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Glycoside hydrolases (GHs, also called glycosidases) catalyze the hydrolysis of glycosidic bonds in polysaccharides. Numerous GH genes have been identified from various organisms and are classified into 188 families, abbreviated GH1 to GH188. Enzymes in the GH32 family hydrolyze fructans, which are present in approximately 15% of flowering plants and are widespread across microorganisms. GH32 genes are rarely found in animals, as fructans are not a typical carbohydrate source utilized in animals. Here, we report the discovery of 242 GH32 genes identified in 84 animal species, ranging from nematodes to crabs. Genetic analyses of these genes indicated that the GH32 genes in various animals were derived from different bacteria via multiple, independent horizontal gene transfer events. The GH32 genes in animals appear functional based on the highly conserved catalytic blades and triads in the active center despite the overall low (35–60%) sequence similarities among the predicted proteins. The acquisition of GH32 genes by animals may have a profound impact on sugar metabolism for the recipient organisms. Our results together with previous reports suggest that the acquired GH32 enzymes may not only serve as digestive enzymes, but also may serve as effectors for manipulating host plants, and as metabolic enzymes in the non-digestive tissues of certain animals. Our results provide a foundation for future studies on the significance of horizontally transferred GH32 genes in animals. The information reported here enriches our knowledge of horizontal gene transfer, GH32 functions, and animal–plant interactions, which may result in practical applications. For example, developing crops via targeted engineering that inhibits GH32 enzymes could aid in the plant’s resistance to animal pests.

Abstract Next‐generation sequencing (NGS) technology advancements continue to reduce the cost of high‐throughput genome‐wide genotyping for breeding and genetics research. Skim sequencing, which surveys the entire genome at low coverage, has become feasible for quantitative trait locus (QTL) mapping and genomic selection in various crops. However, the genome complexity of allopolyploid crops such as wheat (Triticum aestivum L.) still poses a significant challenge for genome‐wide genotyping. Targeted sequencing of the protein‐coding regions (i.e., exome) reduces sequencing costs compared to whole genome re‐sequencing and can be used for marker discovery and genotyping. We developed a method called skim exome capture (SEC) that combines the strengths of these existing technologies and produces targeted genotyping data while decreasing the cost on a per‐sample basis compared to traditional exome capture. Specifically, we fragmented genomic DNA using a tagmentation approach, then enriched those fragments for the low‐copy genic portion of the genome using commercial wheat exome baits and multiplexed the sequencing at different levels to achieve desired coverage. We demonstrated that for a library of 48 samples, ∼7–8× target coverage was sufficient for high‐quality variant detection. For higher multiplexing levels of 528 and 1056 samples per library, we achieved an average coverage of 0.76× and 0.32×, respectively. Combining these lower coverage SEC sequencing data with genotype imputation using a customized wheat practical haplotype graph database that we developed, we identified hundreds of thousands of high‐quality genic variants across the genome. The SEC method can be used for high‐resolution QTL mapping, genome‐wide association studies, genomic selection, and other downstream applications.

Introduction: In the context of increasing rates of methamphetamine (meth) overdose in Los Angeles County, California, USA, in 2021 the Los Angeles County Department of Public Health relaunched Meth Free LA County, a methamphetamine public education campaign built on Rescue Agency’s Decision BlocksTM Strategic Framework. Methods: To assess campaign reach and reception, we examined media data and an online cross-sectional post-campaign survey in 2021 with a convenience sample of 750 county residents ages 18–54 who had used meth, were at-risk, or had personal relationships with people at-risk. We assessed campaign awareness, receptivity, and perceived effectiveness. Participants also reported concern about meth use in their community, recall of 10 facts from campaign content, and if they had taken promoted actions such as seeking help. We compared proportions between campaign-aware and unaware participants and explored if recall predicted these measures. Results: Most participants recalled the campaign (84.1%), felt it offered a new way of looking at meth (73.8%), and reported it made them feel that help is available (84.0%) or that it could help reduce their own use (82.4%). Video advertisements received high perceived effectiveness scores. Greater campaign recall was associated with significantly increased odds of being concerned about meth use in the community, recalling facts, and taking promoted actions. Conclusion: In 2021, Meth Free LA County reached its high-risk audiences. Unlike fear-based campaigns its message resonated with high-risk individuals, providing an example of how public education campaigns on meth and similar illicit substances can connect with their audiences.

