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Introduction: Facial asymmetry, present in all human faces at varying degrees, plays a critical role in clinical fields such as orthodontics, orthognathic and plastic surgeries, and craniofacial reconstruction. Accurate quantification of facial asymmetry is essential for diagnosis, treatment planning, and post-surgical evaluation., Material and Methods: This article examines contemporary methods for quantifying facial asymmetry, including two-dimensional (2D) and three-dimensional (3D) landmark-based approaches, surface curvature analysis, and advanced image-based techniques., Discussion: Emphasis is placed on the advantages, limitations, and applications of each method, to meet clinical or research needs., Conclusion: By integrating technological advancements, future developments are expected to enhance precision and applicability in clinical practice.

The craniocervical junction (CCJ) forms the bridge between the skull and the spine, a highly mobile group of joints that allows the mobility of the head in every direction. The CCJ plays a major role in protecting the inferior brainstem (bulb) and spinal cord, therefore also requiring some stability. Children are subjected to multiple constitutive or acquired diseases involving the CCJ: primary bone diseases such as in FGFR-related craniosynostoses or acquired conditions such as congenital torticollis, cervical spine luxation, and neurological disorders. To design efficient treatment plans, it is crucial to understand the relationship between abnormalities of the craniofacial region and abnormalities of the CCJ. This can be approached by the study of control and abnormal growth patterns. Here we report a model of normal skull base growth by compiling a collection of geometric models in control children. Focused analyses highlighted specific developmental patterns for each CCJ bone, emphasizing rapid growth during infancy, followed by varying rates of growth and maturation during childhood and adolescence until reaching stability by 18 years of age. The focus was on the closure patterns of synchondroses and sutures in the occipital bone, revealing distinct closure trajectories for the anterior intra-occipital synchondroses and the occipitomastoid suture. The findings, although based on a limited dataset, showcased specific age-related changes in width and closure percentages, providing valuable insights into growth dynamics within the first 2 years of life. Integration analyses revealed intricate relationships between skull and neck structures, emphasizing coordinated growth at different stages. Specific bone covariation patterns, as found between the first and second cervical vertebrae (C1 and C2), indicated synchronized morphological changes. Our results provide initial data for designing inclusive CCJ geometric models to predict normal and abnormal growth dynamics., (© 2024 The Author(s). Journal of Anatomy published by John Wiley & Sons Ltd on behalf of Anatomical Society.)

Background: Preoperative severity of unicoronal synostosis varies greatly and involves the frontal bone, skull base and orbits. Degree of deformity affects long-term morphological and functional outcomes after surgery. The aim of this study was to describe the morphological heterogenicity and investigate its relation to patient-specific factors., Materials and Methods: In this retrospective cohort study, non-syndromic unicoronal synostosis patients treated between 2006 and 2022 at Necker Hospital, France or Uppsala University Hospital, Sweden, were included and matched to controls. Severity of skull base, orbital and posterior skull asymmetry, degree of anterior plagiocephaly and Harlequin deformity, lateralisation, head circumference, age, timing of metopic fusion and fusion of peri-pterionic sutures were investigated., Results: Ninety-five patients and ninety-three controls were included. Skull base asymmetry was linearly related to orbital asymmetry (p < 0.001), correlated with earlier CT scans (p = 0.004) and anterior (p < 0.001) and posterior (p = 0.03) plagiocephaly. Posterior plagiocephaly was more common in patients (31%) compared with controls (5%) (p < 0.001). A patent metopic suture above nine months of age was associated with severe Harlequin deformity (p = 0.04) and a lower head circumference when fused (p = 0.03). Fronto-sphenoidal suture fusion was associated with later CT scans (p < 0.001) and less skull base asymmetry (p = 0.002). Spheno-parietal fusion was correlated with decreased skull base asymmetry (p = 0.03). Right lateralisation was more common in females., Conclusions: Heterogenicity of unicoronal synostosis seems to be predominantly explained by variability in skull base morphology. Peri-pterionic fusions might limit deformity., (Copyright © 2024 British Association of Plastic, Reconstructive and Aesthetic Surgeons. Published by Elsevier Ltd. All rights reserved.)

Craniofacial development involves processes leading to the positioning and early growth of the frontal ossification centers. Growth then occurs, mostly secondary to the activity of the sutures, with major interactions with the functional environment, mostly consisting in the growing brain, based on mechanosensation and mechanotransduction mechanisms. Here, we review these processes and assess their relevance in the understanding of craniofacial malformations., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Genetic diseases with craniofacial malformations can be associated with anomalies of the craniocervical joint (CCJ). The functions of the CCJ are thus impaired, as mobility may be either limited by abnormal bone fusion causing headaches, or exaggerated in the case of hypermobility, which may cause irreparable damage to the spinal cord. Restoring the balance between mobility and stability requires surgical correction in children. The anatomy and biomechanics of the CCJ are quite unique, yet have been overlooked in the past decades. Pediatric evidence is so scarce, that investigating the adult CCJ is our best shot to disentangle the form-function relationships of this anatomical region. The motivation of the present study was to understand the morphological and functional basis of motion in the CCJ, in the hope to find morphological features accessible from medical imaging able to predict mobility. To do so, we have quantified the in-vitro kinematics of the CCJ in nine cadaveric asymptomatic adults, and estimated a wide range of mobility variables covering the complexity of spinal motion. We compared these variables with the shape of the occipital, the atlas and the axis, obtained using a dense geometric morphometric approach. Morphological joint congruence was also quantified. Our results suggest a strong relationship between bone shape and motion, with the overall geometry predicting best the primary movements, and the joint facets predicting best the secondary movements. We propose a functional hypothesis stating that the musculoligamental system determines movements of great amplitude, while the shape and congruence of joint facets determine the secondary and coupled movements, especially by varying the geometry of bone stops and the way ligaments are tensioned. We believe this work will provide valuable insights in understanding the biomechanics of the CCJ. Furthermore, it should help surgeons treating CCJ anomalies by enabling them to translate objectives of functional and clinical outcome into clear objectives of morphological outcome., (© 2024 The Author(s). Journal of Morphology published by Wiley Periodicals LLC.)

Background: Trigonocephaly occurs due to the premature fusion of the metopic suture, leading to a triangular forehead and hypotelorism. This condition often requires surgical correction for morphological and functional indications. Metopic ridges also originate from premature metopic closure but are only associated with mid-frontal bulging; their surgical correction is rarely required. Differential diagnosis between these two conditions can be challenging, especially in minor trigonocephaly., Methods: Two hundred seven scans of patients with trigonocephaly (90), metopic rigdes (27), and controls (90) were collected. Geometric morphometrics were used to quantify skull and orbital morphology as well as the interfrontal angle and the cephalic index. An innovative method was developed to automatically compute the frontal curvature along the metopic suture. Different machine-learning algorithms were tested to assess the predictive power of morphological data in terms of classification., Results: We showed that control patients, trigonocephaly and metopic rigdes have distinctive skull and orbital shapes. The 3D frontal curvature enabled a clear discrimination between groups (sensitivity and specificity > 92%). Furthermore, we reached an accuracy of 100% in group discrimination when combining 6 univariate measures., Conclusion: Two diagnostic tools were proposed and demonstrated to be successful in assisting differential diagnosis for patients with trigonocephaly or metopic ridges. Further clinical assessments are required to validate the practical clinical relevance of these tools., (© 2024. The Author(s).)

Understanding how phenotypic divergence arises among natural populations remains one of the major goals in evolutionary biology. As part of competitive exclusion experiment conducted in 1971, 10 individuals of Italian wall lizard (Podarcis siculus (Rafinesque-Schmaltz, 1810)) were transplanted from Pod Kopište Island to the nearby island of Pod Mrčaru (Adriatic Sea). Merely 35 years after the introduction, the newly established population on Pod Mrčaru Island had shifted their diet from predominantly insectivorous towards omnivorous and changed significantly in a range of morphological, behavioural, physiological and ecological characteristics. Here, we combine genomic and quantitative genetic approaches to determine the relative roles of genetic adaptation and phenotypic plasticity in driving this rapid phenotypic shift. Our results show genome-wide genetic differentiation between ancestral and transplanted population, with weak genetic erosion on Pod Mrčaru Island. Adaptive processes following the founder event are indicated by highly differentiated genomic loci associating with ecologically relevant phenotypic traits, and/or having a putatively adaptive role across multiple lizard populations. Diverged traits related to head size and shape or bite force showed moderate heritability in a crossing experiment, but between-population differences in these traits did not persist in a common garden environment. Our results confirm the existence of sufficient additive genetic variance for traits to evolve under selection while also demonstrating that phenotypic plasticity and/or genotype by environment interactions are the main drivers of population differentiation at this early evolutionary stage., (© 2023 John Wiley & Sons Ltd.)

