Pitfalls and challenges associated with phenoconversion in forensic toxcicology.

PURPOSE: In recent years, several publications have demonstrated the interest and the usefulness of pharmacogenetics in forensic toxicology. However, this approach remains namely focused on DNA-based phenotype, which may potentially lead to misinterpretation. Other determinants such as co-medication or physiological parameters may also impact the phenotype. This article aims to highlight the importance of considering such determinants in forensic toxicology, through the original case of a heroin-related fatality. METHOD: Ethanol concentration determination and toxicological screening were performed using gas chromatography with flame ionization detection, liquid chromatography with diode array detection and gas chromatography with mass spectrometry detection. CYP2C19 and CYP2D6 genotypes were determined by Taqman® real-time PCR analyses. RESULTS: Femoral blood analyses revealed the presence of ethanol, morphine, codeine, venlafaxine (VEN), O-desmethylvenlafaxine (ODV) and N-desmethylvenlafaxine (NDV), paroxetine, and risperidone. 6-acetylmorphine was also identified in urine. VEN, paroxetine and risperidone were quantified at supra-therapeutic or toxic blood concentrations. NDV was not quantified. The metabolic ratio of VEN (ODV to VEN) was exceptionally low (about 0.7). Pharmacogenetics testing showed that the patient was heterozygous for the CYP2C19*2 loss-of-function allele, which predict an intermediate metabolism for CYP2C19. None of the deficient CYP2D6 alleles investigated were identified. Those results suggest an extensive CYP2D6-metabolism phenotype. CONCLUSION: A discrepancy was seen between the results of the genomic evaluation and the observed metabolic ratio of VEN. This tends to exclude a genetic origin and lead us to formulate other hypotheses, such as phenoconversion that may have been induced by drug interaction involving patients' regular medications. Phenoconversion is as a complex phenomenon that leads to genotype-phenotype mismatch without any genetic abnormality particularly described for cytochromes P450 2D6 and 2C19. Although transient, phenoconversion can have a significant impact on the analysis and interpretation of genotype-focused clinical outcomes correlation and in forensic toxicology conclusions.

Author Correction: Climatic controls of decomposition drive the global biogeography of forest-tree symbioses.

In this Letter, a middle initial and additional affiliation have been added for author G. J. Nabuurs; two statements have been added to the Supplementary Acknowledgements; and a citation to the French National Institute has been added to the Methods; see accompanying Author Correction for further details.

Climatic controls of decomposition drive the global biogeography of forest-tree symbioses.

The identity of the dominant root-associated microbial symbionts in a forest determines the ability of trees to access limiting nutrients from atmospheric or soil pools1,2, sequester carbon3,4 and withstand the effects of climate change5,6. Characterizing the global distribution of these symbioses and identifying the factors that control this distribution are thus integral to understanding the present and future functioning of forest ecosystems. Here we generate a spatially explicit global map of the symbiotic status of forests, using a database of over 1.1 million forest inventory plots that collectively contain over 28,000 tree species. Our analyses indicate that climate variables-in particular, climatically controlled variation in the rate of decomposition-are the primary drivers of the global distribution of major symbioses. We estimate that ectomycorrhizal trees, which represent only 2% of all plant species7, constitute approximately 60% of tree stems on Earth. Ectomycorrhizal symbiosis dominates forests in which seasonally cold and dry climates inhibit decomposition, and is the predominant form of symbiosis at high latitudes and elevation. By contrast, arbuscular mycorrhizal trees dominate in aseasonal, warm tropical forests, and occur with ectomycorrhizal trees in temperate biomes in which seasonally warm-and-wet climates enhance decomposition. Continental transitions between forests dominated by ectomycorrhizal or arbuscular mycorrhizal trees occur relatively abruptly along climate-driven decomposition gradients; these transitions are probably caused by positive feedback effects between plants and microorganisms. Symbiotic nitrogen fixers-which are insensitive to climatic controls on decomposition (compared with mycorrhizal fungi)-are most abundant in arid biomes with alkaline soils and high maximum temperatures. The climatically driven global symbiosis gradient that we document provides a spatially explicit quantitative understanding of microbial symbioses at the global scale, and demonstrates the critical role of microbial mutualisms in shaping the distribution of plant species.

Structural patterns of the human ABCC4/MRP4 exporter in lipid bilayers rationalize clinically observed polymorphisms.

The ABCC4/MRP4 exporter has a clinical impact on membrane transport of a broad range of xenobiotics. It is expressed at key locations for drug disposition or effects such as in the liver, the kidney and blood cells. Several polymorphisms and mutations (e.g., p.Gly187Trp) leading to MRP4 dysfunction are associated with an increased risk of toxicity of some drugs. So far, no human MRP4 structure has been elucidated, precluding rationalization of these dysfunctions at a molecular level. We constructed an atomistic model of the wild type (WT) MRP4 and the p.Gly187Trp mutant embedded in different lipid bilayers and relaxed them for hundreds of nanoseconds by molecular dynamics simulations. The WT MRP4 molecular structure confirmed and ameliorated the general knowledge about the transmembrane helices and the two nucleotide binding domains. Moreover, our model elucidated positions of three generally unresolved domains: L1 (linker between the two halves of the exporter); L0 (N-terminal domain); and the zipper helices (between the two NBDs). Each domain was thoroughly described in view of its function. The p.Gly187Trp mutation induced a huge structural impact on MRP4, mainly affecting NBD 1 structure and flexibility. The structure of transporter enabled rationalization of known dysfunctions associated with polymorphism of MRP4. This model is available to the pharmacology community to decipher the impact of any other clinically observed polymorphism and mutation on drug transport, giving rise to in silico predictive pharmacogenetics.

