Statistical methods for discrimination of STR genotypes using high resolution melt curve data.

Despite the improvements in forensic DNA quantification methods that allow for the early detection of low template/challenged DNA samples, complicating stochastic effects are not revealed until the final stage of the DNA analysis workflow. An assay that would provide genotyping information at the earlier stage of quantification would allow examiners to make critical adjustments prior to STR amplification allowing for potentially exclusionary information to be immediately reported. Specifically, qPCR instruments often have dissociation curve and/or high-resolution melt curve (HRM) capabilities; this, coupled with statistical prediction analysis, could provide additional information regarding STR genotypes present. Thus, this study aimed to evaluate Qiagen's principal component analysis (PCA)-based ScreenClust® HRM® software and a linear discriminant analysis (LDA)-based technique for their abilities to accurately predict genotypes and similar groups of genotypes from HRM data. Melt curves from single source samples were generated from STR D5S818 and D18S51 amplicons using a Rotor-Gene® Q qPCR instrument and EvaGreen® intercalating dye. When used to predict D5S818 genotypes for unknown samples, LDA analysis outperformed the PCA-based method whether predictions were for individual genotypes (58.92% accuracy) or for geno-groups (81.00% accuracy). However, when a locus with increased heterogeneity was tested (D18S51), PCA-based prediction accuracy rates improved to rates similar to those obtained using LDA (45.10% and 63.46%, respectively). This study provides foundational data documenting the performance of prediction modeling for STR genotyping based on qPCR-HRM data. In order to expand the forensic applicability of this HRM assay, the method could be tested with a more commonly utilized qPCR platform.

Interactions Between Heavy Metal Exposure and Blood Biochemistry in an Urban Population of the Black Swan (Cygnus atratus) in Australia.

There is growing recognition of the threat posed to wildlife by pollutants. Waterbirds are robust bioindicators of ecosystem health, and metal toxicity is a threat to these species in waterways worldwide. Urban waterbirds are likely to be at the highest risk of heavy metal exposure, but this issue has not been widely explored in Australia. Our aim was to estimate contemporary heavy metal exposure in a sedentary urban waterbird population: black swans (Cygnus atratus) inhabiting an inner-city wetland in one of Australia's largest cities, Melbourne. To investigate the physiological implications of legacy heavy metal exposure in these birds, we quantified blood biochemistry profiles and examined their relationships with metal concentrations in feathers. We caught 15 swans in 2021 and took feather samples to measure the concentration of eight heavy metals (chromium (Cr), manganese (Mn), iron (Fe), nickel (Ni), copper (Cu), zinc (Zn), lead (Pb), and mercury (Hg)), and blood samples to measure the concentration of 13 plasma analytes. Multivariate regression analysis revealed few associations between heavy metals and biochemistry markers, and no differences between sexes or age classes. This study presents a baseline dataset of these contaminants and blood biochemical profiles of swans at this wetland that can be used for future monitoring and is an important step toward a better understanding of the threat posed by heavy metals to Australian urban waterbirds.

A quantifiler™ trio-based HRM screening assay for the accurate prediction of single source versus mixed biological samples.

At present, the forensic DNA workflow is not capable of providing information about the contributor status (single source vs. multiple contributors) of evidentiary samples prior to end-point analysis. This exacerbates the challenges inherent to mixtures and low-template DNA samples. If additional sample information could be provided earlier in the workflow, protocols could be implemented to mitigate these challenges. An integrated Quantiplex®- high resolution melt (HRM) assay was shown to be effective in distinguishing between single source and mixture DNA samples; however, integration of the HRM assay into a more commonly used chemistry would be beneficial to the practitioner community. Thus, the assay was redesigned as an integrated Quantifiler™ Trio-HRM assay, which included the identification of a new DNA-binding dye, an increased reaction volume, and the establishment of new data analysis and standard curve metrics for all targets. This redesigned assay produced quantification values and qualitative values that were comparable to those produced when the same samples were tested using the standard Quantifiler™ Trio chemistry and settings. Further, STR profiles generated with quantification values produced from the integrated Quantifiler™ Trio-HRM assay and standard Quantifiler™ Trio chemistry were complete and fully concordant. Most importantly, the integrated Quantifiler™ Trio-HRM assay was able to accurately predict whether a sample was single source or a mixture 79.2% of the time, demonstrating the potential of this approach. With the incorporation of an expanded training set for prediction modeling, and completion of critical developmental validation studies, this assay could prove useful to the forensic DNA practitioner community.

Surgeon volumes: preserving appropriate surgical outcomes in higher-risk patient populations undergoing abdominal wall reconstruction.

