Skeletal Morphology Research Articles

Effects of age and diet consistency on the expression of myosin heavy-chain isoforms on jaw-closing and jaw-opening muscles in a rat model.

Skeletal craniofacial morphology can be influenced by changes in masticatory muscle function, which may also change the functional profile of the muscles. To investigate the effects of age and functional demands on the expression of Myosin Heavy-Chain (MyHC) isoforms in representative jaw-closing and jaw-opening muscles, namely the masseter and digastric muscles respectively. Eighty-four male Wistar rats were divided into four age groups, namely an immature (n = 12; 4-week-old), early adult (n = 24; 16-week-old), adult (n = 24; 26-week-old) and mature adult (n = 24; 38-week-old) group. The three adult groups were divided into two subgroups each based on diet consistency; a control group fed a standard (hard) diet, and an experimental group fed a soft diet. Rats were sacrificed, and masseter and digastric muscles dissected. Real-time quantitative polymerase chain reaction was used to compare the mRNA transcripts of the MyHC isoforms-Myh7 (MyHC-I), Myh2 (MyHC-IIa), Myh4 (MyHC-IIb) and Myh1 (MyHC-IIx)-of deep masseter and digastric muscles. In the masseter muscle, hypofunction increases Myh1 (26, 38 weeks; p < .0001) but decreases Myh4 (26 weeks; p = .046) and Myh2 (26 weeks; p < .0001) expression in adult rats. In the digastric muscle, hypofunction increases Myh1 expression in the mature adult rats (38 weeks; p < .0001), while Myh2 expression decreases in adult rats (26 weeks; p = .021) as does Myh4 (26 weeks; p = .001). Myh7 expression is increased in the digastric muscle of mature adult rats subjected to hypofunction (38 weeks; p = <.0001), while it is very weakly expressed in the masseter. In jaw-opening and jaw-closing muscles, differences in myosin expression between hard- and soft-diet-fed rats become evident in adulthood, suggesting that long-term alteration of jaw function is associated with changes in the expression of MyHC isoforms and potential fibre remodelling. This may give insight into the role of function on masticatory muscles and the resultant craniofacial morphology.

The common marmoset as a translational model of age-related osteoarthritis.

Age-related osteoarthritis (OA) is a degenerative joint disease characterized by pathological changes in nearly every intra- and peri-articular tissue that contributes to disability in older adults. Studying the etiology of age-related OA in humans is difficult due to an unpredictable onset and insidious nature. A barrier in developing OA modifying therapies is the lack of translational models that replicate human joint anatomy and age-related OA progression. The purpose of this study was to determine whether the common marmoset is a faithful model of human age-related knee OA. Semi-quantitative microCTscoring revealed greater radiographic OA in geriatric versus adult marmosets, and the age-related increase in OA prevalence was similar between marmosets and humans. Quantitative assessments indicate greater medial tibial cortical and trabecular bone thickness and heterogeneity in geriatric versus adult marmosets which is consistent with an age-related increase in focal subchondral bone sclerosis. Additionally, marmosets displayed an age-associated increase in synovitis and calcification of the meniscus and patella. Histological OA pathology in the medial tibial plateau was greater in geriatric versus adult marmosets driven by articular cartilage damage, proteoglycan loss, and alteredchondrocyte cellularity. The age-associated increase in medial tibial cartilage OA pathology and meniscal calcification was greater in female versus male geriatric marmosets. Overall, marmosets largely replicate human OA as evident by similar 1) cartilage and skeletal morphology, 2) age-related progression in OA pathology, and 3) sex differences in OA pathology with increasing age. Collectively, these data suggest that the common marmoset is a highly translatable model ofthe naturally occurring, age-related OA seenin humans.

