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Douglas DR Naudie,1,2 Silvio Ndoja,1 Thomas J Wood,3 Lyndsay E Somerville,1 James L Howard,1,2 Richard W McCalden,1,2 Steven J MacDonald,1,2 Brent A Lanting1,2 1Division of Orthopaedic Surgery, Department of Surgery, London Health Sciences, London, Ontario, Canada; 2Bone and Joint Institute, Western University, London, Ontario, Canada; 3Division of Orthopaedic Surgery, Department of Surgery, Hamilton Health Sciences, McMaster University, Hamilton, Ontario, CanadaCorrespondence: Douglas DR NaudieDivision of Orthopaedic Surgery, Department of Surgery, Western University, Schulich School of Medicine, London Health Sciences Centre, University Campus, A9-028,339 Windermere Road, London, Ontario N6A 5A5, CanadaEmail Douglas.Naudie@lhsc.on.caIntroduction: Corrosion at the head–neck interface of modular components in total hip arthroplasty (THA) has been reported as a cause of failure of modern total hip replacement implants. While this method of failure has been well described, it remains poorly understood. The purpose of this study is to review the three most commonly used uncemented femoral stems at our institution over the last fifteen years and to correlate any established risk factors with rates of revision, particularly corrosion.Methods: We reviewed 2095 patients from March 2000 to September 2015 who underwent total hip arthroplasty with one of three uncemented femoral stem designs. All stems were made of a Ti6Al4V alloy with a 12/14 taper design. We included only those stems coupled with a CoCr head and a highly crosslinked polyethylene liner. We evaluated age, gender, body mass index (BMI), femoral head size, head length, neck angle and offset and correlated these to the incidence of all cause revision, as well as revision excluding infection.Results: There were no recognized corrosion-related revisions identified. There was no association between age, BMI, gender, head length, neck angle and offset to all cause revision or revision with infection excluded (p> 0.05). Femoral head size less than 32mm was associated with higher all cause revision rates (OR 4.60 (95% CI 1.8, 11.8)) and when excluding infection as a reason for revision (OR 4.94 (95% CI 1.7, 14.41)).Conclusion: Over the last fifteen years, we have not identified any cases of corrosion with the three most commonly used femoral stems used at out institution. While we acknowledge that no femoral stem is immune to corrosion, certain femoral stem designs may be uniquely resistant to this mode of failure.Level of Evidence: III.Keywords: corrosion, revision, total hip arthroplasty, femoral stems

Mohammad M Alzahrani,1 Thomas J Wood,2 Lyndsay E Somerville,3 Steven J MacDonald,3 James L Howard,3 Edward M Vasarhelyi,3 Brent A Lanting3 1Department of Orthopaedic Surgery, College of Medicine, Imam Abdulrahman Bin Faisal University, Dammam, Saudi Arabia; 2Division of Orthopaedic Surgery, Department of Surgery, Hamilton Health Sciences, McMaster University, Hamilton, ON, Canada; 3Division of Orthopaedic Surgery, Department of Surgery, London Health Sciences Centre University Campus, University of Western Ontario, London, Ontario N6A 5A5, CanadaCorrespondence: Mohammad M AlzahraniDepartment of Orthopaedic Surgery, College of Medicine, Imam Abdulrahman Bin Faisal University, 2835 King Faisal Road, Dammam 34212, Saudi ArabiaEmail mmalzahrani@iau.edu.saIntroduction: Outcomes and longevity of total knee arthroplasty (TKA) depend mainly on restoring knee function, through precise bony resection and appropriate soft tissue balancing. The current literature lacks evidence regarding the degree of radiographic change after intra-operative knee balancing. The purpose of our study was to assess the degree of change in coronal lower extremity alignment by comparing pre-operative to post-operative full-length radiographs (FLR) after quantifying the degree of intra-operative knee balancing and correlate patient-reported outcomes to the extent of balancing required.Patients and Methods: One hundred and fifty-four patients undergoing primary TKA for varus knee osteoarthritis were included in the study. The performed soft tissue releases and bony adjustments to obtain a balanced TKA intra-operatively were prospectively documented and were grouped into minimal, moderate and extensive release groups. Hip-knee-ankle angle (HKA), anatomical femoral-tibial angle (FTA), condylar hip angle (CH), medial proximal tibial angle (MPTA) and condylar plateau angle (CPA) were measured on full-length radiographs both pre-operatively and post-operatively. Frequencies of the soft tissue releases and bony resections in addition to descriptive statistics of the measured angles on the radiographs were recorded. In addition, patient-reported outcome scores (PROMs) were compared between the grouped patients.Results: Of those that were included in the study, 66 knees (42.9%) required minimal release to adequately balance the knee, while 70 (45.5%) required moderate release and 18 (11.7%) required extensive release. No statistically significant differences were found in change of alignment between the groups for all the measured angles. In addition, no difference in PROMs was seen between the groups both pre- and post-operatively.Conclusion: Although we found no association between post-operative coronal alignment of a TKA and the degree of soft tissue release and bony resection, this likely represents the fact that a balanced TKA is dynamic and not dependent on change of single radiographic parameters. Patient-reported outcomes were not impacted by the extent of releases.Keywords: knee arthroplasty, soft tissue, balancing, alignment, outcomes

Background: TKA and unicompartmental knee arthroplasty (UKA) are both utilized to treat unicompartmental knee arthrosis. While some surgeons assume UKA provides better function than TKA, this assumption is based on greater final outcome scores rather than on change in scores and many patients with UKA have higher preoperative scores.Questions/purposes: We therefore asked whether TKA would demonstrate (1) better change in clinical outcome scores from preoperative to postoperative states and (2) better survivorship than UKA.Methods: We evaluated 4087 patients with 5606 TKAs and 179 patients with 279 UKAs performed between 1978 and 2009. Patients with TKA were older and heavier than patients with UKA (mean age, 68 versus 66 years; mean BMI, 32 versus 29). We compared preoperative, latest postoperative, and change in Knee Society Clinical Rating System (KSCRS), SF-12, and WOMAC scores. Minimum followup was 2 years (UKA: mean, 7 years; range, 2.0-23 years; TKA: mean, 6.5 years; range, 2.0-33 years). Preoperative outcome measure scores (WOMAC, SF-12, KSCRS) were higher in the UKA group.Results: Patients with UKA had higher postoperative KSCRS and SF-12 mental scores. Changes in score for all WOMAC domains were similar between groups. Total KSCRS changes in score were similar between groups, although patients with TKA had higher knee scores (49 versus 43) but lower function scores than UKA (21 versus 26). Cumulative revision rate was higher for UKA than for TKA (13% versus 7%). Kaplan-Meier survivorship at 5 and 10 years was 95% and 90%, respectively, for UKA and 98% and 95%, respectively, for TKA.Conclusions: While patients with UKA had higher pre- and postoperative scores than patients with TKA, the changes in scores were similar in both groups and survival appeared higher in patients with TKA. [ABSTRACT FROM AUTHOR]

Unlabelled: There are two broad-based categories of cementless femoral component designs: proximally porous-coated and fully porous-coated. While both have been widely used, there remains debate regarding differences in clinical outcome scores, relative incidence of thigh pain, and development of stress shielding. We investigated these variables in a multicenter prospective randomized blinded clinical trial of 388 patients from three centers: 198 patients had a proximally porous-coated tapered cementless femoral component and 190 patients had a fully porous-coated cementless femoral component. A minimum followup of 2 years (mean, 6.7 years; range, 2.0-8.65 years) was available in 367 of the 388 patients (95%). We observed no differences in age at surgery, body mass index, or preoperative clinical outcome scores (WOMAC, SF-12, Harris hip score, UCLA activity, thigh pain) with the two groups. We determined serial bone density changes in a subcohort of 72 patients from two of the three centers. The postoperative clinical outcome scores were similar at all followup intervals, and we observed no differences in the incidence of thigh pain at any time. Bone density reduction in Gruen Zone 7 was greater with the fully coated stem than the proximally coated stem (24% versus 15% reduction, respectively). Both fully and proximally coated stems performed well, with no clinical differences at 2 years' followup, except in bone mineral density evaluations.Level Of Evidence: Level I, therapeutic study. See Guidelines for Authors for a complete description of levels of evidence. [ABSTRACT FROM AUTHOR]