BACKGROUND: It is increasingly recognized that complex abdominal wall reconstruction (cAWR) necessitates specialized training. No studies have been conducted to assess whether a volume-outcomes relationship is present in cAWR. We sought to determine if outcomes for patients undergoing cAWR varied based on surgeon volume among participants in the Abdominal Core Health Quality Collaborative (ACHQC). METHODS: All patients with ventral hernias undergoing elective cAWR with component separation (lateral component release) were selected from ACHQC database. Surgeons were ranked based on annual number of cAWR procedures performed and then grouped in tertiles. Patient characteristics, hernia risk factors, operative details, and 30-days outcomes were evaluated. RESULTS: A total of 9206 patients were identified, of which 310 (3.4%), 723 (7.9%) and 8173 (88.7%) cAWRs were performed by low (105 surgeons), medium (49) and high-volume (66) surgeons, respectively. Patients operated upon by high-volume surgeons tended to have more comorbidities and higher ASA class (72.5% of class ≥ III, vs 53.5%). Hernia characteristics demonstrated that high-volume surgeons more commonly operated on patients presenting with recurrent hernias (50.2% vs 42%), wider hernias (13.5 cm vs 10.5 cm), associated ostomies (13% vs 3.6%), and prior of surgical site infections (32% vs 26%, P = 0.035). High-volume surgeons more commonly performed posterior component separation procedures (92% vs 84%), utilized permanent mesh (92% vs 88%), and placed mesh in sublay position. In spite of operating on more advanced hernias, high-volume surgeons achieved comparable rates (all P > 0.4) of 30-day surgical site infections (SSI: 6.9% vs 7.1%) and surgical site occurrences requiring procedural intervention (SSO-PI: 8.9% vs 10%). CONCLUSIONS: High-volume surgeons maintain comparable outcomes following cAWR despite performing operations on patients with more comorbidities and advanced hernia disease. These findings should be integrated into the debates related to regionalizing abdominal wall reconstruction procedures among high-volume surgeons.

Perioperative Telemedicine Utilization Among Geriatric Patients Being Evaluated for Abdominal Wall Reconstruction and Hernia Repair.

Introduction: Perioperative telemedicine services have increasingly been utilized for ambulatory care, although concerns exist regarding the feasibility of virtual consultations for older patients. We sought to review telemedicine encounters for geriatric patients evaluated at a hernia repair and abdominal wall reconstruction center. Methods: A retrospective review of telemedicine encounters between May 2020 and May 2021 was performed. Patient characteristics and encounter-specific outcomes were compared among geriatric (older than65 years old) and nongeriatric patients. Clinical care plans for encounters were reviewed to determine potential downstream care utilization. Patient-derived benefits related to time saved in travel time was calculated using geo-mapping. Outcomes for postoperative encounters were assessed to determine if complication rates differed between geriatric and nongeriatric populations. Results: A total of 313 telemedicine encounters (geriatric: 41.9%) were conducted among 251 patients. Reviewing preoperative factors for hernia care, geriatric patients presented with higher rates of recurrent or incisional hernias (87.9% vs. 70.7%, p < 0.01). Potential travel time was longer for geriatric patients (104 min vs. 42 min, p = 0.03) in the preoperative setting. No differences in clinical care plans were found. Only 8.6% of preoperative encounters resulted in recommendations for supplemental in-person evaluation. Operative plans were coordinated for 42.5% of all preoperative telemedicine encounters. There was no difference in complication rate between geriatric and nongeriatric patients (p > 0.05) in the postoperative setting, with no complications directly attributable to telemedicine-based care. Conclusions: Telemedicine-based evaluations appear to function well among geriatric patients seeking hernia repair and abdominal wall reconstruction. Clinical care plans rendered following telemedicine-based encounters are appropriate with a low rate of supplemental in-person evaluations. Telemedicine use resulted in significantly more time saved in commuting to and from clinic for geriatric patients.

Changes in plasma electrolytes, minerals, and hepatic markers of health across the transition period in dairy cows divergent in genetic merit for fertility traits and postpartum anovulatory intervals.

