Advancing Carbon Fiber Composite Inspection: Deep Learning-Enabled Defect Localization and Sizing via 3-Dimensional U-Net Segmentation of Ultrasonic Data.

In Non-Destructive Evaluation (NDE), accurately characterizing defects within components relies on accurate sizing and localization to evaluate the severity or criticality of defects. This study presents for the first time a deep learning methodology using 3-Dimensional (3D) U-Net to localize and size defects in Carbon Fibre Reinforced Polymer (CFRP) composites through volumetric segmentation of ultrasonic testing data. Using a previously developed approach, synthetic training data closely representative of experimental data was used for the automatic generation of ground truth segmentation masks. The model's performance was compared to the conventional amplitude 6 dB drop analysis method used in industry against ultrasonic defect responses from 40 defects fabricated in CFRP components. The results showed good agreement with the 6 dB drop method for in-plane localization and excellent through-thickness localization, with Mean Absolute Errors (MAE) of 0.57 mm and 0.08 mm, respectively. Initial sizing results consistently oversized defects with a 55% higher mean average error than the 6 dB drop method. However, when a correction factor was applied to account for variation between the experimental and synthetic domains the final sizing accuracy resulted in a 35% reduction in MAE compared to the 6 dB drop technique. By working with volumetric ultrasonic data (as opposed to 2D images) this approach reduces pre-processing (such as signal gating) and allows for the generation of 3D defect masks which can be used for the generation of computer aided design files; greatly reducing the qualification reporting burden of NDE operators.

Unsupervised machine learning for flaw detection in automated ultrasonic testing of carbon fibre reinforced plastic composites.

The use of Carbon Fibre Reinforced Plastic (CFRP) composite materials for critical components has significantly surged within the energy and aerospace industry. With this rapid increase in deployment, reliable post-manufacturing Non-Destructive Evaluation (NDE) is critical for verifying the mechanical integrity of manufactured components. To this end, an automated Ultrasonic Testing (UT) NDE process delivered by an industrial manipulator was developed, greatly increasing the measurement speed, repeatability, and locational precision, while increasing the throughput of data generated by the selected NDE modality. Data interpretation of UT signals presents a current bottleneck, as it is still predominantly performed manually in industrial settings. To reduce the interpretation time and minimise human error, this paper presents a two-stage automated NDE evaluation pipeline consisting of a) an intelligent gating process and b) an autoencoder (AE) defect detector. Both stages are based on an unsupervised method, leveraging density-based spatial clustering of applications with noise clustering method for robust automated gating and undefective UT data for the training of the AE architecture. The AE network trained on ultrasonic B-scan data was tested for performance on a set of reference CFRP samples with embedded and manufactured defects. The developed model is rapid during inference, processing over 2000 ultrasonic B-scans in 1.26 s with the area under the receiver operating characteristic curve of 0.922 in simple and 0.879 in complex geometry samples. The benefits and shortcomings of the presented methods are discussed, and uncertainties associated with the reported results are evaluated.

Three-Dimensional Residual Neural Architecture Search for Ultrasonic Defect Detection.

This study presents a deep-learning (DL) methodology using 3-D convolutional neural networks (CNNs) to detect defects in carbon fiber-reinforced polymer (CFRP) composites through volumetric ultrasonic testing (UT) data. Acquiring large amounts of ultrasonic training data experimentally is expensive and time-consuming. To address this issue, a synthetic data generation method was extended to incorporate volumetric data. By preserving the complete volumetric data, complex preprocessing is reduced, and the model can utilize spatial and temporal information that is lost during imaging. This enables the model to utilize important features that might be overlooked otherwise. The performance of three architectures was compared. The first architecture is prevalent in the literature for the classification of volumetric datasets. The second demonstrated a hand-designed approach to architecture design, with modifications to the first architecture to address the challenges of this specific task. A key modification was the use of cuboidal kernels to account for the large aspect ratios seen in ultrasonic data. The third architecture was discovered through neural architecture search (NAS) from a modified 3-D residual neural network (ResNet) search space. In addition, domain-specific augmentation methods were incorporated during training, resulting in significant improvements in model performance, with a mean accuracy improvement of 22.4% on the discovered architecture. The discovered architecture demonstrated the best performance with a mean accuracy increase of 7.9% over the second-best model. It was able to consistently detect all defects while maintaining a model size smaller than most 2-D ResNets. Each model had an inference time of less than 0.5 s, making them efficient for the interpretation of large amounts of data.

A Newly Discovered Dural Venous Sinus of the Skull Base: The Anterior Petroclinoid Sinus.

INTRODUCTION: Neurosurgeons must master microanatomy and its variants that occur at the skull base. To the best of our knowledge, we describe a previously unreported dural venous sinus (DVS) within the anterior petroclinoid fold and discuss its potential surgical relevance. METHODS: Fifteen latex-injected human cadaveric heads (30 sides) underwent skull base dissection using a surgical microscope. The anterior petroclinoid fold was opened in search of evidence of an embedded DVS. When identified, this sinus was documented, photographed, and measured with microcalipers. RESULTS: An anterior petroclinoid sinus (APCS) was identified in 67% of sides. These DVSs had a mean diameter of 0.65 mm and a length ranging from 8 to 15 mm (mean: 12.5 mm). The APCS ran from the anterior aspect of the cavernous sinus, arising just lateral (60%), posterior (20%), or medial (20%) to the anterior clinoid process, to the superior petrosal sinus (65%) or the posterior aspect of the cavernous sinus (35%), at the level of the posterior clinoid process. Along the course of the APCS, the oculomotor nerve was located medially at the oculomotor porus. Although slightly more common and larger on the right side, there was no statistically significant side difference. CONCLUSIONS: An improved knowledge of variant DVSs can lower the risk of intraoperative complications and increase our understanding of the venous outflow from the cavernous sinus. Future neuroimaging techniques might also seek to identify the APCS. This is the first description of a DVS contained within the anterior petroclinoid fold.