Hemifacial myohyperplasia (HFMH) is a rare cause of facial asymmetry exclusively involving facial muscles. The underlying cause and the mechanism of disease progression are unknown. Here, we identified a somatic gain-of-function mutation of PIK3CA in five pediatric patients with HFMH. To understand the physiopathology of muscle hypertrophy in this context, we created a mouse model carrying specifically a PIK3CA mutation in skeletal muscles. PIK3CA gain-of-function mutation led to striated muscle cell hypertrophy, mitochondria dysfunction, and hypoglycemia with low circulating insulin levels. Alpelisib treatment, an approved PIK3CA inhibitor, was able to prevent and reduce muscle hypertrophy in the mouse model with correction of endocrine anomalies. Based on these findings, we treated the five HFMH patients. All patients demonstrated clinical, esthetical, and radiological improvement with proof of target engagement. In conclusion, we show that HFMH is due to somatic alteration of PIK3CA and is accessible to pharmacological intervention., (© 2023 Bayard et al.)

Objective: This study aimed to assess the relevance of using multi-positional MRI (mMRI) to identify cranio-vertebral junction (CVJ) instability in pediatric patients with CVJ anomalies while determining objective mMRI criteria to detect this condition., Material and Methods: Data from children with CVJ anomalies who underwent a mMRI between 2017 and 2021 were retrospectively reviewed. Mobility assessment using mMRI involved: (1) morphometric analysis using hierarchical clustering on principal component analysis (HCPCA) to identify clusters of patients by considering their mobility similarities, assessed through delta (Δ) values of occipito-cervical parameters measured on mMRI; and (2) morphological analysis based on dynamic geometric CVJ models and analysis of displacement vectors between flexion and extension. Receiver operating characteristics (ROC) curves were generated for occipito-cervical parameters to establish instability cut-off values. (3) Additionally, an anatomical qualitative analysis of the CVJ was performed to identify morphological criteria of instability., Results: Forty-seven patients with CVJ anomalies were included (26 females, 21 males; mean age: 10.2 years [3-18]). HCPCA identified 2 clusters: cluster №1 (stable patients, n = 39) and cluster №2 (unstable patients, n = 8). ΔpB-C2 (pB-C2 line delta) at ≥2.5 mm (AUC 0.98) and ΔBAI (Basion-axis Interval delta) ≥ 3 mm (AUC 0.97) predicted instability with 88% sensibility and 95% specificity and 88% sensitivity and 85% specificity, respectively. Geometric CVJ shape analysis differentiated patients along a continuum, from a low to a high CVJ motion that was characterized by a subluxation of C1 in the anterior direction. Qualitative analysis found correlations between instability and C2 anomalies, including fusions with C3 (body p = 0.032; posterior arch p = 0.045; inferior articular facets p = 0.012; lateral mass p = 0.029)., Conclusions: We identified a cluster of pediatric patients with CVJ instability among a cohort of CVJ anomalies that were characterized by morphometric parameters with corresponding cut-off values that could serve as objective mMRI criteria. These findings warrant further validation through prospective case-control studies.

Introduction: Maxillary surgery alone can be proposed for the surgical management of class III malocclusion, but anticipating outcomes for the labiomental muscle complex is challenging due to the mandibular autorotation phenomenon. The objective of this study was to quantify the mandibular and labiomental movements induced by maxillary osteotomy alone in the management of class III malocclusion according to different clinical and surgical variables., Methods: The post-operative changes in mandibular and labiomental shapes were studied by geometric morphometry from the pre- and post-operative lateral cephalometric radiograph of 25 patients. The explanatory variables tested were maxillary advancement, maxillary rotation, and divergence., Results: Soft tissues repositioning are different from postoperative mandibular repositioning after maxillary osteotomy. Neuromuscular adjustments of mandible depend on divergence and the maxillary rotation. Labiomental response only depends on divergence., Conclusion: The surgical procedure does not have the same bone-related and musculocutaneous effects on patients with the same class III malocclusion. It is therefore essential for surgeons to understand the effects of their procedure on musculocutaneous tissues in order to best anticipate post-operative outcomes., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Passalboni et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

We present a novel method for the morphometric analysis of series of 3D shapes, and demonstrate its relevance for the detection and quantification of two craniofacial anomalies: trigonocephaly and metopic ridges, using CT-scans of young children. Our approach is fully automatic, and does not rely on manual landmark placement and annotations. Our approach furthermore allows to differentiate shape classes, enabling successful differential diagnosis between trigonocephaly and metopic ridges, two related conditions characterized by triangular foreheads. These results were obtained using recent developments in automatic nonrigid 3D shape correspondence methods and specifically spectral approaches based on the functional map framework. Our method can capture local changes in geometric structure, in contrast to methods based, for instance, on global shape descriptors. As such, our approach allows to perform automatic shape classification and provides visual feedback on shape regions associated with different classes of deformations. The flexibility and generality of our approach paves the way for the application of spectral methods in quantitative medicine., (© 2023 Wiley Periodicals LLC.)

Introduction: The spaceflight environment presents unique demands on human physiology; among those demands, is sleep. Sleep loss and circadian desynchronization is a major concern for future deep- exploration plans, including long-term crewed missions to the Moon and Mars., Aims: Analyze evidence of sleep disruption in crewmembers during low-Earth orbit missions, identify the use of sleep-promoting medication among crewmembers and deepen the comprehension of challenges to sleep physiology for future missions to the Moon and Mars., Results: Evidence consistently indicates a loss of sleep and circadian rhythm disruption during low-Earth orbit missions. Sleep duration is shortened especially the night before a critical operation and during circadian-misaligned sleep episodes. The prevalence of sleep-promoting medication ranges between 71% and 78%; medication is more frequently taken on circadian-misaligned sleep episodes. Regarding the Moon, Apollo astronauts had variable sleep duration. For some, sleep was restful while others had poor-quality sleep. Many reported fatigue and errors due to the lack of rest. A loss of the 24-h light/dark might be expected due to the Moon's complex illumination characteristics. Regarding Mars, one main challenge will consist in synchronizing the circadian clock to a Martian day (24.65 h)., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Authors.)

Macroevolutionary changes such as variation in habitat use or diet are often associated with convergent, adaptive changes in morphology. However, it is still unclear how small-scale morphological variation at the population level can drive shifts in ecology such as observed at a macroevolutionary scale. Here, we address this question by investigating how variation in cranial form and feeding mechanics relate to rapid changes in diet in an insular lizard ( Podarcis siculus ) after experimental introduction into a new environment. We first quantified differences in the skull shape and jaw muscle architecture between the source and introduced population using three-dimensional geometric morphometrics and dissections. Next, we tested the impact of the observed variation in morphology on the mechanical performance of the masticatory system using computer-based biomechanical simulation techniques. Our results show that small differences in shape, combined with variation in muscle architecture, can result in significant differences in performance allowing access to novel trophic resources. The confrontation of these data with the already described macroevolutionary relationships between cranial form and function in these insular lizards provides insights into how selection can, over relatively short time scales, drive major changes in ecology through its impact on mechanical performance.

Resource-limited environments may drive the rapid evolution of phenotypic traits and ecological preferences optimizing the exploitation of resources. Very small islands are often characterized by reduced food availability, seasonal fluctuations in resources and strong unpredictability. These features may drive the evolution of phenotypic traits such as high bite forces, allowing animals to exploit a wider variety of the available resources. They may also lead to more generalist dietary patterns in response to food scarcity. However, the lack of predators and competitors on such small islands often also leads to high densities and the evolution of strong sexual dimorphism, which may also drive the evolution of bite force. Here, we take advantage of a unique replicated introduction experiment to test whether lizards introduced into very small islands alter their feeding ecology and use different resources, resulting in the evolution of a large body size, large head size and large bite forces. Our results show that three years after their introduction, the island lizards were larger and had greater bite forces and more pronounced sexual dimorphism. However, the diets were only marginally different between animals from the source population on a very large nearby island and those on the islets. Moreover, distinct differences in diet between animals on the different islets were observed, suggesting that the local environment is a strong driver of resource use. Overall, lizards with absolutely and relatively (adjusted for body size) large bite forces did eat larger and harder prey. Taken together, our data suggest that intraspecific competition is an important driver of the rapid evolution of bite force, which may allow these lizards to exploit the scarce and fluctuating resources on the islets. Whether or not lizards will evolve to include other types of food such as plants in their diet, facilitated by their large bite forces, remains to be explored in future studies.

