Hox-A2 protein expression in avian jaws cartilages and muscle primordia development.

OBJECTIVE: To elucidate the branchial origin of the articular and the square (homology of the malleus and the incus of mammals), we used immunohistochemistry to analyse the expression of the Hox-A2 protein during cephalogenesis in chickens. MATERIALS AND METHODS: Immunohistochemistry on paraffin sections of embryos from stage HH16 to HH40. RESULTS: In addition to the columella (equivalent to the mammalian stapes), the joint between the articular and the quadrate bones, and the retro-articular process of the articular (homologous to the short process of the malleus) express Hox-A2, suggesting an intervention of the 2nd arch in their formation. However, we fortuitously observed very intense expression within the early muscle plate of the second arch, which then generalized to all cephalic muscles, and extended to the trunk's myotomes. In the cartilage, the presence of the protein disappeared at stage 35. DISCUSSION AND CONCLUSION: The present results, while confirming the contribution of the second arch to the development of avian equivalents of the mammalian ear ossicles, strongly suggest that the Hox-A2 gene plays a role in muscle development, which remains to be elucidated by more sophisticated techniques.

Standardized statement for the ethical use of human cadaveric tissues in anatomy research papers: Recommendations from Anatomical Journal Editors-in-Chief.

Human cadaveric donors are essential for research in the anatomical sciences. However, many research papers in the anatomical sciences often omit a statement regarding the ethical use of the donor cadavers or, as no current standardized versions exist, use language that is extremely varied. To rectify this issue, 22 editors-in-chief of anatomical journals, representing 17 different countries, developed standardized and simplified language that can be used by authors of studies that use human cadaveric tissues. The goal of these editor recommendations is to standardize the writing approach by which the ethical use of cadaveric donors is acknowledged in anatomical studies that use donor human cadavers. Such sections in anatomical papers will help elevate our discipline and promote standardized language use in others non anatomy journals and also other media outlets that use cadaveric tissues.

Body mass prediction from femoral volume and sixteen other femoral variables in the elderly: BMI and adipose tissue effects.

OBJECTIVES: The frequently used prediction equations of body mass do not seem appropriate for elderly individuals. Here, we establish the relationship between femoral dimensions and known body mass in elderly individuals in order to develop prediction formulas and identify the factors affecting their accuracy. MATERIALS AND METHODS: The body mass linear least-squares regression is based on 17 femoral dimensions, including femoral volume, and 66 individuals. Body proportion and composition effects on accuracy are analyzed by means of the body mass index (BMI) and on a subset sample (n = 25), by means of the masses of adipose, bone and muscle tissues. RESULTS: Most variables significantly reflect body mass. Among them, six dimensions (e.g., biepicondylar breadth, femoral volume, and head femoral diameter) present percent standard errors of estimate ranging from 9.5 to 11% (r = 0.72-0.81) in normal BMI samples. Correlations are clearly lower in samples with normal and abnormal BMI [r = 0.38-0.58; % of standard error of estimate (SEE) = 17.3-19.6%] and not significantly correlated in females (femoral volume) who present high proportions of abnormal BMI and adipose tissue. In the subset, femoral volume is well correlated with bone mass (r = 0.88; %SEE = 7.9%) and lean body mass (r = 0.67; %SEE = 17.2%). DISCUSSION: Our body mass estimation equations for elderly individuals are relevant since relatively low correlations are recurrent in studies using younger individuals of known body mass. However, age, sex, lifestyle, and skeleton considerations of studied populations can provide information about the relevance of the body mass estimation, which is dependent on the BMI classification and the proportion of adipose tissue. Our general considerations can be used for studies of younger individuals.

Further consideration of the curvature of the Neandertal Femur.

OBJECTIVES: Neandertal femora are particularly known for having a marked sagittal femoral curvature. This study examined femoral curvature in Neandertals in comparison to a modern human population from Belgium by the use of three-dimensional (3D) quadric surfaces modeled from the bone surface. 3D models provide detailed information and enabled femoral curvature to be analyzed in conjunction with other morphological parameters. MATERIALS AND METHODS: 3D models were created from CT scans of 75 modern human femora and 7 Neandertal femora. Quadric surfaces (QS) were created from the triangulated surface vertices in all areas of interest (neck, head, diaphyseal shaft, condyles) extracted from previously placed anatomical landmarks. The diaphyseal shaft was divided into five QS shapes and curvature was measured by degrees of difference between QS shapes. Each bone was placed in a local coordinate system enabling each bone to be analyzed in the same way. RESULTS: The use of 3D quadric surface fitting allowed the distribution of curvature with similarly curved femora to be analyzed and the different patterns of curvature between the two groups to be determined. The Neandertals were shown to have a higher degree of femoral curvature and a more distal point of femoral curvature than the modern human population from Belgium. CONCLUSIONS: Morphological aspects of the Neandertal femur are different from this modern human population although mainly seem unrelated to femoral curvature. The relative lack of correlations with other femoral bony morphological factors suggests femoral curvature variations may be related to other aspects.