Unlabelled: The range of motion after TKA depends on many patient, surgical technique, and implant factors. Recently, high-flexion designs have been introduced as a means of ensuring or gaining flexion after TKA. We therefore evaluated factors affecting postoperative flexion to determine whether implant design influences longterm flexion. We prospectively collected data on patients receiving a primary Genesis II total knee replacement with a minimum of 1-year followup (mean, 5.4 years; range, 1-13 years). We recorded pre- and postoperative outcome measures, patient demographics, and implant design (cruciate retaining [CR, n = 160], posterior stabilized [PS, n = 1177], high-flex posterior stabilized [HF-PS, n = 197]). Backward stepwise linear regression modeling identified the following factors affecting postoperative flexion: preoperative flexion, gender, body mass index, and implant design. Independent of gender, body mass index, and preoperative flexion, patients who received a HF-PS and PS design implant had a mean of 8 degrees and 5 degrees more flexion, respectively, than those who received a CR implant. Patients with low flexion preoperatively (<100 degrees) were more likely to gain flexion, whereas those with high flexion preoperatively (>120 degrees) were most likely to maintain or lose flexion postoperatively. Controlling for implant design, patients with high flexion preoperatively (>120 degrees) were more likely to gain flexion with the HF-PS design implant (HF-PS = 32.0%; PS = 15.1%; CR = 4.5%).Level Of Evidence: Level II, prognostic study. See Guidelines for Authors for a complete description of levels of evidence. [ABSTRACT FROM AUTHOR]

Unlabelled: The inset biconvex patella component is an alternative form of patella resurfacing in knee arthroplasty. We retrospectively reviewed 433 patients in whom 521 patella prostheses were implanted before April 1997 to determine survivorship, factors associated with failure of the implant, incidence of anterior knee pain, and factors that may be associated with the latter. We had clinical results for 204 surviving patients (242 knees) without failure of their implants with a minimum 10-year followup (mean, 11.4 years; range, 10-17 years). For the remaining 229 patients we used chart or radiographic review to determine if failure of their implant or other complications had occurred. At latest followup, 14 patella components had been revised for aseptic reasons or were radiographically loose. The 10-year Kaplan-Meier survivorship for the entire cohort for aseptic failure was 97.0%. Aseptic failure of the patella component was associated with the presence of osteonecrosis and the absence of a superior rim of bone radiographically. The incidence of anterior knee pain in surviving patients without failure of their implants was 7.8%. No factor examined was associated with anterior knee pain. Survivorship and clinical and radiographic results are equivalent, but not clearly superior, to those reported for other forms of patella resurfacing.Level Of Evidence: Level IV, therapeutic study. [ABSTRACT FROM AUTHOR]

BACKGROUND: The demand for total hip and total knee arthroplasties is increasing as are the waiting times for these procedures. Because of the differences between rural and urban areas in terms of the provision of arthroplasty services and between the 2 patient groups, patient perspectives of waiting times may also be different. METHODS: To compare waiting times for initial orthopedic consultation and total hip and knee arthroplasties in rural Ontario (Stratford) and in urban Ontario (London), and to compare patient perspectives of these waiting times, we mailed a survey to all 260 patients who underwent total hip or total knee arthroplasty between June 1, 2000, and June 1, 2001. The survey asked for the length of wait for consultation and for surgery, acceptability of waiting time for surgery, the effect of waiting on health and what an acceptable waiting time would be. Of the 260 surveys mailed 202 (78%) were returned. We reviewed the charts of the respondents to determine the actual waiting times. RESULTS: The actual waiting times (mean [and standard deviation]) for initial consultation were significantly (p < 0.001) shorter in the rural (RUR) group (1.10 [0.53] mo) than the urban (URB) group (3.40 [1.34] mo). There was no significant difference in waiting times for surgery between RUR (8.45 [3.32] mo) and URB (9.32 [3.61] mo) groups. Surgical waiting times for both groups showed that 56% of all the patients had to wait longer than 9 months from the date surgery was recommended. Perceived waiting times for consultation were found to be 56.9% longer (p < 0.001) than the actual waiting times, but there was no significant difference between perceived and actual waiting times for surgery (p = 0.40). Fifty percent of the patients stated they were unhappy with the wait for surgery or found the wait unacceptable (56% of URB and 44% of RUR patients). There was no significant difference between RUR and URB in patients' acceptance of their wait for surgery (p = 0.09), but URB patients believed their wait for surgery made a greater contribution to health deterioration (p = 0.003). Thirty-eight percent of RUR and 54% of URB patients believed their surgical wait contributed to 'a lot' or a 'moderate' amount of deterioration in their health. CONCLUSIONS: Waiting times for hip or knee replacement surgery have increased to the point at which over 50% of surgical patients in 2000-2001 in RUR and URB orthopedic practices had waited longer than 9 months for surgery. In comparing these practices, there was a significantly longer wait in urban than rural practices for the initial consultation but no significant difference in waiting times for surgery. [ABSTRACT FROM AUTHOR]

Objective: We quantified the effect of acute exposure to a high dosage of inorganic mercury on gene expression in Drosophila melanogaster using RNA-sequencing of whole adult females., Results: We found 119 genes with higher gene expression following treatment (including all 5 Drosophila metallothionine genes and a number of heat shock protein genes), and 31 with lower expression (several of which are involved in egg formation). Our results highlight biological processes and genetic pathways impacted by exposure to this toxic metal, and provide motivation for future studies to understand the genetic basis of response to mercury., (© 2024. The Author(s).)

Copper is one of a handful of biologically necessary heavy metals that is also a common environmental pollutant. Under normal conditions, copper ions are required for many key physiological processes. However, in excess, copper results in cell and tissue damage ranging in severity from temporary injury to permanent neurological damage. Because of its biological relevance, and because many conserved copper-responsive genes respond to nonessential heavy metal pollutants, copper resistance in Drosophila melanogaster is a useful model system with which to investigate the genetic control of the heavy metal stress response. Because heavy metal toxicity has the potential to differently impact specific tissues, we genetically characterized the control of the gene expression response to copper stress in a tissue-specific manner in this study. We assessed the copper stress response in head and gut tissue of 96 inbred strains from the Drosophila Synthetic Population Resource using a combination of differential expression analysis and expression quantitative trait locus mapping. Differential expression analysis revealed clear patterns of tissue-specific expression. Tissue and treatment specific responses to copper stress were also detected using expression quantitative trait locus mapping. Expression quantitative trait locus associated with MtnA, Mdr49, Mdr50, and Sod3 exhibited both genotype-by-tissue and genotype-by-treatment effects on gene expression under copper stress, illuminating tissue- and treatment-specific patterns of gene expression control. Together, our data build a nuanced description of the roles and interactions between allelic and expression variation in copper-responsive genes, provide valuable insight into the genomic architecture of susceptibility to metal toxicity, and highlight candidate genes for future functional characterization., Competing Interests: Conflicts of interest The authors declare no conflicts of interest., (© The Author(s) 2024. Published by Oxford University Press on behalf of The Genetics Society of America.)

We use ATAC-seq to examine chromatin accessibility for four different tissues in Drosophila melanogaster: adult female brain, ovaries, and both wing and eye-antennal imaginal discs from males. Each tissue is assayed in eight different inbred strain genetic backgrounds, seven associated with a reference quality genome assembly. We develop a method for the quantile normalization of ATAC-seq fragments and test for differences in coverage among genotypes, tissues, and their interaction at 44099 peaks throughout the euchromatic genome. For the strains with reference quality genome assemblies, we correct ATAC-seq profiles for read mis-mapping due to nearby polymorphic structural variants (SVs). Comparing coverage among genotypes without accounting for SVs results in a highly elevated rate (55%) of identifying false positive differences in chromatin state between genotypes. After SV correction, we identify 1050, 30383, and 4508 regions whose peak heights are polymorphic among genotypes, among tissues, or exhibit genotype-by-tissue interactions, respectively. Finally, we identify 3988 candidate causative variants that explain at least 80% of the variance in chromatin state at nearby ATAC-seq peaks., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Huynh et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Introduction: Heavy metal pollutants can have long lasting negative impacts on ecosystem health and can shape the evolution of species. The persistent and ubiquitous nature of heavy metal pollution provides an opportunity to characterize the genetic mechanisms that contribute to metal resistance in natural populations. Methods: We examined variation in resistance to copper, a common heavy metal contaminant, using wild collections of the model organism Drosophila melanogaster . Flies were collected from multiple sites that varied in copper contamination risk. We characterized phenotypic variation in copper resistance within and among populations using bulked segregant analysis to identify regions of the genome that contribute to copper resistance. Results and Discussion: Copper resistance varied among wild populations with a clear correspondence between resistance level and historical exposure to copper. We identified 288 SNPs distributed across the genome associated with copper resistance. Many SNPs had population-specific effects, but some had consistent effects on copper resistance in all populations. Significant SNPs map to several novel candidate genes involved in refolding disrupted proteins, energy production, and mitochondrial function. We also identified one SNP with consistent effects on copper resistance in all populations near CG11825 , a gene involved in copper homeostasis and copper resistance. We compared the genetic signatures of copper resistance in the wild-derived populations to genetic control of copper resistance in the Drosophila Synthetic Population Resource (DSPR) and the Drosophila Genetic Reference Panel (DGRP), two copper-naïve laboratory populations. In addition to CG11825 , which was identified as a candidate gene in the wild-derived populations and previously in the DSPR, there was modest overlap of copper-associated SNPs between the wild-derived populations and laboratory populations. Thirty-one SNPs associated with copper resistance in wild-derived populations fell within regions of the genome that were associated with copper resistance in the DSPR in a prior study. Collectively, our results demonstrate that the genetic control of copper resistance is highly polygenic, and that several loci can be clearly linked to genes involved in heavy metal toxicity response. The mixture of parallel and population-specific SNPs points to a complex interplay between genetic background and the selection regime that modifies the effects of genetic variation on copper resistance., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Everman, Macdonald and Kelly.)