A Protocol for Prolonged Surgical Anaesthesia with Recovery in Fire Salamanders Using Tricaine Mesylate (MS-222): A Case Series.

The objective of this study was to describe prolonged surgical anaesthesia and recovery in fire salamanders (Salamandra salamandra) using tricaine methanesulfonate (MS-222). A total of 14 salamanders were anaesthetised for electromyography wire implantation. Sodium bicarbonate buffered solutions (0.5-4 g l-1) of MS-222 were prepared (adjusted to pH 7.0). Anaesthesia was induced by partial immersion in pre-oxygenated 3 g l-1 solution for 20 min. Buprenorphine (0.5 mg kg-1) was administered subcutaneously. During microsurgery, heart rate (HR), solution pH and temperature were recorded. Reflectance pulse oximeter (SpO2) (Masimo Rad-57) was recorded in two salamanders. Anaesthetic plane and MS-222 pH stability (pH 7.6) were maintained by renewing administration of oxygenated MS-222 solution (0.5-3 g l-1) onto swabs that partially covered the body. Recovery started at the end of surgery (MS-222 0 g l-1). Postoperatively, salamanders were given oral meloxicam (0.2 mg kg-1). Mean time for loss of righting reflex during induction was 13.7 ± 2.2 min. Duration of anaesthesia and time to recovery were 111 ± 24.2 and 31 ± 10.3 min, respectively. Due to complications, two salamanders did not recover. Baseline HR was 67.4 ± 34.5 beats/min, and it decreased significantly until recovery (p ≤ 0.0001). In two salamanders, baseline SpO2 was 85.5% ± 14.5, SpO2 during surgery was 61% ± 6.4, improving to 80.5% ± 2.1 on recovery.In conclusion, prolonged recovery anaesthesia is achievable with MS-222 dilutions in salamander. Reflectance SpO2 could prove valuable during immersion anaesthesia.

AutoBend: An Automated Approach for Estimating Intervertebral Joint Function from Bone-Only Digital Models.

Deciphering the biological function of rare or extinct species is key to understanding evolutionary patterns across the tree of life. While soft tissues are vital determinants of joint function, they are rarely available for study. Therefore, extracting functional signals from skeletons, which are more widely available via museum collections, has become a priority for the field of comparative biomechanics. While most work has focused on the limb skeleton, the axial skeleton plays a critical role in body support, respiration, and locomotion, and is therefore of central importance for understanding broad-scale functional evolution. Here, we describe and experimentally validate AutoBend, an automated approach to estimating intervertebral joint function from bony vertebral columns. AutoBend calculates osteological range of motion (oROM) by automatically manipulating digitally articulated vertebrae while incorporating multiple constraints on motion, including both bony intersection and the role of soft tissues by restricting excessive strain in both centrum and zygapophyseal articulations. Using AutoBend and biomechanical data from cadaveric experiments on cats and tegus, we validate important modeling parameters required for oROM estimation, including the degree of zygapophyseal disarticulation, and the location of the center of rotation. Based on our validation, we apply a model with the center of rotation located within the vertebral disk, no joint translation, around 50% strain permitted in both zygapophyses and disks, and a small amount of vertebral intersection permitted. Our approach successfully reconstructs magnitudes and craniocaudal patterns of motion obtained from ex vivo experiments, supporting its potential utility. It also performs better than more typical methods that rely solely on bony intersection, emphasizing the importance of accounting for soft tissues. We estimated the sensitivity of the analyses to vertebral model construction by varying joint spacing, degree of overlap, and the impact of landmark placement. The effect of these factors was small relative to biological variation craniocaudally and between bending directions. We also present a new approach for estimating joint stiffness directly from oROM and morphometric measurements that can successfully reconstruct the craniocaudal patterns, but not magnitudes, derived from experimental data. Together, this work represents a significant step forward for understanding vertebral function in difficult-to-study (e.g., rare or extinct) species, paving the way for a broader understanding of patterns of functional evolution in the axial skeleton.

