Morphometric analysis of embryonic development in Alligator mississippiensis, Crocodylus johnstoni and Crocodylus porosus

The size of embryos at various stages of development was determined in three species of crocodilian ( Alligator mississippiensis, Crocodylus johnstoni and C. porosus). Various morphometric measurements were taken of embryos throughout development and were described for each stage of development. Increase in size from stage to stage was faster in A. mississippiensis than in C. porosus and C. johnstoni. Hatchlings of A. mississippiensis were large in length but light in mass compared with the hatchlings of C. johnstoni and C. porosus which were heavier per unit length. These morphometric parameters can be used to determine the stage of embryonic development by size. The use of principle component analysis improves this technique further by dampening any anomalous data points.
The rate of embryonic growth in A. mississippiensis appeared to be under greater genetic control than in the two species of Crocodylus. The evolutionary advantages of this phenomenon probably relate to the biology of A. mississippiensis. Due to the northerly range of this species it is advantageous for alligators to hatch as soon as possible, as large as possible, to maximize the period prior to winter hibernation and reduce predation. Tropical crocodiles have fewer selection pressures for rapid development and have slower rates of embryonic growth. Genetic aspects of crocodilian embryonic development have been largely ignored but may help explain some aspects of crocodilian growth under farming conditions.     

Ultrastructure of the spermatozoon of the American Alligator,Alligator mississippiensis (Reptilia: Alligatoridae)

This study details the ultrastructure of the spermatozoa of the American Alligator, Alligator mississippiensis. American Alligator spermatozoa are filiform and slightly curved. The acrosome is tapered at its anterior end and surrounded by the acrosome vesicle and an underlying subacrosomal cone, which rests just cephalic to the nuclear rostrum. One endonuclear canal extends from the subacrosomal cone through the rostral nucleus and deep into the nuclear body. The neck region separates the nucleus and midpiece and houses the proximal centriole and pericentriolar material. The distal centriole extends through the midpiece and has 9 × 3 sets of peripheral microtubules with a central doublet pair within the axoneme that is surrounded by a dense sheath. The midpiece is composed of seven to nine rings of mitochondria, which have combinations of concentrically and septate cristae. The principal piece has a dense fibrous sheath that surrounds an axoneme with a 9 + 2 microtubule arrangement. The sheath becomes significantly reduced in size caudally within the principal piece and is completely missing from the endpiece. Dense peripheral fibers, especially those associated with microtubule doublets 3 and 8, penetrate into the anterior portion of the principal piece axoneme. The data reported here hypothesize that sperm morphology is highly conserved in Crocodylia; however, specific morphological differences can exist between species. J. Morphol. 2011. © 2011 Wiley‐Liss, Inc.     

Using 3D finite element models verified the importance of callus material and microstructure in biomechanics restoration during bone defect repair

Background: There is lack of further observations on the microstructure and material property of callus during bone defect healing and the relationships between callus properties and the mechanical strength. Methods: Femur bone defect model was created in rabbits and harvested CT data to reconstruct finite element models at 1 and 2 months. Three types of assumed finite element models were compared to study the callus properties, which assumed the material elastic property as heterogeneous (R-model), homogenous (H-model) or did not change from 1 to 2 months (U-model). Results: The apparent elastic moduli increased at 2 months (from 355.58 ± 132.67 to 1139.30 ± 967.43 MPa) in R-models. But there was no significant difference in apparent elastic moduli between R-models (355.58 ± 132.67 and 1139.30 ± 967.43 MPa) and H-models (344.79 ± 138.73 and 1001.52 ± 692.12 MPa) in 1 and 2 months. A significant difference of apparent elastic moduli was found between the R-model (1139.30 ± 967.43 MPa) and U-model group (207.15 ± 64.60 MPa) in 2 months. Conclusions: This study showed that the callus structure stability remodeled overtime to achieve a more effective structure, while the material quality of callus tissue is a very important factor for callus strength. At the meantime, this study showed an evidence that the material heterogeneity maybe not as important as it is in bone fracture model.     

