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With the advent of next-generation sequencing (NGS), identifying and better understanding genetic mutations in cancer pathways has become more feasible. A mutation now commonly reported in NGS panels is the SETD2 gene (H3K36 trimethyltransferase). However, its contributions to colorectal cancer (CRC) are not well described. In this study, we describe the clinicopathologic characteristics of SETD2-mutated CRC, determine common mutation sites on the SETD2 gene, and correlate these mutations with the loss of H3K36 trimethylation and the aberrant expression of beta-catenin. By searching pathology reports at our institution which included the 161-gene NGS panel from 2019 to 2021, we identify 24 individuals with SETD2-mutated CRC. All samples were evaluated for microsatellite status, H3K36 trimethylation, and beta-catenin via immunohistochemistry. In this cohort of 24 SETD2-mutated CRC individuals (a median age of 62.4 years [interquartile range: 49.1-73.6]), 10 (41.7%) patients presented at American Joint Committee on Cancer (AJCC) tumor stage II, seven (29.2%) at stage III, six (25%) at stage IV, and one (4.2%) at stage I. Most tumors studied were adenocarcinomas with no further specification (22, 92%), and most tumors were microsatellite stable (18, 82.5%). Thirty-three mutation locations were represented by 24 patients, with one patient having six mutations in the SETD2 gene and two patients having three mutations. The dominant mutation type is missense mutations (N = 29, 87.9%), and no mutation hotspots were found. Only two samples lost trimethylation of histone H3K36, both from individuals with multiple SETD2 mutations and aberrant nuclear beta-catenin expression. SETD2-mutated CRC is similar in clinical and histologic presentation to other commonly reported CRC. SETD2 mutations were missense dominantand showed no hotspots, and multiple mutations are likely necessary for loss of H3K36 trimethylation. These results warrant further study on determining a role of SETD2-histone H3K36 pathway in CRC.

Anal squamous cell carcinoma (SCC) is a human papillomavirus (HPV)-mediated malignancy with increasing incidence. Human immunodeficiency virus (HIV) infection is a significant risk factor for anal SCC; however, it is unknown if HIV infection alters anal lesion progression and HPV strain profile. This study aims to determine whether HIV coinfection is associated with progression of HPV-mediated anal lesions and on their HPV strain diversity. This is a retrospective cohort study of adults with anal squamous intraepithelial lesion (SIL) who presented for anorectal sampling between 2010 and 2019. Using the full cohort, we performed clinicopathologic epidemiologic analysis of HIV coinfection on lesion progression. Using a subset of patients, we conducted molecular analysis of HPV strain diversity as related to HIV status and progression. Our cohort included 2203 individuals, of which 940 (43%) were HIV+. HIV+ status was associated with faster progression at all levels of dysplasia. Our molecular cohort included 329 adults, of which 190 (57.8%) were HIV+. HIV+ status was associated with higher HPV strain diversity (median: 7 [5-9] versus median: 4 [4-6], P .001). Latent class analysis identified specific HPV strain signatures associated with progression. We demonstrate that HIV+ individuals had faster rates of anal SIL progression and that almost all HPV strains were more prevalent in anal samples from HIV+ adults. Our results imply that HIV

Purpose CCHS is an extremely rare congenital disorder requiring artificial ventilation as life support. Typically caused by heterozygous polyalanine repeat expansion mutations (PARMs) in the PHOX2B gene, identification of a relationship between PARM length and phenotype severity has enabled anticipatory management. However, for patients with non-PARMs in PHOX2B (NPARMs, ~10% of CCHS patients), a genotype–phenotype correlation has not been established. This comprehensive report of PHOX2B NPARMs and associated phenotypes, aims at elucidating potential genotype–phenotype correlations that will guide anticipatory management. Methods An international collaboration (clinical, commercial, and research laboratories) was established to collect/share information on novel and previously published PHOX2B NPARM cases. Variants were categorized by type and gene location. Categorical data were analyzed with chi-square and Fisher's exact test; further pairwise comparisons were made on significant results. Results Three hundred two individuals with PHOX2B NPARMs were identified, including 139 previously unreported cases. Findings demonstrate significant associations between key phenotypic manifestations of CCHS and variant type, location, and predicted effect on protein function. Conclusion This study presents the largest cohort of PHOX2B NPARMs and associated phenotype data to date, enabling genotype–phenotype studies that will advance personalized, anticipatory management and help elucidate pathological mechanisms. Further characterization of PHOX2B NPARMs demands longitudinal clinical follow-up through international registries.

Information on bronchoalveolar lavage (BAL) in patients with COVID-19 is limited, and clinical correlation has not been reported. This study investigated the key features of BAL fluids from COVID-19 patients and assessed their clinical significance. A total of 320 BAL samples from 83 COVID-19 patients and 70 non-COVID-19 patients (27 patients with other respiratory viral infections) were evaluated, including cell count/differential, morphology, flow cytometric immunophenotyping, and immunohistochemistry. The findings were correlated with clinical outcomes. Compared to non-COVID-19 patients, BAL from COVID-19 patients was characterized by significant lymphocytosis (p

Aminoacyl-tRNA synthetases (ARSs) are ubiquitous, ancient enzymes that charge amino acids to cognate tRNA molecules, the essential first step of protein translation. Here, we describe 32 individuals from 21 families, presenting with microcephaly, neurodevelopmental delay, seizures, peripheral neuropathy, and ataxia, with de novo heterozygous and bi-allelic mutations in asparaginyl-tRNA synthetase (NARS1). We demonstrate a reduction in NARS1 mRNA expression as well as in NARS1 enzyme levels and activity in both individual fibroblasts and induced neural progenitor cells (iNPCs). Molecular modeling of the recessive c.1633C>T (p.Arg545Cys) variant shows weaker spatial positioning and tRNA selectivity. We conclude that de novo and bi-allelic mutations in NARS1 are a significant cause of neurodevelopmental disease, where the mechanism for de novo variants could be toxic gain-of-function and for recessive variants, partial loss-of-function.