Resumo [Portuguese] Decifrar a função biológica de espécies raras ou extintas é fundamental para se compreender os padrões evolutivos na árvore da vida. Embora os tecidos moles sejam determinantes vitais das funções articulares, estes raramente estão disponíveis para estudo. Portanto, extrair dados funcionais provenientes de esqueletos, que são mais amplamente disponíveis por meio de coleções de museus, tornou-se uma prioridade para o campo da biomecânica comparada. Embora a maioria dos trabalhos biomecânicos tenham focado no esqueleto apendicular, o esqueleto axial também desempenha um papel crítico para o suporte corporal, respiração e locomoção e, portanto, é de importância central para a compreensão da evolução funcional em escalas amplas. Nesse trabalho, nós descrevemos e validamos experimentalmente o AutoBend, uma abordagem automatizada para estimar a função da articulação intervertebral de colunas vertebrais ósseas. O AutoBend calcula a amplitude do movimento osteológico (AMO) manipulando automaticamente as vértebras reconstruídas digitalmente e incorporando várias restrições de movimento, incluindo restrições das interseções ósseas e o papel de restrição dos tecidos moles na tensão excessiva sobre as articulações dos centros vertebrais e zigapofisárias. Usando AutoBend e dados biomecânicos de experimentos cadavéricos em gatos e lagartos tegus, validamos parâmetros de modelagem importantes e necessários para as estimativa do AMO, incluindo o grau de desarticulação zigapofisária e a localização do centro de rotação. Com base nessa validação, aplicamos um modelo com o centro de rotação localizado dentro do disco vertebral, sem translação articular, com cerca de 50% de tensão permitida nas zigapófises e discos vertebrais, além de uma pequena quantidade de intersecção vertebral permitida. Nossa abordagem reconstrói com sucesso magnitudes e padrões de movimento craniocaudais obtidos a partir de experimentos ex vivo, corroborando a sua potencial utilidade. Esse modelo também tem um desempenho melhor do que os métodos mais típicos que dependem apenas das interseções ósseas, enfatizando a importância de se levar em conta o papel dos tecidos moles. Estimamos a sensibilidade das análises à reconstrução do modelo vertebral, variando o espaçamento entre articulações, o grau de sobreposição e o impacto da localização dos pontos de referência. O efeito desses fatores foi pequeno em relação à variação biológica craniocaudal e entre as direções de flexão. Apresentamos aqui também uma nova abordagem para se estimar a rigidez articular diretamente à partir do AMO e medidas morfométricas que podem reconstruir com sucesso os padrões craniocaudais (embora não as magnitudes) derivados de dados experimentais. Este trabalho representa um passo significativo para a melhor compreensão da função vertebral em espécies difíceis de estudar (por exemplo, raras ou extintas), abrindo caminho para uma compreensão mais ampla dos padrões de evolução funcional no esqueleto axial.

Resumen [Spanish] Descifrar la función biológica de especies raras o extintas es fundamental para comprender los patrones evolutivos del árbol de la vida. Aunque los tejidos blandos son determinantes vitales de la función articular, raramente están disponibles para su estudio. Por lo tanto, la extracción de datos funcionales de esqueletos, que están más comumente disponibles a través de colecciones de museos, se ha convertido en una prioridad para el campo de la biomecánica comparada. Aunque la mayor parte del trabajo biomecánico se ha centrado en el esqueleto apendicular, el esqueleto axial también desempeña un papel fundamental para el soporte del cuerpo, la respiración y la locomoción y, por lo tanto, es de vital importancia para comprender la evolución funcional a gran escala. En este trabajo, describimos y validamos experimentalmente AutoBend, una herramienta automatizada para estimar la función de la articulación intervertebral en las columnas vertebrales óseas. AutoBend calcula el rango de movimiento osteológico (RMOo) manipulando automáticamente las vértebras reconstruidas digitalmente e incorporando varias restricciones de movimiento, incluidas las restricciones de intersección ósea y el papel de la restricción de tejidos blandos en la tensión excesiva sobre las articulaciones cigapofisarias y centros vertebrales. Utilizando AutoBend y datos biomecánicos de experimentos cadavéricos en gatos y lagartos tegus, validamos importantes parámetros de modelado necesarios para estimar el RMOo, incluido el grado de desarticulación cigapofisaria y la ubicación del centro de rotación. Con base en esta validación, aplicamos un modelo con el centro de rotación ubicado dentro del disco vertebral, sin traslación articular, con 50% de tensión permisible en la cigapófisis y los discos vertebrales, además de una pequeña cantidad de intersección vertebral permitida. Nuestra herramienta reconstruye con éxito magnitudes craneocaudales y patrones de movimiento obtenidos de experimentos ex vivo, corroborando su potencial utilidad. Este modelo también funciona mejor que los métodos más típicos que se basan solo en las intersecciones óseas, enfatizando la importancia de tener en cuenta el papel de los tejidos blandos. Estimamos la sensibilidad de los análisis a la reconstrucción del modelo vertebral, variando el espaciamiento entre articulaciones, el grado de superposición y el impacto de la ubicación de los puntos de referencia. El efecto de estos factores fue pequeño en relación a la variación biológica craneocaudal y entre las direcciones de flexión. También presentamos aquí un nuevo enfoque para estimar la rigidez articular directamente de la RMOo y mediciones morfométricas que pueden reconstruir con éxito los patrones craneocaudales (aunque no las magnitudes) derivados de datos experimentales. Este trabajo representa un paso significativo hacia una mejor comprensión de la función vertebral en especies difíciles de estudiar (por ejemplo, raras o extintas), allanando el camino para una comprensión más amplia de los patrones de evolución funcional en el esqueleto axial.

The Digestive Diverticula in the Carnivorous Nudibranch, Melibe leonina, Do Not Contain Photosynthetic Symbionts.

