Exploring legal age estimation using DNA methylation.

Minors (subjects under the legal age, established at this study at 18 years) benefit from a series of legal rights created to protect them and guarantee their welfare. However, throughout the world there are many minors who have no way to prove they are underaged, leading to a great interest in predicting legal age with the highest possible accuracy. Current methods, mainly involving X-ray analysis, are highly invasive, so new methods to predict legal age are being studied, such as DNA methylation. To further such studies, we created two age prediction models based on five epigenetic markers: cg21572722 (ELOVL2), cg02228185 (ASPA), cg06639320 (FHL2), cg19283806 (CCDC102B) and cg07082267 (no associated gene), that were analysed in blood samples to determine possible limitations regarding DNA methylation as an effective tool for legal age estimation. A wide age range prediction model was created using a broad set of samples (14-94 years) yielding a mean absolute error (MAE) of ±4.32 years. A second model, the constrained age prediction model, was created using a reduced range of samples (14-25 years) yielding an MAE of ±1.54 years. Both models, in addition to Horvath's Skin & Blood epigenetic clock, were evaluated using a test set comprising 732 pairs of 18-year-old twins (N=426 monozygotic (MZ) and N=306 dizygotic (DZ) pairs), representing a relevant age of study. Through analysis of the two former age prediction models, we found that constraining the age of the samples forming the training set around the desired age of study significantly reduced the prediction error (from MAE: ±4.07 and ±4.27 years for MZ and DZ twins, respectively; to ±1.31 and ±1.3 years). However, despite low prediction errors, DNA methylation models are still prone to classify same-aged individuals in different categories (minors or adults), despite each sample belonging to the same twin pair. Additional evaluation of Horvath's Skin & Blood model (391 CpGs) led to similar results in terms of age prediction errors than if using only five epigenetic markers (MAE: ±1.87 and ±1.99 years for MZ and DZ twins, respectively).

Performance comparison of a previously validated microhaplotype panel and a forensic STR panel for DNA mixture analysis.

Short Tandem Repeats (STRs) are the most widespread markers in forensic genetics. However, STR stutter peaks can mask alleles from a minor contributor when analysing mixtures, hindering the interpretation of complex profiles. In this study we compared the performance of a previously described panel of microhaplotypes (MHs), an alternative type of forensic marker, against a standard STR kit. The parameters evaluated included: capability of determining the minimum number of contributors in the mixture; percentages of allele drop-outs and drop-ins; retrieval of alleles belonging to the minor contributor, and estimation of likelihood ratio (LR) values. In addition, the capacity of EuroForMix software to estimate each donor's percentage of contribution was tested, as well as the impact on results when using manually, or automatically prepared libraries. The MH panel showed better performance than STRs for the detection of 2-contributor mixtures, but the lower degree of polymorphism per MH marker hindered the task of deconvolution with multiple contributors. MHs presented higher drop-in rates and lower drop-out rates, a higher capability to recover the minor contributor's alleles and provided higher LR values than STRs, likely due to the much higher number of loci combined in the panel. Estimations of contributor ratios using EuroForMix showed promising results and marginal differences were found in these values between manually and automatically prepared libraries. Overall, results showed that the mixture detection performance of the MH panel was better or equal to the standard forensic autosomal STR panel, indicating microhaplotypes are informative markers for this purpose.

[Translated article] Epidemiology of Primary Cutaneous Melanoma in the Migjorn Health Sector of Mallorca, Spain From 2003 Through 2021.

The study of the increasing incidence of melanoma over the past few decades is essential regarding prevention and optimization of health resources. We collected cases of melanoma from Hospital son Llàtzer from the Migjorn health sector of Mallorca, Spain from 2003 through 2021, and calculated the incidence of melanoma adjusted to the standard European population. In addition, other demographic and clinicopathological data were descriptively analyzed too. A total of 690 new cases of melanoma were detected with a progressive increase in the age-standardized incidence from 7.47 cases per 100,000 inhabitants/year in 2003 up to 23.84 in 2021 mainly due to early stages of the disease. The incidence of melanoma has increased significantly in Mallorca probably due to the increasing population coming from northern Europe (low phototypes), sun exposure habits (tourism, fishing, agriculture), and improved early diagnosis.

Impact of interfacial curvature on molecular properties of aqueous interfaces.