Background: Achondroplasia is the most frequent FGFR3-related chondrodysplasia, leading to rhizomelic dwarfism, craniofacial anomalies, stenosis of the foramen magnum, and sleep apnea. Craniofacial growth and its correlation with obstructive sleep apnea syndrome has not been assessed in achondroplasia. In this study, we provide a multimodal analysis of craniofacial growth and anatomo-functional correlations between craniofacial features and the severity of obstructive sleep apnea syndrome., Methods: A multimodal study was performed based on a paediatric cohort of 15 achondroplasia patients (mean age, 7.8 ± 3.3 years), including clinical and sleep study data, 2D cephalometrics, and 3D geometric morphometry analyses, based on CT-scans (mean age at CT-scan: patients, 4.9 ± 4.9 years; controls, 3.7 ± 4.2 years)., Results: Craniofacial phenotype was characterized by maxillo-zygomatic retrusion, deep nasal root, and prominent forehead. 2D cephalometric studies showed constant maxillo-mandibular retrusion, with excessive vertical dimensions of the lower third of the face, and modifications of cranial base angles. All patients with available CT-scan had premature fusion of skull base synchondroses. 3D morphometric analyses showed more severe craniofacial phenotypes associated with increasing patient age, predominantly regarding the midface-with increased maxillary retrusion in older patients-and the skull base-with closure of the spheno-occipital angle. At the mandibular level, both the corpus and ramus showed shape modifications with age, with shortened anteroposterior mandibular length, as well as ramus and condylar region lengths. We report a significant correlation between the severity of maxillo-mandibular retrusion and obstructive sleep apnea syndrome (p < 0.01)., Conclusions: Our study shows more severe craniofacial phenotypes at older ages, with increased maxillomandibular retrusion, and demonstrates a significant anatomo-functional correlation between the severity of midface and mandible craniofacial features and obstructive sleep apnea syndrome., (© 2023. The Author(s).)

Objective: Chiari malformation type I (CM-I) is frequent in children and remains a surgical challenge. Several techniques have been described for posterior fossa decompression. No decision algorithm has been validated, and strategies are highly variable between institutions. The goal of this study was to define therapeutic guidelines that take into consideration patient specificities., Methods: The authors retrospectively collected data from patients who were < 18 years of age, were diagnosed with CM-I, and were treated surgically between 2008 and 2018 in 8 French pediatric neurosurgical centers. Data on clinical features, morphological parameters, and surgical techniques were collected. Clinical outcomes at 3 and 12 months after surgery were assessed by the Chicago Chiari Outcome Scale. The authors used a hierarchical clustering method to define clusters of patients by considering their anatomical similarities, and then compared outcomes between surgical strategies in each of these clusters., Results: Data from 255 patients were collected. The mean age at surgery was 9.6 ± 5.0 years, syringomyelia was reported in 60.2% of patients, the dura mater was opened in 65.0% of patients, and 17.3% of patients underwent a redo surgery for additional treatment. The mean Chicago Chiari Outcome Scale score was 14.4 ± 1.5 at 3 months (n = 211) and 14.6 ± 1.9 at 12 months (n = 157). The hierarchical clustering method identified three subgroups with potentially distinct mechanisms underlying tonsillar herniation: bony compression, basilar invagination, and foramen magnum obstruction. Each cluster matched with specific outcomes., Conclusions: This French multicenter retrospective cohort study enabled the identification of three subgroups among pediatric patients who underwent surgery for CM-I, each of which was associated with specific outcomes. This morphological classification of patients might help in understanding the underlying mechanisms and providing personalized treatment.

The kinematics of lizard feeding are the result of complex interactions between the craniocervical, the hyolingual, and the locomotor systems. The coordinated movement of these elements is driven by sensory feedback from the tongue and jaws during intraoral transport. The kinematics of jaw movements have been suggested to be correlated with the functional characteristics of the prey consumed, such as prey mobility and hardness. However, whether and how dietary breadth correlates with the flexibility in the behavioral response has rarely been tested, especially at the intraspecific level. Here we tested whether an increase in dietary breadth was associated with a greater behavioral flexibility by comparing two recently diverged populations of insular Podarcis lizards differing in dietary breadth. To do so, we used a stereoscopic high-speed camera set-up to analyze the jaw kinematics while offering them different prey types. Our results show that prey type impacts kinematics, especially maximum gape, and maximum opening and closing speed. Furthermore, the behavioral flexibility was greater in the population with the greater dietary breadth, suggesting that populations which naturally encounter and feed on more diverse prey items show a greater ability to modulate their movements to deal with variation in functionally relevant prey properties. Finally, the more generalist population showed more stereotyped movements suggesting a finer motor control., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Phenotypictraits have been shown to evolve in response to variation in the environment. However, the evolutionary processes underlying the emergence of phenotypic diversity can typically only be understood at the population level. Consequently, how subtle phenotypic differences at the intraspecific level can give rise to larger-scale changes in performance and ecology remains poorly understood. We here tested for the covariation between ecology, bite force, jaw muscle architecture, and the three-dimensional shape of the cranium and mandible in 16 insular populations of the lizards Podarcis melisellensis and P. sicula. We then compared the patterns observed at the among-population level with those observed at the interspecific level. We found that three-dimensional head shape as well as jaw musculature evolve similarly under similar ecological circumstances. Depending on the type of food consumed or on the level of sexual competition, different muscle groups were more developed and appeared to underlie changes in cranium and mandible shape. Our findings show that the local selective regimes are primary drivers of phenotypic variation resulting in predictable patterns of form and function. Moreover, intraspecific patterns of variation were generally consistent with those at the interspecific level, suggesting that microevolutionary variation may translate into macroevolutionary patterns of ecomorphological diversity., (© 2021 The Authors. Evolution © 2021 The Society for the Study of Evolution.)

Three-dimensional nanostructured magnetic materials have recently been the topic of intense interest since they provide access to a host of new physical phenomena. Examples include new spin textures that exhibit topological protection, magnetochiral effects and novel ultrafast magnetic phenomena such as the spin-Cherenkov effect. Two-photon lithography is a powerful methodology that is capable of realising 3D polymer nanostructures on the scale of 100 nm. Combining this with postprocessing and deposition methodologies allows 3D magnetic nanostructures of arbitrary geometry to be produced. In this article, the physics of two-photon lithography is first detailed, before reviewing the studies to date that have exploited this fabrication route. The article then moves on to consider how non-linear optical techniques and post-processing solutions can be used to realise structures with a feature size below 100 nm, before comparing two-photon lithography with other direct write methodologies and providing a discussion on future developments.

Access to resources is a dynamic and multicausal process that determines the success and survival of a population. It is therefore often challenging to disentangle the factors affecting ecological traits like diet. Insular habitats provide a good opportunity to study how variation in diet originates, in particular in populations of mesopredators such as lizards. Indeed, high levels of population density associated with low food abundance and low predation are selection pressures typically observed on islands. In the present study, the diet of eighteen insular populations of two closely related species of lacertid lizards ( Podarcis sicula and Podarcis melisellensis ) was assessed. Our results reveal that despite dietary variability among populations, diet taxonomic diversity is not impacted by island area. In contrast, however, diet disparity metrics, based on the variability in the physical (hardness) and behavioral (evasiveness) properties of ingested food items, are correlated with island size. These findings suggest that an increase in intraspecific competition for access to resources may induce shifts in functional components of the diet. Additionally, the two species differed in the relation between diet disparity and island area suggesting that different strategies exist to deal with low food abundance in these two species. Finally, sexual dimorphism in diet and head dimensions is not greater on smaller islands, in contrast to our predictions., Competing Interests: The authors declare no conflict of interest., (© 2019 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

The aim of this study was to create a fetal heart rate (FHR) reference curve for singleton bovine fetuses in the first trimester of gestation and to determine its possible relationship with the outcome of pregnancy. Forty-eight Holstein-Friesian cows with one fetus and five cows with twins were used. Fetal heart beatings were recorded on videotape during transrectal scanning with a 5 and/or 7.5 MHz linear array transducer on a weekly basis between Days 40 and 95 of gestation. FHR was calculated by averaging the results of five counts of the same record by the same observer. For singleton pregnancies, a reference curve was created using the mean, the standard deviation (SD) and the 5th and 95th percentiles. The FHR increased from Days 40-46 (173 beats/min) to Days 61-67 (183 beats/min). After a peak, the FHR decreased slowly until Days 89-95 (175 beats/min), while the SD increased. There was no significant difference between singleton and twin fetuses. in the aborted and lost fetuses in twin gestation due to fetal reduction, both bradycardia and tachycardia were detected compared to the singleton pregnancy reference curve.