Biomineralisation of corals inhabiting extreme and marginal environments

ABSTRACT Climate change is the primary threat to coral reefs, causing catastrophic coral mortality on a global scale. Stressors such as ocean acidification limit calcium carbonate availability needed to build skeletons. A wealth of research has shown that climate stressors impair the ability of coral to produce and maintain their skeletons, thereby disrupting functions critical to maintaining coral health as well as threatening the ecologically significant framework for reef building. There are natural marginal and extreme environments however, where coral reefs thrive despite unfavourable conditions. These habitats provide a natural long-term setting to examine the impact of multiple interacting climate change factors on corals and understand the adaptations required by corals to inhabit these conditions. We review studies of coral skeletogenesis in extreme and marginal environments, with a particular focus on mangrove habitats due to their natural fluctuations in temperature, pH and oxygen, analogous to those projected under climate change models. The changes in coral skeletal morphology under stress are described as are the techniques used to visualise these changes. In marginal and extreme environments, corals experience lower calcification rates and produce more porous, less robust skeletons whilst maintaining normal rates of linear extension, suggesting the integrity of coral skeletons are likely to be threatened under future ocean conditions. The biomineralisation processes of corals inhabiting extreme and marginal environments remains an underexplored area of research and has the potential to yield valuable insights into how corals might adapt to climate change and the mechanisms that underlie their resilience to global environmental change.

Radiographic anatomical description of the skeletal system of adult and subadult axolotls (Ambystoma mexicanum): a pilot study

In this study, the use of radiographic imaging to examine the skeletal structures of living axolotls (Ambystoma mexicanum) is described. Dorsoventral and left lateral radiographs were taken of both healthy adult and subadult axolotls without restraint, directly on the X-ray table. The aim was to comprehensively describe their skeletal anatomy using radiographic assessment. The method accurately depicted bone shapes and structures, identifying bone names based on comparisons with similar salamanders. However, internal organs within the coelomic cavity were not clearly visible except for the lungs. Interpreting subadult axolotl radiographs was harder, likely due to less developed skeletal ossification or X-ray parameter variations. Variations were observed in the number of certain bones and vertebrae types among individuals. This method offers a non-invasive way to understand healthy animal skeletal appearance, eliminating the need for autopsies. The radiographic approach contributes to better understand axolotl skeletal morphology, benefiting comparative anatomy, veterinary medicine and conservation efforts.

Cortical bone architecture of hominid intermediate phalanges reveals functional signals of locomotion and manipulation.

Reconstruction of fossil hominin manual behaviors often relies on comparative analyses of extant hominid hands to understand the relationship between hand use and skeletal morphology. In this context, the intermediate phalanges remain understudied. Thus, here we investigate cortical bone morphology of the intermediate phalanges of extant hominids and compare it to the cortical structure of the proximal phalanges, to investigate the relationship between cortical bone structure and inferred loading during manual behaviors. Using micro-CT data, we analyze cortical bone structure of the intermediate phalangeal shaft of digits 2-5 in Pongo pygmaeus (n = 6 individuals), Gorilla gorilla (n = 22), Pan spp. (n = 23), and Homo sapiens (n = 23). The R package morphomap is used to study cortical bone distribution, cortical thickness and cross-sectional properties within and across taxa. Non-human great apes generally have thick cortical bone on the palmar shaft, with Pongo only having thick cortex on the peaks of the flexor sheath ridges, while African apes have thick cortex along the entire flexor sheath ridge and proximal to the trochlea. Humans are distinct in having thicker dorsal shaft cortex as well as thick cortex at the disto-palmar region of the shaft. Variation in cortical bone distribution and properties of the intermediate phalanges is consistent with differences in locomotor and manipulative behaviors in extant great apes. Comparisons between the intermediate and proximal phalanges reveals similar patterns of cortical bone distribution within each taxon but with potentially greater load experienced by the proximal phalanges, even in knuckle-walking African apes. This study provides a comparative context for the reconstruction of habitual hand use in fossil hominins and hominids.

Characterizing the effects of muscle-specific GSK3α/β reduction on murine muscle contractility and metabolism in female mice.

