P311 Promotes IL-4 Receptor‒Mediated M2 Polarization of Macrophages to Enhance Angiogenesis for Efficient Skin Wound Healing.

The transition from the proinflammatory phase to the prohealing phase in wound healing is essential for effective skin wound repair, which involves the balance of M1 and M2 polarization of wound-infiltrating macrophages. P311 plays an essential role in promoting wound closure by enhancing the biological function of epidermal stem cells, endothelial cells, and fibroblasts. Nevertheless, whether and how P311 regulates macrophage polarization remains unclear. In this study, we showed that P311 deficiency reduced the M2 polarization of macrophages, thereby attenuating the secretion of M2-like cytokines. The P311 deficiency prolonged the transition from the proinflammatory phase to the prohealing phase, accompanied by weakened angiogenesis and retarded granulation tissue formation, both of which coordinately hinder the healing of skin wounds. Mechanistically, P311 deficiency downregulated the expression of IL-4 receptor on macrophages, followed by less activation of the IL-4 receptor‒signal transducer and activator of transcription 6 signaling pathway, resulting in impaired M2 macrophage polarization. We further revealed that the mTOR signaling pathway was associated with the regulation of P311 on the expression of IL-4 receptor in macrophages. Thus, our study has highlighted the pivotal role of P311 in promoting the M2 polarization of macrophages for effective skin wound healing.

An ancestral hard-shelled sea turtle with a mosaic of soft skin and scutes.

The transition from terrestrial to marine environments by secondarily aquatic tetrapods necessitates a suite of adaptive changes associated with life in the sea, e.g., the scaleless skin in adult individuals of the extant leatherback turtle. A partial, yet exceptionally preserved hard-shelled (Pan-Cheloniidae) sea turtle with extensive soft-tissue remains, including epidermal scutes and a virtually complete flipper outline, was recently recovered from the Eocene Fur Formation of Denmark. Examination of the fossilized limb tissue revealed an originally soft, wrinkly skin devoid of scales, together with organic residues that contain remnant eumelanin pigment and inferred epidermal transformation products. Notably, this stem cheloniid-unlike its scaly living descendants-combined scaleless limbs with a bony carapace covered in scutes. Our findings show that the adaptive transition to neritic waters by the ancestral pan-chelonioids was more complex than hitherto appreciated, and included at least one evolutionary lineage with a mosaic of integumental features not seen in any living turtle.

The promising significance of liraglutide combined with dapagliflozin or empagliflozin in the prevention of early diabetic nephropathy.

OBJECTIVE: To explore the significance of liraglutide combined with dapagliflozin or empagliflozin in the prevention of early diabetic nephropathy (DN) and its effects on renal function indices. METHODS: Three hundred patients with type 2 diabetes mellitus (T2DM) treated in our hospital from April 2019 to April 2020 were retrospectively included and divided into two groups according to different treatment regimens. Among them, 150 patients who received liraglutide alone were included in the single-drug group, and 150 patients treated with liraglutide combined with dapagliflozin or empagliflozin were included in the combination group. The baseline data and the improvement of inflammatory indices, blood glucose indices and renal function-related indices were compared between the two groups of patients. RESULTS: The baseline data such as age, body mass index, retinopathy, and course of disease had no significant difference between the two groups (P > 0.05). After treatment, the waist-to-hip ratio, total body fat percentage, total body fat mass, total body lean mass, and A/G ratio were significantly decreased in both groups (P < 0.05) compared with before treatment, and were significantly lower in the combination group than in the single-drug group (P < 0.05). The combination group had significantly lower urinary transferrin (Tf), neutrophil gelatinase-associated lipocalin (NGAL) and tumor necrosis factor (TNF)-α, insulin-like growth factor 1 (IGF-1), retinol-binding protein (RBP), homocysteine (Hcy), brain natriuretic peptide (BNP), 24 h urinary albumin excretion ratio (UAER), urine albumin to creatinine ratio (UACR) and urinary liver-type fatty acid binding protein (L-FABP) levels, and higher secretory frizzled-related protein 5 levels than the single-drug group after treatment (P < 0.05). CONCLUSION: Liraglutide combined with dapagliflozin or empagliflozin treatment can effectively reduce the levels of Tf, NGAL and TNF-α in patients with T2DM, and improve the renal function in terms of IGF-1, RBP, Hcy, BNP, UAER, UACR, L-FABP, showing high treatment safety.