Femoral curvature variability in modern humans using three-dimensional quadric surface fitting.

INTRODUCTION: This study analysed femoral curvature in a population from Belgium in conjunction with other morphological characteristics by the use of three-dimensional (3D) quadric surfaces (QS) modelled from the bone surface. METHODS: 3D models were created from computed tomography data of 75 femoral modern human bones. Anatomical landmarks (ALs) were palpated in specific bony areas of the femur (shaft, condyles, neck and head). QS were then created from the surface vertices which enclose these ALs. The diaphyseal shaft was divided into five QS shapes to analyse curvature in different parts of the shaft. RESULTS: Femoral bending differs in different parts of the diaphyseal shaft. The greatest degree of curvature was found in the distal shaft (mean 4.5° range 0.2°-10°) followed by the proximal (mean 4.4° range 1.5°-10.2°), proximal intermediate (mean 3.7° range 0.9°-7.9°) and distal intermediate (mean 1.8° range 0.2°-5.6°) shaft sections. The proximal and distal angles were significantly more bowed than the intermediate proximal and the intermediate distal angle. There was no significant difference between the proximal and distal angle. No significant correlations were found between morphological characteristics and femoral curvature. An extremely large variability of femoral curvature with several bones displaying very high or low degrees of femoral curvature was also found. CONCLUSION: 3D QS fitting enables the creation of accurate models which can discriminate between different patterns in similar curvatures and demonstrates there is a clear difference between curvature in different parts of the shaft.

[Application of the musculo-skeletal modelling software lhpFusionBox to a paleoanthropological problem: the Spyrou Neandertal moves!]. / Application du logiciel de modélisation musculo-squelettique lhpFusionBox à une problématique paléo-anthropologique - Spyrou le Néandertalien marche !

LhpFusionBox is a program originally designed for biomechanical and clinical studies relating to the musculoskeletal system of anatomically modern humans (AMH). The program has recently been adapted for paleontological purposes and used to reconstruct and biomechanically analyse a fossil hominid. There is no complete Neandertal skeleton in the fossil record. The aim of the study was to reconstruct a complete three-dimensional (3D) model of a Neandertal using the relatively complete Spy II Neandertal and to conduct biomechanical feasibility studies on the knee and hamstring moment arms of the skeleton. Different Neandertal specimens were scaled to the size of Spy II to replace incomplete or missing bones. Biomechanical feasibility studies performed on the knee seem to show that Neandertal and AMHh gait is similar and Neandertals were shown to have larger moment arms in the hamstring muscles, which would have given them a mechanical advantage. The complete Neandertal was printed in 3D and used as the base to create the artistic model of "Spyrou" housed at l'Espace de l'Homme de Spy (EHoS) museum.

Impact of the mandibular divergence on the position of the inferior alveolar nerve and mylohyoid nerve: a computed tomography study and its relevance to bilateral sagittal split osteotomy.

PURPOSE: Bilateral sagittal split osteotomy (BSSO) is the most common procedure used to treat mandibular deformities. BSSO procedures include the Epker technique and the Dal Pont technique which are the most frequently used. Because of the intramandibular pathway of the inferior alveolar nerve (IAN), neurosensory disturbance of the lower lip and chin is the most common complication of BSSO. This study performed quantitative measurements from computed tomographic (CT) data obtained on dry human mandibles. The main aim of the study was to evaluate if mandibular divergence can predict the position of the IAN and the mylohyoid nerve (MHN) to prevent nerve injury. METHODS: After CT and 3D reconstruction of 65 dry mandibles, 30 measurements were made on 3 planes for each hemi-mandible. This allowed analysis of the IAN and MHN pathways. Three groups of hemi-mandibles were created depending on their divergence, and a statistical analysis was performed. RESULTS: Eight out of the 30 measurements showed a significant difference among the 3 groups. There was no significant difference for the remaining 22 measurements. CONCLUSIONS: The IAN seems to have a more superior position in the groups of mandibular hypo- and hyper-divergence. Orthognathic surgeons should use a more superficial retromolar bone incision in these cases. Finally, the Epker technique would be safer for preserving the MHN in normo- and hypo-divergent patients.