Sinonasal glomangiopericytoma (SNGP) is a neoplasm arising in the nasal cavity and paranasal sinuses that shows perivascular myoid differentiation. The diagnosis of SNGP may be diagnostically challenging due to a large number of potential mimics. In the present study, we sought to characterize the histological and molecular features of six cases of SNGP found in prior surgical pathology records over a 15-year period. The average age at diagnosis was 48.5 years (range: 31-78 years), and the male-to-female ratio was 1:1. Imaging studies in all six cases demonstrated avidly enhancing, lobulated soft tissue masses in the nasal cavity, extending into the sinuses and nasopharynx. Histologically, the tumors were unencapsulated and composed of a proliferation of closely packed, bland, and uniform spindle cells growing deep to an intact surface respiratory epithelium. The cells were separated by a distinctive vascular network ranging from capillaries to large vascular spaces. All cases demonstrated strong positivity for smooth muscle actin, cyclin D1, CD99, and β-catenin (100%). Targeted sequencing revealed recurrent CTNNB1 missense mutations in all cases tested. Additionally, TLE1 was positive in all cases which has not been previously reported. No tested cases harbored SS18 translocations. We found that while no single marker resolves immunohistochemical overlap between SNGP and its histologic mimics, an extended immunohistochemical panel that includes β-catenin, cyclin D1, STAT6, smooth muscle actin, pan-cytokeratin cocktails, S100, and SOX10 helps to support the diagnosis of SNGP in diagnostically challenging cases without the need for molecular studies.

Background: Ripretinib, a broad-spectrum KIT and platelet-derived growth factor receptor A switch-control tyrosine kinase inhibitor, is approved for the treatment of adult patients with advanced gastrointestinal stromal tumor as ≥ fourth-line therapy. We present the efficacy and safety of ripretinib in patients with KIT-altered metastatic melanoma enrolled in the expansion phase of the ripretinib phase I study. Patients and methods: Patients with KIT-altered metastatic melanoma were enrolled and treated with ripretinib at the recommended phase II dose of 150 mg once daily in 28-day cycles. Investigator-assessed responses according to Response Evaluation Criteria In Solid Tumors version 1.1 were carried out on day 1 of cycles 3, 5, 7, every three cycles thereafter, and at a final study visit. Results: A total of 26 patients with KIT-altered metastatic melanoma (25 with KIT mutations, 1 with KIT-amplification) were enrolled. Patients had received prior immunotherapy (n = 23, 88%) and KIT inhibitor therapy (n = 9, 35%). Confirmed objective response rate (ORR) was 23% [95% confidence interval (CI) 9%-44%; one complete and five partial responses] with a median duration of response of 9.1 months (range, 6.9-31.3 months). Median progression-free survival (mPFS) was 7.3 months (95% CI 1.9-13.6 months). Patients without prior KIT inhibitor therapy had a higher ORR and longer mPFS (n = 17, ORR 29%, mPFS 10.2 months) than those who had received prior KIT inhibitor treatment (n = 9, ORR 11%, mPFS 2.9 months). The most common treatment-related treatment-emergent adverse events (TEAEs) of any grade in ≥15% of patients were increased lipase, alopecia, actinic keratosis, myalgia, arthralgia, decreased appetite, fatigue, hyperkeratosis, nausea, and palmar-plantar erythrodysesthesia syndrome. There were no grade ≥4 treatment-related TEAEs. Conclusions: In this phase I study, ripretinib demonstrated encouraging efficacy and a well-tolerated safety profile in patients with KIT-altered metastatic melanoma, suggesting ripretinib may have a clinically meaningful role in treating these patients. CA extern

Objective 1.) Describe demographic and clinical characteristics of pediatric nasal septal perforations (NSP), 2.) Analyze efficacy of treatment modalities in symptom management and resolution of pediatric NSP, 3.) Describe the surgical technique of external rhinoplasty with vascularized nasal septal flaps in the treatment of nasal septal perforations. Methods IRB-approved retrospective chart review of pediatric patients ages 0–18 years with nasal septal perforations treated at a tertiary care pediatric otolaryngology practice. Demographic and clinical characteristics including gender, age, race, and presenting symptoms, and location, size, and etiology of perforation were collected. Outcomes including persistence of perforation and symptoms at 1 year and most recent visit were recorded. A total of 20 patients were included. Statistical analysis used Fisher's t-test for categorical variables and Wilcoxon rank-sum for continuous variables. Results Median age was 167.5 months (1.5–221.0). The most common etiology was iatrogenic (40%), followed by button battery (20%). Thirty percent of patients underwent surgical repair. Fifty percent of patients who underwent surgical repair achieved closure of their perforation at most recent follow up. Conclusion Pediatric NSP is a challenging issue with limited literature to date. Iatrogenic causes (40%) and button batteries (20%) were the most common etiologies of nasal septal perforation in our study. We introduce an advancement in our center's surgical technique with a case illustration with repair via external rhinoplasty and bilateral vascularized nasal septal flaps. Future larger studies may further elucidate characteristics and treatment modalities associated with successful closure.