The curvature of soft interfaces plays a crucial role in determining their mechanical and thermodynamic properties, both at macroscopic and microscopic scales. In the case of air/water interfaces, particular attention has recently focused on water microdroplets, due to their distinctive chemical reactivity. However, the specific impact of curvature on the molecular properties of interfacial water and interfacial reactivity has so far remained elusive. Here, we use molecular dynamics simulations to determine the effect of curvature on a broad range of structural, dynamical, and thermodynamical properties of the interface. For a droplet, a flat interface, and a cavity, we successively examine the structure of the hydrogen-bond network and its relation to vibrational spectroscopy, the dynamics of water translation, rotation, and hydrogen-bond exchanges, and the thermodynamics of ion solvation and ion-pair dissociation. Our simulations show that curvature predominantly impacts the hydrogen-bond structure through the fraction of dangling OH groups and the dynamics of interfacial water molecules. In contrast, curvature has a limited effect on solvation and ion-pair dissociation thermodynamics. For water microdroplets, this suggests that the curvature alone cannot fully account for the distinctive reactivity measured in these systems, which are of great importance for catalysis and atmospheric chemistry.

Phenogenetic profile and agronomic contribution of Azospirillum argentinense Az39T, a reference strain for the South American inoculant industry.

Azospirillum sp. is a plant growth-promoting rhizobacteria largely recognized for its potential to increase the yield of different important crops. In this work, we present a thorough genomic and phenotypic analysis of A. argentinense Az39T to provide new insights into the beneficial mechanisms of this microorganism. Phenotypic analyses revealed the following in vitro abilities: growth at 20-38 °C (optimum, 28 °C), pH 6.0-8.0 (optimum, pH 6.8), and in the presence of 1% (w/v) NaCl; production of variable amounts of PHB as intracellular granules; nitrogen fixation under microaerophilic conditions; IAA synthesis in the presence of L-tryptophan. Through biochemical (API 20NE) and carbon utilization profiling (Biolog) assays, we proved that A. argentinense Az39T is able to use 15 substrates and metabolize 19 different carbon substrates. Lipid composition indicated a predominance of medium and long-chain saturated fatty acids. A total of 6 replicons classified as one main chromosome, three chromids, and two plasmids, according to their tRNA and core essential genes contents, were identified. Az39T genome includes genes associated with multiple plant growth-promoting (PGP) traits such as nitrogen fixation and production of auxins, cytokinin, abscisic acid, ethylene, and polyamines. In addition, Az39T genome harbor genetic elements associated with physiological features that facilitate its survival in the soil and competence for rhizospheric colonization; this includes motility, secretion system, and quorum sensing genetic determinants. A metadata analysis of Az39T agronomic performance in the pampas region, Argentina, demonstrated significant grain yield increases in wheat and maize, proving its potential to provide better growth conditions for dryland cereals. In conclusion, our data provide a detailed insight into the metabolic profile of A. argentinense Az39T, the strain most widely used to formulate non-legume inoculants in Argentina, and allow a better understanding of the mechanisms behind its field performance.

Inference of tobacco and alcohol consumption habits from DNA methylation analysis of blood.

DNA methylation has become a biomarker of great interest in the forensic and clinical fields. In criminal investigations, the study of this epigenetic marker has allowed the development of DNA intelligence tools providing information that can be useful for investigators, such as age prediction. Following a similar trend, when the origin of a sample in a criminal scenario is unknown, the inference of an individual's lifestyle such as tobacco use and alcohol consumption could provide relevant information to help in the identification of DNA donors at the crime scene. At the same time, in the clinical domain, prediction of these trends of consumption could allow the identification of people at risk or better identification of the causes of different pathologies. In the present study, DNA methylation data from the UK AIRWAVE study was used to build two binomial logistic models for the inference of smoking and drinking status. A total of 348 individuals (116 non-smokers, 116 former smokers and 116 smokers) plus a total of 237 individuals (79 non-drinkers, 79 moderate drinkers and 79 drinkers) were used for development of tobacco and alcohol consumption prediction models, respectively. The tobacco prediction model was composed of two CpGs (cg05575921 in AHRR and cg01940273) and the alcohol prediction model three CpGs (cg06690548 in SLC7A11, cg0886875 and cg21294714 in MIR4435-2HG), providing correct classifications of 86.49% and 74.26%, respectively. Validation of the models was performed using leave-one-out cross-validation. Additionally, two independent testing sets were also assessed for tobacco and alcohol consumption. Considering that the consumption of these substances could underlie accelerated epigenetic ageing patterns, the effect of these lifestyles on the prediction of age was evaluated. To do that, a quantile regression model based on previous studies was generated, and the potential effect of tobacco and alcohol consumption with the epigenetic age was assessed. The Wilcoxon test was used to evaluate the residuals generated by the model and no significant differences were observed between the categories analyzed.

