Magnum metal-on-metal uncemented total hip replacement: 8- to 18-year outcomes of 211 cases.

BACKGROUND: Reports of adverse reactions to metal debris contributed in part to a decline in use of large-bearing metal-on-metal total hip devices. We hypothesize an optimal trunnion design may reduce risk of this failure mode in large-bearing total hip arthroplasty systems. The purpose of this study is to report mid- to long-term outcomes for a single-surgeon series of 211 total hip arthroplasties using the large-bearing Biomet Magnum metal-on-metal system. MATERIALS AND METHODS: Between December 2004 and January 2016, the primary surgeon performed 211 uncemented Magnum total hip arthroplasties in 181 patients. The average length of follow-up was 10.1 ± 3.5 years (range 8-18 years). RESULTS: Using failure of any component as the endpoint, the overall survivorship rate was 98.1% at 10 years and 97.4% at 18 years. These eight failures (3.8% of cohort) included one case of adverse wear-related failure (0.5%), two cases of acetabular ingrowth failure (0.9%), three cases of trunnion corrosion (1.4%), one failure of late infection (0.5%), and one inappropriate revision of components for trochanteric nonunion without instability (0.5%). Excluding failed cases, all components were radiographically stable with no radiolucencies. Except for the one wear failure, ion testing revealed that 97.2% of cases were within optimal whole blood metal ion levels with the remaining ion test results within acceptable levels. CONCLUSIONS: With the uncemented Magnum metal-on-metal total hip, we achieved 97.4% 18-year implant survivorship, exceeding the NICE criteria and registry benchmarks for implant survivorship. We observed a trunnion corrosion rate of 1.4% and no cases of instability. The single case of adverse wear-related failure was caused by acetabular component malposition.

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.

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.

Patient-reported outcomes version of the common terminology criteria for adverse events and quality-of-life linear analogue self-assessment in breast cancer patients receiving radiation therapy: single-institution prospective registry.

PURPOSE/OBJECTIVES: We sought to investigate the impact of patient-reported outcomes version of the common terminology criteria for adverse events (PRO-CTCAE) on overall quality-of-life (QOL) employing linear analogue self-assessment (LASA) in breast cancer (BC) patients undergoing radiation therapy (RT). MATERIALS/METHODS: All patients treated with RT for BC with curative intent from 2015 to 2019 at our institution were included. Breast specific PRO-CTCAE and overall QOL LASA questionnaires were administered at baseline, end-of-treatment, 3, 6, 12 months, and then annually. Minimal clinically important difference in overall QOL was a 10-point change in LASA. Hypofractionation was any treatment > 2 Gy per fraction. Mixed models for repeated measures were used to determine the association of PRO-CTCAE and overall QOL LASA. RESULTS: Three hundred thirty-one (331) patients with a median follow-up of 3.1 years (range 0.4-4.9) were included. Average overall QOL LASA scores were 78.5 at baseline, 79.8 at end-of-treatment, 79.8 at 3 months, 77.1 at 6 months, 79.4 at 12 months, and 79.7 at 24 months. On univariate analysis, patients reporting a grade ≥ 3 PRO-CTCAE had, on average, a 10.4-point reduction in overall LASA QOL (p < 0.0001). On multivariate analysis, not being treated with hypofractionation and higher BMI were predictive for worse overall LASA QOL with a 10-point reduction in LASA for patients reporting a grade ≥ 3 PRO-CTCAE (p < 0.0001). CONCLUSIONS: Patients reporting a grade ≥ 3 PRO-CTCAE experienced statistically significant and clinically meaningful deterioration in overall QOL LASA. Hypofractionation improved QOL while higher BMI predicted for worse QOL. PRO-CTCAE should be integrated into future clinical trials.

[Metastatic Pulmonary Calcification: A Rare Entity in the Differential Diagnosis Examination of Interstitial Lung Diseases]. / Metastatische pulmonale Kalzifikation ­ eine selten diagnostizierte Entität in der differenzialdiagnostischen Abklärung interstitieller Lungenerkrankungen.