Three experiments examined whether second-order conditioning resulted in the formation of a fully-featured temporal map, as proposed by the temporal coding hypothesis. Experiments 1 and 2 examined second-order conditioning with different first- and second-order relationships. Measures of the strength of second-order conditioning were mostly consistent with the temporal coding hypothesis; second-order conditioning was best with arrangements in which CS2 occurred prior to the time that the US normally occurred during CS1-US presentations. However, there was no evidence of anticipatory timing during CS2 during second-order conditioning. A third experiment directly examined whether a fully-featured temporal map was formed during second-order conditioning by examining the acquisition of anticipatory timing in subsequent reinforced second-order trials. The results of Experiment 3 suggested that the effects obtained in Experiments 1 and 2 were due to learning of the temporal order and coincidence of events that resulted in the formation of an ordinal temporal map, but that precise durations were not encoded.

The highly polymorphic human leukocyte antigen (HLA) genes, located in the human major histocompatibility complex, encode the class I and II antigen-presenting molecules, which are centrally involved in the immune response. HLA typing is used for several clinical applications, such as transplantation, pharmacogenetics, and diagnosis of autoimmune disease. HLA typing is highly complex because of the homology of HLA genes and pseudogenes and the extensive polymorphism in the population. The Centers for Disease Control and Prevention established the Genetic Testing Reference Materials Coordination Program (GeT-RM) in partnership with the genetics community to improve the availability of genomic DNA reference materials necessary for quality assurance of genetic laboratory testing. The GeT-RM together with three clinical laboratories and the Coriell Cell Repositories have characterized genomic DNA obtained from a panel of 108 cell lines for all HLA classic polymorphic loci: HLA-A, B, C, DRB1, DRB3, DRB4, DRB5, DQA1, DQB1, DPA1, and DPB1. The goal was to develop a publicly available and renewable source of well-characterized genomic DNA reference materials to support molecular HLA typing assay development, validation, and verification, quality control, and proficiency testing. These genomic DNA samples are publicly available from the National Institutes of General Medical Science Repository at the Coriell Cell Repositories.

Specialised DDH (developmental dysplasion of the hip) clinics are developing around Ireland but are, however, variable in how they are operated. A DDH clinic was set up in the South-east of Ireland in 2002 with the goal of achieving an integrated care pathway between the orthopaedic surgical team and nursing team, working to an explicit protocol while also fostering a strong collaboration with the ultrasound department. This paper aims to explore the effectiveness of this dedicated clinic in the Southeast of Ireland.

Next-generation sequencing (NGS) methods for cancer testing have been rapidly adopted by clinical laboratories. To establish analytical validation best practice guidelines for NGS gene panel testing of somatic variants, a working group was convened by the Association of Molecular Pathology with liaison representation from the College of American Pathologists. These joint consensus recommendations address NGS test development, optimization, and validation, including recommendations on panel content selection and rationale for optimization and familiarization phase conducted before test validation; utilization of reference cell lines and reference materials for evaluation of assay performance; determining of positive percentage agreement and positive predictive value for each variant type; and requirements for minimal depth of coverage and minimum number of samples that should be used to establish test performance characteristics. The recommendations emphasize the role of laboratory director in using an error-based approach that identifies potential sources of errors that may occur throughout the analytical process and addressing these potential errors through test design, method validation, or quality controls so that no harm comes to the patient. The recommendations contained herein are intended to assist clinical laboratories with the validation and ongoing monitoring of NGS testing for detection of somatic variants and to ensure high quality of sequencing results.

Metanephric stromal tumors (MSTs) are rare renal stromal tumors that predominantly affect children. They belong to the metanephric family of tumors, along with metanephric adenofibroma and metanephric adenoma. The previous documentation of BRAF exon 15 mutations in 88% of metanephric adenomas and in isolated cases of metanephric adenofibroma prompted us to investigate the prevalence of these mutations in MSTs and in other pediatric renal stromal tumors. In this study, 17 MSTs, 22 congenital mesoblastic nephromas, and 6 ossifying renal tumors of infancy were selected for BRAF exon 15 testing. Tumor genomic DNA was extracted from formalin-fixed paraffin-embedded tissue, followed by polymerase chain reaction amplification and Sanger dideoxy sequencing with primers flanking the BRAF exon 15 gene. BRAF exon 15 mutations were found in 11 (65%) of the 17 cases of MST, all corresponding to a thymidine-to-adenine substitution at codon 600 (BRAF V600E). All other renal stromal tumors tested were negative for BRAF exon 15 mutations. In conclusion, BRAF V600E mutations are encountered in most MSTs, supporting a link with other metanephric tumors and suggesting a clonal event possibly affecting primordial renal cells. In addition, BRAF V600E mutations have been associated with oncogene-induced senescence in other benign tumors, providing clues to the pathogenesis of metanephric neoplasms in keeping with their overall benign behavior. Our results also suggest a potential diagnostic use for BRAF exon 15 mutations in differentiating MSTs from other pediatric renal stromal tumors, particularly in limited samples.