Optical tweezers are a highly versatile tool for exploration of the mesoscopic world, permitting non-contact manipulation of nanoscale objects. However, direct illumination with intense lasers restricts their use with live biological specimens, and limits the types of materials that can be trapped. Here we demonstrate an indirect optical trapping platform which circumvents these limitations by using hydrodynamic forces to exert nanoscale-precision control over aqueous particles, without directly illuminating them. Our concept is based on optically actuated micro-robotics: closed-loop control enables highly localised flow-fields to be sculpted by precisely piloting the motion of optically-trapped micro-rotors. We demonstrate 2D trapping of absorbing particles which cannot be directly optically trapped, stabilise the position and orientation of yeast cells, and demonstrate independent control over multiple objects simultaneously. Our work expands the capabilities of optical tweezers platforms, and represents a new paradigm for manipulation of aqueous mesoscopic systems.

For an efficient single-photon source a high-count rate into a well-defined spectral and spatial mode is desirable. Here we have developed a hybrid planar Fabry-Pérot microcavity by using a two-photon polymerization process (2PP) where coupling between single-photon sources (diamond colour centres) and resonance modes is observed. The first step consists of using the 2PP process to build a polymer table structure around previously characterized nitrogen-vacancy (NV) centres on top of a distributed Bragg reflector (DBR) with a high reflectivity at the NV zero-phonon line (ZPL). Afterwards, the polymer structure is covered with a silver layer to create a weak (low Q) cavity where resonance fluorescence measurements from the NVs are shown to be in good agreement with analytical and Finite Difference Time Domain (FDTD) results.

The breakdown of semantic memory appears early in Alzheimer's disease (AD). Lexico-semantic difficulties (e.g. difficulties in retrieving words and their meaning) are among the first symptoms of the disease. In order to screen this lexico-semantic decline, we propose the mini-SKQ, a semantic knowledge questionnaire. Mini-SKQ explores semantic impairment in AD patients with 12 questions. We administer mini-SKQ to 77 participants, 39 AD and 38 healthy old people. Results showed a significant difference between the groups: AD patients made significantly more errors than healthy old people. The Receiver operating characteristic (ROC) analysis established the optimal cut-off score of the mini-SKQ at 3 errors (sensitivity 61,5%, specificity 89,5%) in differentiating between AD patients and healthy old people. The mini-SKQ is a fast and easily administered questionnaire. Results indicated favorable indicators to screen semantic knowledge. These first observations underline that mini-SKQ could potentially be attractive for screening semantic memory deterioration.

Long bone inner structure and cross-sectional geometry display a strong functional signal, leading to convergences, and are widely analyzed in comparative anatomy at small and large taxonomic scales. Long bone microanatomical studies have essentially been conducted on transverse sections but also on a few longitudinal ones. Recent studies highlighted the interest in analyzing variations of the inner structure along the diaphysis using a qualitative as well as a quantitative approach. With the development of microtomography, it has become possible to study three-dimensional (3D) bone microanatomy and, in more detail, the form-function relationships of these features. This study focused on the selection of quantitative parameters to describe in detail the cross-sectional shape changes and distribution of the osseous tissue along the diaphysis. Two-dimensional (2D) virtual transverse sections were also performed in the two usual reference planes and results were compared with those obtained based on the whole diaphysis analysis. The sample consisted in 14 humeri and 14 femora of various mammalian taxa that are essentially terrestrial. Comparative quantitative analyses between different datasets made it possible to highlight the parameters that are strongly impacted by size and phylogeny and the redundant ones, and thus to estimate their relevance for use in form-function analyses. The analysis illustrated that results based on 2D transverse sections are similar for both sectional planes; thus if a strong bias exists when mixing sections from the two reference planes in the same analysis, it would not problematic to use either one plane or the other in comparative studies. However, this may no longer hold for taxa showing a much stronger variation in bone microstructure along the diaphysis. Finally, the analysis demonstrated the significant contribution of the parameters describing variations along the diaphysis, and thus the interest in performing 3D analyses; this should be even more fruitful for heterogeneous diaphyses. In addition, covariation analyses showed that there is a strong interest in removing the size effect to access the differences in the microstructure of the humerus and femur. This methodological study provides a reference for future quantitative analyses on long bone inner structure and should make it possible, through a detailed knowledge of each descriptive parameter, to better interpret results from the multivariate analyses associated with these studies. This will have direct implications for studies in vertebrate anatomy, but also in paleontology and anthropology., (© 2018 Anatomical Society.)

In this article we present the results of a study to explore if cross-border DNA matches between the Netherlands and Belgium are relatively more likely to occur in areas near the Dutch-Belgian border than in areas at some distance from this border. For this study we used the results of the transnational DNA profile exchange and comparison between the Belgian and Dutch DNA databases, which first took place in 2014. It appears that the Dutch regions adjacent to Belgium, i.e., Zeeland-West-Brabant, Oost-Brabant and Limburg, have relatively more DNA matches with Belgium than the other Dutch regions. In other words, a DNA profile obtained from a crime scene close to the Dutch border with Belgium is more likely to match with the profile of a person whose DNA profile is stored in the Belgian database than a DNA profile that originates from a crime scene further afield. Our data suggest that crimes committed by repeat offenders show a spatial pattern despite the presence of a national border, with crime scenes clustering in relatively close proximity to each other. The results of this study provide a better understanding of geographical patterns of cross-border criminal mobility., (Copyright © 2016 The Chartered Society of Forensic Sciences. Published by Elsevier Ireland Ltd. All rights reserved.)

Rangeland-based livestock systems have to deal with the significant instability and uncertainty of the agricultural context (policy changes, volatility of input prices, etc.), and especially of the climatic context. Thus, they are particularly concerned by adaptive management strategies. To support the development of such strategies, we developed a board game including a computer model called "Rangeland Rummy". It is to be used by groups of farmers and agricultural consultants in the context of short workshops (about 3 h). Rangeland Rummy builds upon five types of material object: (i) a game board; (ii) a calendar stick indicating the starting date of the game board; (iii) sticks marked with the feed resources available for combinations of vegetation types and their management practices; (iv) cards to define animal groups and their feeding requirements throughout the year; (v) cards related to types of feed that can be attributed to animal groups throughout the year. Using these material objects, farmers collectively design a rangeland-based livestock system. This system is immediately evaluated using a computer model, i.e. a spreadsheet providing graphs and indicators providing information on, among other things, the extent to which quantitative and qualitative animal feeding requirements are covered across the year. Playing the game thus consists in collectively and iteratively designing and evaluating rangeland-based livestock systems, while confronting the players with new contextual challenges (e.g. interannual variability of weather, volatility of input prices) or new farmers' objectives (e.g. being self-sufficient for animal feeding). An example of application of Rangeland Rummy with 3 farmers in southern France is reported. Applications show that it tends to develop farmers' adaptive capacity by stimulating their discussions and the exchange of locally-relevant knowledge on management strategies and practices in rangeland-based livestock systems., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