Dysregulation of skeletal muscle morphology and metabolism is associated with chronic diseases such as obesity and type 2 diabetes. The enzyme glycogen synthase kinase 3 (GSK3) is highly involved in skeletal muscle physiology and metabolism, acting as a negative regulator of muscle size, strength, adaptive thermogenesis, and glucose homeostasis. Correspondingly, we have shown that partial knockdown (∼40%) of GSK3 specifically in skeletal muscle increases lean mass, reduces fat mass, and activates muscle-based adaptive thermogenesis via sarco(endo)plasmic reticulum Ca2+ (SERCA) uncoupling in male mice. However, the effects of GSK3 knockdown in female mice have yet to be investigated. Here, we examined the effects of muscle-specific GSK3 knockdown on body composition, muscle size and strength, and whole body metabolism in female C57BL/6J mice. Our results show that GSK3 content is higher in the female soleus versus the male soleus; however, there were no differences in the extensor digitorum longus (EDL). Furthermore, muscle-specific GSK3 knockdown did not alter body composition in female mice, nor did it alter daily energy expenditure, glucose/insulin tolerance, mitochondrial respiration, or the expression of the SERCA uncouplers sarcolipin and neuronatin. We also did not find any differences in soleus muscle size, strength, or fatigue resistance. In the EDL, we found that an increase in absolute and specific force production, but there were no differences in fatigability. Therefore, our study highlights sex differences in the response to genetic reduction of gsk3, with most of the effects previously observed in male mice being absent in females.NEW & NOTEWORTHY Here we show that partial GSK3 knockdown has minimal effects on whole body metabolism and muscle contractility in female mice. This is partly inconsistent with previous results found in male mice, which reveal a potential influence of biological sex.

Effects of long-term voluntary wheel running and selective breeding for wheel running on femoral nutrient canals.

The nutrient artery provides ~50%-70% of the total blood volume to long bones in mammals. Studying the functional characteristics of this artery invivo can be difficult and expensive, so most researchers have measured the nutrient foramen, an opening on the outer surface of the bone that served as the entry point for the nutrient artery during development and bone ossification. Others have measured the nutrient canal (i.e., the passage which the nutrient artery once occupied), given that the external dimensions of the foramen do not necessarily remain uniform from the periosteal surface to the medullary cavity. The nutrient canal, as an indicator of blood flow to long bones, has been proposed to provide a link to studying organismal activity (e.g., locomotor behavior) from skeletal morphology. However, although external loading from movement and activity causes skeletal remodeling, it is unclear whether it affects the size or configuration of nutrient canals. To investigate whether nutrient canals can exhibit phenotypic plasticity in response to physical activity, we studied a mouse model in which four replicate high runner (HR) lines have been selectively bred for high voluntary wheel-running behavior. The selection criterion is the average number of wheel revolutions on days 5 and 6 of a 6-day period of wheel access as young adults (~6-8 weeks old). An additional four lines are bred without selection to serve as controls (C). For this study, 100 female mice (half HR, half C) from generation 57 were split into an active group housed with wheels and a sedentary group housed without wheels for 12 weeks starting at ~24 days of age. Femurs were collected, soft tissues were removed, and femora were micro-computed tomography scanned at a resolution of 12 μm. We then imported these scans into AMIRA and created 3D models of femoral nutrient canals. We tested for evolved differences in various nutrient canal traits between HR and C mice, plastic changes resulting from chronic exercise, and the selection history-by-exercise interaction. We found few differences between the nutrient canals of HR versus C mice, or between the active and sedentary groups. We did find an interaction between selection history and voluntary exercise for the total number of nutrient canals per femur, in which wheel access increased the number of canals in C mice but decreased it in HR mice. Our results do not match those from an earlier study, conducted at generation 11, which was prior to the HR lines reaching selection limits for wheel running. The previous study found that mice from the HR lines had significantly larger total canal cross-sectional areas compared to those from C lines. However, this discrepancy is consistent with studies of other skeletal traits, which have found differences between HR and C mice to be somewhat inconsistent across generations, including the loss of some apparent adaptations with continued selective breeding after reaching a selection limit for wheel-running behavior.

Semi-Automated MicroCT Analysis of Bone Anatomy and Mineralization in Mouse Models.