Factors defining wheat stem rust pathogen (Pgt) virulence remain poorly characterized. Comparative population genomics based on Pgt haplotypes suggest that structural variation and admixture through somatic hybridization and sexual recombination play an important role in broadening Pgt virulence.

It is widely accepted that DNA replication fork stalling is a common occurrence during cell proliferation, but there are robust mechanisms to alleviate this and ensure DNA replication is completed prior to chromosome segregation. The SMC5/6 complex has consistently been implicated in the maintenance of replication fork integrity. However, the essential role of the SMC5/6 complex during DNA replication in mammalian cells has not been elucidated. In this study, we investigate the molecular consequences of SMC5/6 loss at the replication fork in mouse embryonic stem cells (mESCs), employing the auxin-inducible degron (AID) system to deplete SMC5 acutely and reversibly in the defined cellular contexts of replication fork stall and restart. In SMC5-depleted cells, we identify a defect in the restart of stalled replication forks, underpinned by excess MRE11-mediated fork resection and a perturbed localization of fork protection factors to the stalled fork. Previously, we demonstrated a physical and functional interaction of SMC5/6 with the COP9 signalosome (CSN), a cullin deneddylase that enzymatically regulates cullin ring ligase (CRL) activity. Employing a combination of DNA fiber techniques, the AID system, small-molecule inhibition assays, and immunofluorescence microscopy analyses, we show that SMC5/6 promotes the localization of fork protection factors to stalled replication forks by negatively modulating the COP9 signalosome (CSN). We propose that the SMC5/6-mediated modulation of the CSN ensures that CRL activity and their roles in DNA replication fork stabilization are maintained to allow for efficient replication fork restart when a replication fork stall is alleviated.

Up to 50% of patients with severe congenital heart disease (CHD) develop life-altering neurodevelopmental disability (NDD). It has been presumed that NDD arises in CHD cases because of hypoxia before, during, or after cardiac surgery. Recent studies detected an enrichment in de novo mutations in CHD and NDD, as well as significant overlap between CHD and NDD candidate genes. However, there is limited evidence demonstrating that genes causing CHD can produce NDD independent of hypoxia. A patient with hypoplastic left heart syndrome and gross motor delay presented with a de novo mutation in SMC5. Modeling mutation of smc5 in Xenopus tropicalis embryos resulted in reduced heart size, decreased brain length, and disrupted pax6 patterning. To evaluate the cardiac development, we induced the conditional knockout (cKO) of Smc5 in mouse cardiomyocytes, which led to the depletion of mature cardiomyocytes and abnormal contractility. To test a role for Smc5 specifically in the brain, we induced cKO in the mouse central nervous system, which resulted in decreased brain volume, and diminished connectivity between areas related to motor function but did not affect vascular or brain ventricular volume. We propose that genetic factors, rather than hypoxia alone, can contribute when NDD and CHD cases occur concurrently.

The connection between expression variation and phenotypic diversity in the populations of polyploid crops remains elusive. Here, the authors reveal the impact of genetic variants leading to biased expression of homoeologous genes in hexaploid wheat on agronomic traits.