A 70-year-old patient who received a kidney transplant about 20 years ago due to anti-neutrophil cytoplasmic antibody (ANCA)-positive, rapidly progressive glomerulonephritis was assigned to us for an evaluation of fibrosing interstitial lung disease with computer tomography (CT)-radiological changes assigned to acute alveolitis. She complained about long-term exertional dyspnea. In terms of lung function, there was some slight restriction (FVC 78 % of the target), and pronounced severe diffusion disorder (DLCO 41 % of the target). There was no evidence of exogenous allergic alveolitis in either the history or serology. In the bronchoscopically obtained samples, changes in the sense of a non-specific interstitial lung disease of the fibrotic type, e. g., matching a reaction in the context of immunosuppressive therapy, were observed. Herpes simplex virus was detected microbiologically in the bronchoalveolar lavage (BAL) fluid using the polymerase chain reaction (PCR). Antiviral therapy was carried out under the working diagnosis of herpes pneumonitis. Immunosuppressive therapy was continued. The herpes virus could no longer be detected in a control. The patient initially reported subjective improvement of dyspnea. Repeated control CT imaging was carried out and after about one year, the initial radiological changes were still present and dyspnea was persistent. A new transbronchial lung biopsy revealed metastatic pulmonary calcification. Fortunately, the disease was not active. CONCLUSION: In the differential diagnostic evaluation of interstitial lung diseases, especially in patients with a vulnerable calcium-phosphate balance and acid-base balance, as occurs, for example, in the context of chronic kidney diseases, the possibility of metastatic pulmonary calcification (MPC) must also be considered especially if the radiological picture shows persistent, upper lobe-accentuated ground glass opacities. The diagnosis requires multidisciplinary cooperation between pulmonologists, radiologists and pathologists.

Hip resurfacing as an outpatient procedure: a comparison of overall cost and review of safety.

Recent advancements in arthroplasty surgical techniques and perioperative protocols have reduced the duration of hospitalization and length of recovery, allowing surgeons to perform joint replacement as an outpatient procedure. This study aims to evaluate the cost-effectiveness and safety of outpatient hip resurfacing. Two experienced surgeons performed 485 resurfacing surgeries. We retrospectively compared clinical outcomes and patient satisfaction with published outpatient total hip results. Furthermore, we compared average insurance reimbursement with that of local inpatient hip replacement. No major complications occurred within 6 weeks. Of the 39 patients with previous inpatient experience, 37 (95%) believed their outpatient experience was superior. The average reimbursement for hip arthroplasty at local hospitals was $50,000, while the average payment for outpatient resurfacing at our surgery center was $26,000. We conclude that outpatient hip resurfacing can be accomplished safely, with high patient satisfaction, and at a tremendous financial savings to the insurer/patient. LEVEL OF EVIDENCE: III.

[Cardiac congestion due to compression of the left atrium by an endoleak following endovascular aortic repair (EVAR)]. / Kardiale Kongestion als Folge einer Kompression des linken Atriums durch eine Endoleckage einer endovaskulären Aortenreparatur (EVAR).

In rare cases, global cardiac decompensation is the only sign of a relevant endoleak after endovascular aortic repair (EVAR) therapy from a thoracic aortic aneurysm due to a mechanical compression of the left atrium. In this case report, we describe such an unusual course of disease and we point out the importance of early bed-side echocardiography in the emergency department for immediate diagnostic and therapeutic planning.

MAPlex - A massively parallel sequencing ancestry analysis multiplex for Asia-Pacific populations.

Current forensic ancestry-informative panels are limited in their ability to differentiate populations in the Asia-Pacific region. MAPlex (Multiplex for the Asia-Pacific), a massively parallel sequencing (MPS) assay, was developed to improve differentiation of East Asian, South Asian and Near Oceanian populations found in the extensive cross-continental Asian region that shows complex patterns of admixture at its margins. This study reports the development of MAPlex; the selection of SNPs in combination with microhaplotype markers; assay design considerations for reducing the lengths of microhaplotypes while preserving their ancestry-informativeness; adoption of new population-informative multiple-allele SNPs; compilation of South Asian-informative SNPs suitable for forensic AIMs panels; and the compilation of extensive reference and test population genotypes from online whole-genome-sequence data for MAPlex markers. STRUCTURE genetic clustering software was used to gauge the ability of MAPlex to differentiate a broad set of populations from South and East Asia, the West Pacific regions of Near Oceania, as well as the other globally distributed population groups. Preliminary assessment of MAPlex indicates enhanced South Asian differentiation with increased divergence between West Eurasian, South Asian and East Asian populations, compared to previous forensic SNP panels of comparable scale. In addition, MAPlex shows efficient differentiation of Middle Eastern individuals from Europeans. MAPlex is the first forensic AIM assay to combine binary and multiple-allele SNPs with microhaplotypes, adding the potential to detect and analyze mixed source forensic DNA.