The germline mobilization of transposable elements (TEs) by small RNA mediated silencing pathways is conserved across eukaryotes and critical for ensuring the integrity of gamete genomes. However, genomes are recurrently invaded by novel TEs through horizontal transfer. These invading TEs are not targeted by host small RNAs, and their unregulated activity can cause DNA damage in germline cells and ultimately lead to sterility. Here we use hybrid dysgenesis-a sterility syndrome of Drosophila caused by transposition of invading P-element DNA transposons-to uncover host genetic variants that modulate dysgenic sterility. Using a panel of highly recombinant inbred lines of Drosophila melanogaster, we identified two linked quantitative trait loci (QTL) that determine the severity of dysgenic sterility in young and old females, respectively. We show that ovaries of fertile genotypes exhibit increased expression of splicing factors that suppress the production of transposase encoding transcripts, which likely reduces the transposition rate and associated DNA damage. We also show that fertile alleles are associated with decreased sensitivity to double-stranded breaks and enhanced DNA repair, explaining their ability to withstand high germline transposition rates. Together, our work reveals a diversity of mechanisms whereby host genotype modulates the cost of an invading TE, and points to genetic variants that were likely beneficial during the P-element invasion., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2022 Lama et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Drosophila melanogaster has proved an effective system with which to understand the evolutionary genetics and molecular mechanisms of insecticide resistance. Insecticide use has left signatures of selection in the fly genome, and both functional and quantitative genetic studies in the system have identified genes and variants associated with resistance. Here, we use D. melanogaster and leverage a bulk phenotyping and pooled sequencing "extreme quantitative trait loci" approach to genetically dissect variation in resistance to malathion, an organophosphate insecticide. We resolve 2 quantitative trait loci, one of which implicates allelic variation at the cytochrome P450 gene Cyp6g1, a strong candidate based on previous work. The second shows no overlap with hits from a previous genome-wide association study for malathion resistance, recapitulating other studies showing that different strategies for complex trait dissection in flies can yield apparently different architectures. Notably, we see no genetic signal at the Ace gene. Ace encodes the target of organophosphate insecticide inhibition, and genome-wide association studies have identified strong Ace-linked associations with resistance in flies. The absence of quantitative trait locus implicating Ace here is most likely because our mapping population does not segregate for several of the known functional polymorphisms impacting resistance at Ace, perhaps because our population is derived from flies collected prior to the widespread use of organophosphate insecticides. Our fundamental approach can be an efficient, powerful strategy to dissect genetic variation in resistance traits. Nonetheless, studies seeking to interrogate contemporary insecticide resistance variation may benefit from deriving mapping populations from more recently collected strains., (© The Author(s) 2022. Published by Oxford University Press on behalf of Genetics Society of America.)

We examined the effect of developmental exposure to three heavy metals - cadmium, copper, and lead - on gene expression in adult head tissue in the model organism Drosophila melanogaster . All metals affected development time and/or gene expression level. While variation in the response to each metal was apparent, two differentially-expressed genes were upregulated in response to all three metal treatments, and 11 genes were downregulated in two of the three treatments. Our work reveals that developmental metal exposure has the potential to have long-lasting, metal-specific effects on gene expression in adults, even after the metal stress has been removed., (Copyright: © 2022 by the authors.)

Despite the value of recombinant inbred lines for the dissection of complex traits, large panels can be difficult to maintain, distribute, and phenotype. An attractive alternative to recombinant inbred lines for many traits leverages selecting phenotypically extreme individuals from a segregating population, and subjecting pools of selected and control individuals to sequencing. Under a bulked or extreme segregant analysis paradigm, genomic regions contributing to trait variation are revealed as frequency differences between pools. Here, we describe such an extreme quantitative trait locus, or extreme quantitative trait loci, mapping strategy that builds on an existing multiparental population, the Drosophila Synthetic Population Resource, and involves phenotyping and genotyping a population derived by mixing hundreds of Drosophila Synthetic Population Resource recombinant inbred lines. Simulations demonstrate that challenging, yet experimentally tractable extreme quantitative trait loci designs (≥4 replicates, ≥5,000 individuals/replicate, and selecting the 5-10% most extreme animals) yield at least the same power as traditional recombinant inbred line-based quantitative trait loci mapping and can localize variants with sub-centimorgan resolution. We empirically demonstrate the effectiveness of the approach using a 4-fold replicated extreme quantitative trait loci experiment that identifies 7 quantitative trait loci for caffeine resistance. Two mapped extreme quantitative trait loci factors replicate loci previously identified in recombinant inbred lines, 6/7 are associated with excellent candidate genes, and RNAi knock-downs support the involvement of 4 genes in the genetic control of trait variation. For many traits of interest to drosophilists, a bulked phenotyping/genotyping extreme quantitative trait loci design has considerable advantages., (© The Author(s) 2022. Published by Oxford University Press on behalf of Genetics Society of America. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Planobispora rosea is the natural producer of the potent thiopeptide antibiotic GE2270A. Here, we present the results of a metabolomics and transcriptomics analysis of P. rosea during production of GE2270A. The data generated provides useful insights into the biology of this genetically intractable bacterium. We characterize the details of the shutdown of protein biosynthesis and the respiratory chain associated with the end of the exponential growth phase. We also provide the first description of the phosphate regulon in P. rosea . Based on the transcriptomics data, we show that both phosphate and iron are limiting P. rosea growth in our experimental conditions. Additionally, we identified and validated a new biosynthetic gene cluster associated with the production of the siderophores benarthin and dibenarthin in P. rosea . Together, the metabolomics and transcriptomics data are used to inform and refine the very first genome-scale metabolic model for P. rosea , which will be a valuable framework for the interpretation of future studies of the biology of this interesting but poorly characterized species. IMPORTANCE Planobispora rosea is a genetically intractable bacterium used for the production of GE2270A on an industrial scale. GE2270A is a potent thiopeptide antibiotic currently used as a precursor for the synthesis of two compounds under clinical studies for the treatment of Clostridium difficile infection and acne. Here, we present the very first systematic multi-omics investigation of this important bacterium, which provides a much-needed detailed picture of the dynamics of metabolism of P. rosea while producing GE2270A.

Background: The escalating socioeconomic burden of knee osteoarthritis (OA) underscores the need for innovative strategies to reduce wait times for total knee arthroplasty (TKA). The purpose of this study was to evaluate resource use, costs and health-related quality of life (HRQoL) across the continuum of care for patients with knee OA., Methods: This was a prospective study of 383 patients recruited from a high-volume teaching hospital at different stages of care (referral, consultation and presurgery). Outcomes included health care resource use; costs captured from the health care payer, private sector and societal perspectives; HRQoL measured using the Western Ontario and McMaster Universities Osteoarthritis Index, the 12-Item Short Form Health Survey, and EuroQoL 5-Dimension 5-Level tool; wait times; and the proportion of referrals deemed suitable candidates for surgery., Results: The most commonly used conservative treatments were pharmacotherapy, exercise and lifestyle modification. Forty percent of patients referred for TKA were deemed not to be suitable candidates for surgery. The greatest proportion of costs was borne by the patient or private insurer; a small proportion was borne by the public payer. Across all stages of care, more than 60% of the total costs was attributed to productivity losses. HRQoL remained relatively stable throughout the waiting period (mean wait time from referral to TKA 13.2 mo) but improved postoperatively., Conclusion: The suboptimal primary care management of knee OA calls for the development of innovative models of care. This study may provide valuable guidance on the design and implementation of a new online educational platform to improve referral efficiency and expedite wait times for TKA., Competing Interests: S. MacDonald reports receiving personal fees from DePuy Synthes and holding stock or stock options from Hip Innovations Technology, JointVue, CurvaFix, Parvizi Surgical Innovation, Smith & Nephew and Stryker, outside the submitted work. No other competing interests were declared., (© 2021 CMA Joule Inc. or its licensors.)