The aim of this analysis was to determine whether pregnancy loss (PL) after embryo transfer (ET) in cattle was related to maternal progesterone (P4) concentrations during and shortly after ET, and maternal bovine pregnancy-associated glycoprotein-1 (bPAG-1) concentrations in plasma at days 25-35 of gestation. Embryos (n=260) were produced either in vivo after superovulation (n=115), or in vitro from oocytes (obtained with ovum pick-up) in co-culture (n=44) or cultured in a synthetic medium (n=101). Overall, PL was 56.9% (148) and no significant differences occurred in calving rate among the three embryo production groups. There was no difference in P4 concentrations on days 7-14 of gestation in the three groups, nor between ongoing and interrupted pregnancies. Between days 25 and 35 of pregnancy, bPAG-1 concentrations were unaffected by embryo production, but in cattle that had PL between days 26 and 120, four bPAG-1 profiles could be detected. Between days 25 and 32, bPAG-1 concentrations were influenced by PL, and concentrations were significantly lower in animals in which PL occurred between days 26 and 120 than in those animals that aborted later or calved at term. Early P4 concentrations suggested that maternal luteal factors were not responsible for PL which appeared to be caused by impaired conceptus development (regardless of embryo type) as reflected by low maternal bPAG-1 concentrations prior to embryonic death., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Various parenteral treatment forms of oxytocin, as often used under praxis circumstances, may act differently on contractility of the uterus during the first days of the puerperium. Various patterns of such induced uterotonic responses may lead to alterations in the emptying characteristics of the uterine lumen, thus influencing, as a late consequence, the process of involution. Therefore, this study was designed to test whether two different parenteral administration forms of oxytocin induce changes in peripheral plasma concentrations of 15-ketodihydro-prostaglandin F(2α) (PGF(2α) metabolite) in early post-partum cows. Between 13 and 15 h after uncomplicated calving, healthy dairy cows without retained foetal membranes were treated with 50 IU oxytocin, either intramuscularly (OT-IM group; n = 15) or intravenously (OT-IV group; n = 16). Saline solution was administered intramuscularly as controls (CON group; n = 15). Jugular blood samples were taken at 10-min intervals from 1 h before to 2 h after treatment. Plasma PGF(2α) metabolite levels were measured by radioimmunoassay. No significant differences in peripheral plasma PGF(2α) metabolite concentrations occurred in the OT-IM and CON groups, but mean values significantly increased in the OT-IV group, peaking at 20 min after treatment and reaching pre-treatment baseline values again at 120 min. Although the source of prostaglandins was not investigated in this study, our results suggest that exogenous oxytocin may enhance secretion of prostaglandins by the uterus during the first day after normal calving. These prostaglandins might contribute, by an endocrine or paracrine route, to the stimulation of myometrial contractility when exogenous oxytocin is given during this early post-partum stage., (© 2011 Blackwell Verlag GmbH.)

Repeated ultrasonographic observation of fetal movements was used to distinguish movement patterns and to investigate the rate of occurrence and temporal organisation of these patterns (rest-activity cycles) during the last three weeks of gestation in the pig. By means of transabdominal ultrasonography with a 3.5MHz linear array transducer, motility in ten different fetuses (one per sow) was studied. Six (median; range 4-6) 1h recordings were made per fetus at 3-5 day intervals. Fifty-five 1h recordings were available for analysis. The occurrence of fetal general movements (GM), isolated head (HM), forelimb movements (LM), and rotations (ROT) was analysed from video tapes. For each movement pattern, the trend in occurrence over time was assessed by multilevel analysis. The temporal association between different movement patterns was studied by calculation of the kappa value. ROT occurred very infrequently and showed no particular trend over time. GM, HM, and LM showed a significant decreasing trend towards parturition (P<0.01). Total fetal activity (i.e., the sum of the four movement incidences) declined from an average of 25% of recording time to 9% over the last three weeks of pregnancy. Periods of fetal quiescence gradually increased with progressing gestation (P<0.05). There was no evidence of concordant association between the periods of rest and activity of GM, HM, and LM or of improved temporal linkage between these movement patterns with time. Fetal bodily activity decreases towards parturition mainly due to prolonged periods of rest. Fetal movement patterns show rest-activity cycles, but each pattern appears to cycle independently from the other throughout late gestation. The present results of spontaneous fetal movements in the pig provide reference data for future studies of fetal activity under different zoo technical conditions or pharmacological interventions.

Daily separation of sows and piglets during lactation, intermittent suckling (IS), improves feed intake and postweaning adaptation in piglets. The aim of the current study was to determine how, in piglets that have been subjected to IS, age at weaning and the duration of the preceding IS period contribute to postweaning adaptation through effects on feed intake, growth, and gut characteristics. All piglets had ad libitum access to creep feed from d 7. Litters were subjected to conventional weaning (CW) or to 1 of 3 IS regimens. In CW, litters (n = 29) had continuous access to the sow until weaning (d 26, d 0 = farrowing). During IS, litters had access to the sow between 1600 and 0600 h. Litters in the IS treatments were subjected to IS 1) from d 19 onward and weaned at d 26 (IS19-7D, n = 33), 2) from d 19 onward and weaned at d 33 (IS19-14D, n = 28), or 3) from d 26 onward and weaned at d 33 (IS26-7D, n = 33). The IS19-7D regimen resulted in a relative growth check within the first 2 d after weaning similar to CW litters (72 +/- 13 and 90 +/- 7%, respectively), but in a greater piglet growth (P = 0.014) and feed intake (P = 0.001) between d 2 and 7 postweaning. Moreover, IS19-7D was not associated with a (further) reduction in villus height as observed at d 2 postweaning in CW litters. In IS piglets weaned after an extended lactation (d 33), a markedly smaller weaning-associated relative growth check was observed shortly postweaning (11 +/- 18 and 32 +/- 19% for IS19-14D and IS26-7D litters, respectively). In these litters, feed intake and growth within the first 2 d after weaning were slightly greater when piglets were subjected to IS for 2 wk (IS19-14D) rather than for 1 wk (IS26-7D; P = 0.032 and P = 0.037 for feed intake and growth, respectively). Irrespective of duration of IS, weaning at d 33 with IS was not associated with a reduction in villus height. Irrespective of treatment, plasma citrulline concentrations were reduced at d 2 and 8 postweaning compared with the values at weaning (P < or = 0.01). No correlation was observed between postweaning plasma citrulline concentrations and postweaning small intestinal villus height. This study indicates that 1 wk of IS before weaning at d 26 of lactation improves feed intake and growth between d 2 and 7 postweaning and does not result in a reduction of villus height as observed in CW piglets, although it did not prevent a profound growth check shortly after weaning. However, combining 1 wk of IS with an extended lactation improved postweaning adaptation markedly in terms of growth, feed intake, and gut characteristics. Increasing the duration of IS from 1 to 2 wk slightly improved growth and feed intake shortly after weaning, but the contribution to postweaning adaptation seemed to be relatively small compared with extending lactation.

The final stages of cervical ripening and parturition resemble an inflammatory process. Although the role of cytokines in both spontaneous and experimentally induced parturitions has been described in several small laboratory animals and humans, the involvement of pro-inflammatory and regulatory cytokines in physiologic parturition in cows has not been determined. In this study, the cytokine expression profiles were assessed in bovine cervical tissue at several stages of pregnancy and at parturition. Serial biopsy samples of the cervix were obtained from 10 cows on day 185 and day 275 of pregnancy (which was on average 5.4 days before parturition) and at parturition. Messenger RNA expression levels of interleukin (IL)-1beta, IL-6, IL-8, IL-10 and tumour necrosis factor (TNF)alpha were determined using real-time polymerase chain reaction and the number of neutrophils and eosinophils was estimated by Luna and Sirius Red staining. At parturition, IL-8 expression had increased 430-fold (p < 0.001) when compared with that of the day 185 of pregnancy, large numbers of neutrophils had invaded the cervix while eosinophils remained scarce, IL-1beta had increased eightfold (p < 0.05) and IL-6 had not changed significantly. Additionally, IL-10 was increased by 10-fold (p < 0.001) and TNFalpha decreased by 57% (p < 0.05) when compared with that of the day 185 of pregnancy. The large increase in expression of IL-8, enabling the influx of neutrophils, is indicative of its important role in the final stage of cervical ripening and at parturition. As previous studies have shown that neutrophils excrete matrix metalloproteinases (MMP), this might contribute to softening of the cervix. In contrast, the only slightly increased levels of IL-1, steady concentrations of IL-6 and decreased TNFalpha, the potential consequences of increased IL-10 expression, indicate that final cervical of cows in ripening at term parturition is an inflammatory process influenced by regulatory cytokines.