A number of nudibranchs, including Melibe engeli and Melibe pilosa, harbor symbiotic photosynthetic zooxanthellae. Melibe leonina spends most of its adult life on seagrass or kelp, capturing planktonic organisms in the water column with a large, tentacle-lined oral hood that brings food to its mouth. M. leonina also has an extensive network of digestive diverticula, located just beneath its translucent integument, that are typically filled with pigmented material likely derived from ingested food. Therefore, the focus of this project was to test the hypothesis that M. leonina accumulates symbiotic photosynthetic dinoflagellates in these diverticula. First, we conducted experiments to determine if M. leonina exhibits a preference for light, which would allow chloroplasts that it might be harboring to carry out photosynthesis. We found that most M. leonina preferred shaded areas and spent less time in direct sunlight. Second, we examined the small green circular structures in cells lining the digestive diverticula. Like chlorophyll, they exhibited autofluorescence when illuminated at 480 nm, and they were also about the same size as chloroplasts and symbiotic zooxanthellae. However, subsequent electron microscopy found no evidence of chloroplasts in the digestive diverticula of M. leonina; the structures exhibiting autofluorescence at 480 nm were most likely heterolysosomes, consistent with normal molluscan digestion. Third, we did not find evidence of altered oxygen consumption or production in M. leonina housed in different light conditions, suggesting the lack of any significant photosynthetic activity in sunlight. Fourth, we examined the contents of the diverticula, using HPLC, thin layer chromatography, and spectroscopy. The results of these studies indicate that the diverticula did not contain any chlorophyll, but rather harbored other pigments, such as astaxanthin, which likely came from crustaceans in their diet. Together, all of these data suggest that M. leonina does sequester pigments from its diet, but not for the purpose of symbiosis with photosynthetic zooxanthellae. Considering the translucent skin of M. leonina, the pigmented diverticula may instead provide camouflage.

Varios nudibranquios, incluidos Melibe engeli y Melibe pilosa, albergan zooxantelas fotosintéticas simbióticas. Melibe leonina pasa la mayoría de su vida adulta en pastos marinos o quelpo, donde captura organismos planctónicos en la columna de agua con una gran capucha oral forrada por tentáculos que llevan comida a su boca. Melibe leonina también tiene una extensa red de divertículos digestivos, ubicados justo debajo de su tegumento translúcido, que generalmente están llenos de material pigmentado probablemente derivado de alimentos ingeridos. Por lo tanto, el objetivo de este proyecto fue evaluar la hipótesis de que M. leonina acumula dinoflagelados fotosintéticos simbióticos en estos divertículos. Primero, realizamos experimentos para determinar si M. leonina se orienta hacia la luz, lo cual permitiría a los cloroplastos que podría albergar el realizar la fotosíntesis. Descubrimos que la mayoría de M. leonina prefería las áreas sombreadas y pasaba menos tiempo bajo la luz solar directa. En segundo lugar, examinamos las pequeñas estructuras circulares verdes en las células que recubren los divertículos digestivos. Al igual que la clorofila, exhibieron autofluorescencia cuando se iluminaban a 480 nm, y también tenían aproximadamente el mismo tamaño que los cloroplastos y las zooxantelas simbióticas. No obstante, la microscopía electrónica no produjo evidencia de cloroplastos en los divertículos digestivos de M. leonina. Es probable que las estructuras que exhibieron autofluorescencia en 480 nm fuesen heterolisosomas, lo cual es consistente con la digestión normal de moluscos. En tercer lugar, no encontramos evidencia de un consumo o producción de oxígeno alterado en M. leonina alojadas varias condiciones lumínicas, lo cual sugiere la ausencia de actividad fotosintética significativa en la presencia de luz solar. En cuarto lugar, examinamos el contenido de los divertículos mediante HPLC, cromatografía en capa fina, y espectroscopia. Los resultados de estos estudios indican que los divertículos no contenían clorofila, pero si otros pigmentos como la astaxantina que probablemente provenía de crustáceos en su dieta. Nuestros datos sugieren que M. leonina secuestra pigmentos de su dieta, pero no con el propósito de la simbiosis con zooxantelas fotosintéticas. Teniendo en cuenta la piel translúcida de M. leonina, los divertículos pigmentados podrían quizás proporcionar camuflaje.

Evolution of forelimb musculoskeletal function across the fish-to-tetrapod transition.

One of the most intriguing questions in vertebrate evolution is how tetrapods gained the ability to walk on land. Although many hypotheses have been proposed, few have been rigorously tested using the fossil record. Here, we build three-dimensional musculoskeletal models of the pectoral appendage in Eusthenopteron, Acanthostega, and Pederpes and quantitatively examine changes in forelimb function across the fin-to-limb transition. Through comparison with extant fishes and tetrapods, we show that early tetrapods share a suite of characters including restricted mobility in humerus long-axis rotation, increased muscular leverage for humeral retraction, but not depression/adduction, and increased mobility in elbow flexion-extension. We infer that the earliest steps in tetrapod forelimb evolution were related to limb-substrate interactions, whereas specializations for weight support appeared later. Together, these results suggest that competing selective pressures for aquatic and terrestrial environments produced a unique, ancestral "early tetrapod" forelimb locomotor mode unlike that of any extant animal.

Machine learning at the interface of structural health monitoring and non-destructive evaluation.

While both non-destructive evaluation (NDE) and structural health monitoring (SHM) share the objective of damage detection and identification in structures, they are distinct in many respects. This paper will discuss the differences and commonalities and consider ultrasonic/guided-wave inspection as a technology at the interface of the two methodologies. It will discuss how data-based/machine learning analysis provides a powerful approach to ultrasonic NDE/SHM in terms of the available algorithms, and more generally, how different techniques can accommodate the very substantial quantities of data that are provided by modern monitoring campaigns. Several machine learning methods will be illustrated using case studies of composite structure monitoring and will consider the challenges of high-dimensional feature data available from sensing technologies like autonomous robotic ultrasonic inspection. This article is part of the theme issue 'Advanced electromagnetic non-destructive evaluation and smart monitoring'.