Response to Comment on "The extent of forest in dryland biomes".

De la Cruz et al question the reliability of our results, claiming that we do not refer to the most appropriate spatial extent of drylands. In our response, we explain why we chose an existing and internationally recognized delineation of drylands among several options, and why our findings are due to a difference of remote sensing technique and not to the definition of drylands we have selected.

Response to Comment on "The extent of forest in dryland biomes".

Griffith et al do not question the quality of our analysis, but they question our results with respect to the definition of forest we employed. In our response, we explain why the differences we report result from a difference of technique and not of definition, and how anyone can adapt-as we did-our data set to any forest definition and tree cover threshold of interest.

Response to Comment on "The extent of forest in dryland biomes".

Schepaschenko et al question our findings, claiming that we did not refer to all existing maps and that we did not account for all sources of uncertainty. In our response, we detail our selection criteria for reference maps, which clarify why the work of Schepaschenko et al was not used, and we explain why our uncertainty assessment is complete and how it was misunderstood by Schepaschenko et al.

Short implants compared to implants in vertically augmented bone: a systematic review.

OBJECTIVES: To assess relevant data comparing short implants or implants associated with vertical ridge augmentation derived from RCT's and CCT's. MATERIAL AND METHODS: A PubMed and hand search was performed to identify all RCT's and CCT's published in English language comparing short implants to implants associated with vertical ridge augmentation. RESULTS: The initial search resulted in 3387 articles. A total of 17 articles were eligible for full-text analysis and four were finally included. This review tends to demonstrate similar implant survival rates between implants placed in vertically augmented bone and short implants (95.09% vs. 96.24%, respectively) with a follow-up ranging from 1 to 5 years. In terms of prosthetic survival rates, there were no differences between the treatments. More surgical complications were reported when using implants placed in vertically augmented bone compared to short implants (56 patients with surgical complications compared to 18 patients, respectively). CONCLUSIONS: This evidence should, however, be interpreted with caution as it is derived from four RCT's with limited sample size (ranging from 15 to 30 per group), limited follow-up and performed by the same research group.

Spatio-temporal variation of biting flies, Stomoxys spp. (Diptera: Muscidae), along a man-made disturbance gradient, from primary forest to the city of Makokou (North-East, Gabon).

Understanding the pattern of abundance of vector populations is important to control the potential of transmission of associated pathogens. The pattern of abundance of Stomoxys Geoffroy, an ubiquitous blood-sucking fly, is poorly known in tropical Africa. In this study, we investigated the spatio-temporal pattern of abundance of the Stomoxys genus along a gradient of man-made disturbance in north-eastern Gabon. Three sites (one in primary forest, one in secondary forest and one in a man-made environment) were monitored during 13 months using Vavoua traps. Seven species and subspecies were found to live in sympatry, but with distinct patterns of abundance with respect to space and time. The most abundant species was Stomoxys transvittatus Villeneuve, whereas the rarest species was S. xanthomelas Roubaud. Stomoxys calcitrans Linné was preferentially found in man-made environments, whereas S. xanthomelas was preferentially found in primary forest. Stomoxys abundance was the greatest in secondary forest, then in man-made environments and finally in primary forest. A seasonal variation in Stomoxys abundance was also found. In conclusion, forest degradation and deforestation are likely both to favour the concentration of populations of Stomoxys, and to change the specific composition of the Stomoxys community.

Coordinate regulation of estrogen receptor ß degradation by Mdm2 and CREB-binding protein in response to growth signals.

The biological actions of estrogen are mediated via estrogen receptors ERα and ERß. Yet, other cellular signaling events that also impact ER functions have an important role in breast carcinogenesis. Here, we show that activation of ErbB2/ErbB3 tyrosine kinase receptors with growth factor heregulin-ß prompts ERß degradation by the 26S proteasome, a mechanism that requires the coactivator cAMP response element-binding (CREB)-binding protein (CBP). We found that CBP promotes ERß ubiquitination and degradation through enhancement of the PI3-K/Akt pathway by heregulin-ß, an effect potentiated by a negatively charged hinge region of ERß. Activated Akt triggered the recruitment of E3 ubiquitin ligase Mdm2 to ERß, which was further stabilized by CBP, resulting in ERß poly-ubiquitination. Mutation of CBP Thr-1872 or Mdm2 Ser-186/188 Akt sites resulted in a dissociation of the ERß-CBP-Mdm2 complex and reduced ERß turnover. We found that the decrease in ERß induced by heregulin-ß was associated with reduced target gene promoter occupancy and enhanced proliferation of breast cancer cells. However, knockdown of Mdm2 restored endogenous ERß levels resulting in reduction of breast cancer cell growth. These studies identify a tripartite Akt-regulated phosphorylation mechanism that functions to hamper normal ERß activity and turnover through the concerted actions of CBP and Mdm2 in response to growth factor signaling pathways in breast cancer cells.