Robot-assisted abdominal wall excision of a PSMA-detected prostate cancer metastasis.

Recent advances in novel functional imaging techniques such as PSMA PET may now offer the ability to identify small volume metastases which may otherwise go undetected. The treatment of these lesions is controversial, particularly in the oligometastatic state. We report the case of an abdominal wall metastasis detected with PSMA imaging which we treated with surgical excision. This resulted in an undetectable PSA 6 weeks post excision.

Does statin therapy impact the proximal aortopathy in aortic valve disease?

BACKGROUND: Studies have demonstrated that statin therapy decreases the growth rate of abdominal aneurysms. However, the effect of statin therapy on the proximal aortic disease has not been sufficiently elucidated. AIM: We aimed to analyse the association between statin treatment and the severity of proximal aortopathy in patients with aortic valve disease. DESIGN: Cross-sectional study. METHODS: We prospectively identified 458 patients who were referred for aortic valve surgery from 2008 to 2014. Pre-operative measurement of the proximal aorta was performed by TEE, CT or MRT scan. Data of dyslipidemia treatment was obtained by questionnaire. RESULTS: The mean ascending aortic diameter in the whole study population was comparable in patients with vs. without statin therapy (i.e. 42.7 mm vs. 43.6 mm, P = 0.46). Logistic regression analysis showed no significant association between statin therapy and proximal aortopathy ≥ 40 mm in the whole study group (OR = 0.69, P = 0.10). For the BAV sub-group, HDL (OR = 0.54, P = 0.038) and cholesterol levels (OR = 2.00, P = 0.038) were found significantly associated with the proximal aortic disease. In the BAV cohort, the statin users with target HDL levels presented a significantly smaller proximal aortic diameter (40.1 mm vs. 46 mm, P = 0.02). CONCLUSION: Pre-operative statin therapy demonstrated no significant association with the expression of proximal aortopathy. However, more than 40% of the statin users presented uncontrolled lipid levels at the time of the study. In the BAV sub-group, the statins users with target HDL levels showed a significantly smaller ascending aorta diameter. Target HDL and cholesterol levels were strongly associated with proximal aortic dilation in BAV patients.

Body fluid identification using a targeted mRNA massively parallel sequencing approach - results of a EUROFORGEN/EDNAP collaborative exercise.

In a previous study we presented an assay for targeted mRNA sequencing for the identification of human body fluids, optimised for the Illumina MiSeq/FGx MPS platform. This assay, together with an additional in-house designed assay for the Ion Torrent PGM/S5 platform, was the basis for a collaborative exercise within 17 EUROFORGEN and EDNAP laboratories, in order to test the efficacy of targeted mRNA sequencing to identify body fluids. The task was to analyse the supplied dried body fluid stains and, optionally, participants' own bona fide or mock casework samples of human origin, according to specified protocols. The provided primer pools for the Illumina MiSeq/FGx and the Ion Torrent PGM/S5 platforms included 33 and 29 body fluid specific targets, respectively, to identify blood, saliva, semen, vaginal secretion, menstrual blood and skin. The results demonstrated moderate to high count values in the body fluid or tissue of interest with little to no counts in non-target body fluids. There was some inter-laboratory variability in read counts, but overall the results of the laboratories were comparable in that highly expressed markers showed high read counts and less expressed markers showed lower counts. We performed a partial least squares (PLS) analysis on the data, where blood, menstrual blood, saliva and semen markers and samples clustered well. The results of this collaborative mRNA massively parallel sequencing (MPS) exercise support targeted mRNA sequencing as a reliable body fluid identification method that could be added to the repertoire of forensic MPS panels.

[Impact of weather, time of day and season on the admission and outcome of major trauma patients]. / Wie hängen Eintreffen und Outcome schwerer Verletzter im Traumazentrum von Wetter, Tages- und Jahreszeit ab?