Peripartum metabolism and subsequent reproductive performance of dairy cows are linked, with maladaptation over the transition period associated with poor reproductive success. A herd of seasonal calving, grazing dairy cows was established that differed in their genetic merit for fertility traits. The heifers were produced by a customized mating program to achieve a 10-percentage point divergence in the New Zealand fertility breeding value (FertBV) as follows: +5 FertBV (POS) versus -5% FertBV (NEG), while also limiting divergence in other breeding values, including body weight, body condition score, and milk production. In this study, we aimed to characterize differences in metabolic, mineral, and metabolic stress marker profiles during their first postpartum transition period as primiparous heifers and to examine if animals with longer postpartum anestrous intervals (PPAI; more than 66 d compared with less than 35 d) had greater metabolic dysfunction. Blood was sampled at -21, -14, -7, 0, 4, 7, 10, 14, 17, 21, 28, and 35 d relative to calving in 455 primiparous cows and plasma analyzed. The NEG cows had lower concentrations of both plasma nonesterified fatty acids and ß-hydroxybutyrate at d 7 compared with POS cows. Detailed temporal profiling of various metabolic, mineral, and metabolic stress markers was undertaken in a subset of cows (n = 70). Cows were selected retrospectively to create 4 groups in a 2 × 2 factorial design with either a POS or NEG FertBV and either a short (19-35 d) or long (66-131 d) PPAI. The NEG cows tended, on average, to have lower nonesterified fatty acids and ß-hydroxybutyrate concentrations compared with POS cows across the transition period. Mean body weight and body condition score was greatest in NEG cows when compared with the POS cows and an interaction with day demonstrated this only occurred precalving. They also had indications of improved liver health precalving, with higher albumin-to-globulin ratios and lower bilirubin concentrations. Concentrations of aspartate aminotransferase were lower, and the Na-to-Cl ratio was greater in cows with a long versus a short PPAI at d 28 and d 35 after calving, potentially because of cows with a short PPAI (19-35 d) returning to estrous during this time. Magnesium concentrations were lower in NEG cows with a short PPAI from d 21 onwards, indicating NEG cows may metabolically respond to estrous differently than POS cows. The NEG-long PPAI cows had greater gamma-glutamyl transferase concentrations from calving until d 28 and lower bilirubin concentrations throughout the transition period. Together, the results demonstrate significant effects of FertBV on peripartum metabolic status. However, most of the markers tested returned to reference intervals within 4 d after calving or remained within those intervals for the whole transition period, indicating relatively minor biological effects of FertBV on transition period adaptation. The profound differences in reproductive performance among the groups was not explained by underlying differences in metabolic responses during the transition period.

Reconsidering humaneness.

Animal welfare is increasingly important in the understanding of how human activity affects wildlife, but the conservation community is still grappling with meaningful terminology when communicating this aspect of their work. One example is the use of the terms "humane" and "inhumane." These terms are used in scientific contexts, but they also have legal and social definitions. Without reference to a defined technical standard, describing an action or outcome as humane (or inhumane) constrains science communication because the terms have variable definitions; establish a binary (something is either humane or inhumane); and imply underlying values reflecting a moral prescription. Invoking the term "humane," and especially the strong antithesis "inhumane," can infer a normative judgment of how animals ought to be treated (humane) or ought not to be treated (inhumane). The consequences of applying this terminology are not just academic. Publicizing certain practices as humane can create blurred lines around contentious animal welfare questions and, perhaps intentionally, defer scrutiny of actual welfare outcomes. Labeling other practices as inhumane can be used cynically to erode their public support. We suggest that, if this normative language is used in science, it should always be accompanied by a clear, contextual definition of what is meant by humane.

Reconsideración de la Condición Humana Resumen El bienestar animal es cada vez más importante para el entendimiento de cómo la actividad humana afecta a la fauna, pero la comunidad conservacionista todavía lucha con la terminología significativa cuando comunica este aspecto de su trabajo. Un ejemplo es el uso de los términos humano e inhumano. Estos términos se usan dentro de contextos científicos, pero también cuentan con definiciones sociales y legales. Sin la referencia a un estándar técnico definido, la descripción de una acción o resultado como humano (o inhumano) restringe la comunicación de la ciencia pues estos términos tienen definiciones variables; establecen una opción binaria (algo sólo es humano o sólo es inhumano); e implican valores subyacentes que reflejan una prescripción moral. La invocación del término humano, y especialmente su fuerte antítesis inhumano, puede inferir un juicio normativo sobre cómo los animales deberían ser tratados (humano) o no deberían ser tratados (inhumano). Las consecuencias de aplicar esta terminología no son sólo académicas. La divulgación de ciertas prácticas como humanas puede generar confusión entorno a preguntas polémicas sobre el bienestar animal y, tal vez de manera intencional, aplazar el escrutinio de los resultados actuales para el bienestar. Al contrario, la denominación de otras prácticas como inhumanas puede usarse cínicamente para menoscabar el apoyo público que reciben. Sugerimos que, si se utiliza este lenguaje normativo en la ciencia, siempre debería estar acompañado de una definición clara y contextual de a lo que se refiere con el término humano.