Evolution and development of parrot pseudoteeth.

Parrot embryos carry peculiar appendages at their developing beak that have been described as pseudoteeth. To better characterize the pattern of development responsible for the emergence of these dental appendages, we examined parrot embryos combining conventional histology and microtomography approaches. Using immunohistochemistry, we observed the epithelial and mesenchymal expression of several proteins involved in tooth development in mammals. Parrot pseudoteeth arose by epithelial and mesenchymal evagination, and their early development was similar to the ontogeny of scales and feathers. There was no enamel tissue, and the evaginations were surrounded by the rhamphotheca. In adults, the rhamphotheca covers entirely the appendages, now represented by bone evaginations, which were more numerous in the lower than in the upper beak, being similar to the osseous teeth of the fossil Pelagornithidae. These embryonic pseudoteeth resembled reptile's first-generation teeth and dental appendages of chicken talpid2 mutants. Proteins involved in mammalian odontogenesis, such as SHH, BMP4, PITX2, and PAX9, were found to be generally expressed in beak epithelium and mesenchyme during parrot pseudoteeth development, with clusters of high-level expression in the pseudoteeth rudiments. This suggests that a similar, highly conserved gene expression program gives rise to the appearance of odontode derivatives in numerous species, despite their divergent developmental paths. These results provide new insights into the development and evolution of odontode-derived structures in vertebrates.

Expression of Msx1 and Dlx1 during Dumbo rat head development: Correlation with morphological features.

The Dumbo rat possesses some characteristics that evoke several human syndromes, such as Treacher-Collins: shortness of the maxillary, zygomatic and mandibular bones, and low position of the ears. Knowing that many homeobox genes are candidates in craniofacial development, we investigated the involvement of the Msx1 and Dlx1 genes in the Dumbo phenotype with the aim of understanding their possible role in abnormal craniofacial morphogenesis and examining the possibility of using Dumbo rat as an experimental model for understanding abnormal craniofacial development. We studied the expression of these genes during craniofacial morphogenesis by RT-PCR method. We used Dumbo embryos at E12 and E14 and included the Wistar strain as a control. Semi-quantitative PCR analysis demonstrated that Msx1 and Dlx1 are expressed differently between Dumbo and Wistar rats, indicating that their low expression may underly the Dumbo phenotype.

Development of the Goose Tongue Filiform Papillae: Could it be Tooth-Like Sense Organs? / Desarrollo de las Papilas Filiformes de la Lengua de Ganso: ¿Podrían ser Órganos Sensoriales Parecidos a los Dientes?

SUMMARY: The geese's tongue filiform papillae are particularly long, and exhibit the same morphology of a tooth, evoking the lingual teeth of several fishes. In adult animals, they contain numerous mechanical Herbst's corpuscles but no taste buds. In the embryo, they appear since stage 38 and acquire their definitive shape between stages 38 and 42. They express several proteins associated with mammalian tooth development (BMP4, β-catenin, SHH, PITX2, PAX9), also known to be linked to parrot's pseudoteeth and goose's denticulations development. Neurofilaments are early present in the papillae primordia, and appear particularly numerous in adult papillae. Our results suggest that these papillae constitute a mechanical organ with a « tooth shape » derived from ancestral odontodes, whose development is controlled by numerous genes involved in classical odontogenesis.

Las papilas filiformes de la lengua de los gansos son particularmente largas y exhiben la morfología de un diente, evocando los dientes linguales presentes en varios peces. En los animales adultos, contienen numerosos corpúsculos de Herbst mecánicos, aunque una ausencia de papilas gustativas. En el embrión, aparecen a partir del estadio 38 y adquieren su forma definitiva entre los estadios 38 y 42. Expresan varias proteínas asociadas al desarrollo dentario de los mamíferos (BMP4, β-catenina, SHH, PITX2, PAX9), también conocidas por estar asociadas al desarrollo de pseudodientes en el loro y denticulaciones en el ganso. Los neurofilamentos están presentes tempranamente en los primordios de las papilas y aparecen particularmente numerosos en las papilas adultas. Nuestros resultados sugieren que estas papilas constituyen un órgano mecánico con «forma de diente» derivado de odontoides ancestrales, cuyo desarrollo está controlado por numerosos genes implicados en la odontogénesis clásica.

How different are the Kebara 2 ribs to modern humans?