Soft-Tissue, Rare Earth Element, and Molecular Analyses of Dreadnoughtus schrani, an Exceptionally Complete Titanosaur from Argentina.

Evidence that organic material preserves in deep time (>1 Ma) has been reported using a wide variety of analytical techniques. However, the comprehensive geochemical data that could aid in building robust hypotheses for how soft-tissues persist over millions of years are lacking from most paleomolecular reports. Here, we analyze the molecular preservation and taphonomic history of the Dreadnougtus schrani holotype (MPM-PV 1156) at both macroscopic and microscopic levels. We review the stratigraphy, depositional setting, and physical taphonomy of the D. schrani skeletal assemblage, and extensively characterize the preservation and taphonomic history of the humerus at a micro-scale via: (1) histological analysis (structural integrity) and X-ray diffraction (exogenous mineral content); (2) laser ablation-inductively coupled plasma mass spectrometry (analyses of rare earth element content throughout cortex); (3) demineralization and optical microscopy (soft-tissue microstructures); (4) in situ and in-solution immunological assays (presence of endogenous protein). Our data show the D. schrani holotype preserves soft-tissue microstructures and remnants of endogenous bone protein. Further, it was exposed to LREE-enriched groundwaters and weakly-oxidizing conditions after burial, but experienced negligible further chemical alteration after early-diagenetic fossilization. These findings support previous hypotheses that fossils that display low trace element uptake are favorable targets for paleomolecular analyses.

Soft Tissue and Biomolecular Preservation in Vertebrate Fossils from Glauconitic, Shallow Marine Sediments of the Hornerstown Formation, Edelman Fossil Park, New Jersey.

Endogenous biomolecules and soft tissues are known to persist in the fossil record. To date, these discoveries derive from a limited number of preservational environments, (e.g., fluvial channels and floodplains), and fossils from less common depositional environments have been largely unexplored. We conducted paleomolecular analyses of shallow marine vertebrate fossils from the Cretaceous-Paleogene Hornerstown Formation, an 80-90% glauconitic greensand from Jean and Ric Edelman Fossil Park in Mantua Township, NJ. Twelve samples were demineralized and found to yield products morphologically consistent with vertebrate osteocytes, blood vessels, and bone matrix. Specimens from these deposits that are dark in color exhibit excellent histological preservation and yielded a greater recovery of cells and soft tissues, whereas lighter-colored specimens exhibit poor histology and few to no cells/soft tissues. Additionally, a well-preserved femur of the marine crocodilian Thoracosaurus was found to have retained endogenous collagen I by immunofluorescence and enzyme-linked immunosorbent assays. Our results thus not only corroborate previous findings that soft tissue and biomolecular recovery from fossils preserved in marine environments are possible but also expand the range of depositional environments documented to preserve endogenous biomolecules, thus broadening the suite of geologic strata that may be fruitful to examine in future paleomolecular studies.

Capture of fire smoke particles by leaves of Cunninghamia lanceolata and Schima superba, and importance of leaf characteristics.

Emission of particulate matter (PM) during forest fires is a major source of air pollution and hence purification of atmospheric pollution has gained increasing importance. Trees can absorb polluting gases and fine particles by their leaves from the atmosphere and act as a sustainable air purification filter. However, the capture efficiency varies among tree species; thus exploring the ability of forest trees to capture smoke PM released during forest fires provides a basis for assessing net emissions from forest fires and the impact of smoke on forest ecosystems. In this study, the main afforestation tree species, Cunninghamia lanceolata (Lamb.) Hook, and a fire-resistant tree species, Schima superba Gardn.et Champ, in southern China were exposed to different smoke concentrations by simulating forest fire. The amount of PM per unit leaf area, absorption of nutrient element, leaf surface characteristics and antioxidant enzyme activities were determined. The main findings were: (1) The total quantity of PM captured by unit leaf area (µg·cm-2) of C. lanceolata was 28.25 ± 1.12, 30.52 ± 3.43 and 33.14 ± 3.00 in low, intermediate and high smoke concentrations, respectively. The corresponding values for S. superba was 5.96 ± 0.56, 10.09 ± 1.13 and 12.27 ± 0.39, respectively. (2) Both species had weak absorption capacity for inorganic ions in the PM. (3) The purification of smoke PM by leaves was mainly related to leaf surface roughness, where it was higher for C. lanceolata than S. superba leaves. (4) Smoke treatment positively affected the contents of chlorophyll and soluble protein as well as increased antioxidant enzyme activities. In conclusion, the findings highlight the importance of leaf structural characteristics in capturing smoke particles and C. lanceolata is better suited for purification of atmospheric smoke particles following forest fire than S. superba.