The skeletal system mirrors several processes in the vertebrate body that impact developmental malfunctions, hormonal disbalance, malfunction of calcium metabolism and turn over, and inflammation processes such as arthrosis. X-ray micro computed tomography is a useful tool for 3D in situ evaluation of the skeletal system in a time-related manner, but results depend highly on resolution. Here, we provide the methodological background for a graduated evaluation from whole-body analysis of skeletal morphology and mineralization to high-resolution analysis of femoral and vertebral microstructure. We combine an expert-based evaluation with a machine-learning-based computational approach, including pre-setup analytical task lists. © 2024 Wiley Periodicals LLC. Basic Protocol 1: In vivo microCT scanning and skeletal analysis in mice Basic Protocol 2: Ex vivo high-resolution microCT scanning and microstructural analysis of the femur and L4 vertebra.

Loss of 24-hydroxylated catabolism increases calcitriol and fibroblast growth factor 23 and alters calcium and phosphate metabolism in fetal mice.

Calcitriol circulates at low levels in normal human and rodent fetuses, in part due to increased 24-hydroxylation of calcitriol and 25-hydroxyvitamin D by 24-hydroxylase (CYP24A1). Inactivating mutations of CYP24A1 cause high postnatal levels of calcitriol and the human condition of infantile hypercalcemia type 1, but whether the fetus is disturbed by the loss of CYP24A1 is unknown. We hypothesized that loss of Cyp24a1 in fetal mice will cause high calcitriol, hypercalcemia, and increased placental calcium transport. The Cyp24a1+/- mice were mated to create pregnancies with wildtype, Cyp24a1+/-, and Cyp24a1 null fetuses. The null fetuses were hypercalcemic, modestly hypophosphatemic (compared to Cyp24a1+/- fetuses only), with 3.5-fold increased calcitriol, 4-fold increased fibroblast growth factor 23 (FGF23), and unchanged parathyroid hormone. The quantitative RT-PCR confirmed the absence of Cyp24a1 and 2-fold increases in S100g, sodium-calcium exchanger type 1, and calcium-sensing receptor in null placentas but not in fetal kidneys; these changes predicted an increase in placental calcium transport. However, placental 45Ca and 32P transport were unchanged in null fetuses. Fetal ash weight and mineral content, placental weight, crown-rump length, and skeletal morphology did not differ among the genotypes. Serum procollagen 1 intact N-terminal propeptide and bone expression of sclerostin and Blgap were reduced while calcitonin receptor was increased in nulls. In conclusion, loss of Cyp24a1 in fetal mice causes hypercalcemia, modest hypophosphatemia, and increased FGF23, but no alteration in skeletal development. Reduced incorporation of calcium into bone may contribute to the hypercalcemia without causing a detectable decrease in the skeletal mineral content. The results predict that human fetuses bearing homozygous or compound heterozygous inactivating mutations of CYP24A1 will also be hypercalcemic in utero but with normal skeletal development.

Macroscopic differences in adult human femora are linked to body mass index.

Bone functional adaptation is routinely invoked to interpret skeletal morphology despite ongoing debate regarding the limits of the bone response to mechanical stimuli. The wide variation in human body mass presents an opportunity to explore the relationship between mechanical load and skeletal response in weight-bearing elements. Here, we examine variation in femoral macroscopic morphology as a function of body mass index (BMI), which is used as a metric of load history. A sample of 80 femora (40 female; 40 male) from recent modern humans was selected from the Texas State University Donated Skeletal Collection. Femora were imaged using x-ray computed tomography (voxel size ~0.5 mm), and segmented to produce surface models. Landmark-based geometric morphometric analyses based on the Coherent Point Drift algorithm were conducted to quantify shape. Principal components analyses were used to summarize shape variation, and component scores were regressed on BMI. Within the male sample, increased BMI was associated with a mediolaterally expanded femoral shaft, as well as increased neck-shaft angle and decreased femoral neck anteversion angle. No statistically significant relationships between shape and BMI were found in the female sample. While mechanical stimulus has traditionally been applied to changes in long bong diaphyseal shape it appears that bone functional adaptation may also result in fundamental changes in the shape of skeletal elements.

Quantitative Skeletal Imaging and Image-Based Modeling in Pediatric Orthopaedics.