This study analyses rib geometric parameters of individual ribs of 14 modern human subjects (7 males and 7 females) in comparison to the reconstructed ribs of the Kebara 2 skeleton which was taken from the reconstruction of a Neandertal thorax by Sawyer & Maley (2005). Three-dimensional (3D) models were segmented from CT scans and each rib vertex cloud was placed into a local coordinate system defined from the rib principal axes. Rib clouds were then analysed using best fitting ellipses of the external contours of the cross-section areas. The centroid of each ellipse was then used to measure the centroidal pathway between each slice (rib midline). Curvature of the ribs was measured from the mid-line of the ribs as the sum of angles between successive centroids in adjacent cross sections. Distinct common patterns were noted in all rib geometric parameters for modern humans. The Kebara 2 reconstructed ribs also followed the same patterns. This study demonstrated that there are differences between the sexes in rib geometrical parameters, with females showing smaller rib width, chord length and arc length, but greater curvature (rib torsion, rib axial curvature, rib anterior-posterior bending) than males. The Kebara 2 reconstructed ribs were within the modern human range for the majority of geometrical parameters.

Effect of magnetic field and iron content on NMR proton relaxation of liver, spleen and brain tissues.

Iron accumulation is observed in liver and spleen during hemochromatosis and important neurodegenerative diseases involve iron overload in brain. Storage of iron is ensured by ferritin, which contains a magnetic core. It causes a darkening on T2 -weighted MR images. This work aims at improving the understanding of the NMR relaxation of iron-loaded human tissues, which is necessary to develop protocols of iron content measurements by MRI. Relaxation times measurements on brain, liver and spleen samples were realized at different magnetic fields. Iron content was determined by atomic emission spectroscopy. For all samples, the longitudinal relaxation rate (1/T1 ) of tissue protons decreases with the magnetic field up to 1 T, independently of iron content, while their transverse relaxation rate (1/T2 ) strongly increases with the field, either linearly or quadratically, or a combination thereof. The extent of the inter-echo time dependence of 1/T2 also varies according to the sample. A combination of theoretical models is necessary to describe the relaxation of iron-containing tissues. This can be due to the presence, inside tissues, of ferritin clusters of different sizes and densities. When considering all samples, a correlation (r(2) = 0.6) between 1/T1 and iron concentration is observed at 7.0 T. In contrast the correlation between 1/T2 and iron content is poor, even at high field (r(2) = 0.14 at 7.0 T). Our results show that MRI methods based on T1 or T2 measurements will easily detect an iron overloading at high magnetic field, but will not provide an accurate quantification of tissue iron content at low iron concentrations.

Heat shock protein HSP86 expression during mouse embryo development, especially in the germ-line.

In addition to appearing in response to biological stresses, heat shock proteins are expressed as 'chaperones' by some cells living in physiological conditions. Among these proteins, the HSP90 family seems to function under normal growth conditions in the pathways of numerous signal transducers, cell-cycle and developmental regulators. The aim of this study was to investigate the immunohistochemical expression and the distribution of the two mammalian cytosolic isoforms of HSP90 (HSP84 and HSP86) during mouse development between E6 and E16. Our immunohistochemical investigations revealed a diffuse and ubiquitous expression of HSP84 distributed throughout the embryo in all stages. On the other hand, HSP86 was specifically noticed in diverse somatic cells such as hepatoblasts and epithelia that could traduce cellular differentiation phenomenon. More interestingly, a strong cytoplasm expression of HSP86 could be demonstrated in primordial germ cells from their appearance on the yolk sac membrane until reaching the genital crest. These results suggested a protective mechanism against the phenotypic expression of mutations induced by environmental changes in vertebrates as has been demonstrated in Drosophila.

A Mesolithic case of odontoma?

An ovoid yellowish object has been discovered in an Early Mesolithic collective burial located in the Meuse Basin: the Autours rockshelter (Prov. of Namur, Belgium). It was found among commingled hand and foot bones in a small crack of the rockshelter wall. Mineralogical, radiological and microscopic analyses showed that it was most probably a complex odontoma.

Development of multimedia learning modules for teaching human anatomy: application to osteology and functional anatomy.