Assessment of fetal motility is an approach to evaluate the development and function of the nervous system before birth. Reference values for the time of first occurrence and the incidence of normal fetal movements are indispensable for studies in which prenatal motor activity is applied as a model to study the central and peripheral nervous systems. Studies on fetal motility have been performed in a few species, particularly in the human. The aim of the present study is to describe the ontogeny of fetal motility in the guinea pig, a precocious polytocous species. After a pilot study to establish procedures for repeated ultasonographic scanning of guinea pigs, 10 domesticated animals were scanned (5.0 or 7.5 MHz convex transducer) at 2-4 day intervals between day 24 and 63 of gestation (term age 68 days). Per animal two selected fetuses were each scanned for 15 min. Images were stored on videotape and analyzed off-line for the first onset, presence and quality of fetal movement patterns, and quantity of sideway bendings, general movements, breathing movements and periods of fetal rest. Twenty-five different movement patterns could be characterized, 6 emerging at the onset of motor activity were performed only temporarily. The very first fetal movement was observed on day 24 gestational age, and subsequently most other movements developed during a period of only 5 days. Interfetal difference in onset of the frequently occurring sideway bendings, general movements, and front and hind limb movements was only 2 days. Sideway bendings and general movements co-existed during days 29 to 43. There were developmental trends in the course of pregnancy. Sideway bendings increased rapidly between 24 and 30 days and declined hereafter. General movements and fetal breathing increased during midpregnancy and declined towards parturition. Conversely, fetal rest was observed for approximately 60% of time at midgestation and a marked increase was found towards parturition. There were no significant differences in developmental trend of the various movement patterns between individual fetuses. Fetal motility in the guinea pig followed a specific temporal pattern, like in the human, but at a different time scale. The present quantitative data will enable functional investigations into the role of the neuromuscular system. They may also facilitate studies on the effect of environmental influences, such as stress, drugs, toxic substances, and food conditions, on fetal neurobehavioural development in this species.

The aims of this study were (i) to describe the changes in the volume of large ovarian follicles (diameter >3 cm) during the 48 h egg laying cycle in farmed ostriches, and (ii) to quantify factors affecting the volume of the largest measured follicle and the plasma concentrations of progesterone (P(4)) and estradiol-17beta (E(2)beta). In eight egg-producing birds, which all ovulated during the study period, transcutaneous ultrasound scanning and blood sampling was performed at 3 h intervals. The average volume of the total number of visualized large follicles (V(total)), the largest measured follicle (V(F1)), the second largest follicle (V(F2)) and of all follicles smaller than F2 (V(F3-Fn)) were each higher before than after oviposition. V(total), V(F2) and V(F3-Fn) nearly doubled in the 24-h period before oviposition, while V(F1) remained at an equal, rather high level until oviposition. Immediately after oviposition V(total), as well as the volume of the other follicle categories, decreased within 6 h, i.e. around the moment of ovulation. By performing statistical analysis on the basis of linear mixed-effects modelling, we quantified that: (i) V(F1) was 13.2% higher before than after oviposition and increased with 6.5% when LH increased with 1 ng/ml; (ii) P(4) levels were 93.2% higher before than after oviposition and increased with 43.1% for every 3 h closer to oviposition; when LH and E(2)beta levels and V(F1) increased with 1 ng/ml, 10 pg/ml and 10 ml, respectively, P(4) increased with 116.6%, 50% and 6.1%; and (iii) E(2)beta levels were 35.6% higher before than after oviposition, increased with 2.7% for every 3 h closer to oviposition and increased with 14.6% when LH increased with 1 ng/ml. It is concluded that during the egg-laying cycle in ostriches: (i) follicular mass, as estimated by the volume of visualized follicles larger than 3 cm, increases before and decreases after ovulation, and (ii) follicular dynamics and its accompanying endocrine plasma hormone profiles during the egg-laying cycle in ostriches follow a pattern similar to that in chickens.

The bovine cervix contains a large amount of smooth muscle cells distributed over an outer muscular layer and within a stromal layer. The stromal layer exhibits no electromyographic (EMG) activity at parturition. This leads to the question whether the stromal smooth muscle cells of the bovine cervix are prepared to contract with parturition, or whether they have another function. To this end, cervical biopsies were repeatedly taken from 10 pregnant cows at day-185 and -275 of gestation, at spontaneous, uncomplicated calving and at 30 days after calving. The smooth muscle bundles of the stroma were immunohistochemically analysed (n = 5) with regard to their integrity and cellular density, and the degree of staining for connexin-43, smooth muscle actin alpha (SMA), desmin and vimentin. Additionally, the mRNA expression for connexin-43, SMA, desmin and vimentin was determined with RT-PCR (n = 5). The smooth muscle tissue was arranged in bundles, also at parturition. However, the cellular density of these bundles and the SMA mRNA expression were decreased at parturition. Additionally, the SMA staining and connexin-43 expression and staining remained constant during pregnancy and at parturition. This might indicate that stromal smooth muscle cells are not prepared to contract with parturition, in contrast to the myometrial smooth muscle cells. The smooth muscle cells, stained for SMA, also expressed vimentin, and the proportion of co-expression was increased at day-275 of pregnancy. This suggests that the stromal smooth muscle cells predominantly have a secretory function in cows.

Previously we demonstrated that pre-ovulatory LH and post-ovulatory progesterone (P4) concentrations in plasma were low and embryo development was retarded when sows were induced to ovulate during lactation by submitting them to intermittent suckling (IS). The present study investigated whether this was due to: (1) stage of lactation when IS was initiated, and (2) continuation of IS post-ovulation. Multiparous Topigs40 sows were studied under three conditions: conventional weaning at Day 21 of lactation (C21; n=30), intermittent suckling from Day 14 of lactation (IS14; n=32), and intermittent suckling from Day 21 of lactation (IS21; n=33). Sows were separated from piglets for 12h daily during IS. IS sows were either weaned at ovulation or 20d following ovulation. One-third (21/63) of the IS21 and C21 sows had already ovulated or had large pre-ovulatory follicles at Day 21 and were excluded from further study. Initiation of IS at Day 14 instead of Day 21 of lactation tended to reduce P4 at 7d post-ovulation (P=0.07), did not affect pregnancy rate, and tended to reduce embryo survival (P=0.06). Continuation of IS during pregnancy resulted in lower P4 at 7 and 12d post-ovulation, tended to reduce embryo weight and pregnancy rate (P<0.10), whereas embryo survival was not affected. This study presents data for a population of sows in which follicle growth and ovulation are easily triggered under suckling conditions. Further, when these sows are bred during lactation, initiation of IS at 21 rather than 14d of lactation with weaning at ovulation yields the most desirable reproductive performance.

The objective of the current study was to investigate whether plasma citrulline or intestinal fatty acid-binding protein (I-FABP) concentrations might be used as longitudinal markers for small intestinal function in piglets after weaning. Plasma citrulline and I-FABP concentrations were measured longitudinally in weaned and unweaned piglets, and related to intestinal absorption values (i.e., plasma mannitol and 3-xylose concentrations in a sugar absorption test). Within each litter (n = 10), 2 piglets with a close-to-litter-average BW were selected. At 20.8 +/- 0.4 d of age, the selected piglets per litter were either weaned conventionally (CW) or remained with the sow (UNW). One day before, and 0.5, 2, 4, and 7 d after weaning of the CW piglets, the selected piglets of both groups were subjected to a sugar absorption test. After a 2-h fast, piglets were administered an oral dose of 2 mL/kg of sugar solution, containing 50 mg/kg of mannitol and 100 mg/kg of 3-xylose. One hour after administration, a blood sample was collected from a jugular vein for determination of plasma I-FABP, citrulline, mannitol, and 3-xylose concentrations. Plasma I-FABP concentration showed great variation within treatments, and no difference was observed in plasma I-FABP concentrations between the CW and UNW treatments (P = 0.63). The absorption of 3-xylose was not different between treatments (P = 0.83). Mannitol absorption, however, was less in the weaned CW piglets compared with the UNW piglets (P = 0.003), with the nadir on d 4 postweaning. Weaning also reduced plasma citrulline concentrations in the CW treatment compared with the UNW treatment (P < 0.001). On d 4 and 7 postweaning, plasma citrulline concentrations of CW piglets were less (P < 0.001 and P = 0.0013) than preweaning values. Furthermore, in the CW treatment, plasma citrulline concentrations correlated with plasma mannitol concentrations at d 4 postweaning (r = 0.89, P = 0.008) and overall (r = 0.76, P = 0.001). Based on these results, plasma citrulline concentration seems to be a possible marker for monitoring intestinal function in pigs after weaning.