This study employs a rhetorical analysis of Planned Parenthood’s rhetoric in response to the crisis emerging from the release of controversial videos by the Center for Medical Progress. Using Bostdorff and Vibbert’s concept of values advocacy and Heath’s discussion of external rhetoric, we examine how Planned Parenthood appeals to core American values through social media in order to enhance its image and reinforce its legitimacy. The values of truth, health, agency, education, unity, and equality are ubiquitous throughout Planned Parenthood’s strategic communication with the public. This research contributes to ongoing academic discussions of the theoretical and practical implications of external rhetoric, values advocacy, and legitimacy in public relations.

Undesirable truncated recombinant protein products pose a special expression and purification challenge because such products often share similar chromatographic properties as the desired full length protein. We describe here our observation of both full length and a truncated form of a yeast protein (Gcn5) expressed in Escherichia coli, and the reduction or elimination of the truncated form by mutating a cryptic Shine-Dalgarno or START codon within the Gcn5 coding region. Unsuccessful attempts to engineer in a cryptic translation initiation site into other recombinant proteins suggest that cryptic Shine-Dalgarno or START codon sequences are necessary but not sufficient for cryptic translation in E. coli.

The H-bond donating ability for 127 compounds including drug fragments and isosteres have been quantified using a simple and rapid method with 31P NMR spectroscopy. Functional groups important to medicinal chemistry were evaluated including carboxylic acids, alcohols, phenols, thioic acids and nitrogen group H-bond donors. 31P NMR shifts for binding to a phosphine oxide probe have a higher correlation with equilibrium constants for H-bonding (log KHA) than acidity (pKa), indicating that these binding experiments are representative of H-bonding ability and not proton transfer. Additionally, 31P NMR binding data for carboxylic acid isosteres correlates with physicochemical properties such as lipophilicity, membrane permeability and plasma protein binding. This method has been used to evaluate the H-bond donating ability of small molecule drug compounds such as NSAIDs and antimicrobials.

Sandwich-structured composites are in high demand in various industries, and additive manufacturing has proven its ability to meet this demand. As a result of the advances in three-dimensional (3D) printing techniques, 3D-printed polymers have received considerable attention in fabrication of sandwich structures with complex geometries. This paper is concerned with design, manufacturing, and analysis of the 3D-printed sandwich-structured components which experienced various loadings and environmental conditions. The core structure plays a major role in the in-plane behavior of lattice composites, therefore in this study, sandwich specimens with two types of core topologies made of two common and similar 3D printing filaments, acrylonitrile butadiene styrene (ABS) and acrylonitrile styrene acrylate (ASA), were manufactured. Based on the applications of sandwich-structured parts, they might experience different temperatures in their service life. In order to determine effects of thermal environment, we conducted accelerated thermal aging within temperatures of 22-60 ° C , which is below glass temperature of the examined materials. Based on a series of three-point bending tests, the failure behavior of the original and aged components are determined, and the effects of temperature change on the bending behavior of 3D-printed sandwich parts are discussed. The experimental practice revealed that ASA with honeycomb core specimens indicated highest stability under bending load after thermal aging. The current study sheds lights on durability of 3D-printed sandwich structural elements, and the obtained results demonstrate feasibility of 3D printing technology in fabrication of thermal-stable sandwich structures.

The relationship between process parameters and structural transformations in the fibres at each stage of the carbon fibre manufacturing process play a crucial role in developing high performance carbon fibres. Here we report a systematic method which uses the combination of Taguchi approach and scientific evaluation techniques to establish these relationships for the initial stage of thermal stabilization. Density, cyclization index and fraction of reacted nitriles of a precursor containing acrylonitrile, methacrylate and itaconic acid (AN/MA/IA) were used to assess the progress of stabilization in the fibres with respect to various combinations of process parameters. The extent of progress of stabilization improved with increase in temperature (from 225 to 235 °C) and time (from 12 to 24 min) whereas an opposite trend was observed with increase in the tension on the fibres from (1600–2550 cN). According to optical microscopy, radial heterogeneity was observed in the fibres treated at 235 °C. Interestingly, we were able to identify the existence of heterogeneous modulus distribution from skin to core of the precursor fibres which was further transferred to treated fibres. The overall radial modulus of treated fibres was higher than the precursor fibres. In contrast to the literature, the fracture morphology of the fibre samples indicated that initiation of crack is caused by surface defects rather than radial heterogeneity.

Key points Patients in intensive care experience distress because of a multitude of factors and a significant proportion is attributable to pain.1 A vast majority of patients report moderate to severe pain at some point during their intensive care stay. Pain can be caused or provoked by many pre-existing conditions: acute medical, surgical, or routine aspects of intensive care (Table 1). View this table: Table 1 Causes of pain in intensive care. SCI, spinal cord injuries; GBS, Guillain-Barre syndrome; MS, multiple sclerosis Pain is widely regarded as the fifth vital sign, and it induces a myriad of deleterious physiological changes in most organ systems. Severe pain induces a stress response and sympatho-adrenergic stimulation causing tachycardia, hypertension, increased myocardial oxygen consumption, and can induce myocardial ischaemia in susceptible patients. Poorly managed pain from abdominal incisions reduces diaphragmatic function, causes hyperventilation and atelectasis. Pain especially in sedated patients can present as agitation, delirium and when badly managed can have psychological sequelae like post-traumatic stress disorder,2 depression, and anxiety or it can progress to chronic pain. Systemic deleterious effects of pain include systemic inflammatory response syndrome, hyperglycaemia, immunosuppression, impaired wound healing, hypercoagulability, and increased catabolism. All these detrimental effects can …