A range of heavy metals are required for normal cell function and homeostasis. However, the anthropogenic release of metal compounds into soil and water sources presents a pervasive health threat. Copper is one of many heavy metals that negatively impacts diverse organisms at a global scale. Using a combination of quantitative trait locus (QTL) mapping and RNA sequencing in the Drosophila Synthetic Population Resource, we demonstrate that resistance to the toxic effects of ingested copper in D. melanogaster is genetically complex and influenced by allelic and expression variation at multiple loci. QTL mapping identified several QTL that account for a substantial fraction of heritability. Additionally, we find that copper resistance is impacted by variation in behavioral avoidance of copper and may be subject to life-stage specific regulation. Gene expression analysis further demonstrated that resistant and sensitive strains are characterized by unique expression patterns. Several of the candidate genes identified via QTL mapping and RNAseq have known copper-specific functions (e.g., Ccs, Sod3, CG11825), and others are involved in the regulation of other heavy metals (e.g., Catsup, whd). We validated several of these candidate genes with RNAi suggesting they contribute to variation in adult copper resistance. Our study illuminates the interconnected roles that allelic and expression variation, organism life stage, and behavior play in copper resistance, allowing a deeper understanding of the diverse mechanisms through which metal pollution can negatively impact organisms., (© The Author(s) 2020. Published by Oxford University Press on behalf of Genetics Society of America.)

Evolution by natural selection occurs when the frequencies of genetic variants change because individuals differ in Darwinian fitness components such as survival or reproductive success. Differential fitness has been demonstrated in field studies of many organisms, but it remains unclear how well we can quantitatively predict allele frequency changes from fitness measurements. Here, we characterize natural selection on millions of Single Nucleotide Polymorphisms (SNPs) across the genome of the annual plant Mimulus guttatus. We use fitness estimates to calibrate population genetic models that effectively predict allele frequency changes into the next generation. Hundreds of SNPs experienced "male selection" in 2013 with one allele at each SNP elevated in frequency among successful male gametes relative to the entire population of adults. In the following generation, allele frequencies at these SNPs consistently shifted in the predicted direction. A second year of study revealed that SNPs had effects on both viability and reproductive success with pervasive trade-offs between fitness components. SNPs favored by male selection were, on average, detrimental to survival. These trade-offs (antagonistic pleiotropy and temporal fluctuations in fitness) may be essential to the long-term maintenance of alleles. Despite the challenges of measuring selection in the wild, the strong correlation between predicted and observed allele frequency changes suggests that population genetic models have a much greater role to play in forward-time prediction of evolutionary change., Competing Interests: The authors have declared that no competing interests exist.

Imaginal disc morphogenesis during metamorphosis in Drosophila melanogaster provides an excellent model to uncover molecular mechanisms by which hormonal signals effect physical changes during development. The broad ( br ) Z2 isoform encodes a transcription factor required for disc morphogenesis in response to 20-hydroxyecdysone, yet how it accomplishes this remains largely unknown. Here, we use functional studies of amorphic br 5 mutants and a transcriptional target approach to identify processes driven by br and its regulatory targets in leg imaginal discs. br 5 mutants fail to properly remodel their basal extracellular matrix (ECM) between 4 and 7 hr after puparium formation. Additionally, br 5 mutant discs do not undergo the cell shape changes necessary for leg elongation and fail to elongate normally when exposed to the protease trypsin. RNA-sequencing of wild-type and br 5 mutant leg discs identified 717 genes differentially regulated by br , including a large number of genes involved in glycolysis, and genes that encode proteins that interact with the ECM. RNA interference-based functional studies reveal that several of these genes are required for adult leg formation, particularly those involved in remodeling the ECM. Additionally, br Z2 expression is abruptly shut down at the onset of metamorphosis, and expressing it beyond this time results in failure of leg development during the late prepupal and pupal stages. Taken together, our results suggest that br Z2 is required to drive ECM remodeling, change cell shape, and maintain metabolic activity through the midprepupal stage, but must be switched off to allow expression of pupation genes., (Copyright © 2020 by the Genetics Society of America.)

Objective: Segregating genetic variants contribute to the response to toxic, xenobiotic compounds, and identifying these causative sites can help describe the mechanisms underlying metabolism of toxic compounds. In previous work we implicated the detoxification gene Ugt86Dd in the genetic control of larval nicotine resistance in Drosophila melanogaster. Furthermore, we suggested that a naturally-occurring 22-bp deletion that leads to a stop codon in exon 2 of the gene markedly reduces resistance. Here we use homology directed CRISPR/Cas9 gene editing to specifically test this hypothesis., Results: We edited chromosome three from an inbred strain named A4 which carries the insertion allele at Ugt86Dd, successfully generated four alleles carrying the 22-bp Ugt86Dd deletion, and substituted edited chromosomes back into the A4 background. The original A4 strain, and an un-edited control strain in the same A4 background, show no significant difference in egg-to-adult or larva-to-adult viability on either control media or nicotine-supplemented media, and only slightly delayed development in nicotine media. However, strains carrying the 22-bp deletion showed reduced viability in nicotine conditions, and significantly longer development. Our data strongly suggest that the naturally-occurring 22-bp insertion/deletion event in Ugt86Dd directly impacts variation in nicotine resistance in D. melanogaster.

There is considerable variation in sleep duration, timing and quality in human populations, and sleep dysregulation has been implicated as a risk factor for a range of health problems. Human sleep traits are known to be regulated by genetic factors, but also by an array of environmental and social factors. These uncontrolled, non-genetic effects complicate powerful identification of the loci contributing to sleep directly in humans. The model system, Drosophila melanogaster , exhibits a behavior that shows the hallmarks of mammalian sleep, and here we use a multitiered approach, encompassing high-resolution QTL mapping, expression QTL data, and functional validation with RNAi to investigate the genetic basis of sleep under highly controlled environmental conditions. We measured a battery of sleep phenotypes in >750 genotypes derived from a multiparental mapping panel and identified several, modest-effect QTL contributing to natural variation for sleep. Merging sleep QTL data with a large head transcriptome eQTL mapping dataset from the same population allowed us to refine the list of plausible candidate causative sleep loci. This set includes genes with previously characterized effects on sleep and circadian rhythms, in addition to novel candidates. Finally, we employed adult, nervous system-specific RNAi on the Dopa decarboxylase , dyschronic , and timeless genes, finding significant effects on sleep phenotypes for all three. The genes we resolve are strong candidates to harbor causative, regulatory variation contributing to sleep.

It has been hypothesized that individually-rare hidden structural variants (SVs) could account for a significant fraction of variation in complex traits. Here we identified more than 20,000 euchromatic SVs from 14 Drosophila melanogaster genome assemblies, of which ~40% are invisible to high specificity short-read genotyping approaches. SVs are common, with 31.5% of diploid individuals harboring a SV in genes larger than 5kb, and 24% harboring multiple SVs in genes larger than 10kb. SV minor allele frequencies are rarer than amino acid polymorphisms, suggesting that SVs are more deleterious. We show that a number of functionally important genes harbor previously hidden structural variants likely to affect complex phenotypes. Furthermore, SVs are overrepresented in candidate genes associated with quantitative trait loci mapped using the Drosophila Synthetic Population Resource. We conclude that SVs are ubiquitous, frequently constitute a heterogeneous allelic series, and can act as rare alleles of large effect.

Specific characteristics of the male Achroia grisella acoustic mating signal determine a male's attractiveness toward females. These features are genetically variable in populations, and mapping experiments have been used to identify loci contributing to song variation, and understand the evolutionary forces acting on this important sexual trait. Here we built on this foundation and carried out QTL (Quantitative Trait Locus) mapping using >1,000 recombinant individuals, genotyping this large cohort at thousands of sequence-based markers covering the entire collection of 30 A. grisella chromosomes. This dense marker set, coupled with our development of an annotated, draft genome of A. grisella , allowed us to link >3,000 genome scaffolds, >10,000 predicted genes, and close to 275Mb of genome sequence to chromosomes. Our QTL mapping confirmed a fraction of the QTL identified in a previous study, and additionally revealed novel loci. Collectively, QTL explained only small fractions of the phenotypic variance, suggesting many more causative factors remain below the detection threshold of our study. A surprising, and ultimately challenging feature of our study was the low level of intrachromosomal recombination present in our mapping population. This led to difficulty ordering markers along linkage groups, necessitating a chromosome-by-chromosome mapping approach, rather than true interval mapping, and precluded confident ordering/orienting of scaffolds along each chromosome. Nonetheless, our study increased the genomic resources available for the A. grisella system. Enabled by ever more powerful technologies, future investigators will be able to leverage our data to provide more detailed genetic dissection of male song variation in A. grisella ., (Copyright © 2019 Koseva et al.)

This study examined whether individuals undergoing total hip arthroplasty who are self-rated as "highly active", demonstrated higher pre-operative activity levels than others., Methods: 483 patients were retrospectively identified; 241 were "highly active" and 242 were not. They were compared using WOMAC, self-reported activity and 6-Minute Walk Test (6MWT)., Results: Self-reported highly active adults demonstrated statistically significant higher WOMAC scores (48.85 vs 43.79), increased mean number of activities performed and mean 6MWT (348.14 m vs 274.64 m)., Conclusions: The positive correlation between subjective and objective measures of mobility increase our confidence in the latter as valid indicator of patient's preoperative activity level.