Compassionate versus consequentialist conservation.

Ethical treatment of wildlife and consideration of animal welfare have become important themes in conservation, but ethical perspectives on how best to protect wild animals and promote their welfare are diverse. There are advantages to the consequentialist harms ethical framework applied in managing wild herbivores for conservation purposes. To minimize harms while achieving conservation goals, we argue that overabundant wild herbivores should in many cases be managed through consumptive in situ killing. Advantages of this policy are that the negative welfare states imposed on animals last only a short time; remaining animals are not deprived of positive welfare states (e.g., linked to rearing offspring); poor welfare states of animals in overabundant populations are avoided (e.g., starvation); negative welfare impacts on heterospecifics through resource depletion (i.e., competition) are prevented; harvesting meat reduces the number of (agricultural) animals raised to supply meat; and minimal costs maximize funding for other wildlife management and conservation priorities. Alternative ethical approaches to our consequentialist framework include deontology (containing animal rights) and virtue ethics, some of which underpin compassionate conservation. These alternative ethical approaches emphasize the importance of avoiding intentional killing of animals but, if no population reduction occurs, are likely to impose considerable unintentional harms on overabundant wildlife and indirectly harm heterospecifics through ineffective population reduction. If nonlethal control is used, it is likely that overabundant animals would be deprived of positive welfare states and economic costs would be prohibitive. We encourage conservation stakeholders to consider animal-welfare consequentialism as an ethical approach to minimize harms to the animals under their care as well as other animals that policies may affect while at the same time pursuing conservation goals.

Conservación Compasiva versus Conservación Consecuencialista Resumen El trato ético de la fauna y la consideración por el bienestar animal se han convertido en temas importantes para la conservación, pero las perspectivas éticas sobre cuáles son las mejores maneras para proteger a los animales silvestres y promover su bienestar son diversas. Existen ventajas en el marco de trabajo ético de los daños consecuencialistas aplicados en el manejo de herbívoros silvestres por motivos de conservación. Para minimizar los daños a la vez que se alcanzan los objetivos de conservación argumentamos que los herbívoros silvestres sobreabundantes deberían, en muchos casos, ser manejados por medio de muertes in situ consuntivas. Las ventajas de esta política son que los estados negativos de bienestar impuestos sobre los animales duran muy poco tiempo; a los animales que permanecen no se les priva de un estado positivo de bienestar (p. ej.: conectados a la crianza de descendencia); se evitan los estados pobres de bienestar en las poblaciones sobreabundantes (p. ej.: hambruna); se previenen los impactos negativos de bienestar por medio de la disminución sobre los heteroespecíficos (es decir, competencia); la crianza para carne reduce el número de animales (agrícolas) criados para abastecer de carne; y los costos mínimos maximizan el financiamiento para otras prioridades de manejo y conservación de fauna. Las estrategias éticas alternativas a nuestro marco de trabajo consecuencialista incluyen la deontología (que contiene los derechos de los animales) y la ética de virtudes, algunas de las cuales apoyan la conservación compasiva. Estas estrategias éticas alternativas enfatizan la importancia de evitar la muerte intencional de los animales pero, si no ocurre una reducción de la población, es probable que impongan daños considerables accidentales sobre la fauna sobreabundante y dañen indirectamente a los heteroespecíficos por medio de la reducción infructuosa de la población. Si se utiliza un control no letal, es probable que los animales sobreabundantes quedarían privados de estados positivos de bienestar y los costos económicos serían prohibitivos. Alentamos a los accionistas de la conservación a considerar el consecuencialismo del bienestar animal como una estrategia ética para minimizar los daños que sufren los animales bajo su cuidado así como otros animales que podrían ser afectados por las políticas mientras se busca alcanzar objetivos de conservación.

Underaddressed animal-welfare issues in conservation.

Much progress has been made toward assessing and improving animal welfare in conservation. However, several glaring knowledge gaps remain where animal-welfare concerns exist but animal-welfare studies have not been performed in politically sensitive contexts. Based on contemporary issues in Australia, we identified 4 topics that require more research: animal-welfare oversight for operations designated as management (as opposed to research); animal-welfare impacts of biological agents used to control invasive animals; welfare of animals hunted recreationally; and animal-welfare impacts associated with indigenous wildlife use. Animal-welfare science may be applied to these sensitive topics through simple quantitative studies (e.g., quantifying the frequency of adverse animal-welfare events). Several such studies have effectively addressed animal-welfare concerns in similarly contentious contexts, including feral camel (Camelus dromedarius) culling in Australia, recreational hunting in Scandinavia, and indigenous whale hunting in the United States. For discussions of animal welfare in conservation to be evidence-based, courageous research is required in the 4 key areas we identified.

