An exploration of sacral morphology using geometric morphometrics and three-dimensionally derived interlandmark distances.

INTRODUCTION: Accurate sex estimation is an important component of a biological profile in forensic anthropology. The pelvis is widely accepted as the most dimorphic osseous structure, and thus, this dimorphism is also reflected by the sacrum. AIM: This study aimed to explore sacral morphology and to derive a practically applicable discriminant function formula for sex estimation. MATERIALS: A total of 20 three-dimensional landmarks were digitised on a sample of 200 sacra from a sample of South Africans of African descent (Black South Africans) with ages ranging between 20 and 90 years, equally distributed for sex. METHODS: Geometric morphometric methods were used to analyse sacral morphology and sexual dimorphism as it captures size-independent shape variation and three-dimensional morphology. RESULTS: Size-independent shape analysis revealed four sacral structures and metrics that account for most of its shape variation. When these were compared between the sexes, we found that sacral curvature pattern, rather than depth, differed between sexes and that males have greater anterior sacral heights. Females have larger alae relative to the body of S1. In addition, the anterior posterior breadth of the sacral canal is larger in males, as is the relative size and projection of the superior articular processes. Discriminant analyses of these data produced average accuracies of only 72.5%, but this improved to 84.5% when using novel interlandmark distances derived from the raw coordinate data. CONCLUSION: Our results demonstrate that landmark-based techniques allow for a more nuanced understanding of structural variation. In addition, accuracies were achieved that surpass traditional metrics using an equal number of variables. These results contribute to our understanding of sacral dimorphism and will assist in forensic casework.

Assessing the effects of tooth loss in adult crania using geometric morphometrics.

With high numbers of unidentified skeletonised remains recovered annually in South Africa and an increased number of edentate individuals being reported, the question arises as to whether tooth loss would result in craniofacial changes which might alter the accuracy of osteological analyses. Forty-five fixed landmarks together with sliding semilandmarks were collected from 229 white South African crania and were used to capture curve data pertaining to the basicranium, alveoli, zygomatic arches, nasal aperture and orbits. Geometric morphometric methods were employed to assess the effects of tooth loss on these structures. Although a number of effects were seen when the skull was analysed in its entirety, only the alveoli proved to be significantly affected when regions were analysed individually. Both upper facial height and palate shape were affected by tooth loss, which may influence various osteometric measurements and qualitative traits that are used during the assessment of ancestry and sex.

Visualising trends in dentition to lip mouth morphology using geometric morphometrics.

Linear measurements taken from bony landmarks are often utilised in facial approximation (FA) to estimate and plan the placement of overlying soft tissue features. This process similarly guides craniofacial superimposition (CFS) practices. Knowledge of how hard and soft tissue features spatially relate around the mouth region is, however, limited. Geometric morphometric techniques have thus been used to investigate size and shape variation in dentition-to-lip mouth morphology in a South African population. Twenty landmarks (twelve dentition, eight lips) were digitised, using cone-beam CT images of the anterior craniofacial complex in a Frankfurt/Frankfort position, for 147 individuals aged between 20 and 75 years. Principal Component Analysis and Canonical Variate Analysis established that much shape variation exists. A two-way ANOVA identified significant (p < 0.0001) population and sex variation with mouth shape. Black individuals presented with thicker lips, with the oral fissure aligning closely to the dental occlusion. Oral fissure position for white individuals corresponded to the inferior one-quarter (females) or one-sixth (males) of the maxillary central incisor crowns. Males presented larger dimensions than females, but females had a greater lip-to-teeth height ratio than their male counterparts. A pooled within-group regression analysis assessed the effect of age on the dentition and lips and found that it had a significant (p < 0.0001) impact on mouth shape. Ageing was associated with a reduced lip and teeth height, increased mouth width, and a lowered oral fissure and cheilion placement. The generated mean shape data, with metric guides, offer a visual and numerical guide that builds on existing FA and CFS standards, enhancing our understanding of hard and soft tissue relationships.

Adult hippocampal neurogenesis in Egyptian fruit bats from three different environments: Are interpretational variations due to the environment or methodology?