We leverage two complementary Drosophila melanogaster mapping panels to genetically dissect starvation resistance-an important fitness trait. Using >1600 genotypes from the multiparental Drosophila Synthetic Population Resource (DSPR), we map numerous starvation stress QTL that collectively explain a substantial fraction of trait heritability. Mapped QTL effects allowed us to estimate DSPR founder phenotypes, predictions that were correlated with the actual phenotypes of these lines. We observe a modest phenotypic correlation between starvation resistance and triglyceride level, traits that have been linked in previous studies. However, overlap among QTL identified for each trait is low. Since we also show that DSPR strains with extreme starvation phenotypes differ in desiccation resistance and activity level, our data imply multiple physiological mechanisms contribute to starvation variability. We additionally exploited the Drosophila Genetic Reference Panel (DGRP) to identify sequence variants associated with starvation resistance. Consistent with prior work these sites rarely fall within QTL intervals mapped in the DSPR. We were offered a unique opportunity to directly compare association mapping results across laboratories since two other groups previously measured starvation resistance in the DGRP. We found strong phenotypic correlations among studies, but extremely low overlap in the sets of genomewide significant sites. Despite this, our analyses revealed that the most highly associated variants from each study typically showed the same additive effect sign in independent studies, in contrast to otherwise equivalent sets of random variants. This consistency provides evidence for reproducible trait-associated sites in a widely used mapping panel, and highlights the polygenic nature of starvation resistance., (Copyright © 2019 Everman et al.)

Background: Revision of TKA as a result of polyethylene wear is decreasing, but long-term wear performance of polyethylene is still a topic of interest to surgeons and device manufacturers seeking to improve longevity. Measuring wear of modern, wear-resistant implants has been described using radiostereometric analysis (RSA). Performing in vivo measurements would establish whether implant retrieval studies are representative of wear in well-performing knees., Questions/purposes: For a single knee implant system, we sought to determine (1) the linear wear rate using RSA; (2) the association between demographic factors and wear rate; and (3) the association between limb alignment and wear rate., Methods: A total of 49 patients with a minimum followup of 10 years (median, 12 years; range, 10-20 years) were retrospectively selected. During the examined period, 4082 TKAs were performed of which 2085 were the implant examined in this study. There were 71 of these patients who met the criteria including an available full-leg radiograph postoperatively, and 34 of these patients returned for examination along with 15 additional from a separate RSA study that also met the criteria. All patients received a posterior-stabilized, cobalt-chromium-on-conventional polyethylene total knee implant from a single implant system, which was the most commonly used at our institution at the time. Patients underwent standing RSA examinations from 0° to 120° of flexion at a single time point without the use of marker beads. Linear wear rates (including creep) were measured based on intersections between the femoral component and tibial insert models. Associations between wear and patient age at surgery, sex, height, weight, body mass index, tibial insert size, and limb alignment were examined., Results: Using the maximum linear wear rate from any flexion angle, the lateral rate was 0.047 mm/year (interquartile range [IQR], 0.034-0.066 mm/year) and the medial rate was 0.052 mm/year (IQR, 0.040-0.069 mm/year). Using the median of the linear wear rates across all flexion angles, the lateral rate was 0.027 mm/year (IQR, 0.017-0.046 mm/year) and the medial rate was 0.038 mm/year (IQR, 0.022-0.054 mm/year). This rate for males was 0.049 mm/year medially (IQR, 0.042-0.077 mm/year) and 0.032 mm/year laterally (IQR, 0.026-0.059 mm/year), and for females was 0.027 mm/year medially (0.016-0.039 mm/year) and 0.020 mm/year laterally (IQR, 0.013-0.032 mm/year). The wear rate for males was greater medially (difference = 0.022 mm/year, p < 0.001) and laterally (difference = 0.012 mm/year, p = 0.008). There were associations between greater wear and increasing height (ρ = 0.48, p < 0.001 medially and ρ = 0.30, p = 0.04 laterally), decreasing body mass index (ρ = -0.31, p = 0.03 medially), and greater implant size (ρ = 0.34, p = 0.02 medially). Increasingly varus leg alignment was associated with greater medial wear (ρ = 0.33, p = 0.02)., Conclusions: Greater wear rates were associated with demographic factors and leg alignment. Further RSA wear studies of other modern implant systems would provide complementary information to retrieval studies and valuable data on wear resistance., Clinical Relevance: Good wear resistance was demonstrated by well-performing implants in patients at long-term followup with wear magnitudes in agreement with reported values from retrieval studies.

In the process of generating herpes simplex virus 1 (HSV-1) mutations in the viral regulatory gene encoding infected cell protein 0 (ICP0), we isolated a viral mutant, termed KOS-NA, that was severely impaired for acute replication in the eyes and trigeminal ganglia (TG) of mice, defective in establishing a latent infection, and reactivated poorly from explanted TG. To identify the secondary mutation(s) responsible for the impaired phenotypes of this mutant, we sequenced the KOS-NA genome and noted that it contained two nonsynonymous mutations in UL39 , which encodes the large subunit of ribonucleotide reductase, ICP6. These mutations resulted in lysine-to-proline (residue 393) and arginine-to-histidine (residue 950) substitutions in ICP6. To determine whether alteration of these amino acids was responsible for the KOS-NA phenotypes in vivo , we recombined the wild-type UL39 gene into the KOS-NA genome and rescued its acute replication phenotypes in mice. To further establish the role of UL39 in KOS-NA's decreased pathogenicity, the UL39 mutations were recombined into HSV-1 (generating UL39 mut ), and this mutant virus showed reduced ocular and TG replication in mice comparable to that of KOS-NA. Interestingly, ICP6 protein levels were reduced in KOS-NA-infected cells relative to the wild-type protein. Moreover, we observed that KOS-NA does not counteract caspase 8-induced apoptosis, unlike wild-type strain KOS. Based on alignment studies with other HSV-1 ICP6 homologs, our data suggest that amino acid 950 of ICP6 likely plays an important role in ICP6 accumulation and inhibition of apoptosis, consequently impairing HSV-1 pathogenesis in a mouse model of HSV-1 infection. IMPORTANCE HSV-1 is a major human pathogen that infects ∼80% of the human population and can be life threatening to infected neonates or immunocompromised individuals. Effective therapies for treatment of recurrent HSV-1 infections are limited, which emphasizes a critical need to understand in greater detail the events that modulate HSV-1 replication and pathogenesis. In the current study, we identified a neuroattenuated HSV-1 mutant (i.e., KOS-NA) that contains novel mutations in the UL39 gene, which codes for the large subunit of ribonucleotide reductase (also known as ICP6). This mutant form of ICP6 was responsible for the attenuation of KOS-NA in vivo and resulted in diminished ICP6 protein levels and antiapoptotic effect. Thus, we have determined that subtle alteration of the UL39 gene regulates expression and functions of ICP6 and severely impacts HSV-1 pathogenesis, potentially making KOS-NA a promising vaccine candidate against HSV-1., (Copyright © 2018 American Society for Microbiology.)