Dynamics of pollutant emissions from wildfires in Mainland China.

We analyzed the dynamics of pollutant emissions from wildfires in mainland China from 2001 to 2019 using MODIS fire products combined with the measurements of emission factors of different vegetation types. The biomass distribution in Mainland China has heterogeneous temporal and spatial pattern, with inter-year variations and a decreasing trend from east to west. Overall, from 2001 to 2019, biomass combustion in Mainland China reached 479.59 Tg (25.24 Tg·a-1), in which northeast, north, east, south, central, northwest, and southwest regions accounted for 20.95%, 31.14%, 8.89%, 9.06%, 3.98%, 0.33% and 25.64% of total biomass combustion, respectively. The emissions of CO, CO2, CxHy, NOx, PM2.5, TC, OC and EC were 47.30, 288.05, 12.90, 0.40, 1.43, 0.83, 0.70, and 0.12 Tg (1 Tg = 1012g), respectively. PM2.5, TC and OC emissions increased in the southwest, while all pollutant emissions declined significantly in the southern region. For particulate matter from wildfires, both the ratio of its emissions to total dust and the ratio of its concentration to atmospheric PM2.5 showed an increasing trend, implying that the relative environmental impacts of particulate emissions from wildfires may be rising. In addition, our results show that the current Chinese wildfire management has successfully reduced on average more than 80% of pollutant emissions from wildfire from 2001 to 2019 compared to the natural wildfire regime (no strict wildfire management). This research on the temporal-spatial changes of pollutant emissions from wildfires in Mainland China provides support for further exploration of wildfire impacts on regional environments, and indicates the effectiveness of Chinese current wildfire policy on the pollutant emission mitigation.

Environmental Factors Affecting Feather Taphonomy.

The exceptional preservation of feathers in the fossil record has led to a better understanding of both phylogeny and evolution. Here we address factors that may have contributed to the preservation of feathers in ancient organisms using experimental taphonomy. We show that the atmospheres of the Mesozoic, known to be elevated in both CO2 and with temperatures above present levels, may have contributed to the preservation of these soft tissues by facilitating rapid precipitation of hydroxy- or carbonate hydroxyapatite, thus outpacing natural degradative processes. Data also support that that microbial degradation was enhanced in elevated CO2, but mineral deposition was also enhanced, contributing to preservation by stabilizing the organic components of feathers.

Progranulin improves neural development via the PI3K/Akt/GSK-3ß pathway in the cerebellum of a VPA-induced rat model of ASD.

Autism spectrum disorder (ASD) is a neurodevelopmental disease featuring social interaction deficits and repetitive/stereotyped behaviours; the prevalence of this disorder has continuously increased. Progranulin (PGRN) is a neurotrophic factor that promotes neuronal survival and differentiation. However, there have not been sufficient studies investigating its effect in animal models of autism. This study investigated the effects of PGRN on autistic phenotypes in rats treated with valproic acid (VPA) and assessed the underlying molecular mechanisms. PGRN was significantly downregulated in the cerebellum at postnatal day 14 (PND14) and PND35 in VPA-exposed rats, which simultaneously showed defective social preference, increased repetitive behaviours, and uncoordinated movements. When human recombinant PGRN (r-PGRN) was injected into the cerebellum of newborn ASD model rats (PND10 and PND17), some of the behavioural defects were alleviated. r-PGRN supplementation also reduced cerebellar neuronal apoptosis and rescued synapse formation in ASD rats. Mechanistically, we confirmed that PGRN protects neurodevelopment via the PI3K/Akt/GSK-3ß pathway in the cerebellum of a rat ASD model. Moreover, we found that prosaposin (PSAP) promoted the internalisation and neurotrophic activity of PGRN. These results experimentally demonstrate the therapeutic effects of PGRN on a rat model of ASD for the first time and provide a novel therapeutic strategy for autism.