We quantified both proliferative (Ki-67 immunohistochemistry) and immature (doublecortin immunohistochemistry) cells within the dentate gyrus of adult Egyptian fruit bats from three distinct environments: (a) primary rainforest, (b) subtropical woodland, and (c) fifth-generation captive-bred. We used four different previously reported methods to assess the effect of the environment on proliferative and immature cells: (a) the comparison of raw totals of proliferative and immature cells; (b) these totals standardized to brain mass; (c) these totals expressed as a density using the volume of the granular cell layer (GCLv) for standardization; and (d) these totals expressed as a percentage of the total number of granule cells. For all methods, the numbers of proliferative cells did not differ statistically among the three groups, indicating that the rate of proliferation, while malleable to experimental manipulation or transiently in response to events of importance in the natural habitat, appears to occur, for the most part, at a predetermined rate within a species. For the immature cells, raw numbers and standardizations to brain mass and GCLv revealed no difference between the three groups studied; however, standardization to total granule cell numbers indicated that the two groups of wild-caught bats had significantly higher numbers of immature neurons than the captive-bred bats. These contrasting results indicate that the interpretation of the effect of the environment on the numbers of immature neurons appears method dependent. It is possible that current methods are not sensitive enough to reveal the effect of different environments on proliferative and immature cells.

The distribution, number, and certain neurochemical identities of infracortical white matter neurons in a lar gibbon (Hylobates lar) brain.

We examined the number, distribution, and immunoreactivity of the infracortical white matter neuronal population, also termed white matter interstitial cells (WMICs), in the brain of a lesser ape, the lar gibbon. Staining for neuronal nuclear marker (NeuN) revealed WMICs throughout the infracortical white matter, these cells being most numerous and dense close to cortical layer VI, decreasing significantly in density with depth in the white matter. Stereological analysis of NeuN-immunopositive cells revealed a global estimate of ~67.5 million WMICs within the infracortical white matter of the gibbon brain, indicating that the WMICs are a numerically significant population, ~2.5% of the total cortical gray matter neurons that would be estimated for a primate brain the mass of that of the lar gibbon. Immunostaining revealed subpopulations of WMICs containing neuronal nitric oxide synthase (nNOS, ~7 million in number, with both small and large soma volumes), calretinin (~8.6 million in number, all of similar soma volume), very few WMICs containing parvalbumin, and no calbindin-immunopositive neurons. These nNOS, calretinin, and parvalbumin immunopositive WMICs, presumably all inhibitory neurons, represent ~23.1% of the total WMIC population. As the white matter is affected in many cognitive conditions, such as schizophrenia, autism and also in neurodegenerative diseases, understanding these neurons across species is important for the translation of findings of neural dysfunction in animal models to humans. Furthermore, studies of WMICs in species such as apes provide a crucial phylogenetic context for understanding the evolution of these cell types in the human brain.

Three-dimensionally derived interlandmark distances for sex estimation in intact and fragmentary crania.

The skull is the element most frequently presented to forensic anthropologists for analysis yet weathering, corpse maiming, and scavenger activity often result in damage and fragmentation. This fragmentation results in a reduction in the number of traditional calliper derived measurements that can be obtained and subjected to discriminant based analyses for sex estimation. In this investigation, we employed three-dimensional geometric morphometric methods to derive novel interlandmark distance measures across six regions of the cranium including the basicranium, basipalate, zygoma, orbits and the cranium globally to create functions to discriminate sex with high efficacy, even in the event of fragmentation. Forty-five homologous landmarks were digitised across each of 227 (114 males and 113 females) South African crania of European descent (white) sampled from the Raymond A Dart Collection of Human Skeletons, housed in the School of Anatomical Sciences, University of the Witwatersrand, South Africa. A total of 990 interlandmark distances (ILDs) were mathematically derived using Pythagorean geometry. These ILDs were then filtered by region and subjected to both direct and stepwise discriminant function analyses. Discriminant equations where derived for each region and achieved the following average cross-validated sex estimation accuracies: basicranium-74%; basipalate-80.2%; zygomatic-82.4; orbits-71.8%; nasomaxilla-83.7%; global cranium-88.2%. A large number of the ILDs used to derive the discriminant functions are novel, demonstrating the efficacy of geometric morphometric methods and illustrating the need to reassess old methods of data collection using modern methods to determine whether they best capture biological differences. The results of this study provide an invaluable contribution to forensic anthropology in South Africa as it provides an accurate, practical means of assessing sex using fragmentary material that may otherwise have been disregarded. These will undeniable aid in accurate sex estimation and ultimately, victim identification.