Background: In 2012, a new Knee Society Knee Scoring System (KSS) was developed and validated to address the needs for a scoring system that better encompasses the expectations, satisfaction, and physical involvement of a younger, more active population of patients undergoing TKA. Revalidating this tool in a separate population by individuals other than the developers of the scoring system seems important, because such replication would tend to confirm the generalizability of this tool., Questions/purposes: The purposes of this study were (1) to validate the KSS using a separate sample of patients undergoing primary TKA; and (2) to evaluate the internal consistency of the KSS., Methods: Intervention and control groups from a randomized controlled trial with no between-group differences were pooled. Preoperative and postoperative (6 weeks and 1 year) data were used. Patients with osteoarthritis undergoing primary TKA completed the patient-reported component of the KSS, Knee Injury and Osteoarthritis Outcome Score (KOOS), SF-12, two independent questions about expectations of surgery, and the Patient Acceptable Symptom State (PASS) single-question outcome. This study included 345 patients with 221 (64%) women, an average (SD) age of 64 (8.6) years, a mean (SD) body mass index of 32.9 (7.5) kg/m, and 225 (68%) having their first primary TKA. Loss to followup in the control group was 18% and loss to followup in the intervention group was 13%. We quantified cross-sectional (preoperative scores) and longitudinal validity (pre- to postoperative change scores) by evaluating associations between the KSS and KOOS subscales using Spearman's correlation coefficient. Preoperative known-group validity of the KSS symptoms and functional activity score was evaluated with a one-way analysis of variance across three levels of physical health status using the SF-12 Physical Component Score. Known-group validity of the KSS expectation score was evaluated with an unpaired t-test by comparing means across known expectation groups. Known-group validity of the KSS satisfaction score was evaluated with an unpaired t-test by comparing means across yes/no response groupings of the PASS single-question outcome. Internal consistency for each KSS subscale was evaluated with Cronbach's α., Results: Cross-sectional validity (ie, associations at a single point in time) was supported because correlation coefficients between KSS symptoms, functional activities, and satisfaction scores and scores on the KOOS pain subscale ranged from 0.60 to 0.73 (all correlations p < 0.01). Values were similar for associations with the KOOS function in the activities of daily living (ADL) subscale (0.66-0.69) and less (0.41-0.58) for correlations with the other three KOOS subscales. Longitudinal validity (ie, associations of change scores between two time points) was also supported because correlation coefficients between KSS symptoms, functional activities, and satisfaction change scores and the KOOS pain and ADL change scores varied from 0.63 to 0.73. Correlation coefficients were lower for the other three KOOS subscale change scores, suggesting a weaker relationship with KOOS symptoms (0.48-0.53), sports (0.47-0.51), and quality of life (0.60-0.65) (all correlations p < 0.01). Known-group validity (ie, differences between groups that are known to differ on a given characteristic) was confirmed by between-group differences for the symptoms and functional activities score comparisons as well as the comparisons with the expectations and satisfaction scores of the KSS (all p < 0.01). Cronbach's α (ie, association among subscale items) varied from 0.68 (discretionary activities) to 0.94 (postoperative expectations) across four KSS subscales., Conclusions: Moderate-sized correlation coefficients and consistent differences between known groups support the validity of the KSS. Internal consistency values were also acceptable. The patient-reported subscales of the KSS are a valid and internally consistent outcome assessment for TKA.

Background: Tibial baseplate roughness and polyethylene-insert micromotion resulting from locking-mechanism loosening can lead to polyethylene backside wear in TKAs. However, many retrieval studies examining these variables have evaluated only older TKA implant designs., Questions: We used implant-retrieval analysis to examine if there were differences in: (1) backside damage scores, (2) backside damage modes, and (3) backside linear wear rates in five TKA implant designs owing to differing baseplate surface roughness and locking mechanisms. Additionally, we examined if (4) patient demographics influence backside damage and wear., Methods: Five TKA implant models (four modern and one historical design) were selected with different tibial baseplate and/or locking mechanism designs. Six tibial inserts retrieved at the time of revision from each TKA model were matched for time in vivo, age of the patient at TKA revision, BMI, sex, revision number, and revision reason. Each insert backside was analyzed for: (1) visual total damage score and (2) individual visual damage modes, both by two observers and with an intraclass correlation coefficient of 0.66 (95% CI, 0.39-0.92), and (3) linear wear rate measured by micro-CT. Median primary outcomes were compared among the five designs. For our given sample size among five groups we could detect with 80% power a 10-point difference in damage score and an 0.11-mm per year difference in wear rate., Results: The polished tibial design with a partial peripheral capture locking mechanism and anterior constraint showed a lower total damage score compared with the nonpolished tibial design with only a complete peripheral-rim locking mechanism (median, 12.5; range, 9.5-18.0; 95% CI, 9.58-16.42 versus median, 22.3; range, 15.5-27.0; 95% CI, 17.5-26.5; p = 0.019). The polished baseplate with a tongue-in-groove locking mechanism showed more abrasions than the nonpolished baseplate with a peripheral-rim capture and antirotational island (median, 7.25; range, 0.5-8.0; 95% CI, 2.67-8.99 versus median, 0.75; range, 0-1.5; 95% CI, 0.20-1.47; p = 0.016)). Dimpling was a unique wear mode to the nonpolished baseplates with the peripheral-rim capture and antirotational island (median, 5.5; range, 2.0-9.0; 95% CI, 2.96-8.38) and the peripheral-rim capture alone (median, 9.0; range, 6.0-10.0; 95% CI, 7.29-10.38). Overall, the linear wear rate for polished designs was lower than for nonpolished designs (0.0102 ± 0.0044 mm/year versus 0.0224 ± 0.0119 mm/year; p < 0.001). Two of the polished baseplate designs, the partial peripheral capture with anterior constraint (median, 0.083 mm/year; range, 0.0037-0.0111 mm/year; 95% CI, 0.0050-0.0107 mm versus median, 0.0245 mm/year; range, 0.014-0.046 mm/year; 95% CI, 0.0130-0.0414 mm; p = 0.008) and the tongue-in-groove locking mechanism (median, 0.0085 mm/year; range, 0.005-0.015 mm/year; 95% CI, 0.0045-0.0138 mm; p = 0.032) showed lower polyethylene linear wear rates compared with the nonpolished baseplate design with only a peripheral-rim capture., Conclusions: Total damage scores and linear wear rates were highest involving the nonpolished design with only a peripheral rim capture. There were no differences among the other TKA designs regarding damage and wear, but this finding should be considered in the setting of a relatively small sample size., Clinical Relevance: Our study showed that in the complex interplay between baseplate surface finish and locking mechanism design, a polished baseplate with a robust locking mechanism had the lowest backside damage and linear wear. However, improvements in locking mechanism design in nonpolished baseplates potentially may offset some advantages of a polished baseplate. Further retrieval analyses need to be done to confirm such findings, especially analyzing current crosslinked polyethylene. Additionally, we need mid- and long-term studies comparing TKA revisions attributable to wear and osteolysis among implants before understanding if such design differences are clinically relevant.

Identifying the sequence polymorphisms underlying complex trait variation is a key goal of genetics research, since knowing the precise causative molecular events allows insight into the pathways governing trait variation. Genetic analysis of complex traits in model systems regularly starts by constructing QTL maps, but generally fails to identify causative sequence polymorphisms. Previously we mapped a series of QTL contributing to resistance to nicotine in a Drosophila melanogaster multiparental mapping resource and here use a battery of functional tests to resolve QTL to the molecular level. One large-effect QTL resided over a cluster of UDP-glucuronosyltransferases, and quantitative complementation tests using deficiencies eliminating subsets of these detoxification genes revealed allelic variation impacting resistance. RNAseq showed that Ugt86Dd had significantly higher expression in genotypes that are more resistant to nicotine, and anterior midgut-specific RNA interference (RNAi) of this gene reduced resistance. We discovered a segregating 22-bp frameshift deletion in Ugt86Dd , and accounting for the InDel during mapping largely eliminates the QTL, implying the event explains the bulk of the effect of the mapped locus. CRISPR/Cas9 editing of a relatively resistant genotype to generate lesions in Ugt86Dd that recapitulate the naturally occurring putative loss-of-function allele, leads to a large reduction in resistance. Despite this major effect of the deletion, the allele appears to be very rare in wild-caught populations and likely explains only a small fraction of the natural variation for the trait. Nonetheless, this putatively causative coding InDel can be a launchpad for future mechanistic exploration of xenobiotic detoxification., (Copyright © 2017 Highfill.)

Premise of the Study: Although asexual taxa are generally seen as evolutionary dead ends, asexuality appears to provide a short-term benefit in some taxa, including a wider geographic distribution compared to sexual relatives. However, this may be an illusion created by multiple, morphologically cryptic, asexual lineages, each occupying a relatively small area. In this study we investigate the role of multiple lineages in the biogeography of Myriopteris gracilis Fée (Pteridaceae), a North American apomictic triploid fern species with a particularly large range., Methods: Range-wide asexuality was assessed by counting spores/sporangium in 606 Myriopteris gracilis specimens from across the species range, and lineage structure was assessed with both plastid DNA sequence and Genotyping By Sequencing (GBS) SNP datasets., Key Results: Spore counting of >600 specimens identified no sexual populations, establishing that Myriopteris gracilis is exclusively asexual. The plastid data estimated the crown age of M. gracilis at ca. 2.5 mya and identified two lineages, each largely confined to the eastern or western portions of the range. These groups were further subdivided by the GBS data, revealing at least seven asexual lineages of varying geographic distributions, each occupying a relatively small portion of the total range of M. gracilis., Conclusions: Although maintained exclusively through asexual reproduction, the broad distribution of Myriopteris gracilis is a compilation of numerous, independently formed asexual lineages. Since no single asexual lineage occupies the full extent of the species distribution, recurrent lineage formation should be considered when evaluating the short-term benefit of asexuality in this taxon and others., (© 2017 Botanical Society of America.)