Aggressive interactions lead to changes in both future behavior and circulating testosterone (T) concentrations in animals across taxa. The specific neural circuitry and neurochemical systems by which these encounters alter neuroendocrine functioning are not well understood. Neurons expressing the inhibitory and stimulatory neuropeptides, RFamide-related peptide (RFRP) and kisspeptin, respectively, project to neural loci regulating aggression in addition to neuroendocrine cells controlling sex steroid production. Given these connections to both the reproductive axis and aggression circuitry, RFRP and kisspeptin are in unique positions to mediate post-encounter changes in both T and behavior. The present study examined the activational state of RFRP and kisspeptin neurons of male C57BL/6 mice following an aggressive encounter. Both winners and losers exhibited reduced RFRP/FOS co-localization relative to handling stress controls. Social exposure controls did not display reduced RFRP neuronal activation, indicating that this effect is due to aggressive interaction specifically rather than social interaction generally. RFRP neuronal activation positively correlated with latencies to display several offensive behaviors within winners. These effects were not observed in the anteroventral periventricular (AVPV) nucleus kisspeptin cell population. Together, these findings point to potential neuromodulatory role for RFRP in aggressive behavior and in disinhibiting the reproductive axis to facilitate an increase in T in response to social challenge.

We examine the accuracy of several approaches to represent the quantum mechanical Schrodinger, Klein–Gordon and Dirac Hamilton operators by optimized spatial matrices. Two of the approaches are based on periodic and reflecting boundaries and have an error scaling with the number of spatial grid points that is significantly better than the ones based on the usual approaches where the momentum operator is approximated by finite-difference schemes. These N × N matrices are optimum in the sense that their eigenvalues and eigenvectors are exact representations on the spatial grid for the continuous solutions of the corresponding force-free Hamiltonian. As an example, we apply these techniques to compute the vacuum’s polarization charge density from the Dirac and Foldy–Wouthuysen theory.

This study evaluated the influence of the Programmed Nutrition Beef Program and exogenous growth promotants (ExGP) on water holding capacity characteristics of enhanced beef strip loins. Sixty, frozen strip loins, arranged in a 2 × 2 factorial treatment arrangement with dietary program serving as the first factor and use of ExGP as the second factor, were thawed, injected with an enhancement solution, and stored for 7 days. Loins from ExGP cattle possessed the ability to bind more (P0.05) water before pumping and bind less (P0.05) water after pumping and storage. Loin pH across treatments was similar (P0.10) before injection, but increased post-injection and after storage (P0.01). Treatments did not affect loin purge loss, steak cook loss, and expressible moisture (P0.10). The Programmed Nutrition Beef Program and use of ExGPs minimally impacted water holding capacity of enhanced frozen/thawed beef strip loins.

Under certain models of animal competition, individuals are expected to gather information about opponent quality in order to determine whether they should fight or withdraw. However, the ability to process complex information differs between individuals and across brain hemispheres: a feature of vertebrate cognition known as lateralization that is not anticipated by contest models. I investigated the relationship between aggressive behaviour and mating success during the fallow deer, Dama dama, rut and a measure of lateralization derived from eye preference during parallel walking. Results show that there was no relationship between the tendency to escalate to fighting or predictability in the tendency to engage in fighting and lateralization. Conversely, there was a quadratic relationship between third-party intervention behaviour and lateralization: the greater the tendency to intervene in ongoing fights the lower the degree of lateralization. However, individuals that showed lateralization for right-eye use were least likely to be targeted by the intervening male; thus lateralization is beneficial in this context because targeted males are highly likely to lose this subsequent encounter. The relationship between lateralization and mating success was also nonlinear: males that showed little evidence for an eye bias during lateral displays had the greatest mating success. Taken together, individuals that showed lateralization benefited from avoiding being targeted after third-party intervention; conversely, individuals that showed little evidence for lateralization actively intervened during ongoing fights and had higher mating success. These results suggest that, although lateralization does appear to confer a fitness advantage on individuals, this is not as extensive as anticipated.

This Perspectives article describes methods-based proficiency testing (MBPT), the benefits and limitations of MBPT, why the time is right for MBPT in molecular diagnostics, and how MBPT for next-generation sequencing is being developed by the College of American Pathologists.

We describe the case of a Ewing sarcoma with prominent myxoid stroma of the temporal bone in a 26-year-old female. Histologically, the tumor exhibited a fascicular growth pattern of round to spindled cells with a minimal amount of pale eosinophilic to clear cytoplasm and oval or spindled nuclei with finely dispersed chromatin and small nucleoli. Myxoid changes were prominent (>50%), with reticular or pseudoacinar growth of the loosely cohesive cells. The tumor showed strong expression of CD99, FLI1, and CD56 and was positive for the EWSR1-FLI1 fusion transcript. The diagnosis of Ewing sarcoma with myxoid stroma (myxoid variant) is particularly problematic owing to the large number of potential mimics. The tumor extends the morphologic spectrum of Ewing sarcoma beyond the previously described histological variants, and broadens the differential diagnosis. For any round/spindle cell sarcoma, prominent myxoid stroma and CD99 immunoreactivity should prompt consideration for molecular studies that include analysis of both EWSR1 and FLI1.