Chromosome movements and licensing of synapsis must be tightly regulated during early meiosis to ensure accurate chromosome segregation and avoid aneuploidy, although how these steps are coordinated is not fully understood. Here we show that GRAS-1, the worm homolog of mammalian GRASP/Tamalin and CYTIP, coordinates early meiotic events with cytoskeletal forces outside the nucleus. GRAS-1 localizes close to the nuclear envelope (NE) in early prophase I and interacts with NE and cytoskeleton proteins. Delayed homologous chromosome pairing, synaptonemal complex (SC) assembly, and DNA double-strand break repair progression are partially rescued by the expression of human CYTIP in gras-1 mutants, supporting functional conservation. However, Tamalin, Cytip double knockout mice do not exhibit obvious fertility or meiotic defects, suggesting evolutionary differences between mammals. gras-1 mutants show accelerated chromosome movement during early prophase I, implicating GRAS-1 in regulating chromosome dynamics. GRAS-1-mediated regulation of chromosome movement is DHC-1-dependent, placing it acting within the LINC-controlled pathway, and depends on GRAS-1 phosphorylation at a C-terminal S/T cluster. We propose that GRAS-1 coordinates the early steps of homology search and licensing of SC assembly by regulating the pace of chromosome movement in early prophase I.

Audience segmentation is necessary in health communications to ensure equitable resource distribution. Peer crowds, which are macro-level teen subcultures, are effective psychographic segments for health communications because each crowd has unique mindsets, values, norms, and health behavior profiles. These mindsets affect behaviors, and can be used to develop targeted health communication campaigns to reach those in greatest need. Though peer crowd research is plentiful, no existing peer crowd measurement tool has been formally validated. As such, we developed and validated Virginia's Mindset Lens Survey (V-MLS), a mindset-based teen peer crowd segmentation survey to support health communication efforts. Using an online convenience sample of teens (N = 1,113), we assessed convergent and discriminant validity by comparing the V-MLS against an existing, widely-used peer crowd survey (I-Base Survey®) utilizing a multi-trait multi-method matrix. We also examined the V-MLS's predictive ability through a series of regressions using peer crowd scores to predict behaviors, experiences, and traits relevant to health communication campaign planning. The V-MLS demonstrated reliability and convergent and discriminant validity. Additionally, the V-MLS effectively distinguished teen peer crowds with unique health behaviors, experiences, and personal traits. When combined with appropriate information processing and campaign development frameworks, this new tool can complement existing instruments to inform message framing, tone, and style for campaigns that target at-risk teens to increase campaign equity and reach.

A new subgenus Myanmamastix subgen. n. and four new species of Rhabdomastix from Cenomanian Burmese amber were described: Rhabdomastix (Myanmamastix) asiatica subgen. and sp. n., Rhabdomastix (Myanmamastix) cretacica subgen. and sp. n., Rhabdomastix (Myanmamastix) krzeminskae subgen. and sp. n., Rhabdomastix (Myanmamastix) myanmae subgen. and sp. n. The species Rhabdomastix jarzembowskii known from Burmese amber was reclassified to the new subgenus Myanmamastix subgen. n. An analysis of the morphological structures and taxonomical differences between the representatives of Cretaceous Rhabdomastix was carried out. The manuscript presents results of original research, including the first 3D reconstruction of a Dipteran preserved as an inclusion in Burmese amber using computer microtomography.http://www.zoobank.org/urn:lsid:zoobank.org:pub:79C82852-2502-4A2F-B684-CCCE10D6C525

Introgression from wild relatives is a valuable source of novel allelic diversity for breeding. We investigated the genomic patterns of introgression from Aegilops tauschii, the diploid ancestor of the wheat D genome, into winter wheat (Triticum aestivum) cultivars. The population of 351 BC1F3:5 lines was selected based on phenology from crosses between six hexaploid wheat lines and 21 wheat-Ae. tauschii octoploids. SNP markers developed for this population and a diverse panel of 116 Ae. tauschii accessions by complexity-reduced genome sequencing were used to detect introgression based on the identity-by-descent analysis. Overall, introgression frequency positively correlated with recombination rate, with a high incidence of introgression at the ends of chromosomes and low in the pericentromeric regions, and was negatively related to sequence divergence between the parental genomes. Reduced introgression in the pericentromeric low-recombining regions spans nearly 2/3 of each chromosome arm, suggestive of the polygenic nature of introgression barriers that could be associated with multilocus negative epistasis between the alleles of wild and cultivated wheat. On the contrary, negative selection against the wild allele of Tg, controlling free-threshing trait and located in the high-recombining chromosomal region, led to reduced introgression only within ∼10 Mbp region around Tg. These results are consistent with the effect of selection on linked variation described by the Hill-Robertson effect, and offer insights into the introgression population development for crop improvement to maximize retention of introgressed diversity across entire genome.