Características clínicas y epidemiológicas de pacientes diagnosticados de escabiosis en España: oportunidades de mejora. Estudio transversal multicéntrico CLINI-AEDV / Clinical and Epidemiological Characteristics of Patients Being Treated for Scabies in Spain: Opportunities for Improvement — A Multicenter Cross-Sectional CLINI-AEDV Study

Antecedentes Diversos trabajos apoyan la hipótesis de que en España se está produciendo un aumento de incidencia de la escabiosis, y existen dudas sobre el posible desarrollo de resistencias y el incremento de formas clínicas atípicas. Los objetivos de este estudio fueron caracterizar el perfil demográfico y clínico de los pacientes de escabiosis atendidos por dermatólogos en España, identificar la posible aparición de escabiosis atípicas, así como describir la frecuencia y los posibles factores de riesgo de los fracasos terapéuticos previos. Métodos Realizamos un estudio observacional, transversal, multicéntrico con recogida de datos prospectiva, en abril y mayo de 2023 dentro de la plataforma CLINI-AEDVp de la Academia Española de Dermatología y Venereología. Resultados Se reclutaron 186 casos de escabiosis activa (51% mujeres) en 31 centros participantes de 15 comunidades autónomas. Se requirió un nivel A, B o C de los criterios de consenso de la International Alliance for the Control of Scabies (IACS) para el diagnóstico. El 92% de los pacientes presentaron formas clínicas típicas de escabiosis y un 66% había recibido tratamiento escabicida previo para el episodio en curso. De los pacientes previamente tratados, solo un 36% había recibido y cumplimentado una pauta terapéutica adecuada que incluyera el tratamiento simultáneo de convivientes, y un 50% careció de un documento escrito y claro con las recomendaciones. Conclusiones Una elevada proporción de los casos de escabiosis atendidos actualmente ha recibido tratamiento previo. En estos se observan defectos corregibles que pueden justificar parte de los fracasos terapéuticos. Trabajar en la mejora de las deficiencias encontradas ayudará a un mejor control de la enfermedad y a evaluar la efectividad actual de los escabicidas disponibles (AU)

Background Several studies support the hypothesis that scabies is on the rise in Spain. There are also concerns about the possible development of resistance to treatment and an increase in atypical presentations. The aims of this study were to describe the demographic and clinical characteristics of patients with scabies seen by dermatologists in Spain, to identify the possible emergence of atypical forms of scabies, and to explore the frequency of treatment failures and associated risk factors. Methods We conducted an observational, cross-sectional, multicenter study of data collected prospectively in April and May 2023 using the CLINI-AEDVp platform created by the Spanish Academy of Dermatology and Venereology (AEDV). Results Participating dermatologists from 31 hospitals in 15 of Spain's autonomous communities recorded 186 cases of active scabies (51% in women) during the study period. A diagnostic certainty level of A, B or C as per the International Alliance for the Control of Scabies Consensus Criteria was required for diagnosis. Overall, 92% of patients had typical scabies and 66% had already been treated with a scabicide for the current episode. Of the treated patients, only 36% had received and completed adequate treatment (including the simultaneous treatment of all household members) and 50% had not received clear written recommendations. Conclusions In a high proportion of the cases of scabies studied, the patient had already received treatment. In those cases, we observed several remediable shortcomings that could explain why some of these treatments had failed. Remedying these deficiencies should lead to better control of scabies and an improved assessment of the actual effectiveness of currently available scabicides (AU)

[Translated article] Clinical and Epidemiological Characteristics of Patients Being Treated for Scabies in Spain: Opportunities for Improvement — A Multicenter Cross-Sectional CLINI-AEDV Study / Características clínicas y epidemiológicas de pacientes diagnosticados de escabiosis en España: oportunidades de mejora. Estudio transversal multicéntrico CLINI-AEDV