One class of model relating to animal contest behaviour assumes that individuals gather information concerning their opponents’ competitive ability; these models argue that such a process allows contestants to avoid engaging in dangerous fighting behaviour with a superior opponent. The brain hemispheres of vertebrates are lateralised in that they are specialised for processing different type of information. Within the context of the current study, we might expect that lateralisation would play a role in facilitating the assessment of opponent quality; nevertheless, the degree of lateralisation shown by individuals can vary suggesting that contest behaviour might also vary based on the ability to process information about competitor quality. The current study tests this hypothesis by predicting that the duration that individuals engage in fighting and the rate of aggressive contest actions should decrease as lateralisation increases. There was a positive relationship between two laterality indices and the duration spent in antler contact; thus lateralised individuals experienced greater time costs. Further, lateralised individuals also experienced a greater disparity in contest actions: there was a negative relationship between lateralisation and the difference in the mean number of backward pushes achieved during fights. When only opponent signal rate was considered there was no effect of lateralisation, therefore, there is support for a mutual assessment process. These results suggest that information gathering via lateral displays may be disadvantageous to lateralised individuals during escalated fighting.

Individuals often differ from each other in how they respond to environmental events: a feature of behaviour often termed animal personality. Furthermore, animals often show unpredictability in how much they respond to these events over time leading to the suggestion that personality and intraindividual variability (IIV) might have important fitness consequences. We investigated this hypothesis by focusing on the tendency for individually identifiable male fallow deer, Dama dama , to escalate low-level (noncontact) agonistic interactions to fighting during the rut. Males differed in their tendency to escalate noncontact interactions to fighting; however, repeatability in escalation rates was unstable over the rut suggesting that escalation rate is a poor measure of personality. There was no difference in the level of IIV in escalation rate shown by males. A comparison of IIV recorded over two consecutive annual ruts showed that IIV was highly correlated between years; therefore, over an extended time period individuals were consistent in their willingness to engage in fighting. There was a nonlinear relationship between IIV and mating success; specifically, individuals that showed either low or high IIV in escalation rate had lower mating success than individuals that showed intermediate levels of IIV. Aggression and the willingness to commit to fighting are important fitness correlates; nevertheless, our understanding of how personality and IIV are related to aggression is poorly understood. This study shows that although escalation rate appears to be a poor measure of personality in the fallow deer, IIV is related to increased fitness in individuals that show intermediate levels of predictability in their willingness to fight over the rut.

In order to evaluate the importance of molecular shape of inhibitor molecules and the charge/H-bond and hydrophobic interactions, we synthesized three types of molecules and tested them against a sialyltransferase. The first type of compounds were designed as substrate mimics in which the phosphate in CMP-Neu5NAc was replaced by a non-hydrolysable, uncharged 1,2,3-triazole moiety. The second type of compound contained a 2-deoxy-2,3-dehydro-acetylneuraminic moiety which was linked to cytidine through its carboxylic acid and amide linkers. In the third type of compound the sialyl phosphate was substituted by an aryl sulfonamide which was then linked to cytidine. Inhibition study of these cytidine conjugates against Campylobacter jejuni sialyltransferase Cst 06 showed that the first type of molecules are competitive inhibitors, whereas the other two could only inhibit the enzyme non-competitively. The results indicate that although the binding specificity may be guided by molecular shape and H-bond interaction, the charge and hydrophobic interactions contributed most to the binding affinity.

The present studies examined whether victims believed that forgiveness requires interpersonal interaction between the victim and offender. Having an interpersonal conceptualization of forgiveness was positively related to a collectivistic worldview. Also, an interpersonal conceptualization of forgiveness was positively related to forgiveness measures that stressed interpersonal interactions with the offender. However, an interpersonal conceptualization of forgiveness was positively related to a general measure of forgiveness only for those participants who were in a continuing relationship with the offender. The current research is an important step in understanding the contexts in which one’s conceptualization of forgiveness may influence its practice.

article i nfo The study aims were twofold: 1) identify the localization and the cytoarchitecture of the retrotrapezoid nucleus (RTN) in the human fetus and infant and 2) ascertain if the RTN, given its essential role in animal studies for the maintenance of breathing and chemoreception, showed abnormalities in victims of sudden perinatal and infant death (sudden intrauterine unexplained death/SIUD — and sudden infant death syndrome/SIDS). We examined SIDS and SIUD cases and Controls (n=58) from 34 gestational weeks to 8 months of postnatal age by complete autopsy, in-depth autonomic nervous system histological examination, and immunohistochemical analysis of the PHOX2B gene, a transcriptional factor involved in Congenital Central Hypoventilation Syndrome that has been defined as a marker of rat RTN neurons. We identified a group of PHOX2B-immunopositive neurons within the caudal pons, contiguous to the facial/ parafacial complex, in 90% of Controls, likely the homologous human RTN (hRTN). We observed structural and/or PHOX2B-expression abnormalities of the hRTN in 71% of SIUD/SIDS cases vs 10% of Controls (pb0.05). In conclusion we suggest that developmental abnormalities of the hRTN may seriously compromise chemore- ception control, playing a critical role in the pathogenesis of both SIUD and SIDS.