Temas de Bienestar Animal Sin Tratar en la Conservación Resumen En la conservación se ha progresado mucho en la evaluación y el mejoramiento del bienestar animal. Sin embargo, todavía permanecen varios vacíos evidentes en donde existe preocupación por el bienestar animal, pero los estudios sobre este bienestar no se han realizado en contextos políticamente sensibles. Con base en temas contemporáneos en Australia, identificamos cuatro temas que requieren de más investigación: omisión del bienestar animal por operaciones designadas como manejo (en lugar de investigación); impactos de los agentes biológicos usados para controlar a animales invasores sobre el bienestar animal; bienestar de los animales cazados por recreación; e impactos sobre el bienestar animal asociados con el uso de la fauna nativa. La ciencia del bienestar animal puede aplicarse a estos temas sensibles por medio de estudios cuantitativos (p. ej.: cuantificación de la frecuencia de eventos adversos para el bienestar animal). Varios de estos estudios han tratado efectivamente las preocupaciones por el bienestar animal en contextos similarmente polémicos, incluyendo el sacrificio de camellos ferales (Camelus dromedarius) en Australia, la cacería recreativa en Escandinavia, y la caza de ballenas por aborígenes en los Estados Unidos. Para que las discusiones sobre el bienestar animal en la conservación estén basadas en evidencias, se requiere de investigaciones atrevidas en las cuatro áreas clave que identificamos.

A novel, integrated forensic microdevice on a rotation-driven platform: Buccal swab to STR product in less than 2 h.

This work describes the development of a novel microdevice for forensic DNA processing of reference swabs. This microdevice incorporates an enzyme-based assay for DNA preparation, which allows for faster processing times and reduced sample handling. Infrared-mediated PCR (IR-PCR) is used for STR amplification using a custom reaction mixture, allowing for amplification of STR loci in 45 min while circumventing the limitations of traditional block thermocyclers. Uniquely positioned valves coupled with a simple rotational platform are used to exert fluidic control, eliminating the need for bulky external equipment. All microdevices were fabricated using laser ablation and thermal bonding of PMMA layers. Using this microdevice, the enzyme-mediated DNA liberation module produced DNA yields similar to or higher than those produced using the traditional (tube-based) protocol. Initial microdevice IR-PCR experiments to test the amplification module and reaction (using Phusion Flash/SpeedSTAR) generated near-full profiles that suffered from interlocus peak imbalance and poor adenylation (significant -A). However, subsequent attempts using KAPA 2G and Pfu Ultra polymerases generated full STR profiles with improved interlocus balance and the expected adenylated product. A fully integrated run designed to test microfluidic control successfully generated CE-ready STR amplicons in less than 2 h (<1 h of hands-on time). Using this approach, high-quality STR profiles were developed that were consistent with those produced using conventional DNA purification and STR amplification methods. This method is a smaller, more elegant solution than current microdevice methods and offers a cheaper, hands-free, closed-system alternative to traditional forensic methods.

Seed architecture shapes embryo metabolism in oilseed rape.

Constrained to develop within the seed, the plant embryo must adapt its shape and size to fit the space available. Here, we demonstrate how this adjustment shapes metabolism of photosynthetic embryo. Noninvasive NMR-based imaging of the developing oilseed rape (Brassica napus) seed illustrates that, following embryo bending, gradients in lipid concentration became established. These were correlated with the local photosynthetic electron transport rate and the accumulation of storage products. Experimentally induced changes in embryo morphology and/or light supply altered these gradients and were accompanied by alterations in both proteome and metabolome. Tissue-specific metabolic models predicted that the outer cotyledon and hypocotyl/radicle generate the bulk of plastidic reductant/ATP via photosynthesis, while the inner cotyledon, being enclosed by the outer cotyledon, is forced to grow essentially heterotrophically. Under field-relevant high-light conditions, major contribution of the ribulose-1,5-bisphosphate carboxylase/oxygenase-bypass to seed storage metabolism is predicted for the outer cotyledon and the hypocotyl/radicle only. Differences between in vitro- versus in planta-grown embryos suggest that metabolic heterogeneity of embryo is not observable by in vitro approaches. We conclude that in vivo metabolic fluxes are locally regulated and connected to seed architecture, driving the embryo toward an efficient use of available light and space.