Background Several studies support the hypothesis that scabies is on the rise in Spain. There are also concerns about the possible development of resistance to treatment and an increase in atypical presentations. The aims of this study were to describe the demographic and clinical characteristics of patients with scabies seen by dermatologists in Spain, to identify the possible emergence of atypical forms of scabies, and to explore the frequency of treatment failures and associated risk factors. Methods We conducted an observational, cross-sectional, multicenter study of data collected prospectively in April and May 2023 using the CLINI-AEDVp platform created by the Spanish Academy of Dermatology and Venereology (AEDV). Results Participating dermatologists from 31 hospitals in 15 of Spain's autonomous communities recorded 186 cases of active scabies (51% in women) during the study period. A diagnostic certainty level of A, B or C as per the International Alliance for the Control of Scabies Consensus Criteria was required for diagnosis. Overall, 92% of patients had typical scabies and 66% had already been treated with a scabicide for the current episode. Of the treated patients, only 36% had received and completed adequate treatment (including the simultaneous treatment of all household members) and 50% had not received clear written recommendations. Conclusions In a high proportion of the cases of scabies studied, the patient had already received treatment. In those cases, we observed several remediable shortcomings that could explain why some of these treatments had failed. Remedying these deficiencies should lead to better control of scabies and an improved assessment of the actual effectiveness of currently available scabicides (AU)

Antecedentes Diversos trabajos apoyan la hipótesis de que en España se está produciendo un aumento de incidencia de la escabiosis, y existen dudas sobre el posible desarrollo de resistencias y el incremento de formas clínicas atípicas. Los objetivos de este estudio fueron caracterizar el perfil demográfico y clínico de los pacientes de escabiosis atendidos por dermatólogos en España, identificar la posible aparición de escabiosis atípicas, así como describir la frecuencia y los posibles factores de riesgo de los fracasos terapéuticos previos. Métodos Realizamos un estudio observacional, transversal, multicéntrico con recogida de datos prospectiva, en abril y mayo de 2023 dentro de la plataforma CLINI-AEDVp de la Academia Española de Dermatología y Venereología. Resultados Se reclutaron 186 casos de escabiosis activa (51% mujeres) en 31 centros participantes de 15 comunidades autónomas. Se requirió un nivel A, B o C de los criterios de consenso de la International Alliance for the Control of Scabies (IACS) para el diagnóstico. El 92% de los pacientes presentaron formas clínicas típicas de escabiosis y un 66% había recibido tratamiento escabicida previo para el episodio en curso. De los pacientes previamente tratados, solo un 36% había recibido y cumplimentado una pauta terapéutica adecuada que incluyera el tratamiento simultáneo de convivientes, y un 50% careció de un documento escrito y claro con las recomendaciones. Conclusiones Una elevada proporción de los casos de escabiosis atendidos actualmente ha recibido tratamiento previo. En estos se observan defectos corregibles que pueden justificar parte de los fracasos terapéuticos. Trabajar en la mejora de las deficiencias encontradas ayudará a un mejor control de la enfermedad y a evaluar la efectividad actual de los escabicidas disponibles (AU)

Clinical and Epidemiological Characteristics of Patients Being Treated for Scabies in Spain: Opportunities for Improvement - A Multicenter Cross-Sectional CLINI-AEDV Study. / Características clínicas y epidemiológicas de pacientes diagnosticados de escabiosis en España: oportunidades de mejora. Estudio transversal multicéntrico CLINI-AEDV.

BACKGROUND: Several studies support the hypothesis that scabies is on the rise in Spain. There are also concerns about the possible development of resistance to treatment and an increase in atypical presentations. The aims of this study were to describe the demographic and clinical characteristics of patients with scabies seen by dermatologists in Spain, to identify the possible emergence of atypical forms of scabies, and to explore the frequency of treatment failures and associated risk factors. METHODS: We conducted an observational, cross-sectional, multicenter study of data collected prospectively in April and May 2023 using the CLINI-AEDVp platform created by the Spanish Academy of Dermatology and Venereology (AEDV). RESULTS: Participating dermatologists from 31 hospitals in 15 of Spain's autonomous communities recorded 186 cases of active scabies (51% in women) during the study period. A diagnostic certainty level of A, B or C as per the International Alliance for the Control of Scabies Consensus Criteria was required for diagnosis. Overall, 92% of patients had typical scabies and 66% had already been treated with a scabicide for the current episode. Of the treated patients, only 36% had received and completed adequate treatment (including the simultaneous treatment of all household members) and 50% had not received clear written recommendations. CONCLUSIONS: In a high proportion of the cases of scabies studied, the patient had already received treatment. In those cases, we observed several remediable shortcomings that could explain why some of these treatments had failed. Remedying these deficiencies should lead to better control of scabies and an improved assessment of the actual effectiveness of currently available scabicides.