Populations maintain considerable segregating variation in the response to toxic, xenobiotic compounds. To identify variants associated with resistance to boric acid, a commonly-used household insecticide with a poorly understood mechanism of action, we assayed thousands of individuals from hundreds of strains. Using the Drosophila Synthetic Population Resource (DSPR), a multi-parental population (MPP) of inbred genotypes, we mapped six QTL to short genomic regions containing few protein-coding genes (3-188), allowing us to identify plausible candidate genes underlying resistance to boric acid toxicity. One interval contains multiple genes from the cytochrome P450 family, and we show that ubiquitous RNAi of one of these genes, Cyp9b2 , markedly reduces resistance to the toxin. Resistance to boric acid is positively correlated with caffeine resistance. The two phenotypes additionally share a pair of QTL, potentially suggesting a degree of pleiotropy in the genetic control of resistance to these two distinct xenobiotics. Finally, we screened the Drosophila Genetic Reference Panel (DGRP) in an attempt to identify sequence variants within mapped QTL that are associated with boric acid resistance. The approach was largely unsuccessful, with only one QTL showing any associations at QTL-specific 20% False Discovery Rate (FDR) thresholds. Nonetheless, these associations point to a potential candidate gene that can be targeted in future validation efforts. Although the mapping data resulting from the two reference populations do not clearly overlap, our work provides a starting point for further genetic dissection of the processes underlying boric acid toxicity in insects., (Copyright © 2017 Najarro et al.)

Background: The purpose of this study was to report the radiographic wear rates from a previous randomized controlled trial of first-generation highly crosslinked versus conventional polyethylene in total hip arthroplasty (THA) at a minimum of 13 years' follow-up., Methods: Patients returned for radiographic imaging and radiostereometric analysis (RSA). Radiographs were reviewed for the presence of osteolysis or component loosening. Femoral head penetration (which includes both wear and creep) was measured using RSA. We compared Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), 12-Item Short Form Health Survey (SF-12) and Harris Hip Scores (HHS) with preoperative values., Results: There was 1 revision in each group. There was no difference in WOMAC, SF-12, or HHS outcome scores between the highly crosslinked and conventional polyethylene groups (all p ≥ 0.13). Wear rate was lower with crosslinked polyethylene than conventional polyethylene (0.04 ± 0.02 mm/year v. 0.08 ± 0.03 mm/year, p = 0.007)., Conclusion: First-generation crosslinked polyethylene demonstrates greater wear resistance than conventional polyethylene after 13 years of implantation. Crosslinked polyethylene continues to outperform conventional polyethylene into the second decade of implantation.

The use of modular total knee arthroplasty (TKA) implants allows surgeons to perform isolated tibial polyethylene insert exchange (IPE) while retaining well-fixed and stable components.The purported advantages of IPE include preservation of bone stock, shorter operating time, less blood loss, faster rehabilitation and lower cost. However, the indications for IPE are limited.IPE for wear and osteolysis has moderate success in the medium term but should be avoided in cases of accelerated wear. In selected cases, debridement and IPE for early infection can result in low morbidity with high success rates in the short term. IPE for arthrofibrosis has poor results.IPE should be undertaken with caution and an institutional algorithm should be followed. Cite this article: EFORT Open Rev 2017;2:66-71 DOI: 10.1302/2058-5241.2.160049., Competing Interests: ICMJE Conflict of Interest Statement: None

Feline dysautonomia (FD) is a multiple system neuropathy of unknown aetiology. An apparently identical disease occurs in horses (equine grass sickness, EGS), dogs, rabbits, hares, sheep, alpacas and llamas. Horses with acute EGS have a marked reduction in plasma concentrations of the sulphur amino acids (SAA) cyst(e)ine and methionine, which may reflect exposure to a neurotoxic xenobiotic. The aim of this study was to determine whether FD cats have alterations in amino acid profiles similar to those of EGS horses. Amino acids were quantified in plasma/serum from 14 FD cats, 5 healthy in-contact cats which shared housing and diet with the FD cats, and 6 healthy control cats which were housed separately from FD cats and which received a different diet. The adequacy of amino acids in the cats' diet was assessed by determining the amino acid content of tinned and dry pelleted foods collected immediately after occurrences of FD. Compared with controls, FD cats had increased concentrations of many essential amino acids, with the exception of methionine which was significantly reduced, and reductions in most non-essential amino acids. In-contact cats also had inadequate methionine status. Artefactual loss of cysteine during analysis precluded assessment of the cyst(e)ine status. Food analysis indicated that the low methionine status was unlikely to be attributable to dietary inadequacy of methionine or cystine. Multi-mycotoxin screening identified low concentrations of several mycotoxins in dry food from all 3 premises. While this indicates fungal contamination of the food, none of these mycotoxins appears to induce the specific clinico-pathologic features which characterise FD and equivalent multiple system neuropathies in other species. Instead, we hypothesise that ingestion of another, as yet unidentified, dietary neurotoxic mycotoxin or xenobiotic, may cause both the characteristic disease pathology and the plasma SAA depletion.

An open question in evolutionary biology is the relationship between standing variation for a trait and the variation that leads to interspecific divergence. By identifying loci underlying phenotypic variation in intra- and interspecific crosses we can determine the extent to which polymorphism and divergence are controlled by the same genomic regions. Sexual traits provide abundant examples of morphological and behavioral diversity within and among species, and here we leverage variation in the Drosophila sex comb to address this question. The sex comb is an array of modified bristles or 'teeth' present on the male forelegs of several Drosophilid species. Males use the comb to grasp females during copulation, and ablation experiments have shown that males lacking comb teeth typically fail to mate. We measured tooth number in >700 genotypes derived from a multiparental advanced-intercross population, mapping three moderate-effect loci contributing to trait heritability. Two quantitative trait loci (QTLs) coincide with previously identified intra- and interspecific sex comb QTL, but such overlap can be explained by chance alone, in part because of the broad swathes of the genome implicated by earlier, low-resolution QTL scans. Our mapped QTL regions encompass 70-124 genes, but do not include those genes known to be involved in developmental specification of the comb. Nonetheless, we identified plausible candidates within all QTL intervals, and used RNA interference to validate effects at four loci. Notably, TweedleS expression knockdown substantially reduces tooth number. The genes we highlight are strong candidates to harbor segregating, functional variants contributing to sex comb tooth number.

Background: In previous studies, 50%-70% of patients referred to orthopedic surgeons for total knee replacement (TKR) were not surgical candidates at the time of initial assessment. The purpose of our study was to identify and cross-validate patient self-reported predictors of suitability for TKR and to determine the clinical utility of a predictive model to guide the timing and appropriateness of referral to a surgeon., Methods: We assessed pre-consultation patient data as well as the surgeon's findings and post-consultation recommendations. We used multivariate logistic regression to detect self-reported items that could identify suitable surgical candidates., Results: Patients' willingness to undergo surgery, higher rating of pain, greater physical function, previous intra-articular injections and patient age were the factors predictive of patients being offered and electing to undergo TKR., Conclusion: The application of the model developed in our study would effectively reduce the proportion of nonsurgical referrals by 25%, while identifying the vast majority of surgical candidates (> 90%). Using patient-reported information, we can correctly predict the outcome of specialist consultation for TKR in 70% of cases. To reduce long waits for first consultation with a surgeon, it may be possible to use these items to educate and guide referring clinicians and patients to understand when specialist consultation is the next step in managing the patient with severe osteoarthritis of the knee.

Closely-related, and otherwise morphologically similar insect species frequently show striking divergence in the shape and/or size of male genital structures, a phenomenon thought to be driven by sexual selection. Comparative interspecific studies can help elucidate the evolutionary forces acting on genital structures to drive this rapid differentiation. However, genetic dissection of sexual trait divergence between species is frequently hampered by the difficulty generating interspecific recombinants. Intraspecific variation can be leveraged to investigate the genetics of rapidly-evolving sexual traits, and here we carry out a genetic analysis of variation in the posterior lobe within D. melanogaster. The lobe is a male-specific process emerging from the genital arch of D. melanogaster and three closely-related species, is essential for copulation, and shows radical divergence in form across species. There is also abundant variation within species in the shape and size of the lobe, and while this variation is considerably more subtle than that seen among species, it nonetheless provides the raw material for QTL mapping. We created an advanced intercross population from a pair of phenotypically-different inbred strains, and after phenotyping and genotyping-by-sequencing the recombinants, mapped several QTL contributing to various measures of lobe morphology. The additional generations of crossing over in our mapping population led to QTL intervals that are smaller than is typical for an F2 mapping design. The intervals we map overlap with a pair of lobe QTL we previously identified in an independent mapping cross, potentially suggesting a level of shared genetic control of trait variation. Our QTL additionally implicate a suite of genes that have been shown to contribute to the development of the posterior lobe. These loci are strong candidates to harbor naturally-segregating sites contributing to phenotypic variation within D. melanogaster, and may also be those contributing to divergence in lobe morphology between species., Competing Interests: The authors have declared that no competing interests exist.