BACKGROUND AND OBJECTIVES: The potential association of external factors such as time of day, lunar phases or specific weather conditions on the daily management and outcome of trauma emergencies remains under debate. We undertook this trauma centre investigation to detect possible worthwhile factors of influence in order to optimize the organizational structure of trauma admissions. MATERIALS AND METHODS: Retrospective cohort analysis over the years 2010-2013, including all emergency trauma admissions with a new injury severity score (NISS) ≥8 (major trauma) treated in a teaching hospital in the lowland of Switzerland (uni- and multivariable analysis; p < 0.05). RESULTS: During the study period, 1178 major trauma patients were admitted to the hospital. The mean age of trauma victims was 53 ± 23 and the average ISS was 14 ± 8. More patients arrived within the summer months than during the rest of the year (p < 0.001). Higher energy trauma was found to correlate with higher daytime temperature, longer duration of sunshine (each p < 0.001), and change in weather conditions (p = 0.008). In contrast, snowfall and lunar phases did not demonstrate any association with the number or characteristics of trauma admissions. Multivariable analysis demonstrated that altogether longer sunshine, higher minimum daytime temperature and lower air humidity, compared to the previous day, accounted for 31 % of major trauma admissions. We could not find any impact of the investigated external factors on the outcome of patients. CONCLUSIONS: The study shows a significant relationship between specific weather conditions, such as higher daytime temperature or change in circulation, and the admission of major trauma patients. Due to the small effect in our setting, our results do not implicate any according change in the management of resources. Nevertheless, for hospitals in other geographic or more exposed weather regions, such effects could indeed be relevant and therefore should be tested.

A GHEP-ISFG collaborative study on the genetic variation of 38 autosomal indels for human identification in different continental populations.

A collaborative effort was carried out by the Spanish and Portuguese Speaking Working Group of the International Society for Forensic Genetics (GHEP-ISFG) to promote knowledge exchange between associate laboratories interested in the implementation of indel-based methodologies and build allele frequency databases of 38 indels for forensic applications. These databases include populations from different countries that are relevant for identification and kinship investigations undertaken by the participating laboratories. Before compiling population data, participants were asked to type the 38 indels in blind samples from annual GHEP-ISFG proficiency tests, using an amplification protocol previously described. Only laboratories that reported correct results contributed with population data to this study. A total of 5839 samples were genotyped from 45 different populations from Africa, America, East Asia, Europe and Middle East. Population differentiation analysis showed significant differences between most populations studied from Africa and America, as well as between two Asian populations from China and East Timor. Low FST values were detected among most European populations. Overall diversities and parameters of forensic efficiency were high in populations from all continents.

Inter-laboratory evaluation of the EUROFORGEN Global ancestry-informative SNP panel by massively parallel sequencing using the Ion PGM™.

The EUROFORGEN Global ancestry-informative SNP (AIM-SNPs) panel is a forensic multiplex of 128 markers designed to differentiate an individual's ancestry from amongst the five continental population groups of Africa, Europe, East Asia, Native America, and Oceania. A custom multiplex of AmpliSeq™ PCR primers was designed for the Global AIM-SNPs to perform massively parallel sequencing using the Ion PGM™ system. This study assessed individual SNP genotyping precision using the Ion PGM™, the forensic sensitivity of the multiplex using dilution series, degraded DNA plus simple mixtures, and the ancestry differentiation power of the final panel design, which required substitution of three original ancestry-informative SNPs with alternatives. Fourteen populations that had not been previously analyzed were genotyped using the custom multiplex and these studies allowed assessment of genotyping performance by comparison of data across five laboratories. Results indicate a low level of genotyping error can still occur from sequence misalignment caused by homopolymeric tracts close to the target SNP, despite careful scrutiny of candidate SNPs at the design stage. Such sequence misalignment required the exclusion of component SNP rs2080161 from the Global AIM-SNPs panel. However, the overall genotyping precision and sensitivity of this custom multiplex indicates the Ion PGM™ assay for the Global AIM-SNPs is highly suitable for forensic ancestry analysis with massively parallel sequencing.

The influence of dispersal on a predator-prey system with two habitats.

Dispersal between different habitats influences the dynamics and stability of populations considerably. Furthermore, these effects depend on the local interactions of a population with other species. Here, we perform a general and comprehensive study of the simplest possible system that includes dispersal and local interactions, namely a 2-patch 2-species system. We evaluate the impact of dispersal on stability and on the occurrence of bifurcations, including pattern forming bifurcations that lead to spatial heterogeneity, in 19 different classes of models with the help of the generalized modelling approach. We find that dispersal often destabilizes equilibria, but it can stabilize them if it increases population losses. If dispersal is nonrandom, i.e. if emigration or immigration rates depend on population densities, the correlation of stability with dispersal rates is positive in part of the models. We also find that many systems show all four types of bifurcations and that antisynchronous oscillations occur mostly with nonrandom dispersal.