Periodontopathic Microbiota and Atherosclerosis: Roles of TLR-Mediated Inflammation Response.

Atherosclerosis is a chronic inflammatory disease with a high prevalence worldwide, contributing to a series of adverse cardiovascular and cerebrovascular diseases. Periodontal disease induced by pathogenic periodontal microbiota has been well established as an independent factor of atherosclerosis. Periodontal microorganisms have been detected in atherosclerotic plaques. The high-risk microbiota dwelling in the subgingival pocket can stimulate local and systematic host immune responses and inflammatory cascade reactions through various signaling pathways, resulting in the development and progression of atherosclerosis. One often-discussed pathway is the Toll-like receptor-nuclear factor-κB (TLR-NF-κB) signaling pathway that plays a central role in the transduction of inflammatory mediators and the release of proinflammatory cytokines. This narrative review is aimed at summarizing and updating the latest literature on the association between periodontopathic microbiota and atherosclerosis and providing possible therapeutic ideas for clinicians regarding atherosclerosis prevention and treatment.

P311 Facilitates the Angiogenesis and Wound Healing Function of MSCs by Increasing VEGF Production.

As a potential clinical therapeutic cell for injured tissue repair, mesenchymal stem cells (MSCs) have attracted increasing attention. Enhancing the pro-healing function of MSCs has gradually become an essential topic in improving the clinical efficacy of MSCs. Recently, studies have shown that neuronal protein 3.1 (P311) plays a crucial role in promoting skin wound healing, suggesting P311 gene modification may improve the pro-healing function of MSCs. In this study, we demonstrated that increasing the in vivo expression of P311 could significantly enhance the ability of MSCs to lessen the number of inflammatory cells, increase the expression of IL10, reduce the levels of TNF-α and IFN-γ, increase collagen deposition, promote angiogenesis, and ultimately accelerate skin wound closure and improve the quality of wound healing. Importantly, we uncovered that P311 enhanced the pro-angiogenesis function of MSCs by increasing the production of vascular endothelial growth factor (VEGF) in vitro and in vivo. Mechanistically, we revealed that the mTOR signalling pathway was closely related to the regulation of P311 on VEGF production in MSCs. Together, our data displayed that P311 gene modification in MSCs augments their capabilities to promote skin wound closure, which might bring the dawn for its clinical application in the future.

P311 promotes type II transforming growth factor-ß receptor mediated fibroblast activation and granulation tissue formation in wound healing.

Background: P311, a highly conserved 8 kDa intracellular protein, has recently been reported to play an important role in aggravating hypertrophic scaring by promoting the differentiation and secretion of fibroblasts. Nevertheless, how P311 regulates the differentiation and function of fibroblasts to affect granulation tissue formation remains unclear. In this work, we studied the underlying mechanisms via which P311 affects fibroblasts and promotes acute skin wound repair. Methods: To explore the role of P311, both in vitro and in vivo wound-healing models were used. Full-thickness skin excisional wounds were made in wild-type and P311-/- C57 adult mice. Wound healing rate, re-epithelialization, granulation tissue formation and collagen deposition were measured at days 3, 6 and 9 after skin injury. The biological phenotypes of fibroblasts, the expression of target proteins and relevant signaling pathways were examined both in vitro and in vivo. Results: P311 could promote the proliferation and differentiation of fibroblasts, enhance the ability of myofibroblasts to secrete extracellular matrix and promote cell contraction, and then facilitate the formation of granulation tissue and eventually accelerate skin wound closure. Importantly, we discovered that P311 acts via up-regulating the expression of type II transforming growth factor-ß receptor (TGF-ßRII) in fibroblasts and promoting the activation of the TGF-ßRII-Smad signaling pathway. Mechanistically, the mammalian target of rapamycin signaling pathway is closely implicated in the regulation of the TGF-ßRII-Smad pathway in fibroblasts mediated by P311. Conclusions: P311 plays a critical role in activation of the TGF-ßRII-Smad pathway to promote fibroblast proliferation and differentiation as well as granulation tissue formation in the process of skin wound repair.