Background: Considerable natural variation for lifespan exists within human and animal populations. Genetically dissecting this variation can elucidate the pathways and genes involved in aging, and help uncover the genetic mechanisms underlying risk for age-related diseases. Studying aging in model systems is attractive due to their relatively short lifespan, and the ability to carry out programmed crosses under environmentally-controlled conditions. Here we investigate the genetic architecture of lifespan using the Drosophila Synthetic Population Resource (DSPR), a multiparental advanced intercross mapping population., Results: We measured lifespan in females from 805 DSPR lines, mapping five QTL (Quantitative Trait Loci) that each contribute 4-5 % to among-line lifespan variation in the DSPR. Each of these QTL co-localizes with the position of at least one QTL mapped in 13 previous studies of lifespan variation in flies. However, given that these studies implicate >90 % of the genome in the control of lifespan, this level of overlap is unsurprising. DSPR QTL intervals harbor 11-155 protein-coding genes, and we used RNAseq on samples of young and old flies to help resolve pathways affecting lifespan, and identify potentially causative loci present within mapped QTL intervals. Broad age-related patterns of expression revealed by these data recapitulate results from previous work. For example, we see an increase in antimicrobial defense gene expression with age, and a decrease in expression of genes involved in the electron transport chain. Several genes within QTL intervals are highlighted by our RNAseq data, such as Relish, a critical immune response gene, that shows increased expression with age, and UQCR-14, a gene involved in mitochondrial electron transport, that has reduced expression in older flies., Conclusions: The five QTL we isolate collectively explain a considerable fraction of the genetic variation for female lifespan in the DSPR, and implicate modest numbers of genes. In several cases the candidate loci we highlight reside in biological pathways already implicated in the control of lifespan variation. Thus, our results provide further evidence that functional genetics tests targeting these genes will be fruitful, lead to the identification of natural sequence variants contributing to lifespan variation, and help uncover the mechanisms of aging.

We announce the draft genome sequence for Chromobacterium violaceum strain CV017, used as a model and tool to understand acyl-homoserine lactone-dependent quorum sensing. The assembly consists of 4,774,638-bp contained in 211 scaffolds., (Copyright © 2016 Wang et al.)

Background: Hard-on-hard bearings for total hip arthroplasty continue to warrant analysis even though crosslinked polyethylene is performing very well. Ceramic-on-metal (CoM) has low in vitro wear and did well in an early clinical trial. We report on a prospective, randomized, multicenter investigational device trial comparing CoM with metal-on-metal (MoM)., Questions/purposes: (1) Is there a difference in the number or type of revisions comparing CoM with MoM? (2) Are cobalt and chromium metal levels different for CoM and MoM THA?, Methods: Between August 2005 and October 2006, of 1015 patients screened, 390 patients were enrolled at 11 centers and randomized to 194 CoM and 196 MoM bearings. There was no difference in the preoperative patient demographics between the study groups. Mean followup was 50 months (range, 22-75 months). Seventy-two patients from two centers had metal level analysis., Results: With the numbers available, there was no difference in the proportion of patients undergoing revisions between the MoM and the CoM cohorts (MOM: 3% [six of 196]; COM: 1.5% [three of 194]; p = 0.50). Four MoM revisions were unrelated to the bearing surface. Two had bearing surface-related reoperations, one for an aseptic lymphocyte-dominated vasculitis-associated lesion and one for elevated metal levels with acetabular malposition. None of the CoM revisions were related to the bearing surface. The metal level analysis revealed that in contrast to the CoM, the MoM bearing group had increasing values of erythrocyte and serum cobalt from 1 to 5 years (CoM erythrocyte 0.45-0.55 ppb, p = 0.11 and CoM serum 0.88-0.85, p = 0.55, and MoM erythrocyte 0.32-0.51 ppb, p < 0.01 and MoM serum 0.65-1.01 ppb, p < 0.01). In addition, the MoM cobalt levels in erythrocytes and serum at 5 years were more variable than at 1 year (erythrocyte interquartile range [IQR], 0.26-0.44 to 0.31-1.21 ppb and serum IQR, 0.42-0.80 to 0.64-2.20 ppb, p < 0.02 for both)., Conclusions: Although both bearings performed well at short-term followup, the CoM bearing group had no wear-related revisions and maintained consistently low metal levels. The MoM cobalt elevations may be important considering recent reports of taper corrosion. This CoM bearing was approved by the FDA but withdrawn from the market because of low sales. If it were available, the authors would not use CoM until long-term data were available. The bearing would have to outperform crosslinked polyethylene because it is unlikely that CoM metal levels will return to normal., Level of Evidence: Level I, therapeutic study.

Background: Although it is understood that backside damage occurs in polyethylene acetabular liners, the effect of highly crosslinked polyethylene, which has completely replaced conventional polyethylene, has yet to be examined., Questions/purposes: The purpose of this study was to answer the following questions: (1) With conventional polyethylene (CPE), did the acetabular design make a difference in backside wear? (2) Is there a difference in backside damage between CPE and crosslinked polyethylene (XLPE) liners? (3) With XLPE, is the difference in backside wear between designs still present? (4) Is there any difference in backside damage in various zones on backside of individual liners?, Methods: This single-institution retrieval analysis involved visual damage scoring on the backside of 233 polyethylene liners implanted between 2002 and 2011. The liners were retrieved from either polished/dovetail cups (PD) or roughened/wire cups (RW) made by two different manufacturers. The inserts were divided into four groups: PD-CPE (n = 105), PD-XLPE (n = 16), RW-CPE (n = 99), and RW-XLPE (n = 13). Aseptic loosening and polyethylene wear were the predominant reasons for revision of CPE liners, whereas instability and infection were the common reasons for revision of XLPE liners. The time in situ (TIS) was shorter for the XLPE liners as compared with the CPE liners (PD-CPE: 8.5 years [SD 4.4]; RD-CPE: 9.5 [SD 4.8]; PD-CPE: 3.8 [SD 3.7]; RD-XLPE: 4.0 [SD 4.3]). The backside of each liner was divided into five zones and graded initially from a scale of 0 to 3 for seven different damage modes by one observer. There was substantial interobserver (kappa 0.769) and intraobserver (0.736) reliability on repeat scoring. To compare the effects of crosslinking, a subset of the conventional polyethylene liners was matched to the crosslinked liners based on TIS, resulting in 16 per group for the two PD types and 13 per group for the two RW types., Results: Total damage scores in the RW-CPE (19 ± 7) group were greater (p < 0.001) than the PD-CPE group (8 ± 4). Damage was reduced with XLPE (p = 0.02) only for the RW-XLPE (9 ± 9) versus RW-CPE-type sockets (4 ± 4). There was no difference (p = 0.16) between the RW-XLPE group and the PD-XLPE group. Damage scores in the peripheral zone of the locking mechanisms were higher for RW-CPE (4 ± 3) compared with PD-CPE (1.4 ± 1.0, p < 0.001) and RW-XLPE (1.3 ± 1.3, p = 0.02). Damage was greater (p < 0.001) within the superior zones (7 ± 4) compared with the inferior zones (3.0 ± 2.8) of all liners., Conclusions: The current study shows greater backside damage for CPE in the roughened, wire lock cups. XLPE was shown to be more resistant to backside damage for both socket designs., Clinical Relevance: Although the polished/dovetail cups seem to cause less backside damage in the CPE liners than roughened/wire cups, the use of XLPE liners has been shown to protect the liner from backside damage in either cup design. If an acetabular shell has a functional locking mechanism, surgeons should not worry about cup design when undertaking isolated liner revision using XLPE liners.

Type 17 helper T-cell cytokines have been implicated in the pathogenesis of inflammatory bowel disease, a chronic condition affecting the gastrointestinal tract, but information regarding their contribution to pathology in different regions of the gut is lacking. By using a murine model of bacteria-induced typhlocolitis, we investigated the role of IL-17A, IL-17F, and IL-22 in cecal versus colonic inflammation. Cecal, but not colonic, pathology in C57BL/6 mice inoculated with Helicobacter hepaticus plus anti-IL-10 receptor (IL-10R) monoclonal antibody was exacerbated by co-administration of anti-IL-17A monoclonal antibody, suggesting a disease-protective role for IL-17A in the cecum. In contrast, anti-IL-17F had no effect on H. hepaticus-induced intestinal pathology. Neutralization of IL-22 prevented the development of colonic, but not cecal, inflammation in H. hepaticus-infected anti-IL-10R-treated mice, demonstrating a pathogenic role for IL-22 in the colon. Analysis of transcript levels revealed differential expression of IL-22R, IL-22 binding protein, and IL-23R between cecum and colon, a finding that may help explain why these tissues respond differently after anti-IL-22 treatment. Analysis of microarray data from healthy human intestine further revealed significant differences in cytokine receptor transcript levels (including IL-22RA1 and IL-23R) in distinct parts of the human gut. Together, our findings demonstrate that individual type 17 helper T-cell cytokines can have proinflammatory or anti-inflammatory effects in different regions of the intestine, an observation that may have implications for interventions against human inflammatory bowel disease., (Copyright © 2015 American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.)