Cre recombinase expression or topical tamoxifen treatment do not affect retinal structure and function, neuronal vulnerability or glial reactivity in the mouse eye.

Mice with a constitutive or tamoxifen-induced Cre recombinase (Cre) expression are frequently used research tools to allow the conditional deletion of target genes via the Cre-loxP system. Here we analyzed for the first time in a comprehensive and comparative way, whether retinal Cre expression or topical tamoxifen treatment itself would cause structural or functional changes, including changes in the expression profiles of molecular markers, glial reactivity and photoreceptor vulnerability. To this end, we characterized the transgenic α-Cre, Lmop-Cre and the tamoxifen-inducible CAGG-CreER™ mouse lines, all having robust Cre expression in the neuronal retina. In addition, we characterized the effects of topical tamoxifen treatment itself in wildtype mice. We performed morphometric analyses, immunohistochemical staining, in vivo ERG and angiography analyses and realtime RT-PCR analyses. Furthermore, the influence of Cre recombinase or topical tamoxifen exposure on neuronal vulnerability was studied by using light damage as a model for photoreceptor degeneration. Taken together, neither the expression of Cre, nor topical tamoxifen treatment caused detectable changes in retinal structure and function, the expression profiles of investigated molecular markers, glial reactivity and photoreceptor vulnerability. We conclude that the Cre-loxP system and its induction through tamoxifen is a safe and reliable method to delete desired target genes in the neural retina.

[Long-term outcome following multiple trauma in working age : A prospective study in a Swiss trauma center]. / Langzeitoutcome nach Polytrauma im erwerbsfähigen Alter : Eine prospektive Datenerhebung an einem Schweizer Traumazentrum.

BACKGROUND: Given the lack of data in the available literature, we were interested in the disability rate and corresponding insurance costs following multiple trauma in Switzerland. The possible impact of demographic, traumatic and hospital process factors as well as subjective and objective longer-term outcome variables on insurance data acquired were examined. MATERIAL AND METHODS: Following multiple trauma the clinical and socioeconomic parameters in 145 survivors of working age were investigated over 2 and 4 years post-injury at a Swiss trauma center (University Hospital Basel). The correlation with the corresponding data provided by the largest Swiss accident insurance company (Suva, n = 63) was tested by univariate and multivariate analysis and patients insured at Suva were compared with those insured elsewhere (n = 82). RESULTS: The mean level of disability in this cohort of multiple trauma patients insured at Suva was 43 %. The insurer expected costs of more than 1 million Swiss Francs per multiply injured patient. In univariate analysis, only discrete correlations (maximum r = 0.37) were found with resulting disability, but significant correlations were found in subsequent multivariate testing most of all for age and the sequential organ failure assessment (SOFA 11 % and 15 % predictive capacity, p = 0.001; corrected R2 = 0.26). Among variables of longer-term outcome the Euro Quality of Life Group health-related quality of life in five dimensions (EQ-5D) correlated almost as highly with the objective extent of disability as did the reduced capacity to work declared by the patients (0.64 and 0.7, respectively). CONCLUSION: The estimation of long-term disability following multiple trauma based on primary data following injury appears to be possible only to a limited extent. Given the clinical and socioeconomic relevance, comparable analyses are necessary by including all insurance providers involved.

Experimental validation of a nonlinear µFE model based on cohesive-frictional plasticity for trabecular bone.

Trabecular bone is a porous mineralized tissue playing a major load bearing role in the human body. Prediction of age-related and disease-related fractures and the behavior of bone implant systems needs a thorough understanding of its structure-mechanical property relationships, which can be obtained using microcomputed tomography-based finite element modeling. In this study, a nonlinear model for trabecular bone as a cohesive-frictional material was implemented in a large-scale computational framework and validated by comparison of µFE simulations with experimental tests in uniaxial tension and compression. A good correspondence of stiffness and yield points between simulations and experiments was found for a wide range of bone volume fraction and degree of anisotropy in both tension and compression using a non-calibrated, average set of material parameters. These results demonstrate the ability of the model to capture the effects leading to failure of bone for three anatomical sites and several donors, which may be used to determine the apparent behavior of trabecular bone and its evolution with age, disease, and treatment in the future.