Analysis of regulatory effect of miR-149-5p on Sphingosine-1-phosphate receptor 2 of pericytes and its neuroprotective molecular mechanism after acute cerebral ischemia reperfusion in rats.

To investigate the effect of miR-149-5p on sphingosine-1-phosphate receptor 2 (S1PR2) expression level and contents of matrix metalloproteinase (MMP-9) and superoxide dismutase (SOD) in the pericytes after acute cerebral ischemia reperfusion in rats, so as to clarify the neuroprotective molecular mechanism induced by miR-149-5p and provide references for the treatment of neurological diseases, 60 male SD rats aged 7-8 weeks were selected and divided randomly into test group (establishing middle cerebral artery occlusion (MCAO) model) and control group (no modeling). Rat pericytes and peripheral cerebral infarction tissues were collected 12 h, 1 d, 3 d, 5 d, and 7 d after MCAO modeling, respectively. The pericytes were identified by immunofluorescence assay (IFA) and transfected with miR-149-5p. Fluorescence quantitative PCR (FQPCR) and Western blot were adopted to detect S1PR2 expression level. The expression of S1PR2 in MCAO model rats was detected by IFA. Immunohistochemistry (IHC) and quantitative real-time PCR (qRT-PCR) were used to detect the changes of MMP9 protein and mRNA levels of SOD1, SOD2, and SOD3 in brain tissue. The results showed that mRNA level and protein expression level of S1PR2 in the test group were higher than those in the control group three days after MCAO modeling (P < 0.05); the expression of S1PR2 increased 12 h after MCAO modeling and returned to the normal level on the 5th day, and the content of MMP9 protein in brain tissue of the test group was significantly lower than that of the control group (P < 0.05); the mRNA levels and SODs activity of SOD1, SOD2, and SOD3 in the test group were higher than those in the control group (P < 0.05). Therefore, miR-149-5p played a neuroprotective role by regulating S1PR2 to change the expression levels of SODS and MMP9.

Analysis of role of rat cerebral pericytes in cerebral vasospasm after subarachnoid hemorrhage and molecular mechanism of neurovascular injury.

To investigate mechanism of pericytes in the early stage of subarachnoid haemorrhage (SAH) and its associated microvascular spasm and neurovascular injury, 100 healthy 8-week-old Sprague-Dawley male rats were taken as subjects and divided into four groups: group A (sham operation, control group), group B (SAH operation group), group C (SAH operation group treated with scutellarin), and group D (SAH operation group treated with L-nitro-arginine). 72 hours after the operation, the rats were conducted assessment of neurological impairment, observation of microangiography, detection of blood-brain barrier permeability, observation of skull base haemorrhage, identification of pericyte culture, and measurement of blood nitric oxide. The results showed that neurological impairment score, degree of micro-vasoconstriction, and BBB permeability of group C were significantly better than those of group B and D (P<0.05), there was no significant difference between group C and group A (P>0.05). There were significantly fewer blood clots in the brain of group C, and the order of expression levels of α-smooth muscle actin (α-SMA) in perioperative cells of the four groups from highest to lowest were D, B, C, and A. Nitric oxide concentration inhibited expression of α-SMA in pericytes after SAH at both protein and mRNA levels. The detection results of nitric oxide in the blood of four groups of rats confirmed that pericyte phenotype conversion and actin α-SMA expression could be prevented by upregulation of nitric oxide in serum, so as to relieve pathological symptoms after SAH operation.

Baicalin Inhibits NLRP3 Inflammasome Activity Via the AMPK Signaling Pathway to Alleviate Cerebral Ischemia-Reperfusion Injury.