Development of the dermal skeleton in Alligator mississippiensis (Archosauria, Crocodylia) with comments on the homology of osteoderms

The dermal skeleton (=exoskeleton) has long been recognized as a major determinant of vertebrate morphology. Until recently however, details of tissue development and diversity, particularly among amniotes, have been lacking. This investigation explores the development of the dermatocranium, gastralia, and osteoderms in the American alligator, Alligator mississippiensis. With the exception of osteoderms, elements of the dermal skeleton develop early during skeletogenesis, with most initiating ossification prior to mineralization of the endoskeleton. Characteristically, circumoral elements of the dermatocranium, including the pterygoid and dentigerous elements, are among the first to form. Unlike other axially arranged bones, gastralia develop in a caudolateral to craniomedial sequence. Osteoderms demonstrate a delayed onset of development compared with the rest of the skeleton, not appearing until well after hatching. Osteoderm development is asynchronous across the body, first forming dorsally adjacent to the cervical vertebrae; the majority of successive elements appear in caudal and lateral positions. Exclusive of osteoderms, the dermal skeleton initiates osteogenesis via intramembranous ossification. Following the establishment of skeletal condensations, some preossified spicules become engorged with many closely packed clusters of chondrocyte-like cells in a bone-like matrix. This combination of features is characteristic of chondroid bone, a tissue otherwise unreported among nonavian reptiles. No secondary cartilage was identified in any of the specimens examined. With continued growth, dermal bone (including chondroid bone) and osteoid are resorbed by multinucleated osteoclasts. However, there is no evidence that these cells contribute to the rugose pattern of bony ornamentation characteristic of the crocodylian dermatocranium. Instead, ornamentation develops as a result of localized concentrations of bone deposited by osteoblasts. Osteoderms develop in the absence of osteoblastic cells, osteoid, and periosteum; bone develops via the direct transformation of the preexisting dense irregular connective tissue. This mode of bone formation is identified as metaplasia. Importantly, it is also demonstrated that osteoderms are not histologically uniform but involve a range of tissues including calcified and uncalcified dense irregular connective tissue. Between taxa, not all osteoderms develop by homologous processes. However, it is concluded that all osteoderms may share a deep homology, connected by the structural and skeletogenic properties of the dermis.     

Three-dimensional finite element stress analysis of the dentate human mandible.

The biomechanical events which accompany functional loading of the human mandible are not fully understood. The techniques normally used to record them are highly invasive. Computer modelling offers a promising alternative approach in this regard, with the additional ability to predict regional stresses and strains in inaccessible locations. In this study, we built two three-dimensional finite element (FE) models of a human mandible reconstructed from tomographs of a dry dentate jaw. The first model was used for a complete mechanical characterization of physical events. It also provided comparative data for the second model, which had an increased vertical corpus depth. In both cases, boundary conditions included rigid restraints at the first right molar and endosteal cortical surfaces of the articular eminences of temporal bones. Groups of parallel multiple vectors simulated individual masticatory muscle loads. The models were solved for displacements, stresses, strains, and forces. The simulated muscle loads in the first model deformed the mandible helically upward and toward its right (working) side. The highest principal stresses occurred at the bite point, anterior aspects of the coronoid processes, symphyseal region, and right and left sides of the mandibular corpus. In general, the observed principal stresses and strains were highest on the periosteal cortical surface and alveolar bone. At the symphyseal region, maximum principal stresses and strains were highest on the lower lingual mandibular aspect, whereas minimum principal stresses and strains were highest on its upper labial side. Subcondylar principal strains and condylar forces were higher on the left (balancing or nonbiting) side than on the right mandibular side, with condylar forces more concentrated on the anteromedial aspect of the working-side condyle and on the central and lateral aspects of the left. When compared with in vivo strain data from macaques during comparable biting events, the predictive strain values from the first model were qualitatively similar. In the second model, the reduced tensile stress on the working-side, and decreased shear stress bilaterally, confirmed that lower stresses occurred on the lower mandibular border with increased jaw depth. Our results suggested that although the mandible behaved in a beam-like manner, its corpus acted more like a combination of open and closed cross sections due to the presence of tooth sockets, at least for the task modelled.(ABSTRACT TRUNCATED AT 400 WORDS)     