Cannabinoid receptor-interacting protein 1a modulates CB1 receptor signaling and regulation.

Cannabinoid CB1 receptors (CB1Rs) mediate the presynaptic effects of endocannabinoids in the central nervous system (CNS) and most behavioral effects of exogenous cannabinoids. Cannabinoid receptor-interacting protein 1a (CRIP1a) binds to the CB1R C-terminus and can attenuate constitutive CB1R-mediated inhibition of Ca(2+) channel activity. We now demonstrate cellular colocalization of CRIP1a at neuronal elements in the CNS and show that CRIP1a inhibits both constitutive and agonist-stimulated CB1R-mediated guanine nucleotide-binding regulatory protein (G-protein) activity. Stable overexpression of CRIP1a in human embryonic kidney (HEK)-293 cells stably expressing CB1Rs (CB1-HEK), or in N18TG2 cells endogenously expressing CB1Rs, decreased CB1R-mediated G-protein activation (measured by agonist-stimulated [(35)S]GTPγS (guanylyl-5'-[O-thio]-triphosphate) binding) in both cell lines and attenuated inverse agonism by rimonabant in CB1-HEK cells. Conversely, small-interfering RNA-mediated knockdown of CRIP1a in N18TG2 cells enhanced CB1R-mediated G-protein activation. These effects were not attributable to differences in CB1R expression or endocannabinoid tone because CB1R levels did not differ between cell lines varying in CRIP1a expression, and endocannabinoid levels were undetectable (CB1-HEK) or unchanged (N18TG2) by CRIP1a overexpression. In CB1-HEK cells, 4-hour pretreatment with cannabinoid agonists downregulated CB1Rs and desensitized agonist-stimulated [(35)S]GTPγS binding. CRIP1a overexpression attenuated CB1R downregulation without altering CB1R desensitization. Finally, in cultured autaptic hippocampal neurons, CRIP1a overexpression attenuated both depolarization-induced suppression of excitation and inhibition of excitatory synaptic activity induced by exogenous application of cannabinoid but not by adenosine A1 agonists. These results confirm that CRIP1a inhibits constitutive CB1R activity and demonstrate that CRIP1a can also inhibit agonist-stimulated CB1R signaling and downregulation of CB1Rs. Thus, CRIP1a appears to act as a broad negative regulator of CB1R function.

Near infrared (NIR) strong field ionization and imaging of C60 sputtered molecules: overcoming matrix effects and improving sensitivity.

Strong field ionization (SFI) was applied for the secondary neutral mass spectrometry (SNMS) of patterned rubrene films, mouse brain sections, and Botryococcus braunii algal cell colonies. Molecular ions of rubrene, cholesterol, C31 diene/triene, and three wax monoesters were detected, representing some of the largest organic molecules ever ionized intact by a laser post-ionization experiment. In rubrene, the SFI SNMS molecular ion signal was ~4 times higher than in the corresponding secondary-ion mass spectroscopy (SIMS) analysis. In the biological samples, the achieved signal improvements varied among molecules and sampling locations, with SFI SNMS, in some cases, revealing analytes made completely undetectable by the influence of matrix effects in SIMS.

Measles virus spread initiated at international mass gatherings in Europe, 2011.

Three parallel transmission chains of measles virus (MV) variant 'D8-Villupuram' (D8-V) originated from two coinciding international mass gathering (MG) events in Rimini, Italy, in June 2011. MV D8-V was independently introduced into Germany by two unvaccinated persons, and into Slovenia by one unvaccinated person who had attended these events. Secondary spread of D8-V was restricted to two generations of transmission in Slovenia as well as in Germany where the virus was further disseminated at another MG. Serological and epidemiological investigation of the D8-V-associated German and Slovenian cases revealed different antibody responses and age distributions. Primary infected young persons between 11 and 27 years-old were affected in Germany, whereas the group of Slovenian cases comprised adults aged from 28 to 47 years and a high proportion (9/14; 64%) of patients with secondary vaccine failure (SVF). Our study demonstrates that monitoring of MV transmission chains in an international context and adequate serological investigation of cases with remote vaccination can contribute to identify susceptibility gaps.

Soccer injuries in children requiring trauma center admission.