At present no standard pharmacological intervention strategy is available to reduce these adverse effects of birth asphyxia. In the present study we aimed to evaluate placental transfer of allopurinol, an inhibitor of XOR. For this purpose, fetal catheterization of the jugular vein was conducted in five late pregnant sows (one fetus per sow). At 24-48 h after surgery, sows received allopurinol (15 mg/kg body weight; i.v.) and pharmacokinetics of allopurinol and its active metabolite oxypurinol were measured in both late pregnant sows and fetuses. Maternal and fetal blood samples were collected during and after allopurinol administration. Maternal C(max) values averaged 41.90 microg/mL (allopurinol) and 3.68 microg/mL (oxypurinol). Allopurinol crossed the placental barrier as shown by the average fetal C(max) values of 5.05 microg/mL at 1.47 h after allopurinol administration to the sow. In only one fetus low plasma oxypurinol concentrations were found. Incubations of subcellular hepatic fractions of sows and 24-h-old piglets confirmed that allopurinol could be metabolized into oxypurinol. In conclusion, we demonstrated that allopurinol can cross the placental barrier, a prerequisite for further studies evaluating the use of allopurinol as a neuroprotective agent to reduce the adverse effects following birth asphyxia in neonatal piglets.

Collagen is denatured in the gradual cervical ripening process during late pregnancy, already before the onset of final cervical ripening at parturition. Matrix Metallo Proteinases (MMPs) might be responsible for this process. To investigate the presence and potential function of MMPs at the different stages of the ripening process, serial cervical biopsies were obtained from 10 cows at Days 185 and 275 of pregnancy (approximately 5 days before calving), at parturition and at 30 days after parturition. The mRNA and protein expression of MMP-1, MMP-2, and MMP-9 and of the tissue inhibitors of MMPs (TIMP)-1 and TIMP-2 were semi-quantitatively determined using RT-PCR, respectively, zymography, Westernblot, and ELISA techniques and the localization of MMP-2 protein and presence of granulocytes by immunohistochemistry and Luna staining. At parturition compared to 185 days pregnancy the MMP-1 protein expression and the numbers of granulocytes were significantly increased by 3 and 26-fold respectively. MMP-2 mRNA and protein expression had already increased 2.5 (P < 0.05) and twofold (P < 0.05) at 5 days before parturition, prior to final ripening. At that time, MMP-2 was present in smooth muscle cells and extra cellular matrix. TIMP-1 mRNA expression was significantly increased at parturition and TIMP-2 mRNA expression peaked at 5 days before parturition. The increased expression of MMP-2 at 5 days before parturition, suggests that in the cow MMP-2 is responsible for collagen denaturation in the last part of gradual cervical ripening, while MMP-1 and MMP-9 are only active during the final cervical ripening process at parturition.

Contents Compared with other parts of the reproductive cycle, the parturition process of domestic animals has received relatively little attention from researchers during the last two decades. Yet, interventions during parturition, either by the owner or the veterinarian, are frequently judged necessary and perinatal losses in most of the domestic species are (still) rather high. The present paper focuses on the birth process in three domestic species, the dog, pig and cow, mainly because most of the experimental and research work of the authors themselves was dealing with these three species. Against the background of specific clinical problems, breeding circumstances and economic performance, selected topics and questions are addressed to identify areas for future research. These are, among others, related to the mechanisms that trigger the birth process (are the foetuses involved in the dog and sow?), the role of hormones and cytokines during ripening of the cervix (activation and/or de-activation of specific matrix metallo-proteinases and their local inhibitors), the functional significance of fetal body movements to prepare the foetus for expulsion, and the exact time course of events involved in the transportation of an individual foetus from its intrauterine location, via the pelvic canal, to the outside world. Research on some of these topics, such as the ripening of the cervix in cows, might also contribute to resolve clinical problems in human obstetrics.

Perinatal asphyxia results in tissue and cellular changes during the reperfusion period and clinical signs like perinatal mortality and decreased vitality at birth in newborn piglets. This study aimed to develop and validate a model of birth asphyxia, mimicking the evolvement of birth asphyxia in natural farrowings by conducting umbilical cord clamping (UCC) in term piglets during caesarean sections under general anaesthesia. In total 23 piglets were subjected to 5-8min of UCC and 24 piglets served as controls. Acid-base balance values and heart rates measured before UCC remained fairly constant throughout the surgical procedure, indicating nearly identical starting conditions of piglets within and between litters. UCC resulted in a significant, mild, mixed respiratory-metabolic acidosis (pH 7.22, pCO(2) 9.8kPa, BE(ecf) 2mmol/L, lactate 6.5mmol/L; controls: pH 7.31, pCO(2) 8.5kPa, BE(ecf) 5mmol/L, lactate 4mmol/L) at 10min after birth (defined as simultaneous cutting of the umbilical cord and removal of a plastic bag that had been placed over the head to avoid air intake). Heart rates were significantly decreased during UCC (range: 83-107beats/min versus 128-134beats/min in controls). Rectal temperatures and changes in body weight until 72h of life were not affected by UCC. Interestingly, four control and seven clamped piglets did not survive as no independent respiration could be attained. Birth weights and duration of UCC of these piglets did not differ significantly from those in surviving control and clamped piglets. In conclusion the mixed respiratory-metabolic acidosis arising in the surviving clamped piglets is not as severe as can be expected in highly asphyxiated, vaginally delivered newborn piglets. Repeatability of the model is compromised by considerable variation in the individual response to UCC.

This presentation will focus on the assessment of the condition of the piglet and calf during and shortly after birth and will review some of the physiological and clinical factors that may have played a role during their delivery and influenced their viability at birth.

Prenatal stress can affect the offspring's behaviour, physiology, and immune parameters. This paper summarises and discusses experimental and field studies on prenatal maternal stress in pigs. Often, elevated maternal corticosteroid concentrations during gestation are used to model prenatal stress. We used prolonged oral administration of cortisol (hydrocortisone acetate, HCA) to pregnant sows, which resulted in elevated maternal plasma and salivary cortisol concentrations. This treatment induced elevated fetal basal and adrenocorticotropic hormone (ACTH)-induced plasma cortisol concentrations, as demonstrated by a pilot study. Postnatally, it reduced birth weight of the piglets, and resulted in more live born piglets and higher preweaning mortality. In addition, it reduced the female offspring's salivary cortisol response to ACTH, and it enhanced the piglets' novelty-induced locomotion and vocalisations, and the piglets were more aggressive in a social test. Some of these effects depended on the period of gestation during which maternal cortisol concentrations were elevated, and on the sex of the offspring. These results demonstrate that piglet physiology and behaviour can indeed be affected when the mother has elevated cortisol concentrations during gestation. Regular mixing of pregnant sows with unfamiliar sows during the last third of gestation did not affect maternal salivary cortisol concentrations. Also, it did not affect the piglets' performance, behaviour, adrenocortical response to ACTH, or wound healing. Regular mixing of pregnant sows during the last third of gestation did not affect the piglets' characteristics as studied in these experiments. However, performance and behaviour of piglets were highly influenced by the social rank of their mother during gestation. Our studies have demonstrated that piglets can be affected by elevated maternal cortisol concentrations during fetal development and by social rank of the pregnant sow during gestation.

To increase our understanding of the endocrine changes associated with parturition in dogs, plasma concentrations of progesterone (P4), 15-ketodihydroprostaglandin F(2alpha) (PGFM), estradiol-17-beta (E2beta), cortisol, ACTH, prolactin (PRL), LH, and FSH were measured in six spontaneously whelping bitches and in six bitches in which parturition was induced with the progesterone-receptor blocker aglépristone on day 58 of pregnancy. Expulsion of pups in the induced group took place in the presence of P4 concentrations that were still elevated. PGFM concentrations increased before parturition in both groups, but levels were lower in the induced bitches. PGFM levels reached a maximum in both groups during parturition and quickly decreased in the spontaneously whelping group after parturition, but remained elevated in the induced group. In both groups, cortisol concentrations reached similar maximum levels during the last 30 h before the onset of expulsion. During the 3 days postpartum, cortisol concentrations were higher in the induced group. The highly variable ACTH concentrations did not differ significantly throughout the study within or between groups. In both groups, E2beta concentrations decreased and PRL concentrations increased between the late gestational period and the 30-h period before parturition. Concentrations of both LH (spontaneously whelping group) and FSH (both groups) decreased between late gestation and the postpartum period. The results of this study illustrate the hormonal changes around parturition in the bitch, and reveal that aglépristone-induced parturition is associated with still incomplete luteolysis, an altered PGFM profile, and elevated postpartum cortisol concentrations as compared with spontaneously whelping dogs.