Computer-assisted learning (CAL) is growing quickly within academic programs. Although the anatomical commercial packages that are available for this learning have attractive advantages, they also have drawbacks: they are frequently not in the local language of the students, they do not perfectly answer the needs of the local academic program, and their cost is frequently more than students can afford. This study describes a relatively inexpensive method to create CAL tutorials, whose content can be fully customized to local academic needs in terms of both program and language. The study describes its use in creating multimedia learning modules (MLMs) about Osteology and joint kinematics. The pedagogical content in these modules was collected from objective experiments to give students the opportunity to access new scientific knowledge during their education. It can be replaced, as desired, by almost any content due to the flexibility of the production method. Each MLM consists of two complementary subelements: a multimedia theoretical lecture and a three-dimensional interactive laboratory. Such MLMs are in use at both the University of Brussels (ULB) and the National University of Rwanda (NUR). The development of this work was part of the VAKHUM project, and the pedagogical validation is currently being performed as part of the MULTIMOD project.

Sex determination using the Probabilistic Sex Diagnosis (DSP: Diagnose Sexuelle Probabiliste) tool in a virtual environment.

The hip bone is one of the most reliable indicators of sex in the human body due to the fact it is the most dimorphic bone. Probabilistic Sex Diagnosis (DSP: Diagnose Sexuelle Probabiliste) developed by Murail et al., in 2005, is a sex determination method based on a worldwide hip bone metrical database. Sex is determined by comparing specific measurements taken from each specimen using sliding callipers and computing the probability of specimens being female or male. In forensic science it is sometimes not possible to sex a body due to corpse decay or injury. Skeletalization and dissection of a body is a laborious process and desecrates the body. There were two aims to this study. The first aim was to examine the accuracy of the DSP method in comparison with a current visual sexing method on sex determination. A further aim was to see if it was possible to virtually utilise the DSP method on both the hip bone and the pelvic girdle in order to utilise this method for forensic sciences. For the first part of the study, forty-nine dry hip bones of unknown sex were obtained from the Body Donation Programme of the Université Libre de Bruxelles (ULB). A comparison was made between DSP analysis and visual sexing on dry bone by two researchers. CT scans of bones were then analysed to obtain three-dimensional (3D) virtual models and the method of DSP was analysed virtually by importing the models into a customised software programme called lhpFusionBox which was developed at ULB. The software enables DSP distances to be measured via virtually-palpated bony landmarks. There was found to be 100% agreement of sex between the manual and virtual DSP method. The second part of the study aimed to further validate the method by analysing thirty-nine supplementary pelvic girdles of known sex blind. There was found to be a 100% accuracy rate further demonstrating that the virtual DSP method is robust. Statistically significant differences were found in the identification of sex between researchers in the visual sexing method although both researchers identified the same sex in all cases in the manual and virtual DSP methods for both the hip bones and pelvic girdles.

Caspase-2 immunohistochemistry in differentiating cells during mouse cephalic development

Caspases are proteases primarily involved in the process of apoptosis; however, caspases can exert non-apoptotic functions. The purpose of this work was to use immunohistochemistry to analyse the expression sites of caspase-2 during normal mouse cephalic development and in embryos exposed to irradiation. Control embryos from embryonic day 9 (E9) to E17 were analysed, and E9 and 10 irradiated embryos were removed and observed after administration of 2 Gy irradiation at embryonic day 9. Surprisingly, not only apoptotic cells expressed caspase-2. In addition, numerous cell populations in normal and experimental embryos displayed transient but intense caspase-2 immunoreactivity, with nuclear and cytoplasmic localisation. This immunoreactivity was not observed with caspase-3 and -9 antibodies. Cranial neural crest cells, premuscular blastemata, cartilage, teeth, the heart, the eye and some other structures displayed caspase-2 expression, with progressive changes during embryonic development. These changing patterns evoke progressive waves of cell differentiation in specific cell populations. Little is known regarding the non-apoptotic functions of caspase-2. Despite the difficulty in understanding the role of this protease during cell differentiation, the fact that caspase-2 is known to prevent DNA damage and to protect the cell cycle could be closely associated with our observations, which point to the need for further research, particularly in caspase-2 knockout mice

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The development of several organs and appendages is impaired in mice lacking Sp6.

SP6 belongs to the SP/KLF family of transcription factors, characterized by a DNA-binding domain composed of three zinc fingers of the C(2)H(2) type. The Sp6 gene generates two different transcripts, termed Sp6 and epiprofin, which differ in the first exon and encode the same SP6 protein. These transcripts are mainly expressed in the skin, the teeth, and the limb buds of embryos and also in the adult lungs. To gain insight into the biological function of the SP6 protein, we knocked out the gene by eliminating the full coding region. The resulting Sp6 null mice are nude, lack functional teeth, and present limb and lung malformations. We also showed that the identified abnormalities are associated with apoptotic misregulations. In conclusion, this work indicates that Sp6 plays a critical role in the development of several epithelium-containing organs or appendages, possibly by regulating apoptosis.