BACKGROUND: Soccer continues to gain popularity among youth athletes, and increased numbers of children playing soccer can be expected to result in increased injuries. OBJECTIVE: We reviewed children with soccer injuries severe enough to require trauma activation at our Level I trauma center to determine injury patterns and outcome. Our goal is to raise awareness of the potential for injury in youth soccer. METHODS: A retrospective review was performed using the trauma registry and electronic medical records at a Level I trauma center to identify children (< 18 years old) treated for soccer injury from 1999-2009. Data reviewed include age, gender, mechanism, injury, procedures, and outcome. RESULTS: Eighty-one children treated for soccer injury were identified; 38 (47%) were male. Of these, 20 had injury severe enough to require trauma team activation and 61 had minor injury. Mean age was 14 years old (range 5-17 years, SD 2.3). Lower extremity was the most common site of injury (57%), followed by upper extremity (17%), head (16%), and torso (10%). Mechanisms were: kicked or kneed in 27 patients (33%), collision with another player in 25 (31%), fall in 18 (22%), struck by ball in 10 (12%), and unknown in 1 (1%). Procedures included reduction of fractures, splenectomy, abdominal abscess drainage, and surgical feeding access. Long hospitalizations were recorded in some cases. There were no deaths. CONCLUSION: Although less common, injury requiring prolonged hospital admission and invasive operative procedures exist in the expanding world of youth soccer. With increasing participation in the sport, we anticipate greater numbers of these child athletes presenting with serious injury.

Uncontrolled pain: a call for better study design.

Studies assessing animal pain in veterinary research are often performed primarily for the benefit of animals. Frequently, the goal of these studies is to determine whether the analgesic effect of a novel treatment is clinically meaningful, and therefore has the capacity to improve the welfare of treated animals. To determine the treatment effect of a potential analgesic, control groups are necessary to allow comparison. There are negative control groups (where pain is unattenuated) and positive control groups (where pain is attenuated). Arising out of animal welfare concerns, there is growing reluctance to use negative control groups in pain studies. But for studies where pain is experimentally induced, the absence of a negative control group removes the opportunity to demonstrate that the study methods could differentiate a positive control intervention from doing nothing at all. For studies that are controlled by a single comparison group, the capacity to distinguish treatment effects from experimental noise is more difficult; especially considering that pain studies often involve small sample sizes, small and variable treatment effects, systematic error and use pain assessment measures that are unreliable. Due to these limitations, and with a focus on farm animals, we argue that many pain studies would be enhanced by the simultaneous inclusion of positive and negative control groups. This would help provide study-specific definitions of pain and pain attenuation, thereby permitting more reliable estimates of treatment effects. Adoption of our suggested refinements could improve animal welfare outcomes for millions of animals globally.

Exposure to Persistent Organic Pollutants in Australian Waterbirds.

There is growing worldwide recognition of the threat posed by persistent organic pollutants (POPs) to wildlife populations. We aimed to measure exposure levels to POPs in a Southern Hemisphere aquatic waterbird species, the nomadic gray teal (Anas gracilis), which is found across Australia. We collected wings from 39 ducks harvested by recreational hunters at two sites (one coastal, one inland) in Victoria, southeastern Australia, in 2021. We examined three groups of POPs: nine congeners of polychlorinated biphenyls (PCBs), 13 organochlorine pesticides (OCPs), and 12 polycyclic aromatic hydrocarbons (PAHs). The PCBs, OCPs, and PAHs were detected at quantifiable levels in 13%, 72%, and 100% of birds, respectively. Of the congeners we tested for in PCBs, OCPs, and PAHs, 33%, 38%, and 100% were detected at quantifiable levels, respectively. The highest levels of exposure to POPs that we found were to the PAH benzo[b]fluoranthene, occurring at a concentration range of 1.78 to 161.05 ng/g wet weight. There were some trends detected relating to differences between geographical sites, with higher levels of several PAHs at the coastal versus inland site. There were several strong, positive associations among PAHs found. We discuss potential sources for the POPs detected, including industrial and agricultural sources, and the likely role of large-scale forest fires in PAH levels. Our results confirm that while Australian waterbirds are exposed to a variety of POPs, exposure levels are currently relatively low. Additional future investigations are required to further characterize POPs within Australian waterbird species. Environ Toxicol Chem 2024;43:736-747. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Lead exposure and source attribution for a mammalian scavenger before and after a culling program.