Influence of bicortical techniques in internal connection placed in premaxillary area by 3D finite element analysis

The aim of study was to evaluate the stress distribution in implant-supported prostheses and peri-implant bone using internal hexagon (IH) implants in the premaxillary area, varying surgical techniques (conventional, bicortical and bicortical in association with nasal floor elevation), and loading directions (0°, 30° and 60°) by three-dimensional (3D) finite element analysis. Three models were designed with Invesalius, Rhinoceros 3D and Solidworks software. Each model contained a bone block of the premaxillary area including an implant (IH, Ø4 × 10 mm) supporting a metal-ceramic crown. 178 N was applied in different inclinations (0°, 30°, 60°). The results were analyzed by von Mises, maximum principal stress, microstrain and displacement maps including ANOVA statistical test for some situations. Von Mises maps of implant, screws and abutment showed increase of stress concentration as increased loading inclination. Bicortical techniques showed reduction in implant apical area and in the head of fixation screws. Bicortical techniques showed slight increase stress in cortical bone in the maximum principal stress and microstrain maps under 60° loading. No differences in bone tissue regarding surgical techniques were observed. As conclusion, non-axial loads increased stress concentration in all maps. Bicortical techniques showed lower stress for implant and screw; however, there was slightly higher stress on cortical bone only under loads of higher inclinations (60°).     

Multi-scale finite element modeling allows the mechanics of amphibian neurulation to be elucidated

The novel multi-scale computational approach introduced here makes possible a new means for testing hypotheses about the forces that drive specific morphogenetic movements. A 3D model based on this approach is used to investigate neurulation in the axolotl (Ambystoma mexicanum), a type of amphibian. The model is based on geometric data from 3D surface reconstructions of live embryos and from serial sections. Tissue properties are described by a system of cell-based constitutive equations, and parameters in the equations are determined from physical tests. The model includes the effects of Shroom-activated neural ridge reshaping and lamellipodium-driven convergent extension. A typical whole-embryo model consists of 10,239 elements and to run its 100 incremental time steps requires 2 days. The model shows that a normal phenotype does not result if lamellipodium forces are uniform across the width of the neural plate; but it can result if the lamellipodium forces decrease from a maximum value at the mid-sagittal plane to zero at the plate edge. Even the seemingly simple motions of neurulation are found to contain important features that would remain hidden, they were not studied using an advanced computational model. The present model operates in a setting where data are extremely sparse and an important outcome of the study is a better understanding of the role of computational models in such environments.     

Estimation of conditions evoking fracture in finger bones under pinch loading based on finite element analysis

A finger finite element (FE) model was created from CT images of a Japanese male in order to obtain a shape-biofidelic model. Material properties and articulation characteristics of the model were taken from the literature. To predict bone fracture and realistically represent the fracture pattern under various loading conditions, the ESI-Wilkins-Kamoulakos rupture model in PAM-CRASH (ESI Group S.A., Paris, France) was utilized in this study with parameter values of the rupture model determined by compression testing and simulation of porcine fibula. A finger pinch simulation was then conducted to validate the finger FE model. The force-displacement curve and fracture load from the pinch simulation was compared to the result of finger pinch test using cadavers. Simulation results are coincident with the test result, indicating that the finger FE model can be used in an analysis of finger bone fracture during pinch accident. With this model, several pinch simulations were conducted with different pinching object’s stiffness and pinching energy. Conditions for evoking finger bone fracture under pinch loading were then estimated based on these results. This study offers a novel method to predict possible hazards of manufactured goods during the design process, thus finger injury due to pinch loading can be avoided.     