[Translated article] Clinical and Epidemiological Characteristics of Patients Being Treated for Scabies in Spain: Opportunities for Improvement - A Multicenter Cross-Sectional CLINI-AEDV Study.

BACKGROUND: Several studies support the hypothesis that scabies is on the rise in Spain. There are also concerns about the possible development of resistance to treatment and an increase in atypical presentations. The aims of this study were to describe the demographic and clinical characteristics of patients with scabies seen by dermatologists in Spain, to identify the possible emergence of atypical forms of scabies, and to explore the frequency of treatment failures and associated risk factors. METHODS: We conducted an observational, cross-sectional, multicenter study of data collected prospectively in April and May 2023 using the CLINI-AEDVp platform created by the Spanish Academy of Dermatology and Venereology (AEDV). RESULTS: Participating dermatologists from 31 hospitals in 15 of Spain's autonomous communities recorded 186 cases of active scabies (51% in women) during the study period. A diagnostic certainty level of A, B or C as per the International Alliance for the Control of Scabies Consensus Criteria was required for diagnosis. Overall, 92% of patients had clinical features of classic scabies and 66% had already been treated with a scabicide for the current episode. Of the treated patients, only 36% had received and completed adequate treatment (including the simultaneous treatment of all household members) and 50% had not received clear written recommendations. CONCLUSIONS: In a high proportion of scabies cases, the patient has already received treatment. In those cases, we observe several remediable shortcomings that could explain why some of these treatments fail. Remedying these deficiencies should lead to better control of scabies and an improved assessment of the actual effectiveness of currently available scabicides.

Adapting an established Ampliseq microhaplotype panel to nanopore sequencing through direct PCR.

We have adapted an established Ampliseq microhaplotype panel for nanopore sequencing with the Oxford Nanopore Technologies (ONT) system, as a cost-effective and highly scalable solution for forensic genetics applications. For this purpose, we designed a protocol combining direct PCR amplification from unextracted DNA with ONT library construction and sequencing using the MinION device and workflow. The analysis of reference samples at input amounts of 5-10 ng of DNA demonstrates stable coverage patterns, allele balance, and strand bias, reaching profile completeness and concordance rates of ∼95%. Similar levels were achieved when using direct-PCR from blood, buccal and semen swabs. Dilution series results indicate sensitivity is maintained down to 250 pg of input DNA, and informative profiles are produced down to 62.5 pg. Finally, we demonstrated the forensic utility of the nanopore workflow by analyzing two third degree pedigrees that showed low likelihood ratio values after the analysis of an extended panel of 38 STRs, achieving likelihood ratios 2-3 orders of magnitude higher when testing with the MinION-based haplotype data.

Development and evaluations of the ancestry informative markers of the VISAGE Enhanced Tool for Appearance and Ancestry.

The VISAGE Enhanced Tool for Appearance and Ancestry (ET) has been designed to combine markers for the prediction of bio-geographical ancestry plus a range of externally visible characteristics into a single massively parallel sequencing (MPS) assay. We describe the development of the ancestry panel markers used in ET, and the enhanced analyses they provide compared to previous MPS-based forensic ancestry assays. As well as established autosomal single nucleotide polymorphisms (SNPs) that differentiate sub-Saharan African, European, East Asian, South Asian, Native American, and Oceanian populations, ET includes autosomal SNPs able to efficiently differentiate populations from Middle East regions. The ability of the ET autosomal ancestry SNPs to distinguish Middle East populations from other continentally defined population groups is such that characteristic patterns for this region can be discerned in genetic cluster analysis using STRUCTURE. Joint cluster membership estimates showing individual co-ancestry that signals North African or East African origins were detected, or cluster patterns were seen that indicate origins from central and Eastern regions of the Middle East. In addition to an augmented panel of autosomal SNPs, ET includes panels of 85 Y-SNPs, 16 X-SNPs and 21 autosomal Microhaplotypes. The Y- and X-SNPs provide a distinct method for obtaining extra detail about co-ancestry patterns identified in males with admixed backgrounds. This study used the 1000 Genomes admixed African and admixed American sample sets to fully explore these enhancements to the analysis of individual co-ancestry. Samples from urban and rural Brazil with contrasting distributions of African, European, and Native American co-ancestry were also studied to gauge the efficiency of combining Y- and X-SNP data for this purpose. The small panel of Microhaplotypes incorporated in ET were selected because they showed the highest levels of haplotype diversity amongst the seven population groups we sought to differentiate. Microhaplotype data was not formally combined with single-site SNP genotypes to analyse ancestry. However, the haplotype sequence reads obtained with ET from these loci creates an effective system for de-convoluting two-contributor mixed DNA. We made simple mixture experiments to demonstrate that when the contributors have different ancestries and the mixture ratios are imbalanced (i.e., not 1:1 mixtures) the ET Microhaplotype panel is an informative system to infer ancestry when this differs between the contributors.