Baicalin has been reported to have ameliorative effects on nerve-induced hypoxic ischemia injury; however, its role in the NLRP3 inflammasome-dependent inflammatory response during cerebral ischemia-reperfusion remains unclear. To investigate the molecular mechanisms involved in baicalin alleviating cerebral ischemia-reperfusion injury, we investigated the AMPK signaling pathway which regulates NLRP3 inflammasome activity. SD rats were treated with baicalin at doses of 100 mg/kg and 200 mg/kg, respectively, after middle cerebral artery occlusion at 2 h and reperfusion for 24 h (MCAO/R). MCAO/R treatment significantly increased cerebral infarct volume, changed the ultrastructure of nerve cells, and activated the NLRP3 inflammasome, manifesting as significantly increased expression of NLRP3, ASC, cleaved caspase-1, IL-1ß, and IL-18. Our results demonstrated that baicalin treatment effectively reversed these phenomena in a dose-dependent manner. Additionally, inhibition of NLRP3 expression was found to promote the neuroprotective effects of baicalin on cortical neurons. Furthermore, baicalin remarkably increased the expression of p-AMPK following oxygen glucose deprivation/reperfusion (OGD/R). The expression of the NLRP3 inflammasome was also increased when the AMPK pathway was blocked by compound C. Taken together, our findings reveal that baicalin reduces the activity of the NLRP3 inflammasome and consequently inhibits cerebral ischemia-reperfusion injury through activation of the AMPK signaling pathway.

Abnormal spatiotemporal expression pattern of progranulin and neurodevelopment impairment in VPA-induced ASD rat model.

Some environmental risk factors have been proven to contribute to the etiology of autism spectrum disorder (ASD). Exposure to the antiepileptic drug valproic acid (VPA) during pregnancy significantly increases the risk of ASD in humans, and consequently is utilized as a validated animal model of ASD in rodents; however, the precise molecular and cellular mechanisms remain ill-defined. In the present study, we investigated the effect of prenatal VPA exposure on the spatiotemporal dynamics of Progranulin (PGRN) expression, neuronal apoptosis, synapse density, and AKT/GSK-3ß pathway activation in the brains of VPA-exposed offspring. Results from behavioral tests were consistent with prior studies showing impaired sociability, restricted interests and increased repetitive behaviors in VPA rats at postnatal days 28-32. Our data also indicated that VPA exposure resulted in abnormal dynamics of PGRN expression in different brain regions at the different development stages. The temporal and spatial patterns of PGRN expression were consistent with the spatiotemporal regularity of abnormalities, which observed in apoptosis-related protein levels, neuron numbers, dendritic spine density, synapse-related protein levels, and AKT/GSK-3ß phosphorylation in VPA rats. It suggests that prenatal VPA exposure may affect the spatiotemporal regularity of neuronal apoptosis and synaptic development/regression via interfering with the spatiotemporal process of PGRN expression and downstream AKT/GSK-3ß pathway activation. This may be a potential mechanism of the abnormal neuroanatomical changes and ASD-like behaviors in VPA-induced ASD.

A case of White-Sutton syndrome arising from a maternally-inherited mutation in POGZ.

POGZ is located on chromosome 1q21.3, encoding a pogo transposable element-derived protein with a zinc finger cluster. White-Sutton syndrome (WHSUS, OMIM:616364) is a genetic disorder resulting from de novo heterozygous pathogenic variants in POGZ, which manifests as intellectual disability, autism spectrum disorder, specific facial features and other phenotypic spectra. To date, a total of twenty-one de novo POGZ mutations in WHSUS have been reported. Here we report the identification of a novel missense variant in the coding region of the POGZ gene (c.4042G>C), which occurred in a 15-year-old male and his mother with WHSUS. We describe their clinical features and compare them with clinical data of patients with WHSUS from the literature. Our finding broadens the spectrum of POGZ mutations and provides a good example of precision medicine through the combination of exome sequencing and clinical testing.

Effect of forest floor fuel moisture content on chemical components of PM2.5 emitted during combustion.