Multiple paternity and mating patterns in the American alligator, Alligator mississippiensis

Eggs were sampled from 22 wild American alligator nests from the Rockefeller Wildlife Refuge in south-west Louisiana, along with the females guarding the nests. Three nests were sampled in 1995 and 19 were sampled in 1997. Females and offspring from all clutches were genotyped using five polymorphic microsatellite loci and the three nests from 1995 were also genotyped using one allozyme locus. Genotypes of the hatchlings were consistent with the guarding females being the mothers of their respective clutches. Multiple paternity was found in seven of the 22 clutches with one being fathered by three males, and the remaining six clutches having genotypes consistent with two males per clutch. Paternal contributions of multiply sired clutches were skewed. Some males sired hatchlings of more than one of the 22 clutches either as one of two sires of a multiple paternity clutch, as the sole sire of two different clutches, or as the sole sire of one clutch and one of two sires of a multiply sired clutch. There was no significant difference between females that had multiple paternity clutches and those that had singly sired clutches with respect to female total length (P = 0.844) and clutch size (P = 0.861). Also, there was no significant correlation between genetic relatedness of nesting females and pairwise nest distances (r2 = 0.003, F1,208 = 0.623, P = 0.431), indicating that females in this sample that nested close to one another were no more related than any two nesting females chosen at random. Eleven mutations were detected among hatchlings at the five loci over the 22 clutches. Most of these mutations (eight of 11) occurred at Ami(mu)-17, the only compound microsatellite locus of the five used in this study, corresponding to a mutation rate of 1.7 x 10-3. Finally, most of the mutations (82%) were homoplasious, i.e., mutating to an allelic state already present in this Louisiana population.     

Fluctuating water depths affect American alligator (Alligator mississippiensis) body condition in the Everglades,Florida, USA

Successful restoration of wetland ecosystems requires knowledge of wetland hydrologic patterns and an understanding of how those patterns affect wetland plant and animal populations. Within the Everglades, Florida, USA restoration, an applied science strategy including conceptual ecological models linking drivers to indicators is being used to organize current scientific understanding to support restoration efforts. A key driver of the ecosystem affecting the distribution and abundance of organisms is the timing, distribution, and volume of water flows that result in water depth patterns across the landscape. American alligators (Alligator mississippiensis) are one of the ecological indicators being used to assess Everglades restoration because they are a keystone species and integrate biological impacts of hydrological operations through all life stages. Alligator body condition (the relative fatness of an animal) is one of the metrics being used and targets have been set to allow us to track progress. We examined trends in alligator body condition using Fulton's K over a 15 year period (2000–2014) at seven different wetland areas within the Everglades ecosystem, assessed patterns and trends relative to restoration targets, and related those trends to hydrologic variables. We developed a series of 17 a priori hypotheses that we tested with an information theoretic approach to identify which hydrologic factors affect alligator body condition. Alligator body condition was highest throughout the Everglades during the early 2000s and is approximately 5–10% lower now (2014). Values have varied by year, area, and hydrology. Body condition was positively correlated with range in water depth and fall water depth. Our top model was the “Current” model and included variables that describe current year hydrology (spring depth, fall depth, hydroperiod, range, interaction of range and fall depth, interaction of range and hydroperiod). Across all models, interaction between range and fall water depth was the most important variable (relative weight of 1.0) followed by spring and fall water depths (0.99), range (0.96), hydroperiod (0.95) and interaction between range and hydroperiod (0.95). Our work provides additional evidence that restoring a greater range in annual water depths is important for improvement of alligator body condition and ecosystem function. This information can be incorporated into both planning and operations to assist in reaching Everglades restoration goals.     

Predicting the permeability of trabecular bone by micro-computed tomography and finite element modeling

The intrinsic permeability of bone plays an important role in the transport of nutrients and minerals within the tissue, and affects the mechanical stimuli that are related to the fate of the stem cells. The objective of this study was to establish a method to assess trabecular bone permeability using experimental and finite element (FE) modeling approaches based on micro computed tomography (µCT) images. Human cadaveric tibia cube specimens (N=23) were scanned with µCT. The permeability was measured experimentally using a custom-developed constant-head permeameter, and computationally by a poroelastic formulation to simulate the fluid flow within the discretized bone matrix and pore phase. The average of the experimentally measured permeability was 4.84×10⁻¹⁰ m² with a standard deviation of 3.70×10⁻¹⁰ m². A regression model of the µCT determined that the maximum bone area to total area ratio (maxBA/TA) for all slices that are perpendicular to the direction of fluid flow explained 84% of the variability of the natural logarithm of the experimentally measured permeability. The 2D measure of maxBA/TA performed better than 3D measures in general, although some parameters were reasonably well associated with permeability such as bone volume ratio (BV/TV, r=−0.71), the bone surface/bone volume (BS/BV, r=0.73), and the trabecular thickness (TbTh, r=−0.71). The correlation between the permeability predicted with FE models and experimentally measured permeability was reasonable (r=0.69), but the FE approach did not accurately represent the wide variability of permeability measured experimentally. The results of this study suggest that the changes in the trabecular bone microarchitecture have an exponential relationship with permeability, and the use of µCT-based 2D measurement of maxBA/TA performs well at predicting permeability, thus providing a convenient approach to measure this important aspect affecting biomechanical functions in the tissue.     