Lead-based ammunition is a significant source of environmental lead and threatens species that scavenge lead-shot carcasses, particularly in areas with intensive shooting. With the impacts of lead on avian scavengers well established, there is increasing focus on the effects of lead on mammalian scavengers. We investigated lead exposure in a morphologically specialized mammalian scavenger, the Tasmanian devil (Sarcophilus harrisii), by analyzing their blood lead levels (BLLs) before and after a marsupial culling program using linear mixed effects models. We compared lead isotope signatures in devil blood to those in the culling ammunition to inform potential source attributions. We sampled 23 devils before culling and 15 after culling, finding no significant difference in mean BLLs pre and post-culling. However, devils captured closer to forestry coupes where culling had occurred had higher BLLs, and a greater proportion of devils displayed elevated BLLs post-culling (33 % compared to 18 % pre-culling). The highest BLL (7.93 µg/dL) was found in a devil post-culling and this individual had lead isotope signatures that matched the ammunition samples analyzed, suggesting the individual was exposed to lead from scavenging on culled carcasses. While 18 % of the devil blood lead samples had isotope signatures consistent with the ammunition samples, most were measurably different, indicating other sources of lead in the landscape. BLLs in our study landscape were similar to published BLLs for wild devils across Tasmania. That said, lead isotope signatures in the blood of individual devils sampled both before and after culling shifted closer to those of ammunition samples post-culling. Our results indicate that while some individual devils may have been exposed to lead from culling, most devils in the landscape did not show evidence of recent exposure. However, even low lead levels can adversely impact wildlife health and immunity, a particular concern for devils, a species endangered by disease.

Feasibility of Left Anterior Descending Coronary Artery Sparing Radiation Therapy for Locally Advanced Lung Cancer.

Efforts to mitigate radiation therapy (RT)-associated cardiotoxicity have focused on constraining mean heart dose. However, recent studies have shown greater predictive power with cardiac substructure dose metrics, such as the left anterior descending (LAD) coronary artery volume (V) receiving 15 Gy (V15Gy) ≥10%. Herein, we investigated the feasibility of LAD radiation sparing in contemporary intensity modulated RT (IMRT)/volumetric modulated arc therapy (VMAT) lung cancer plans. Single institution retrospective analysis of 54 patients with locally advanced lung cancer treated with thoracic RT was conducted between February 2018 and August 2021. After excluding 33 (5 = non-IMRT/VMAT or intentionally LAD-optimized; 28 = LAD V15Gy <10%), 21 plans with LAD V15Gy ≥10% were identified for LAD reoptimization with intent to meet LAD V15Gy <10% while maintaining meeting organ at risk (OAR) metrics and target coverage with original plan parameters. Dosimetric variables were compared using paired t tests. Most patients (57.1%, 12/21) were treated with definitive RT, 8 of 21 patients (38.1%) with postoperative RT, and 1 with neoadjuvant RT. The median prescribed RT dose was 60 Gy (range, 50.4-66 Gy) in 30 fractions (range, 28-33 fractions). LAD reoptimized plans (vs original) led to significant reductions in mean LAD V15Gy (39.4% ± 13.9% vs 9.4% ± 13.0%; P < .001) and mean LAD dose (12.9 Gy ± 4.6 Gy vs 7.6 Gy ± 2.8 Gy; P < .001). Most (85.7%; 18/21) LAD reoptimized plans achieved LAD V15Gy <10%. There were no statistically significant differences in overall lung, esophageal, or spinal cord dose metrics. Only 1 reoptimization (1/21) exceeded an OAR constraint that was initially met in the original plan. To our knowledge, this is the first report describing the feasibility of LAD-optimized lung cancer RT planning using the newly identified LAD V15Gy constraint. We observed that LAD V15Gy <10% is achievable in more than 85% of plans initially exceeding this constraint, with minimal dosimetric tradeoffs. Our results support the feasibility of routine incorporation of the LAD as an OAR in modern thoracic IMRT/VMAT planning.

Lead poisoning of raptors: state of the science and cross-discipline mitigation options for a global problem.

Lead poisoning is an important global conservation problem for many species of wildlife, especially raptors. Despite the increasing number of individual studies and regional reviews of lead poisoning of raptors, it has been over a decade since this information has been compiled into a comprehensive global review. Here, we summarize the state of knowledge of lead poisoning of raptors, we review developments in manufacturing of non-lead ammunition, the use of which can reduce the most pervasive source of lead these birds encounter, and we compile data on voluntary and regulatory mitigation options and their associated sociological context. We support our literature review with case studies of mitigation actions, largely provided by the conservation practitioners who study or manage these efforts. Our review illustrates the growing awareness and understanding of lead exposure of raptors, and it shows that the science underpinning this understanding has expanded considerably in recent years. We also show that the political and social appetite for managing lead ammunition appears to vary substantially across administrative regions, countries, and continents. Improved understanding of the drivers of this variation could support more effective mitigation of lead exposure of wildlife. This review also shows that mitigation strategies are likely to be most effective when they are outcome driven, consider behavioural theory, local cultures, and environmental conditions, effectively monitor participation, compliance, and levels of raptor exposure, and support both environmental and human health.