Musculoskeletal imaging serves a critical role in clinical care and orthopaedic research. Image-based modeling is also gaining traction as a useful tool in understanding skeletal morphology and mechanics. However, there are fewer studies on advanced imaging and modeling in pediatric populations. The purpose of this review is to provide an overview of recent literature on skeletal imaging modalities and modeling techniques with a special emphasis on current and future uses in pediatric research and clinical care. While many principles of imaging and 3D modeling are relevant across the lifespan, there are special considerations for pediatric musculoskeletal imaging and fewer studies of 3D skeletal modeling in pediatric populations. Improved understanding of bone morphology and growth during childhood in healthy and pathologic patients may provide new insight into the pathophysiology of pediatric-onset skeletal diseases and the biomechanics of bone development. Clinical translation of 3D modeling tools developed in orthopaedic research is limited by the requirement for manual image segmentation and the resources needed for segmentation, modeling, and analysis. This paper highlights the current and future uses of common musculoskeletal imaging modalities and 3D modeling techniques in pediatric orthopaedic clinical care and research.

Verification of biological relationships in the Dietrichstein princely family from Mikulov by cranial non-metric traits analysis (Moravia, 17th to 19th centuries)

This study details the verification of documented biological family relationships in the skeletal remains of 9 members of the Dietrichstein princely family from their tomb in Mikulov, Moravia, using skeletal morphology. Another goal was to determine the degree of morphological similarity to other examined family members of the unknown individual from the coffin of Walter Francis Xaver, 4th Prince of Dietrichstein. For these purposes, 93 cranial non-metric traits were evaluated, the degree of similarity of individuals was evaluated using the similarity coefficient. The results support the authenticity of the remains of individual family members and the documented biological family relationships. The variability of biologically related individuals is smaller than that of unrelated individuals. The unknown individual from the coffin of Prince Walter Francis Xaver is most similar to Rosa Barbara Ludovica, Countess of Dietrichstein, née Countess of Wallis, Baroness of Carrighmain (Oct. 8th, 1792 – Jun. 27th 1844). Given that her remains were also found unexpectedly in the tomb, in the coffin of Maximilian II, 1st Prince of Dietrichstein, the possibility that the two finds are related can be admitted.

Effect of electroacupuncture combined with sulforaphane in the treatment of sarcopenia in SAMP8 mice.

Skeletal muscles mitochondrial dysfunction is the main cause of sarcopenia. Both electroacupuncture (EA) and sulforaphane (SFN) have been shown to improve oxidative stress and inflammation levels to maintain mitochondrial function, but the effects and mechanisms of their combination on sarcopenia are unclear. This study aimed to investigate the regulatory effects of EA combined with SFN on sarcopenia. SAMP8 mice were used and intervened with EA or SFN, respectively, and Masson and HE staining were used to observe pathological changes in skeletal muscle tissue. Transmission electron microscopy was used to detect tissue mitochondrial changes. TUNEL staining was used to assess apoptosis. The biochemical and molecular content was tested by ELISA, western blot, and qRT-PCR. The results showed that oxidative stress, apoptosis, and IL-6, TNF-α, Atrogin-1, and MuRF1 levels in skeletal muscles cells were suppressed and mitochondrial damage was repaired after EA or SFN intervention. In addition, we found that the above changes were associated with the activation of the AMPK/Sirt1/PGC-1α pathway in skeletal muscle tissues, and the promotion effect of combined EA and SFN intervention was more significant. In conclusion, this study found that EA combined with SFN mediated the repair of mitochondrial damage through activation of the AMPK/Sirt1/PGC-1α pathway, thereby alleviating skeletal muscles morphology and function in sarcopenia. This study combines EA with SFN, which not only broadens the use of electroacupuncture and SFN but also provides a scientific experimental basis for the treatment of sarcopenia.