Mechanical significance of femoral head trabecular bone structure in Loris and Galago evaluated using micromechanical finite element models

Work on the interspecific and intraspecific variation of trabecular bone in the proximal femur of primates demonstrates important architectural variation between animals with different locomotor behaviors. This variation is thought to be related to the processes of bone adaptation whereby bone structure is optimized to the mechanical environment. Micromechanical finite element models were created for the proximal femur of the leaping Galago senegalensis and the climbing and quadrupedal Loris tardigradus by converting bone voxels from high-resolution X-ray computed tomography scans of the femoral head to eight-noded brick elements. The resulting models had approximately 1.8 million elements each. Loading conditions representing takeoff phase of a leap and more generalized load orientations were applied to the models, and the models were solved using the iterative "row-by-row" matrix-vector multiplication algorithm. The principal strain and Von Mises stress results for the leaping model were similar for both species at each load orientation. Similar hip joint reaction forces in the range of 4.9 x to 12 x body weight were calculated for both species under each loading condition, but the hip reaction values estimated for Loris were higher than predicted based on locomotor behavior. These results suggest that functional adaptation to hip joint loading may not fully explain the differences in femoral head trabecular bone structure in Galago and Loris. The finite element method represents a unique and useful tool for analyzing the functional adaptation of trabecular bone in a diversity of animals and for reconstructing locomotor behavior in extinct taxa.     

Multiyear multiple paternity and mate fidelity in the American alligator, Alligator mississippiensis

We examined multiple paternity during eight breeding events within a 10-year period (1995–2005) for a total of 114 wild American alligator nests in Rockefeller Wildlife Refuge in south-west Louisiana. Our goals included examining (i) within population variation in multiple paternity among years, (ii) variation in multiple paternity in individual females and (iii) the potential for mate fidelity. To accomplish this, in the current study, eggs were sampled from 92 nests over 6 years and analysed along with 22 nests from a previous 2-year study. Genotypes at five microsatellite loci were generated for 1802 alligator hatchlings. Multiple paternity was found in 51% of clutches and paternal contributions to these clutches were highly skewed. Rates of multiple paternity varied widely among years and were consistently higher in the current study than previously reported for the same population. Larger females have larger clutches, but are not more likely to have multiply sired nests. However, small females are unlikely to have clutches with more than two sires. For 10 females, nests from multiple years were examined. Seven (70%) of these females exhibited long-term mate fidelity, with one female mating with the same male in 1997, 2002 and 2005. Five females exhibiting partial mate fidelity (71%) had at least one multiple paternity nest and thus mated with the same male, but not exclusively. These patterns of mate fidelity suggest a potential role for mate choice in alligators.     

Comparative finite element analysis of the cranial performance of four herbivorous marsupials

Marsupial herbivores exhibit a wide variety of skull shapes and sizes to exploit different ecological niches. Several studies on teeth, dentaries, and jaw adductor muscles indicate that marsupial herbivores exhibit different specializations for grazing and browsing. No studies, however, have examined the skulls of marsupial herbivores to determine the relationship between stress and strain, and the evolution of skull shape. The relationship between skull morphology, biomechanical performance, and diet was tested by applying the finite element method to the skulls of four marsupial herbivores: the common wombat (Vombatus ursinus), koala (Phascolarctos cinereus), swamp wallaby (Wallabia bicolor), and red kangaroo (Macropus rufus). It was hypothesized that grazers, requiring stronger skulls to process tougher food, would have higher biomechanical performance than browsers. This was true when comparing the koala and wallaby (browsers) to the wombat (a grazer). The cranial model of the wombat resulted in low stress and high mechanical efficiency in relation to a robust skull capable of generating high bite forces. However, the kangaroo, also a grazer, has evolved a very different strategy to process tough food. The cranium is much more gracile and has higher stress and lower mechanical efficiency, but they adopt a different method of processing food by having a curved tooth row to concentrate force in a smaller area and molar progression to remove worn teeth from the tooth row. Therefore, the position of the bite is crucial for the structural performance of the kangaroo skull, while it is not for the wombat which process food along the entire tooth row. In accordance with previous studies, the results from this study show the mammalian skull is optimized to resist forces generated during feeding. However, other factors, including the lifestyle of the animal and its environment, also affect selection for skull morphology to meet multiple functional demands. J. Morphol. 276:1230–1243, 2015. © 2015 Wiley Periodicals, Inc.     