Abstract Background Our understanding of how the complexity of the wheat genome influences the distribution of chromatin states along the homoeologous chromosomes is limited. Using a differential nuclease sensitivity assay, we investigate the chromatin states of the coding and repetitive regions of the allopolyploid wheat genome. Results Although open chromatin is found to be significantly enriched around genes, the majority of MNase-sensitive regions are located within transposable elements (TEs). Chromatin of the smaller D genome is more accessible than that of the larger A and B genomes. Chromatin states of different TEs vary among families and are influenced by the TEs’ chromosomal position and proximity to genes. While the chromatin accessibility of genes is influenced by proximity to TEs, and not by their position on the chromosomes, we observe a negative chromatin accessibility gradient along the telomere-centromere axis in the intergenic regions, positively correlated with the distance between genes. Both gene expression levels and homoeologous gene expression bias are correlated with chromatin accessibility in promoter regions. The differential nuclease sensitivity assay accurately predicts previously detected centromere locations. SNPs located within more accessible chromatin explain a higher proportion of genetic variance for a number of agronomic traits than SNPs located within more closed chromatin. Conclusions Chromatin states in the wheat genome are shaped by the interplay of repetitive and gene-encoding regions that are predictive of the functional and structural organization of chromosomes, providing a powerful framework for detecting genomic features involved in gene regulation and prioritizing genomic variation to explain phenotypes.

Abstract Background The SMC5/6 complex, cohesin and condensin are the three mammalian members of the structural maintenance of chromosomes (SMC) family, large ring-like protein complexes that are essential for genome maintenance. The SMC5/6 complex is the least characterized complex in mammals; however, it is known to be involved in homologous recombination repair (HRR) and chromosome segregation. Results In this study, a yeast two-hybrid screen was used to help elucidate novel interactions of the kleisin subunit of the SMC5/6 complex, NSMCE4A. This approach discovered an interaction between NSMCE4A and GPS1, a COP9 signalosome (CSN) component, and this interaction was further confirmed by co-immunoprecipitation. Additionally, GPS1 and components of SMC5/6 complex colocalize during interphase and mitosis. CSN is a cullin deNEDDylase and is an important factor for HRR. Depletion of GPS1, which has been shown to negatively impact DNA end resection during HRR, caused an increase in SMC5/6 levels at sites of laser-induced DNA damage. Furthermore, inhibition of the dennedylation function of CSN increased SMC5/6 levels at sites of laser-induced DNA damage. Conclusion Taken together, these data demonstrate for the first time that the SMC5/6 and CSN complexes interact and provides evidence that the CSN complex influences SMC5/6 functions during cell cycle progression and response to DNA damage.