Flight Performance of the Early Cretaceous Bird Confuciusornis sanctus: Evidence from an Exceptionally Preserved Fossil

The Chinese early Cretaceous Confuciusornis sanctus is the most abundant Mesozoic bird and a model species for studies of early avian evolution. While previous investigations were largely focused on aspects of the anatomy, taxonomy and systematics, and life history and ecology of this bird, there has been minimal research on its flight properties. Our study centers on a well-preserved specimen with exquisite details of its plumage. NHMW 1997z0112/0001 affords novel information on aspects of its skeletal morphology, particularly from the axial and appendicular skeletons, and its plumage provides the opportunity to quantify key flight-related variables including, wingtip, wing chord, body mass, wingspan, and wing area. We use these parameters to provide a quantitative assessment of the flight properties of C. sanctus. Most previous studies have suggested that this species was unable to achieve prolonged flights. However, our results indicate that the capacity for this bird to perform prolonged flights cannot be discarded, given that our data shows it might have been able to combine periods of flapping with periods of efficient low-speed gliding. Specifically, our results indicate that while having slightly less capacity than modern gliding birds, the gliding capacity of C. sanctus would have been significantly higher than that of modern short-term fliers such as land fowl. On the basis of these inferences, we conclude that C. sanctus could fly efficiently for prolonged periods of time when used a combination of flapping and gliding periods.

CRISPR/Cas9-mediated nexilin deficiency interferes with cardiac contractile function in zebrafish in vivo

Nexilin (NEXN) plays a crucial role in stabilizing the sarcomeric Z-disk of striated muscle fibers and, when mutated, leads to dilated cardiomyopathy in humans. Due to its early neonatal lethality in mice, the detailed impact of the constitutive homozygous NEXN knockout on heart and skeletal muscle morphology and function is insufficiently investigated. Here, we characterized a constitutive homozygous CRISPR/Cas9-mediated nexn knockout zebrafish model. We found that Nexn deficient embryos developed significantly reduced cardiac contractility and under stressed conditions also impaired skeletal muscle organization whereas skeletal muscle function seemed not to be affected. Remarkably, in contrast to nexn morphants, CRISPR/Cas9 nexn−/− knockout embryos showed a milder phenotype without the development of a pronounced pericardial edema or blood congestion. nexn-specific expression analysis as well as whole transcriptome profiling suggest some degree of compensatory mechanisms. Transcripts of numerous essential sarcomeric proteins were massively induced and may mediate a sarcomere stabilizing function in nexn−/− knockout embryos. Our findings demonstrate the successful generation and characterization of a constitutive homozygous nexn knockout line enabling the detailed investigation of the role of nexn on heart and skeletal muscle development and function as well as to assess putative compensatory mechanisms induced by the loss of Nexn.

Superheater ash deposit ageing – Impact of melt fraction on morphology and chemistry

The initial melting behaviour of synthetic ash deposits, with a focus on how the amount of melt formed, affects the final deposit morphology, was studied. Binary and ternary reciprocal deposits were exposed to a temperature gradient with varying exposure time, steel temperature, and furnace temperature. Two different archetype deposit morphologies emerged. The final deposit morphology was identified to depend on the amount of melt formed during the experiment. A skeletal deposit morphology developed in systems with less than 30 wt-% melt. Short-term experiments proved that the formation of the skeletal structure is accelerated by the presence of melt, accumulating at contact points of distinct particles. If melt fractions exceeded 30 wt-%, a molten morphology developed, as the amount of melt present was sufficient to fill the pores within the deposit. For molten deposits of ternary reciprocal systems, enrichment of K in the formed melt and concurrent movement toward the steel was observed. The results confirmed a deposit ageing mechanism that has been proposed earlier, based on full-scale superheater deposit samples. The experiments showed that deposit melting and ageing behaviour are strongly connected, as the two identified morphologies differed in their ageing behaviour. The presented results improve the understanding of the initial melting behaviour of deposits and illustrate the connection between deposit morphology and deposit ageing. The results can be utilised as input data for deposition models and help in predicting the final deposit morphology and its associated ageing behaviour, aiding boiler operators and manufacturers dealing with deposit-related issues.

A Neanderthal/Denisovan GLI3 variant contributes to anatomical variations in mice.

Changes in genomic structures underlie phenotypic diversification in organisms. Amino acid-changing mutations affect pleiotropic functions of proteins, although little is known about how mutated proteins are adapted in existing developmental programs. Here we investigate the biological effects of a variant of the GLI3 transcription factor (GLI3R1537C) carried in Neanderthals and Denisovans, which are extinct hominins close to modern humans. R1537C does not compromise protein stability or GLI3 activator-dependent transcriptional activities. In contrast, R1537C affects the regulation of downstream target genes associated with developmental processes. Furthermore, genome-edited mice carrying the Neanderthal/Denisovan GLI3 mutation exhibited various alterations in skeletal morphology. Our data suggest that an extinct hominin-type GLI3 contributes to species-specific anatomical variations, which were tolerated by relaxed constraint in developmental programs during human evolution.

