in pediatric patients with R/R AML, particularly those with epigenetic alterations