Temporal niche switching in Arabian oryx (Oryx leucoryx): Seasonal plasticity of 24h activity patterns in a large desert mammal.

The Arabian oryx, a moderately large mammal that inhabits a harsh desert environment, has been shown to exhibit seasonal variations in activity and inactivity patterns. Here we analyzed the continuous year-round activity patterns of twelve free-roaming Arabian oryx under natural conditions from two varying desert environments in Saudi Arabia using abdominally implanted activity meters. We simultaneously recorded weather parameters at both sites to determine whether environmental factors are responsible for temporal niche switching as well as the seasonal structuring and timing of this behavioural plasticity. Our results demonstrate that Arabian oryx undergo temporal niche switching of 24h activity patterns at a seasonal level and exhibit distinct nocturnal/crepuscular activity during summer, diurnal activity during winter and intermittent patterns of behaviour during the transitional seasons of autumn and spring. In addition, the oryx exhibited inter- and intra-seasonal variations in the temporal budgeting of 24h activity patterns. Strong relationships with both photoperiod and ambient temperatures were found and in some instances suggested that increasing ambient temperatures are a primary driving force behind seasonal shifts in activity patterns. These adaptive patterns may be dictated by the availability of food and water, which in turn are strongly influenced by seasonal climate variations. Overall, the adaptive responses of free-roaming Arabian oryx in such harsh and non-laboratorial conditions provide a framework for comparing wild populations as well as aiding conservation efforts.

The first hominin from the early Pleistocene paleocave of Haasgat, South Africa.

Haasgat is a primate-rich fossil locality in the northeastern part of the Fossil Hominid Sites of South Africa UNESCO World Heritage Site. Here we report the first hominin identified from Haasgat, a partial maxillary molar (HGT 500), that was recovered from an ex situ calcified sediment block sampled from the locality. The in situ fossil bearing deposits of the Haasgat paleokarstic deposits are estimated to date to slightly older than 1.95 Ma based on magnetobiostratigraphy. This places the hominin specimen at a critical time period in South Africa that marks the last occurrence of Australopithecus around 1.98 Ma and the first evidence of Paranthropus and Homo in the region between ∼2.0 and 1.8 Ma. A comprehensive morphological evaluation of the Haasgat hominin molar was conducted against the current South African catalogue of hominin dental remains and imaging analyses using micro-CT, electron and confocal microscopy. The preserved occlusal morphology is most similar to Australopithecus africanus or early Homo specimens but different from Paranthropus. Occlusal linear enamel thickness measured from micro-CT scans provides an average of ∼2.0 mm consistent with Australopithecus and early Homo. Analysis of the enamel microstructure suggests an estimated periodicity of 7-9 days. Hunter-Schreger bands appear long and straight as in some Paranthropus, but contrast with this genus in the short shape of the striae of Retzius. Taken together, these data suggests that the maxillary fragment recovered from Haasgat best fits within the Australopithecus-early Homo hypodigms to the exclusion of the genus Paranthropus. At ∼1.95 Ma this specimen would either represent another example of late occurring Australopithecus or one of the earliest examples of Homo in the region. While the identification of this first hominin specimen from Haasgat is not unexpected given the composition of other South African penecontemporaneous site deposits, it represents one of the few hominin localities in the topographically-distinct northern World Heritage Site. When coupled with the substantial differences in the mammalian faunal communities between the northern localities (e.g., Haasgat, Gondolin) and well-sampled Bloubank Valley sites (e.g., Sterkfontein, Swartkrans, Kromdraai), the recovery of the HGT 500 specimen highlights the potential for further research at the Haasgat locality for understanding the distribution and interactions of hominin populations across the landscape, ecosystems and fossil mammalian communities of early Pleistocene South Africa. Such contextual data from sites like Haasgat is critical for understanding the transition in hominin representation at ∼2 Ma sites in the region from Australopithecus to Paranthropus and early Homo.

Quantification of the subpubic angle in South Africans.