Patterns of Limb and Epaxial Muscle Activity During Walking in the Fire Salamander, Salamandra salamandra.

Salamanders and newts (urodeles) are often used as a model system to elucidate the evolution of tetrapod locomotion. Studies range from detailed descriptions of musculoskeletal anatomy and segment kinematics, to bone loading mechanics and inferring central pattern generators. A further area of interest has been in vivo muscle activity patterns, measured through electromyography (EMG). However, most prior EMG work has primarily focused on muscles of the forelimb or hindlimb in specific species or the axial system in others. Here we present data on forelimb, hindlimb, and epaxial muscle activity patterns in one species, Salamandra salamandra, during steady state walking. The data are calibrated to limb stride cycle events (stance phase, swing phase), allowing direct comparisons to homologous muscle activation patterns recorded for other walking tetrapods (e.g., lizards, alligators, turtles, mammals). Results demonstrate that Salamandra has similar walking kinematics and muscle activity patterns to other urodele species, but that interspecies variation does exist. In the forelimb, both the m. dorsalis scapulae and m. latissimus dorsi are active for 80% of the forelimb swing phase, while the m. anconaeus humeralis lateralis is active at the swing-stance phase transition and continues through 86% of the stance phase. In the hindlimb, both the m. puboischiofemoralis internus and m. extensor iliotibialis anterior are active for 30% of the hindlimb swing phase, while the m. caudofemoralis is active 65% through the swing phase and remains active for most of the stance phase. With respect to the axial system, both the anterior and posterior m. dorsalis trunci display two activation bursts, a pattern consistent with stabilization and rotation of the pectoral and pelvic girdles. In support of previous assertions, comparison of Salamandra muscle activity timings to other walking tetrapods revealed broad-scale similarities, potentially indicating conservation of some aspects of neuromuscular function across tetrapods. Our data provide the foundation for building and testing dynamic simulations of fire salamander locomotor biomechanics to better understand musculoskeletal function. They could also be applied to future musculoskeletal simulations of extinct species to explore the evolution of tetrapod locomotion across deep-time.

Padrones de actividad muscular epaxial y apendicular durante la cursorialidad de la salamandra-de-fuego, Salamandra salamandra Las salamandras y los tritones (urodelos) son utilizados con frecuencia como un sistema modelo para dilucidar la evolución de la locomoción en los tetrápodos. Los estudios previos varían de descripciones detalladas de la anatomía musculoesquelética y cinemática de los segmentos del cuerpo, a la mecánica de la capacidad de soporte de carga estructural ósea y la generación de padrones centrales. Otra área de interés ha sido los padrones de actividad muscular in vivo, medidos por electromiografía (EMG). Sin embargo, la mayoría de los trabajos anteriores con EMG se han centrado principalmente en los músculos de los miembros anteriores o posteriores en especies específicas o en el sistema axial de otras. En este trabajo, presentamos datos sobre los padrones de actividad muscular en los músculos de los miembros anteriores, posteriores y de la musculatura epaxial en una especie, Salamandra salamandra, durante la marcha continua. Los datos se calibran para los períodos del ciclo de caminar de los miembros (fase de soporte, fase de movimiento), lo que permite comparaciones directas con padrones de activación muscular homólogos registrados para otros tetrápodos cursoriales (por ejemplo, lagartos, caimanes, tortugas y mamíferos). Los resultados demuestran que Salamandra tiene padrones de cinemática cursorial y actividad muscular similares a otras especies de urodelos, pero con variación interespecífica. En los miembros anteriores, ambos los m. dorsalis scapulae y m. latissimus dorsi están activos en 80% de la fase de movimiento del miembro anterior, mientras que el m. anconaeus humeralis lateralis se activa en la transición de la fase de movimiento-soporte y permanece activo en 86% de la fase de soporte. En los miembros posteriores, ambos m. puboischiofemoralis internus y m. extensor iliotibialis anterior están activos en 30% de la fase de movimiento de los miembros posteriores, mientras que el m. caudofemoralis está activo durante el 65% de la fase de movimiento, permaneciendo activo durante la mayor parte de la fase de soporte. Con respecto al sistema axial, las porciones anterior y posterior del m. dorsalis trunci exhibe dos períodos de activación, un padrón consistente con la estabilización y rotación de la cintura pélvica y pectoral. Como sugirido anteriormente, la comparación de los tiempos de actividad muscular de Salamandra con otros tetrápodos cursoriales reveló similitudes en gran escala, lo que podría indicar la conservación de algunos aspectos de la función neuromuscular entre los tetrápodos. Nuestros datos proporcionan una base para la construcción y prueba de simulaciones dinámicas de la biomecánica locomotora de salamandras-de-fuego para comprender mejor las funciones musculoesqueléticas. Nuestros resultados también se pueden aplicar a futuras simulaciones musculoesqueléticas de especies extintas para explorar la evolución de la locomoción de tetrápodos en el tiempo profundo.