Highlights ► More responding when occasion setter signals transfer CS signalling the same outcome. ► Similar responding when pseudo-occasion setter signals target CS signalling the same outcome. ► Less responding when transfer CS signals target CS signalling the same outcome. ► Results inconsistent with configural theory, consistent with hierarchical theory., Four experiments in rats examined whether occasion setters and target CSs play qualitatively different roles in occasion-setting discriminations. Two visual occasion setters, A and B, signalled reinforcement of two auditory target CSs, x and y, with sucrose and oil (A…x → suc, B…y → oil, A−, B−, x−, y−); in addition two transfer CSs w and z were paired with sucrose and oil (w → suc, z → oil). When w and z were substituted for x and y (A…w, B…w, A…z, B…z) more responding was observed when both stimuli had been paired with the same outcome (Experiments 1 and 3a). No effect was observed when two visual “pseudo-occasion setters”, C and D (paired with sucrose and oil in a trace relation to the US:C… → suc, D… → oil), were substituted for the occasion setters A and B (C…x, D…x, C…y, D…y; Experiments 2, 3b and 4). These results could not be explained in terms of Pavlovian summation: responding to combinations of Pavlovian CSs paired with same or different outcomes was either the same, or lower when both stimuli had been paired with the same outcome (Experiment 4). Implications of these results for theories of occasion setting and configural learning are discussed.

Lateral displays during agonistic interactions are assumed to provide contestants the opportunity to assess the opponent’s quality. Once an individual has acquired sufficient information, a decision to terminate the display can be made and either escalation to fighting or de-escalation and movement away from the opponent occurs. Because cerebral lateralization enhances the processing of complex cognitive tasks in the visual domain, it is possible that lateralization might provide an efficient means by which an assessment of the opponent's quality is facilitated. I investigated whether there is evidence for lateralization in the decision to terminate parallel walks during fights recorded between individually identifiable male fallow deer, Dama dama. Males exhibited a significant bias to terminate parallel walks when displaying their right flank over their left; therefore, the decision to end a parallel walk showed evidence for lateralization. There was an increasing tendency to show a right-sided bias when terminating the parallel walk as body length increased and heavier males terminated parallel walks sooner than lighter males. Therefore, larger individuals displayed a greater tendency towards lateralization than smaller individuals. Moreover, lighter males showed a greater tendency to escalate parallel walks to fighting. These results show evidence for lateralization in the decision to terminate lateral displays during escalated contests in an ungulate species, and suggest that lateralization provides a mechanism by which contestants can resolve contests at a low cost.

Third-party interventions of dyadic contests are often explained by appealing to high-level cognitive processes such as coalition formation between group members. However, alternative accounts that do not appeal to sophisticated cognitive processes have been proposed. We tested predictions from two such models using the fallow deer, Dama dama, as the model taxon: (1) a random target model that argues that intervention is directed randomly towards a target and (2) a specific target model that assumes that targeting is directed at contestants that have low resource-holding potential. Contrary to predictions of the specific target model, we found no evidence that targeting following third-party intervention increased as the resource-holding potential of the target declined. Both models argue that intervention serves to prevent individuals from gaining a winner effect and advancing up the hierarchy. Being targeted did not result in a decline in dominance rank, although targeting was associated with investment in dominance-related fighting tactics. Fight intervention was associated with an increase in rank early in the rut and accounted for increased mating success. Therefore, interveners benefited beyond simply preventing rivals from advancing in the hierarchy. In theoretical terms, a random target as opposed to a specific target model explains intervention behaviour in the fallow deer.

Aim Improve availability of publicly available, renewable, and well-characterized genomic DNA reference materials to support molecular HLA typing assay development, validation and verification, quality control, and proficiency testing. Methods The Centers for Disease Control and Prevention’s (CDC) Genetic Testing Reference Materials Coordination Program (GeT-RM) together with three clinical laboratories and the Coriell Cell Repositories characterized genomic DNA from 108 cell lines for HLA-A, B, C, DRB1, DRB3, DRB4, DRB5, DQA1, DQB1, DPA1, and DPB1. DNA was isolated by the Coriell Cell Repositories and distributed to the 3 laboratories. Methodologies used were: PCR-SSO-LABType® SSO (One Lambda, Canoga Park, CA), SSP – Olerup SSP® (Stockholm, Sweden), SBT-AlleleSEQR® HLA-SBT (Abbott Molecular, Des Plaines, IL) and NGS - HoloType HLA™ kits (Omixon Inc., Budapest, Hungary). Each of the 11 genes was tested by 2 or 3 methods. Consensus genotype was the NGS result, which provided the highest level of resolution, and was consistent with the results obtained by the parallel testing using other methods. Results HLA typing results using SBT, SSP, SSO and NGS were 99.95 % concordant. Of the 1905 alleles typed, only one discrepancy was observed. The discrepant result was found to be due to an error in SBT/SSO typing. Five unique novel alleles with differences in exonic regions were identified and three were submitted to the WHO Nomenclature Committee. Ambiguities persisted in 50/1905 allele calls, most due to alternative cis/trans combinations of exons 2 and 3 of DPB1 (n = 21 pairs: 42 alleles) and 8 caused by polymorphisms in a non-covered region (exon1) of the DRB1and DPB1 genes. Thirty HLA-A, 54-B, 30-C, 36-DRB1, 4-DRB3, 4-DRB4, 3-DRB5, 19-DQA1, 17-DQB1, 10-DPA1 and 29-DPB1 alleles are represented in the panel. These alleles cover a high percentage of HLA specificities present in five ancestry groups in the USA population. Conclusion These genomic DNA samples are publicly available from the National Institutes of General Medical Science (NIGMS) Repository at the Coriell Cell Repositories. M. Bettinotti: 3. Speaker’s Bureau; Company/Organization; One Lambda Thermo Fisher. D. Ferriola: 7. Other (Identify); Company/Organization; Licensing proceeds of the HLA-NGS protocol by CHOP to Omixon. J.L. Duke: 7. Other (Identify); Company/Organization; receiving proceeds from the licensing of the HLA-NGS protocol by CHOP to Omixon. D. Monos: 4. Scientific/Medical Advisor; Company/Organization; Omixon. 7. Other (Identify); Company/Organization; receiving proceeds from the licensing of the HLA-NGS protocol by CHOP to Omixon.