During lactation, daily separation of sow and piglets, intermittent suckling (IS), can induce lactational oestrus and ovulation. This study examined effects of IS on subsequent early embryo survival and development. Multiparous Topigs40 sows were separated from their piglets for either 12 consecutive hours per day (IS12, n = 13) or two times for 6 h per day (IS6, n = 10) from day 14 of lactation onwards until 23 days after ovulation. Control sows (C, n = 17) were weaned at day 21 of lactation. Oestrus was shown in all treatments within 5 days after the start of treatment. Sows were inseminated each day of oestrus and slaughtered at D23 after ovulation. Intermittent suckling did not significantly affect pregnancy rates of sows (75% IS12 vs 78% IS6 vs 94% C; p > 0.10). Embryo survival was not significantly affected by IS (IS12: 57%; IS6: 51%; p > 0.10) although it seemed to be lower than in C sows (70%). Some parameters of embryo, placental and uterine development were affected by IS, especially in the IS6 group. IS6 embryos had shorter placentas (17.5 +/- 1.2 cm; p < 0.05) than C (20.3 +/- 1.4 cm) and IS12 sows (20.9 +/- 0.7 cm) were smaller and less developed than C sows (p < 0.05). In conclusion, embryo survival does not seem significantly affected by IS, although numerical differences were great. Embryo development, however, was negatively affected in IS6 sows possibly due to a combination of high milk production, stress and lactational effects on uterine development.

The objectives of the current study were to determine how intermittent suckling (IS) affects nursing behavior, litter activity, and general behavioral patterns during lactation, and whether IS during an extended lactation period results in behavioral patterns associated with piglet distress. Intermittent suckling was applied either with 6-h separation intervals (IS6) or with 12-h separation intervals (IS12) and was compared with the conventional treatment (CT). In the CT (n = 17 litters), sows were continuously present until weaning (d 21, d 0 = farrowing). In both IS6 and IS12, sows were separated from their litter for 12 h/d, beginning at d 14 and lasting until weaning (d 43 +/- 1 d). In IS6, litters (n = 14) and sows were separated from 0800 to 1400 and from 2000 to 0200; in IS12 litters (n = 14) and sows were separated between 0800 and 2000. In IS litters, the activity pattern over the 24-h cycle was markedly changed by IS; litter activity was lower (P < 0.001) during sow absence and greater (P < 0.001) during sow presence compared with the unweaned CT litters. Moreover, both total nursing frequency (P < 0.001) and the percentage (P < 0.002) of successful nursings were reduced by IS. Although total nursing frequency was greater in IS6 compared with IS12 (on d 21 and 28), no differences in the frequency of successful nursings existed between IS6 and IS12 from d 14 onward. Eating behavior was increased shortly after the onset of IS (d 17) in both IS6 (P = 0.059) and IS12 (P < 0.001) compared with the unweaned CT litters. The IS12 litters showed more eating behavior compared with IS6 and their exploratory behavior increased in time (P < 0.001), whereas IS6 showed more nursing behavior. Aggressive or manipulative behavior of both IS treatments was similar compared with the unweaned CT, and remained relatively unaltered with time in IS12 and IS6. Weaning in the CT resulted in more manipulative (P < 0.001) and aggressive (P = 0.004) behavior compared with pre-weaning values. Intermittent suckling may contribute to adaptation to the postweaning state by stimulating eating behavior, without causing obvious behavioral distress.

While oxytocin is widely used in the treatment of dystocia in dogs, there is little information about its secretion before and during normal unassisted whelping. We therefore measured plasma oxytocin concentrations during late pregnancy and the expulsive stage of parturition. Blood samples were collected from eight dogs at 3-min intervals during a 42-min period between the 2nd and 14th day before whelping and during parturition after the birth of 1-3 pups. The litters consisted of 5-15 pups and the progression of the expulsive stage was linear and nearly parallel in the eight bitches. The overall mean (+/-S.D.) plasma oxytocin concentration during late pregnancy was 3.6+/-2.1pg/ml. Mean values in individual dogs ranged from 1.2 to 7.4 pg/ml, but the intra-animal variation was rather small. During the expulsive stage the overall mean (+/-S.D.) plasma oxytocin concentration was 12.9+/-13.9 pg/ml, with mean values in individual dogs ranging from 3.5 to 46 pg/ml. The mean area under the oxytocin curve for parturient dogs was significantly higher (P<0.05) than for pregnant dogs. During the expulsive stage, the peak plasma oxytocin level in individual dogs ranged between 10 and 117 pg/ml. In six of the eight dogs a pup was born during blood collection and in five of these animals the plasma oxytocin concentration increased temporarily during periods of abdominal straining and expulsion. However, straining efforts and expulsion were not consistently associated with a rise in the circulating oxytocin level. It is concluded that in the dog plasma oxytocin levels are higher and more variable during the expulsive stage of parturition than during late pregnancy. Interrelationships between the secretion pattern of oxytocin, the level of uterine contractility, and the progress of fetal expulsion in dogs need further exploration.

The cervix must regain its normal diameter after parturition. Until now, little has been known about the pattern of cervical closure and the possible influences of myometrial and cervical contractions in this process. We continuously measured the cervical diameter with ultrasound cervimetry during the first 48h after calving in six cows with retained fetal membranes, while uterine (n=6) and cervical outer muscular layer (n=4) electromyographic (EMG) activity was measured with bipolar EMG electrodes. We found that the cervical diameter which was 6.2cm (+/-0.7) at 1.4h after calving, initially increased to 9.0cm (+/-1.0) during the first 14.8h (+/-2.8) postpartum. After this time, the diameter decreased gradually to 5.3cm (+/-1.0) at 48h after calving. The overall EMG activity after parturition decreased by 59% (+/-6) and 35% (+/-17) for the uterus and cervix, respectively. The decrease in EMG activity was due to a 50% (+/-7) decrease in EMG amplitudes of the myometrium; the EMG amplitudes of the cervix decreased by only 8% (+/-21) (P>0.05). At the same time in the cervix, burst frequency decreased by 69% (+/-17), while the decrease in burst frequency of the myometrium was only 11% (+/-5) (P>0.05). Uterine myometrial and cervical EMG activity after parturition showed burst patterns. These contractions of the uterus and cervix were accompanied by and correlated with transient dilatations of the caudal cervix. This could have functional relevance in the evacuation of the uterus.

In this study we described and analysed changes in the numbers of large ovarian follicles (diameter 6.1-9.0 cm) and in the plasma concentrations of luteinizing hormone (LH) and estradiol-17beta (E(2)beta) in relation to individual egg production figures of farmed ostriches (Struthio camelus spp.) throughout one year. Ultrasound scanning and blood sampling for plasma hormone analysis were performed in 9 hens on a monthly basis during the breeding season and in two periods of the non-breeding season. Our data demonstrated that: (1) large follicles were detected and LH concentrations were elevated already 1 month before first ovipositions of the egg production season took place; (2) E(2)beta concentrations increased as soon as the egg production season started; (3) numbers of large follicles, LH and E(2)beta concentrations were elevated during the entire egg production season; and that (4) numbers of large follicles, LH and E(2)beta concentrations decreased simultaneous with or following the last ovipositions of the egg production season. By comparing these parameters during the egg production season with their pre-and post-seasonal values, significant differences were found in the numbers of large follicles and E(2)beta concentrations between the pre-seasonal, seasonal and post-seasonal period; while LH concentrations were significantly different between the seasonal and post-seasonal period. In conclusion, our data demonstrate that changes in numbers of large follicles and in concentrations of LH and E(2)beta closely parallel individual egg production figures and provide some new cues that egg production in ostriches is confined to a marked reproductive season. Moreover, our data provide indications that mechanism, initiating, maintaining and terminating the egg production season in farmed breeding ostriches are quite similar to those already known for other seasonal breeding bird species.