Padrões de atividade muscular epaxial e apendicular durante a cursorialidade da salamandra-de-fogo, Salamandra salamandra Salamandras e tritões (urodelos) são freqüentemente utilizados como um sistema modelo para elucidar a evolução da locomoção em tetrápodes. Estudos anteriores variam de descrições detalhadas da anatomia musculoesquelética e cinemática dos segmentos corporais, a mecânica da capacidade de carga estrutural óssea e geradora de padrões centrais. Uma outra área de interesse tem sido os padrões de atividade muscular in vivo, medidos por eletromiografia (EMG). No entanto, a maioria dos trabalhos anteriores de EMG concentrou-se principalmente nos músculos dos membros anteriores ou posteriores em espécies específicas ou no sistema axial de outras. Nesse trabalho, apresentamos dados sobre os padrões de atividade muscular nos membros anteriores, posteriores e musculatura epaxial em uma espécie, Salamandra salamandra, durante caminhada em modo contínuo. Os dados são calibrados para os períodos do ciclo de caminhada dos membros (fase de apoio, fase de movimento), permitindo comparações diretas com padrões de ativação muscular homólogos registrados para outros tetrápodes cursoriais (por exemplo, lagartos, jacarés, tartarugas e mamíferos). Os resultados demonstram que Salamandra possui padrões de cinemática cursorial e atividade muscular semelhantes à outras espécies de urodelos, mas com variação interespecífica. Nos membros anteriores, ambos os m. dorsalis scapulae e m. latissimus dorsi estão ativos em 80% da fase de movimento do membro anterior, enquanto o m. anconaeus humeralis lateralis é ativado na transição da fase de movimento-apoio e continua ativo em 86% da fase de apoio. Nos membros posteriores, ambos m. puboischiofemoralis internus e m. extensor iliotibialis anterior estão ativos em 30% da fase de movimento dos membros posteriores, enquanto o m. caudofemoralis está ativo por 65% da fase de movimento, permanecendo ativo na maior parte da fase de apoio. No que diz respeito ao sistema axial, as porções anterior e posterior do m. dorsalis trunci exibe dois períodos de ativação, um padrão consistente com a estabilização e rotação da cintura peitoral e pélvica. Como préviamente sugerido, a comparação dos tempos de atividade muscular de Salamandra com outros tetrápodes cursoriais revelou similaridades em larga escala, potencialmente indicando a conservação de alguns aspectos da função neuromuscular entre tetrápodes. Os nossos dados fornecem uma base para a construção e testagem de simulações dinâmicas da biomecânica locomotora de salamandras-de-fogo para se entender melhor as funções músculo-esqueléticas. Nossos resultados também podem ser aplicados a futuras simulações músculo-esqueléticas de espécies extintas para explorar a evolução da locomoção de tetrápodes no tempo profundo.

Cohort-level disease prediction by extrapolation of individual-level predictions in transition dairy cattle.

Dairy cattle experience metabolic stress during the transition from late gestation to early lactation resulting in higher risk for several economically important diseases (e.g. mastitis, metritis, and ketosis). Metabolic stress is described as a physiological state composed of 3 processes: nutrient metabolism, oxidative stress, and inflammation. Current strategies for monitoring transition cow nutrient metabolism include assessment of plasma non-esterified fatty acids and beta-hydroxybutyrate concentrations around the time of calving. Although this method is effective at identifying cows with higher disease risk, there is often not enough time to implement intervention strategies to prevent health disorders from occurring around the time of calving. Previously, we published predictive models for early lactation diseases at the individual cow level at dry-off. However, it is unknown if predictive probabilities from individual-level models can be aggregated to the cohort level to predict cohort-level incidence. Therefore, our objective was to test different data aggregation methods using previously published models that represented the 3 components of metabolic stress (nutrient metabolism, oxidative stress, and inflammation). We included 277 cows from five Michigan dairy herds for this prospective cohort study. On each farm, two to four calving cohorts were formed, totaling 18 cohorts. We measured biomarker data at dry-off and followed the cows until 30 days post-parturition for cohort disease incidence, which was defined as the number of cows: 1) having one or more clinical transition disease outcome, and/or 2) having an adverse health event (abortion or death of calf or cow) within each cohort. We tested three different aggregation methods that we refer to as the p-central, p-dispersion, and p-count methods. For the p-central method, we calculated the averaged predicted probability within each cohort. For the p-dispersion method, we calculated the standard deviation of the predicted probabilities within a cohort. For the p-count method, we counted the number of cows above a specified threshold of predicted probability within each cohort. We built four sets of models: one for each aggregation method and one that included all three aggregation methods (p-combined method). We found that the p-dispersion method was the only method that produced viable predictive models. However, these models tended to overestimate incidence in cohorts with low observed counts and underestimate risk in cohorts with high observed counts.

Cohort-level disease prediction using aggregate biomarker data measured at dry-off in transition dairy cattle: A proof-of-concept study.