The moisture content of forest floor fuels changes continuously with the influence of environmental factors; thus it has an important impact on the concentration and chemical composition of particulate matter emitted during forest fire. However, most previous studies quantify emissions of particulate matter and constituents using dry samples. In this study, we use a self-designed semi closed combustion simulator to quantify emission of total carbon (TC), organic carbon (OC), elemental carbon (EC) and water-soluble ions in fine particulate matter (PM2.5) using fuels of four tree species that differ in moisture content (0, 10, 20 and 30%). The results showed that the emissions of TC, OC and EC and total water-soluble inorganic ions increased significantly (<0.05) with increasing moisture content of fuels, and fuels of coniferous species emitted significantly more pollutants than fuels of broadleaved species. Similarly combustion of leaf samples emitted more carbonaceous components and water-soluble ions than combustion of branches. K+, NH4+ and Cl- were the main components of water-soluble ionic species, and emissions of K+, Ca2+, Na+, Mg2+, NH4+, Cl-, Br-, NO3-, NO2-, SO42- increased with increasing moisture content of fuels. Fuel moisture content had a great impact on the inorganic salt composition in the particulate matter emitted during combustion. The findings have an important implication on the use of prescribed early fire as management tools as the moisture content of the fuels early during the dry season is still high.

Keratan sulfate as a marker for medullary bone in fossil vertebrates.

The ability to determine the sex of extinct dinosaurs by examining the bones they leave behind would revolutionize our understanding of their paleobiology; however, to date, definitive sex-specific skeletal traits remain elusive or controversial. Although living dinosaurs (i.e., extant birds) exhibit a sex-specific tissue called medullary bone that is unique to females, the confident identification of this tissue in non-avian archosaurs has proven a challenge. Tracing the evolution of medullary bone is complicated by existing variation of medullary bone tissues in living species; hypotheses that medullary bone structure or chemistry varied during its evolution; and a lack of studies aimed at distinguishing medullary bone from other types of endosteal tissues with which it shares microstructural and developmental characteristics, such as pathological tissues. A recent study attempted to capitalize on the molecular signature of medullary bone, which, in living birds, contains specific markers such as the sulfated glycosaminoglycan keratan sulfate, to support the proposed identification of medullary bone of a non-avian dinosaur specimen (Tyrannosaurus rex MOR 1125). Purported medullary bone samples of MOR 1125 reacted positively to histochemical analyses and the single pathological control tested (avian osteopetrosis) did not, suggesting the presence of keratan sulfate might serve to definitively discriminate these tissues for future studies. To further test these results, we sampled 20 avian bone pathologies of various etiologies (18 species), and several MB samples. Our new data universally support keratan sulfate as a reliable marker of medullary bone in birds. However, we also find that reactivity varies among pathological bone tissues, with reactivity in some pathologies indistinguishable from MB. In the current sample, some pathologies comprised of chondroid bone (often a major constituent of skeletal pathologies and developing fracture calluses in vertebrates) contain keratan sulfate. We note that beyond chemistry, chondroid bone shares many characteristics with medullary bone (fibrous matrix, numerous and large cell lacunae, potential endosteal origin, trabecular architecture) and medullary bone has even been considered by some to be a type of chondroid bone. Our results suggest that the presence of keratan sulfate is not exclusive evidence for MB, but rather must be used as one in a suite of criteria available for identifying medullary bone (and thus gravid females) in non-avian dinosaur specimens. Future studies should investigate whether there are definite chemical or microstructural differences between medullary bone and reactive chondroid bone that can discriminate these tissues.

Molecular tests support the viability of rare earth elements as proxies for fossil biomolecule preservation.

The rare earth element (REE) composition of a fossil bone reflects its chemical alteration during diagenesis. Consequently, fossils presenting low REE concentrations and/or REE profiles indicative of simple diffusion, signifying minimal alteration, have been proposed as ideal candidates for paleomolecular investigation. We directly tested this prediction by conducting multiple biomolecular assays on a well-preserved fibula of the dinosaur Edmontosaurus from the Cretaceous Hell Creek Formation previously found to exhibit low REE concentrations and steeply-declining REE profiles. Gel electrophoresis identified the presence of organic material in this specimen, and subsequent immunofluorescence and enzyme-linked immunosorbant assays identified preservation of epitopes of the structural protein collagen I. Our results thereby support the utility of REE profiles as proxies for soft tissue and biomolecular preservation in fossil bones. Based on considerations of trace element taphonomy, we also draw predictions as to the biomolecular recovery potential of additional REE profile types exhibited by fossil bones.