扬子鳄与密河鳄基因组DNA的复性动力学和组织结构的研究

通过DNA复性动力学的分析,测定了扬子鳄和密河鳄基因组的组织结构,两者的复性曲线非常相象,高度重复DNA含量很少,而中度重复DNA序列各占基因组的30％左右。中度重复序列部分能再分成三个组分,虽然每个组分的序列复杂性和拷贝数有差别,但每部分的总复杂性是相似的。我们已测得扬子鳄基因组的单拷贝序列为3.68×10’bp,基因组为5.65×10’bp;密河鳄基因组的单拷贝序列为3.67×10’bp,基因组为5.24×10’bp。根据基因组的组织结构的分析,在分子水平上表明两种鳄鱼是亲缘关系十分相近的种。     

Mechanics of the right whale mandible: Full scale testing and finite element analysis

In an effort to better understand the mechanics of ship-whale collision and to reduce the associated mortality of the critically endangered North Atlantic right whale, a comprehensive biomechanical study has been conducted by the Woods Hole Oceanographic Institution and the University of New Hampshire. The goal of the study is to develop a numerical modeling tool to predict the forces and stresses during impact and thereby the resulting mortality risk to whales from ship strikes.Based on post-mortem examinations, jaw fracture was chosen as a fatal endpoint for the whales hit by a vessel. In this paper we investigate the overall mechanical behavior of a right whale mandible under transverse loading and develop a finite element analysis model of the bone. The equivalent elastic modulus of the cortical component of right whale mandible is found by comparing full-scale bending tests with the results of numerical modeling. The finite element model of the mandible can be used in conjunction with a vessel-whale collision event model to predict bone fracture for various ship strike scenarios.     

Biomechanical analysis of foot with different foot arch heights: a finite element analysis

Clinically, different foot arch heights are associated with different tissue injuries to the foot. To investigate the possible factors contributing to the difference in foot arch heights, previous studies have mostly measured foot pressure in either low-arched or high-arched feet. However, little information exists on stress variation inside the foot with different arch heights. Therefore, this study aimed to implement the finite element (FE) method to analyse the influence of different foot arches. This study established a 3D foot FE model using software ANSYS 11.0. After validating the FE model, this study created low-arched, high-arched and normal-arched foot FE models. The FE analysis found that both the stress and strain on the plantar fascia and metatarsal were higher in the high-arched foot, whereas the stress and strain on the calcaneous, navicular and cuboid were higher in low-arched foot. Additionally, forefoot pressure was increased with an increase in arch height.     

Free vibration analysis of microtubules based on the molecular mechanics and continuum beam theory

A molecular structural mechanics (MSM) method has been implemented to investigate the free vibration of microtubules (MTs). The emphasis is placed on the effects of the configuration and the imperfect boundaries of MTs. It is shown that the influence of protofilament number on the fundamental frequency is strong, while the effect of helix-start number is almost negligible. The fundamental frequency is also found to decrease as the number of the blocked filaments at boundaries decreases. Subsequently, the Euler–Bernoulli beam theory is employed to reveal the physics behind the simulation results. Fitting the Euler–Bernoulli beam into the MSM data leads to an explicit formula for the fundamental frequency of MTs with various configurations and identifies a possible correlation between the imperfect boundary conditions and the length-dependent bending stiffness of MTs reported in experiments.