Targeted Electromyographic Biofeedback With Endoanal Electrostimulation for Anal Incontinence.

Purpose. Anal incontinence (AI) is a disabling condition with a variable response to conservative physical therapies. We assess the utility of combining electromyographic biofeedback with endoanal electrostimulation targeted to the weakest areas of the pelvic floor using the MAPLe® probe (Multiple Array Probe Leiden Novuqare). Methods. Patients with AI unresponsive to conservative measures were assessed before and after treatment with anorectal manometry (ARM), electromyography (EMG), Wexner Continence Scoring, Visual Analog Scoring (VAS), FIQL and SF-12 quality of life determination. Results. Of 29 patients in the final analysis, there was an improvement in the mean Wexner continence score from 13.59 to 8.03 and a concomitant improvement in the reported VAS from 3.45 to 6.72. Both Wexner continence and VAS scores were maintained during follow-up. Maximum voluntary manometric contraction significantly improved from 91.76 mmHg to 110.33 mmHg with no changes in resting pressure. The EMG values ​​(µV) that significantly improved included the average and peak resistance, the average general voluntary contraction, and the average and peak voluntary contraction for both the external anal sphincter and the puborectalis. In the FIQL, behavior, depression and shame domains improved after treatment and during follow-up with lifestyle improvements detected at 6 and 12 months. Physical and mental components of the SF-12 improved at 6 and 12 months. Conclusions. Targeted electromyographic biofeedback and endoanal electrostimulation using MAPLe® probe in AI patients sustainably improves objective ARM and EMG parameters along with subjective reporting of continence severity, VAS, and quality of life.

Development and inter-laboratory evaluation of the VISAGE Enhanced Tool for Appearance and Ancestry inference from DNA.

Responding to the growing scientific and practical interest in forensic DNA phenotyping, the VISible Attributes through GEnomics (VISAGE) Consortium was founded in 2017 with the main goal of developing and validating new and reliable molecular and statistical tools to predict appearance, ancestry and age from DNA. Here, we describe the development and inter-laboratory evaluation and validation of the VISAGE Enhanced Tool for Appearance and Ancestry inference from DNA. The VISAGE Enhanced Tool for Appearance and Ancestry is the first forensic-driven genetic laboratory tool that comprises well-established markers for eye, hair and skin color with more recently discovered DNA markers for eyebrow color, freckling, hair shape and male pattern baldness and bio-geographic ancestry informative DNA markers. The bio-geographic ancestry markers include autosomal SNPs (bi- and tri-allelic SNPs), X-SNPs, Y-SNPs and autosomal Microhaplotypes. In total, primers targeting 524 SNPs (representing a 97.6% assay conversion rate) were successfully designed using AmpliSeq into a single primer pool (i.e., one multiplex assay) and sequenced with the Ion S5. In a collaborative framework, five VISAGE laboratories tested the VISAGE Enhanced Tool for Appearance and Ancestry on reproducibility, sensitivity, genotyping concordance, mixtures, species specificity and performance in relevant forensic conditions, including inhibitor-spiked, mock casework and artificially degraded samples. Based on our results, the VISAGE Enhanced Tool for Appearance and Ancestry is a robust, reproducible, and - for the large SNP number - fairly sensitive MPS assay with high concordance rates. With the VISAGE Enhanced Tool for Appearance and Ancestry introduced here, the VISAGE Consortium delivers the first single DNA-test for combined appearance prediction based on seven traits together with bio-geographic ancestry inference based on major continental regions for separated bi-parental and paternal ancestry, which represents the most comprehensive validated laboratory tool currently available for Forensic DNA Phenotyping.

Epigenetic age estimation in saliva and in buccal cells.