Abstract Background Cigarette lifestyle marketing with psychographic targeting has been well documented, but few studies address non-cigarette tobacco products. This study examined how young adults respond to e-cigarette advertisements featuring diverse peer crowds – peer groups with shared identities and lifestyles – to inform tobacco counter-marketing design. Methods Fifty-nine young adult tobacco users in California participated in interviews and viewed four to five e-cigarette advertisements that featured characters from various peer crowd groups. For each participant, half of the advertisements they viewed showed characters from the same peer crowd as their own, and the other half of the advertisements featured characters from a different peer crowd. Advertisements were presented in random order. Questions probed what types of cues are noticed in the advertisements, and whether and how much participants liked or disliked the advertisements. Results Results suggest that participants liked and provided richer descriptions of characters and social situations in the advertisements featuring their own peer crowd more than the advertisements featuring a different peer crowd. Mismatching age or device type was also noted: participants reported advertisements showing older adults were not intended for them. Participants who used larger vaporizers tended to dislike cigalike advertisements even if they featured a matching peer crowd. Conclusion Peer crowd and lifestyle cues, age and device type are all salient features of e-cigarette advertising for young adults. Similarly, educational campaigns about e-cigarettes should employ peer crowd-based targeting to engage young adults, though messages should be carefully tested to ensure authentic and realistic portrayals.

The introgression from wild relatives have a great potential to broaden the availability of beneficial allelic diversity for crop improvement in breeding programs. Here, we assessed the impact of the introgression from 21 diverse accessions of Aegilops tauschii, the diploid ancestor of the wheat D genome, into 6 hard red winter wheat cultivars on yield and yield component traits. We used 5.2 million imputed D genome SNPs identified by the whole-genome sequencing of parental lines and the sequence-based genotyping of introgression population, including 351 BC1F3:5 lines. Phenotyping data collected from the irrigated and non-irrigated field trials revealed that up to 23% of the introgression lines (ILs) produce more grain than the parents and check cultivars. Based on 16 yield stability statistics, the yield of 12 ILs (3.4%) was stable across treatments, years, and locations; 5 of these lines were also high yielding lines, producing 9.8% more grain than the average yield of check cultivars. The most significant SNP- and haplotype-trait associations were identified on chromosome arms 2DS and 6DL for the spikelet number per spike (SNS), on chromosome arms 2DS, 3DS, 5DS, and 7DS for grain length (GL) and on chromosome arms 1DL, 2DS, 6DL, and 7DS for grain width (GW). The introgression of haplotypes from A. tauschii parents was associated with an increase in SNS, which was positively correlated with a heading date (HD), whereas the haplotypes from hexaploid wheat parents were associated with an increase in GW. We show that the haplotypes on 2DS associated with an increase in the spikelet number and HD are linked with multiple introgressed alleles of Ppd-D1 identified by the whole-genome sequencing of A. tauschii parents. Meanwhile, some introgressed haplotypes exhibited significant pleiotropic effects with the direction of effects on the yield component traits being largely consistent with the previously reported trade-offs, there were haplotype combinations associated with the positive trends in yield. The characterized repertoire of the introgressed haplotypes derived from A. tauschii accessions with the combined positive effects on yield and yield component traits in elite germplasm provides a valuable source of alleles for improving the productivity of winter wheat by optimizing the contribution of component traits to yield.

Background: Fatty liver, a major health problem worldwide, is the earliest pathological change in the progression of alcohol-associated (AFL) and non-alcoholic fatty liver disease (NAFL). Though the causes of AFL and NAFL differ, both share similar histological and some common pathophysiological characteristics. In this study, we sought to examine mechanisms responsible for lipid dynamics in liver and adipose tissue in the setting of AFL and NAFL in response to 48 h of fasting.Methods: Male rats were fed Lieber-DeCarli liquid control or alcohol-containing diet (AFL model), chow or high-fat pellet diet (NAFL model). After 6–8 weeks of feeding, half of the rats from each group were fasted for 48 h while the other half remained on their respective diets. Following sacrifice, blood, adipose, and the liver were collected for analysis.Results: Though rats fed AFL and NAFL diets both showed fatty liver, the physiological mechanisms involved in the development of each was different. Here, we show that increased hepatic de novo fatty acid synthesis, increased uptake of adipose-derived free fatty acids, and impaired triglyceride breakdown contribute to the development of AFL. In the case of NAFL, however, increased dietary fatty acid uptake is the major contributor to hepatic steatosis. Likewise, the response to starvation in the two fatty liver disease models also varied. While there was a decrease in hepatic steatosis after fasting in ethanol-fed rats, the control, chow and high-fat diet-fed rats showed higher levels of hepatic steatosis than pair-fed counterparts. This diverse response was a result of increased adipose lipolysis in all experimental groups except fasted ethanol-fed rats.Conclusion: Even though AFL and NAFL are nearly histologically indistinguishable, the physiological mechanisms that cause hepatic fat accumulation are different as are their responses to starvation.