During the transition from late gestation to early lactation, dairy cattle are at increased risk for disease. Herd-level monitoring for disease risk involves evaluating multiple factors, including food intake, cow density, and biomarkers of nutrient metabolism. Biomarkers that are measured include non-esterified fatty acids (NEFA) and beta-hydroxybutyrate (BHB), which are usually measured in a subset of the herd (i.e. cohort). If a certain proportion of cows in the cohort are above a specific threshold for a biomarker, the cohort is considered at high risk of disease. Few previous studies have investigated other methods to aggregate individual cow-level data to the cohort level. We designed a proof-of-concept study to determine if biomarker aggregation methods may be useful to predict cohort incidence of adverse health events including 1) clinical diseases: mastitis, metritis, retained placenta, ketosis, lameness, pneumonia, milk fever, displaced abomasum, 2) and abortion or death of the calf or the cow. The study design was a prospective cohort study that used cows (N = 277) from five Michigan commercial dairy herds. Multiple cohorts of cows (two to four cohorts per farm, 18 total) were enrolled that shared the same dry-off date. We tested three different methods (central, dispersion, and count) to aggregate individual cow data (i.e. biomarkers and covariates) measured at dry-off. The central method consisted of calculating the average value of each variable within a cohort, and the dispersion method involved taking the standard deviation or mean absolute deviation about the median of each variable within a cohort. The count method consisting of counting the number of cows above a specific threshold for each variable within a cohort. We used best subsets selection to select a bouquet of candidate models for each aggregation method and averaged the predictions over the model set. We built 4 sets of Poisson regression models: one for each aggregation method and a combined model that included all three methods. We evaluated the models based on goodness-of-fit, model calibration using scoring rules, and comparison of observed versus predicted counts. The central and the combined method produced models that had good fit and model calibration. These results indicate that it may be possible to use aggregate measures to predict cohort disease incidence as early as dry-off. The next step is to test biomarker aggregation methods in studies with larger sample sizes.

Increasing the germination percentage of a declining native orchid (Himantoglossum adriaticum) by pollen transfer and outbreeding between populations.

The declining native orchid Himantoglossum adriaticum H. Baumann is a European endemic of priority interest (92/43/ EEC, Annex II). Northern Italian populations of H. adriaticum are small and isolated, with depressed seed set. Given the important implications for plant population conservation, we tested the hypothesis that artificial pollen transfer (hand-pollination) and outbreeding between populations increases fruit set and seed germination percentage. The background fruit set and in vitro germination rates were determined for ten reference populations. An artificial cross-pollination experiment included (a) pollen transfer from one large population to two small and isolated populations; (b) pollen transfer between two small but not isolated populations; (c) within-population pollen transfer (control). All seeds were sown on a modified Malmgren's medium and cultured in a controlled environment. Germination percentage was compared using a Kruskal-Wallis anova. The background fruit set (mean = 18%) and germination (<5%) rates were consistently low across populations. Fruit set after hand-pollination was consistently 100%. Pollen transfer from the largest population to smaller populations resulted in an increase in total germination ranging from 0.9% to 2.9%. The largest increase in germination occurred between small-sized and less isolated populations (from 1.7% to 5.1%). The results of pollen transfer between the small populations are particularly encouraging, as the mean increase in germination was almost four times that of the control. Outbreeding can be considered a valuable tool to increase genetic flow and germination in natural populations, limit the accumulation of detrimental effects on fitness driven by repeated breeding with closely-related individuals, thereby increasing the possibility of conservation of rare or endangered species.

Predictive models for early lactation diseases in transition dairy cattle at dry-off.

During the transition period, dairy cattle undergo tremendous metabolic and physiological changes to prepare for milk synthesis and secretion. Failure to sufficiently regulate these changes may lead to metabolic stress, which increases risk of transition diseases. Metabolic stress is defined as a physiological state consisting of 3 components: aberrant nutrient metabolism, oxidative stress, and inflammation. Current monitoring methods to detect cows experiencing metabolic stress involve measuring biomarkers for nutrient metabolism. However, these biomarkers, including non-esterified fatty acids, beta-hydroxybutyrate, and calcium are typically measured a few weeks before to a few days after calving. This is a retroactive approach, because there is little time to integrate interventions that remediate metabolic stress in the current cohort. Our objective was to determine if biomarkers of metabolic stress measured at dry-off are predictive of transition disease risk. We designed a prospective cohort study carried out on 5 Michigan dairy farms (N = 277 cows). We followed cows from dry-off to 30 days post-calving. Diseases and adverse outcomes were grouped in an aggregate outcome that included mastitis, metritis, retained placenta, ketosis, lameness, pneumonia, milk fever, displaced abomasum, abortion, and death of the calf or the cow. We used best subsets selection to select candidate models for four different sets of models: one set for each component of metabolic stress (nutrient metabolism, oxidative stress, and inflammation), and a combined model that included all 3 components. We used model averaging to obtain averaged predicted probabilities across each model set. We hypothesized that the averaged predictions from the combined model set with all 3 components of metabolic stress would be more effective at predicting disease than each individual component model set. The area under the curve estimated using receiver operator characteristic curves for the combined model set (0.93; 95% confidence interval [CI] = 0.90-0.96) was significantly higher compared with averaged predictions from the inflammation (0.87; 95% CI = 0.83-0.91), oxidative stress (0.78; 95% CI = 0.72-0.84), and nutrient metabolism (0.73; 95% CI = 0.67-0.79) model sets (p < 0.05). Our results indicate that it may be possible to detect cattle at risk for some transition diseases as early as dry-off. This has important implications for disease prevention, as earlier identification of cows at risk of health disorders will allow for earlier implementation of intervention strategies. A limitation of the current study is that we did not perform external validation. Future validation studies are needed to confirm our findings.

Enzymatic scarification of Anacamptis morio (Orchidaceae) seed facilitates lignin degradation, water uptake and germination.