First Eocene–Miocene anuran fossils from Peruvian Amazonia: insights into neotropical frog evolution and diversity

AbstractAnurans are one of the most diverse vertebrate groups, particularly in Amazonia, where species richness exceeds that of anywhere else. Amazonian frogs belong to three main lineages (Hyloidea, Microhylidae and Pipidae), each of which diversified during the Cenozoic. However, due to the virtual absence of an anuran fossil record in that area, the evolutionary history of modern lineages has so far remained accessible only via molecular data. In recent decades, a series of field campaigns in Peruvian Amazonia led to the discovery of an unparalleled set of anuran bone fragments, scattered across different sites spanning the Eocene–Miocene time interval. Here, we describe these first Palaeogene and early Neogene anurans from Peru with a focus on humeral and ilial morphology, identifying five humeral and five ilial morphotypes. Humeral morphotypes suggest the presence of different lineages of Brachycephaloidea in Peruvian fossil assemblages, whereas ilial morphotypes suggest the presence of Leptodactylidae, although leptodactylid‐like ilia also occur in some extant brachycephaloids. Pipids were also identified based on both humeral and ilial fragments. This study fills a major temporal and geographical gap in the evolutionary history of South American anurans, while further uncovering a lack of knowledge in the skeletal morphology of extant anuran families, as well as their inter‐ and intra‐species variability.

Old bones new tricks. Review of Behavior in Our Bones: How Human Behavior Influences Skeletal Morphology. Edited by Cara Stella Hirst, Rebecca J. Gilmour, Kimberly A. Plomp, and Francisca Alves Cardoso. Cambridge, MA: Elsevier Inc. 317 pp. ISBN: 9780128213834. $93.99/$125.00 (ebook/hardback).

Old bones new tricks. Review of Behavior in Our Bones: How Human Behavior Influences Skeletal Morphology. Edited by Cara Stella Hirst, Rebecca J. Gilmour, Kimberly A. Plomp, and Francisca Alves Cardoso. Cambridge, MA: Elsevier Inc. 317 pp. ISBN: 9780128213834. $93.99/$125.00 (ebook/hardback).

Label-free quantitative proteomics in serum reveals candidate biomarkers associated with low bone mineral density in Mexican postmenopausal women.

Postmenopausal osteoporosis is a public health problem leading to an increased risk of fractures, negatively impacting women's health. The absence of sensitive and specific biomarkers for early detection of osteoporosis represents a substantial challenge for improving patient management. Herein, we aimed to identify potential candidate proteins associated with low bone mineral density (BMD) in postmenopausal women from the Mexican population. Serum samples from postmenopausal women (40 with normal BMD, 40 with osteopenia (OS), and 20 with osteoporosis (OP)) were analyzed by label-free LC-MS/MS quantitative proteomics. Proteome profiling revealed significant differences between the OS and OP groups compared to individuals with normal BMD. A quantitative comparison of proteins between groups indicated 454 differentially expressed proteins (DEPs). Compared to normal BMD, 14 and 214 DEPs were found in OS and OP groups, respectively, while 226 DEPs were identified between OS and OP groups. The protein-protein interaction and enrichment analysis of DEPs were closely linked to the bone mineral content, skeletal morphology, and immune response activation. Based on their role in bone metabolism, a panel of 12 candidate biomarkers was selected, of which 1 DEP (RYR1) was found upregulated in the OS and OP groups, 8 DEPs (APOA1, SHBG, FETB, MASP1, PTK2B, KNG1, GSN, and B2M) were upregulated in OP and 3 DEPs (APOA2, RYR3, and HBD) were downregulated in OS or OP. The proteomic analysis described here may help discover new and potentially non-invasive biomarkers for the early diagnosis of osteoporosis in postmenopausal women.