Age estimation based on epigenetic markers is a DNA intelligence tool with the potential to provide relevant information for criminal investigations, as well as to improve the inference of age-dependent physical characteristics such as male pattern baldness or hair color. Age prediction models have been developed based on different tissues, including saliva and buccal cells, which show different methylation patterns as they are composed of different cell populations. On many occasions in a criminal investigation, the origin of a sample or the proportion of tissues is not known with certainty, for example the provenance of cigarette butts, so use of combined models can provide lower prediction errors. In the present study, two tissue-specific and seven age-correlated CpG sites were selected from publicly available data from the Illumina HumanMethylation 450 BeadChip and bibliographic searches, to help build a tissue-dependent, and an age-prediction model, respectively. For the development of both models, a total of 184 samples (N = 91 saliva and N = 93 buccal cells) ranging from 21 to 86 years old were used. Validation of the models was performed using either k-fold cross-validation and an additional set of 184 samples (N = 93 saliva and N = 91 buccal cells, 21-86 years old). The tissue prediction model was developed using two CpG sites (HUNK and RUNX1) based on logistic regression that produced a correct classification rate for saliva and buccal swab samples of 88.59 % for the training set, and 83.69 % for the testing set. Despite these high success rates, a combined age prediction model was developed covering both saliva and buccal cells, using seven CpG sites (cg10501210, LHFPL4, ELOVL2, PDE4C, HOXC4, OTUD7A and EDARADD) based on multivariate quantile regression giving a median absolute error (MAE): ± 3.54 years and a correct classification rate ( %CP±PI) of 76.08 % for the training set, and an MAE of ± 3.66 years and a %CP±PI of 71.19 % for the testing set. The addition of tissue-of origin as a co-variate to the model was assessed, but no improvement was detected in age predictions. Finally, considering the limitations usually faced by forensic DNA analyses, the robustness of the model and the minimum recommended amount of input DNA for bisulfite conversion were evaluated, considering up to 10 ng of genomic DNA for reproducible results. The final multivariate quantile regression age predictor based on the models we developed has been placed in the open-access Snipper forensic classification website.

Eurasiaplex-2: Shifting the focus to SNPs with high population specificity increases the power of forensic ancestry marker sets.

To compile a new South Asian-informative panel of forensic ancestry SNPs, we changed the strategy for selecting the most powerful markers for this purpose by targeting polymorphisms with near absolute specificity - when the South Asian-informative allele identified is absent from all other populations or present at frequencies below 0.001 (one in a thousand). More than 120 candidate SNPs were identified from 1000 Genomes datasets satisfying an allele frequency screen of ≥ 0.1 (10 % or more) allele frequency in South Asians, and ≤ 0.001 (0.1 % or less) in African, East Asian, and European populations. From the candidate pool of markers, a final panel of 36 SNPs, widely distributed across most autosomes, were selected that had allele frequencies in the five 1000 Genomes South Asian populations ranging from 0.4 to 0.15. Slightly lower average allele frequencies, but consistent patterns of informativeness were observed in gnomAD South Asian datasets used to validate the 1000 Genomes variant annotations. We named the panel of 36 South Asian-specific SNPs Eurasiaplex-2, and the informativeness of the panel was evaluated by compiling worldwide population data from 4097 samples in four genome variation databases that largely complement the global sampling of 1000 Genomes. Consistent patterns of allele frequency distribution, which were specific to South Asia, were observed in all populations in, or closely sited to, the Indian sub-continent. Pakistani populations from the HGDP-CEPH panel had markedly lower allele frequencies, highlighting the need to develop a statistical system to evaluate the ancestry inference value of counting the number of population-specific alleles present in an individual.

La evaluación de la fragilidad puede mejorar la predicción del APACHE II en pacientes ancianos ingresados en UCI tras cirugía digestiva / Evaluating frailty may complement APACHE II in estimating mortality in elderly patients admitted to the ICU after digestive surgery