The auxin-inducible degron (AID) system is becoming a widely used method for rapid and reversible degradation of target proteins. This system has been successfully used to study gene and protein functions in eukaryotic cells and common model organisms, such as nematode and fruit fly. To date, applications of the AID system in mammalian stem cell research are limited. Furthermore, standard mouse models harboring the AID system have not been established. Here we have explored the utility of the H11 safe-harbor locus for integration of the TIR1 transgene, an essential component of auxin-based protein degradation system. We have shown that the H11 locus can support constitutive and conditional TIR1 expression in mouse and human embryonic stem cells, as well as in mice. We demonstrate that the AID system can be successfully employed for rapid degradation of stable proteins in embryonic stem cells, which is crucial for investigation of protein functions in quickly changing environments, such as stem cell proliferation and differentiation. As embryonic stem cells possess unlimited proliferative capacity, differentiation potential, and can mimic organ development, we believe that these research tools will be an applicable resource to a broad scientific audience.

Purpose: Top surgery (i.e. mastectomy) has been shown to improve gender dysphoria and quality of life in adult transmasculine patients. However, even as an increasing number of adolescents and young adults present for gender-affirming surgery, the impact of top surgery on this population is not well described. Minor patients require parental consent and often face more stringent insurance restrictions. This prospective study aims to increase the body of evidence for gender-affirming top surgery in adolescents and young adults. We will measure the change in self-reported gender dysphoria, gender congruence, body image, and chest dysphoria. Methods: This is a prospective, multi-institutional study. Transmasculine and non-binary, designated female at birth, patients between the age of 13-25 years presenting for top surgery consultation were recruited from: Northwestern Memorial Hospital, The University of Illinois at Chicago, or Ann & Robert H. Lurie Children’s Hospital of Chicago. Patients completed four patient-reported outcomes measures at three time points: pre-operative baseline, three-months postoperative, and one-year postoperative. The questionnaires employed included the Transgender Congruence Scale (TCS), the Utretcht Gender Dysphoria Scale (UGDS), the Chest Dysphoria Measure (CDM), and the Body Image Scale (BIS). Preliminary interim analysis of mean change scores between pre- and three-month postoperative surveys was performed using paired, two-sided t-tests with confidence level at 95%. Results: Thirty nine patients have been enrolled to date. At interim analysis, ten patients, mean age 18.6±2.9, range 15-24 years, had completed 3-month follow-up. Eight identified as transmasculine, one non-binary/genderqueer, and one identified as ‘other.’ Mean change from baseline to three-months of the TCS appearance congruence sub-scale was 7.3 points (p = 0.002), mean change of the internal congruence sub-scale was 0.1 points (p = 0.859), and total score scale was 7.4 points (p = 0.002). The UGDS demonstrated a mean change of -2.1 points at three-months (p = 0.099). The Chest Dysphoria Measure demonstrated mean change of -28.3 points at three-months (p < 0.001). The BIS total score mean change was -12.3 points at three-months (p = 0.011). Among the BIS subscales, the primary sexual characteristics score had a mean change of -5.5 points (p=0.003), secondary sexual characteristics had a mean change of -4.0 points (p = 0.047) and neutral characteristics had a mean change of -1.6 points (p = 0.259) at three months. Conclusion: Our preliminary findings demonstrate that gender-affirming chest surgery improves chest dysphoria, appearance congruence, and overall gender congruence in transmasculine and non-binary adolescents and young adults. We anticipate that the final data will inform clinical practice guidelines for transgender and non-binary patients seeking mastectomy and chest masculinization.