Due to the high crime rate in South Africa, forensic anthropologists are increasingly approached to aid in the identification of skeletonized remains, with sex and population affinity assignment being some of the most critical tasks they face. For over a century, the pelvis has been known to be one of the most sexually dimorphic bones of the human body and the subpubic angle is one of the most accurate, albeit scarcely quantified, features thereof. Hence, the aim of this study was to quantify the size of the subpubic angle and compare it between male and female South Africans of African (black) and European (white) descent. One hundred and forty five (145) pelves were selected, consisting of 68 white (43 male and 25 female) and 77 black South Africans (44 male and 33 female), from the Raymond A. Dart Collection of Human Skeletons housed at the University of the Witwatersrand, Johannesburg, South Africa. Each of the pelves were articulated and placed into a custom built stand for photographing. Measures of the subpubic angle from these digital images were subjected to numerous statistical analyses. Results indicated that significant differences exist between the sexes, as well as between the two population groups. For black individuals it was found that males generally possessed a subpubic angle of 74.9° or less, with larger values being indicative of the female sex. For white individuals, subpubic angles of 81.4° and less indicated males whilst larger values indicated females, with an average accuracy of 86% for both population groups. These results illustrate the advantages of using the subpubic angle to assist in the estimation of sex and population affinity and also reinforce the need for population specific parameters to be applied.

Quantitative analysis of neocortical gyrencephaly in African elephants (Loxodonta africana) and six species of cetaceans: comparison with other mammals.

This study provides quantitative data on the extent of gyrencephaly in the large-brained African elephant and several species of cetaceans (from smaller to larger brained) in comparison with other mammals. Across three mammalian orders (primates, carnivores, and artiodactyls), the species with the larger brains are more gyrencephalic with each order, exhibiting a specific negative allometry. The African elephant, with a 5-kg brain, has a gyrencephalic index (GI) of 3.89, which, though highly gyrencephalic, is not more so than would be predicted for a mammal with a 5-kg brain. The cetaceans had an average GI of 5.43, are the most gyrencephalic mammals studied to date, and are more gyrencephalic than one would predict based on comparison with other mammals. No relationship between brain mass and GI was evident in the cetaceans as seen in other mammals, with all cetaceans showing similar GIs irrespective of brain mass (range of GI 5.23-5.70, range of brain mass 577-5617 g). This is yet another parameter indicating cetaceans to be neuroanatomical outliers. Two species of pinnipeds studied had GIs that were well above those seen for terrestrial carnivores, and the aquatic manatee was close to lissencephalic. Thus, all three groups of marine mammals showed unusual extents of cortical gyrencephaly, indicating a morphological alteration of the telencephalon associated with the return to the marine environment. The analysis suggests that cortical thickness and neuronal density are important factors in determining the extent of gyrencephaly across mammalian species.

Acquisition of brains from the African elephant (Loxodonta africana): perfusion-fixation and dissection.

The current correspondence describes the in situ perfusion-fixation of the brain of the African elephant. Due to both the large size of proboscidean brains and the complex behaviour of these species, the acquisition of good quality material for comparative neuroanatomical analysis from these species is important. Three male African elephants (20-30 years) that were to be culled as part of a larger population management strategy were used. The animals were humanely euthanized and the head removed from the body. Large tubes were inserted into to the carotid arteries and the cranial vasculature flushed with a rapid (20 min) rinse of 100 l of cold saline (4 degrees C). Following the rinse the head was perfusion-fixed with a slower rinse (40 min) of 100 l of cold (4 degrees C) 4% paraformaldehyde in 0.1M phosphate buffer. This procedure resulted in well-fixed neural and other tissue. After perfusion the brains were removed from the skull with the aid of power tools, a procedure taking between 2 and 6h. The brains were immediately post-fixed in the same solution for 72 h at 4 degrees C. The brains were subsequently placed in a sucrose solution and finally an antifreeze solution and are stored in a -20 degrees C freezer. The acquisition of high quality neural material from African elephants that can be used for immunohistochemistry and electron microscopy is of importance in understanding the "hardware" underlying the behaviour of this species. This technique can be used on a variety of large mammals to obtain high quality material for comparative neuroanatomical studies.