Objetivo: Analizar si la fragilidad puede mejorar la predicción de mortalidad en los pacientes ingresados en UCI tras una cirugía digestiva. Diseño: Estudio prospectivo, observacional y con seguimiento a 6 meses de una cohorte de pacientes que ingresaron en UCI entre el 1 de junio de 2018 hasta el 1 de junio de 2019. Ámbito: UCI quirúrgica de un hospital de tercer nivel. Pacientes: Serie de pacientes sucesivos mayores de 70 años que ingresaron en UCI inmediatamente después de una intervención quirúrgica sobre el aparato digestivo. Fueron incluidos 92 pacientes y se excluyeron 2 por pérdida de seguimiento a los 6 meses. Intervenciones: Al ingreso en UCI se estimó gravedad y pronóstico mediante el APACHE II, y fragilidad mediante la Clinical Frailty Scale y el modified Frailty Index. Variables de interés principales: Mortalidad en UCI, intrahospitalaria y a los 6 meses. Resultados: El modelo que mejor predice mortalidad en UCI es el APACHE II, con un área bajo la curva ROC (ABC) de 0,89 y una buena calibración. El modelo que combina APACHE II y Clinical Frailty Scale es el que mejor predice mortalidad intrahospitalaria (ABC: 0,82), mejorando significativamente la predicción del APACHE II aislado (ABC: 0,78; Integrated Discrimination Index: 0,04). La fragilidad es un factor predictor de mortalidad a los 6 meses, siendo el modelo que combina la Clinical Frailty Scale y el modified Frailty Index el que ha demostrado mayor discriminación (ABC: 0,84). Conclusiones: La fragilidad puede complementar al APACHE II mejorando su predicción de mortalidad hospitalaria. Además, ofrece una buena predicción de la mortalidad a los 6 meses de la cirugía. Para la mortalidad en UCI, la fragilidad pierde su poder de predicción mientras que el APACHE II aislado muestra una excelente capacidad predictiva (AU)

Objective: To analyze whether frailty can improve the prediction of mortality in patients admitted to the ICU after digestive surgery. Design: Prospective, observational, 6-month follow-up study of a cohort of patients admitted to the ICU between June 1, 2018, and June 1, 2019. Setting: Surgical ICU of a third level hospital. Patients: Series of successive patients older than 70 years who were admitted to the ICU immediately after a surgical intervention on the digestive system. 92 patients were included and 2 were excluded due to loss of follow-up at 6 months. Interventions: Upon admission to the ICU, severity and prognosis were assessed by APACHE II, and fragility by the Clinical Frailty Scale and the modified Frailty Index. Main variables of interest: ICU, in-hospital and 6-month mortality. Results: The model that best predicts mortality in the ICU is the APACHE II, with an area under the ROC curve (AUC) of 0.89 and a good calibration. The model that combines APACHE II and Clinical Frailty Scale is the one that best predicts in-hospital mortality (AUC: 0.82), significantly improving the prediction of isolated APACHE II (AUC: 0.78; Integrated Discrimination Index: 0.04). Frailty is a predictor of mortality at 6 months, being the model that combines Clinical Frailty Scale and Frailty Index the one that has shown the greatest discrimination (AUC: 0.84). Conclusions: Frailty can complement APACHE II by improving its prediction of hospital mortality. Furthermore, it offers a good prediction of mortality 6 months after surgery. For mortality in ICU, frailty loses its predictive power, whereas isolated APACHE II shows excellent predictive capacity (AU)

Evaluating frailty may complement APACHE II in estimating mortality in elderly patients admitted to the ICU after digestive surgery.

OBJECTIVE: To analyze whether frailty can improve the prediction of mortality in patients admitted to the ICU after digestive surgery. DESIGN: Prospective, observational, 6-month follow-up study of a cohort of patients admitted to the ICU between June 1, 2018, and June 1, 2019. SETTING: Surgical ICU of a third level hospital. PATIENTS: Series of successive patients older than 70 years who were admitted to the ICU immediately after a surgical intervention on the digestive system. 92 patients were included and 2 were excluded due to loss of follow-up at 6 months. INTERVENTIONS: Upon admission to the ICU, severity and prognosis were assessed by APACHE II, and fragility by the Clinical Frailty Scale and the modified Frailty Index. MAIN VARIABLES OF INTEREST: ICU, in-hospital and 6-month mortality. RESULTS: The model that best predicts mortality in the ICU is the APACHE II, with an area under the ROC curve (AUC) of 0.89 and a good calibration. The model that combines APACHE II and Clinical Frailty Scale is the one that best predicts in-hospital mortality (AUC: 0.82), significantly improving the prediction of isolated APACHE II (AUC: 0.78; Integrated Discrimination Index: 0.04). Frailty is a predictor of mortality at 6 months, being the model that combines Clinical Frailty Scale and Frailty Index the one that has shown the greatest discrimination (AUC: 0.84). CONCLUSIONS: Frailty can complement APACHE II by improving its prediction of hospital mortality. Furthermore, it offers a good prediction of mortality 6 months after surgery. For mortality in ICU, frailty loses its predictive power, whereas isolated APACHE II shows excellent predictive capacity.