Mutations of SMC5/6 components cause developmental defects, including primary microcephaly. To model neurodevelopmental defects, we engineered a mouse wherein Smc5 is conditionally knocked out (cKO) in the developing neocortex. Smc5 cKO mice exhibited neurodevelopmental defects due to neural progenitor cell (NPC) apoptosis, which led to reduction in cortical layer neurons. Smc5 cKO NPCs formed DNA bridges during mitosis and underwent chromosome missegregation. SMC5/6 depletion triggers a CHEK2-p53 DNA damage response, as concomitant deletion of the Trp53 tumor suppressor or Chek2 DNA damage checkpoint kinase rescued Smc5 cKO neurodevelopmental defects. Further assessment using Smc5 cKO and auxin-inducible degron systems demonstrated that absence of SMC5/6 leads to DNA replication stress at late-replicating regions such as pericentromeric heterochromatin. In summary, SMC5/6 is important for completion of DNA replication prior to entering mitosis, which ensures accurate chromosome segregation. Thus, SMC5/6 functions are critical in highly proliferative stem cells during organism development.

Background:. More than 75% of major limb amputees experience chronic pain; however, data on severity and experience of pain are inconsistent. Without a benchmark using quantitative patient-reported outcomes, it is difficult to critically assess the efficacy of novel treatment strategies. Our primary objective is to report quantitative pain parameters for a large sample of amputees using the validated Patient-reported Outcomes Measurement System (PROMIS). Secondarily, we hypothesize that certain patient factors will be associated with worse pain. Methods:. PROMIS and Numerical Rating Scales for residual limb pain (RLP) and phantom limb pain (PLP) were obtained from a cross-sectional survey of upper and lower extremity amputees recruited throughout North America via amputee clinics and websites. Demographics (gender, age, race, and education) and clinical information (cause, amputation level, and time since amputation) were collected. Regression modeling identified factors associated with worse pain scores (P < 0.05). Results:. Seven hundred twenty-seven surveys were analyzed, in which 73.4% reported RLP and 70.4% reported PLP. Median residual PROMIS scores were 46.6 [interquartile range (IQR), 41–52] for RLP Intensity, 56.7 (IQR, 51–61) for RLP Behavior, and 55.9 (IQR, 41–63) for RLP Interference. Similar scores were calculated for PLP parameters: 46.8 (IQR, 41–54) for PLP Intensity, 56.2 (IQR, 50–61) for PLP Behavior, and 54.6 (IQR, 41–62) for PLP Interference. Female sex, lower education, trauma-related amputation, more proximal amputation, and closer to time of amputation increased odds of PLP. Female sex, lower education, and infection/ischemia-related amputation increased odds of RLP. Conclusion:. This survey-based analysis provides quantitative benchmark data regarding RLP and PLP in amputees with more granularity than has previously been reported.

Summary:. Management of postoperative pain is a challenge for healthcare providers in all surgical fields, especially in the context of the current opioid epidemic. We developed a cell phone application to monitor pain, medication use, and relevant quality of life domains (eg, mood, mobility, return to work, and sleep) in patients with neurogenic pain, including those with limb loss. A literature review was conducted to define application length and design parameters. The final application includes 12 questions for patients with limb loss and 8 for patients with neurogenic pain without limb loss. Pilot testing with 21 participants demonstrates acceptable time to complete the application (mean = 158 seconds, SD = 81 seconds) and usability, based on the mHealth App Usability Questionnaire. We aim for our application to serve as an outcome measure for evaluation of an evolving group of peripheral nerve procedures, including targeted muscle reinnervation. In addition, the application could be adapted for clinical use in patients undergoing these procedures for neurogenic pain and thus serve as a tool to monitor and manage pain medication use.

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