Forensic ancestry analysis with two capillary electrophoresis ancestry informative marker (AIM) panels: Results of a collaborative EDNAP exercise.

There is increasing interest in forensic ancestry tests, which are part of a growing number of DNA analyses that can enhance routine profiling by obtaining additional genetic information about unidentified DNA donors. Nearly all ancestry tests use single nucleotide polymorphisms (SNPs), but these currently rely on SNaPshot single base extension chemistry that can fail to detect mixed DNA. Insertion-deletion polymorphism (Indel) tests have been developed using dye-labeled primers that allow direct capillary electrophoresis detection of PCR products (PCR-to-CE). PCR-to-CE maintains the direct relationship between input DNA and signal strength as each marker is detected with a single dye, so mixed DNA is more reliably detected. We report the results of a collaborative inter-laboratory exercise of 19 participants (15 from the EDNAP European DNA Profiling group) that assessed a 34-plex SNP test using SNaPshot and a 46-plex Indel test using PCR-to-CE. Laboratories were asked to type five samples with different ancestries and detect an additional mixed DNA sample. Statistical inference of ancestry was made by participants using the Snipper online Bayes analysis portal plus an optional PCA module that analyzes the genotype data alongside calculation of Bayes likelihood ratios. Exercise results indicated consistent genotyping performance from both tests, reaching a particularly high level of reliability for the Indel test. SNP genotyping gave 93.5% concordance (compared to the organizing laboratory's data) that rose to 97.3% excluding one laboratory with a large number of miscalled genotypes. Indel genotyping gave a higher concordance rate of 99.8% and a reduced no-call rate compared to SNP analysis. All participants detected the mixture from their Indel peak height data and successfully assigned the correct ancestry to the other samples using Snipper, with the exception of one laboratory with SNP miscalls that incorrectly assigned ancestry of two samples and did not obtain informative likelihood ratios for a third. Therefore, successful ancestry assignments were achieved by participants in 92 of 95 Snipper analyses. This exercise demonstrates that ancestry inference tests based on binary marker sets can be readily adopted by laboratories that already have well-established CE regimes in place. The Indel test proved to be easy to use and allowed all exercise participants to detect the DNA mixture as well as achieving complete and concordant profiles in nearly all cases. Lastly, two participants successfully ran parallel next-generation sequencing analyses (each using different systems) and achieved high levels of genotyping concordance using the exercise PCR primer mixes unmodified.

Inter-laboratory evaluation of SNP-based forensic identification by massively parallel sequencing using the Ion PGM™.

Next generation sequencing (NGS) offers the opportunity to analyse forensic DNA samples and obtain massively parallel coverage of targeted short sequences with the variants they carry. We evaluated the levels of sequence coverage, genotyping precision, sensitivity and mixed DNA patterns of a prototype version of the first commercial forensic NGS kit: the HID-Ion AmpliSeq™ Identity Panel with 169-markers designed for the Ion PGM™ system. Evaluations were made between three laboratories following closely matched Ion PGM™ protocols and a simple validation framework of shared DNA controls. The sequence coverage obtained was extensive for the bulk of SNPs targeted by the HID-Ion AmpliSeq™ Identity Panel. Sensitivity studies showed 90-95% of SNP genotypes could be obtained from 25 to 100pg of input DNA. Genotyping concordance tests included Coriell cell-line control DNA analyses checked against whole-genome sequencing data from 1000 Genomes and Complete Genomics, indicating a very high concordance rate of 99.8%. Discordant genotypes detected in rs1979255, rs1004357, rs938283, rs2032597 and rs2399332 indicate these loci should be excluded from the panel. Therefore, the HID-Ion AmpliSeq™ Identity Panel and Ion PGM™ system provide a sensitive and accurate forensic SNP genotyping assay. However, low-level DNA produced much more varied sequence coverage and in forensic use the Ion PGM™ system will require careful calibration of the total samples loaded per chip to preserve the genotyping reliability seen in routine forensic DNA. Furthermore, assessments of mixed DNA indicate the user's control of sequence analysis parameter settings is necessary to ensure mixtures are detected robustly. Given the sensitivity of Ion PGM™, this aspect of forensic genotyping requires further optimisation before massively parallel sequencing is applied to routine casework.