The drag flick was introduced to field hockey in the early 1990’s by Dutch player Taco Hajo van den Honert. It has become a major offensive weapon as a set play in penalty corners, with skilled drag flickers highly sought after at all levels of the game. This study investigated the forces between ball and stick and the location of the ball on the stick during a drag flick. Force sensors were mounted on the surface of the stick to determine both the forces applied and ball position with high temporal resolution during drag flicking. A high g accelerometer was used to measure the acceleration of the stick head. These devices were synchronised in a common wireless data acquisition system. Three drag flickers with different skill levels were used as subjects and were instructed to direct the ball at a particular target region, a small sector of the field hockey goal. The force sensor data showed that the force and location of the ball during a drag flick was important for controlling drag flicks. The use of the force sensors also affected the friction of the stick surface and made it more difficult for the subjects to impart curvature onto the ball’s trajectory consistent with elite competition. Further development of the sensors is required to improve measurement and analysis.

According to life history theory, males of iteroparous species are expected to trade off investment between current and future reproduction based on age (mating strategy or terminal investment hypotheses) or body condition (individual quality hypothesis). However, although central to this latter model, the question concerning whether and to what extent condition regulates competitive investment in polygynous species is unknown. Consequently, we investigated this issue with reference to fight structure in fallow deer contests. Support for the individual quality hypothesis was limited: males with larger necks as determined by prerut neck girth fought for longer than males with smaller necks. However, prime-aged males had higher investment in fighting than preprime- or postprime-aged males indicating that investment in fighting might be age related. Other aspects of our results also failed to support condition-related predictions; although we found that jump clashing and vocal rate were related to weight loss and decline in neck girth, respectively, there was no relationship between investment in fighting and prerut measures of body size. Moreover, we also found that rank was predicted by investment in fighting (backward pushing) rather than body condition. Our results show that, in addition to body condition and age, variation in competitive investment between individuals also influences reproductive effort in the fallow deer.

The binding of RCC1 (regulator of chromosome condensation 1) to chromatin is critical for cellular processes such as mitosis, nucleocytoplasmic transport, and nuclear envelope formation because RCC1 recruits the small GTPase Ran (Ras-related nuclear protein) to chromatin and sets up a Ran-GTP gradient around the chromosomes. However, the molecular mechanism by which RCC1 binds to nucleosomes, the repeating unit of chromatin, is not known. We have used biochemical approaches to test structural models for how the RCC1 beta-propeller protein could bind to the nucleosome. In contrast to the prevailing model, RCC1 does not appear to use the beta-propeller face opposite to its Ran-binding face to interact with nucleosomes. Instead, we find that RCC1 uses a conformationally flexible loop region we have termed the switchback loop in addition to its N-terminal tail to bind to the nucleosome. The juxtaposition of the RCC1 switchback loop to its Ran binding surface suggests a novel mechanism for how nucleosome-bound RCC1 recruits Ran to chromatin. Furthermore, this model accounts for previously unexplained observations for how Ran can interact with the nucleosome both dependent and independent of RCC1 and how binding of the nucleosome can enhance RCC1's Ran nucleotide exchange activity.

Male ungulates engage in intense competition for access to females during the breeding season. Although fights are generally dyadic level encounters, they are on occasion disrupted by the intervention of third-party males. We investigated these third-party interventions using predictions derived from Dugatkin's model (Dugatkin 1998, Proceedings of the Royal Society of London, Series B , 265, 433–437) of intervention behaviour. The model argues that when an individual successfully defeats an opponent there is an increase in the probability of winning a subsequent contest: a winner effect. Third-party intervention behaviour is predicted to occur as it serves to prevent either member of a competing dyad from successfully defeating his opponent, achieving a winner effect and subsequently becoming a threat to the intervener. Consistent with model predictions, our results show that intervening males held significantly higher rank than males that did not intervene and were also more likely to be dominant to both of the competing males. Intervening males did not selectively target competitors based on rank, nor did they target males based on overall dyadic rates of aggression between the intervener and competing males. Furthermore, interveners were more likely to have won their interaction immediately prior to intervention and were also likely to win their interaction subsequent to intervention when compared with contest success of the two competing males. Our results are consistent with predictions that support a winner effect for intervention behaviour in fallow deer fights.