The seed coat of many species contains hydrophobic lignins, and in soil the action of microbial ligninases may contribute to release from dormancy. Laboratory use of ligninases to stimulate germination is promising because of the specific action on the seed coat, whereas chemical scarification agents may also corrode the embryo. We hypothesised that exposure of Anacamptis morio (Orchidaceae) seeds to fungal laccase would stimulate germination, and that the mechanism involves lignin degradation and increased imbibition. Germination capacity in vitro was quantified with 1 U filter-sterilised laccase added to agar medium following autoclaving, compared to a 10% bleach solution (standard bleach surface sterilisation/scarification method used in orchid seed sowing). Lignin degradation was quantified using an optical method (phloroglucinol-HCl staining) combined with image analysis, following experimental pre-treatments involving immersion in laccase solution, distilled water (negative control) or bleach (positive control). Water uptake after experimental treatments was quantified as the proportion of seeds exhibiting visible uptake of an aqueous fluorochrome under UV excitation. Laccase stimulated a doubling of germination in vitro with respect to bleach surface sterilisation/scarification alone, from 23.7 to 49.8% (P = 0.007). Laccase and bleach methods both significantly decreased the optical signal of phloroglucinol (for laccase, to 79.9 ± 1.3% of controls; anova: F = 10.333, P = 0.002). Laccase resulted in a modest but highly significant (P < 0.0001) increase in water uptake with respect to the control (11.7%; cf 99.4% for bleach). Laccase scarification can stimulate germination of A. morio through a mechanism of targeted seed coat degradation. The results demonstrate the potential of this relatively non-invasive enzymatic scarification technique.

Fossils reveal the complex evolutionary history of the mammalian regionalized spine.

A unique characteristic of mammals is a vertebral column with anatomically distinct regions, but when and how this trait evolved remains unknown. We reconstructed vertebral regions and their morphological disparity in the extinct forerunners of mammals, the nonmammalian synapsids, to elucidate the evolution of mammalian axial differentiation. Mapping patterns of regionalization and disparity (heterogeneity) across amniotes reveals that both traits increased during synapsid evolution. However, the onset of regionalization predates increased heterogeneity. On the basis of inferred homology patterns, we propose a "pectoral-first" hypothesis for region acquisition, whereby evolutionary shifts in forelimb function in nonmammalian therapsids drove increasing vertebral modularity prior to differentiation of the vertebral column for specialized functions in mammals.

Characteristics, outcomes, prognostic factors and treatment of patients with T-cell prolymphocytic leukemia (T-PLL).

BACKGROUND: T-cell prolymphocytic leukemia (T-PLL) is a rare and aggressive disease. In this study, we report our experience from 119 patients with T-PLL. PATIENTS AND METHODS: We reviewed the clinico-pathologic records of 119 consecutive patients with T-PLL, who presented to our institution between 1990 and 2016. RESULTS: One hundred and nineteen patients with T-PLL were analysed. Complex karyotype and aberrations in chromosome 14 were seen in 65% and 52% patients, respectively. Seventy-five patients (63%) were previously untreated and 43 (37%) were initially treated outside our institution. Sixty-three previously untreated patients (84%) received frontline therapies. Overall, 95 patients (80%) have died. Median overall survival (OS) from diagnosis was 19 months [95% confidence interval (CI) 16-26 months]. Using recursive partitioning (RP), we found that patients with hemoglobin < 9.3 g/dl, lactate dehydrogenase (LDH) ≥ 1668 IU/l, white blood cell ≥ 208 K/l and ß2M ≥ 8 mg/l had significantly inferior OS and patients with hemoglobin < 9.3 g/dl had inferior progression-free survival (PFS). In multivariate analysis, we identified that presence of pleural effusion [hazard ratio (HR) 2.08 (95% CI 1.11-3.9); P = 0.02], high LDH (≥ 1668 IU/l) [HR 2.5 (95% CI 1.20-4.24); P < 0.001)], and low hemoglobin (< 9.3 g/dl) [HR 0.33 (95% CI 0.14-0.75); P = 0.008] were associated with shorter OS. Fifty-five previously untreated patients received treatment with an alemtuzumab-based regimen (42 monotherapy and 13 combination with pentostatin). Overall response rate, complete remission rate (CR) for single-agent alemtuzumab and alemtuzumab combined with pentostatin were 83%, 66% and 82%, 73% respectively. In patients who achieved initial CR, stem cell transplantation was not associated with longer PFS and OS. CONCLUSION: Outcomes in T-PLL remain poor. Multicenter collaborative effort is required to conduct prospective studies.

Clinical Monitoring of Tooth Wear Progression in Patients over a Period of One Year Using CAD/CAM.

PURPOSE: The aim of this study was to clinically monitor the progression of tooth wear over a period of 1 year in a cohort of referred tooth wear patients through the use of a computer-aided design/computer-assisted manufacture (CAD/CAM) scanner and a standardized scanning/assessment methodology. MATERIALS AND METHODS: Polyether impressions were made of 11 participants (130 teeth) at baseline and at 1 year. Impressions were poured in type IV dental stone and the anterior teeth were 3D scanned. A surface-matching software was used to compare 1-year and baseline scans and identify any dimensional differences. RESULTS: Parafunctional habits were reported by all patients. All participants exhibited tooth wear ≥ 140 µm in depth and extending to ≥ 280 µm in at least one tooth. Maxillary central incisors were the most commonly and severely affected teeth. CONCLUSION: The ability of the developed CAD/CAM scanning methodology in clinical monitoring of tooth wear was demonstrated. Further research is needed to assess its practicality in large-scale epidemiologic tooth wear studies.