The Crocodylian proatlas functions to redistribute venous blood and cerebrospinal fluid.

The proatlas, a bone located between the skull and the neural spines of the cervical vertebrae, is best known from reptiles. Most previous studies of the proatlas have centered on its developmental, debating the relationship between the proatlas and the cervical neural arches. The present study was intended as a description of the proatlas in the American alligator (Alligator mississippiensis) and an experimental test of its hypothesized role in venous blood and cerebrospinal fluid (CSF) distribution. In Alligator, the proatlas is chevron-shaped; ventrally it has a loose connection to the dorsal surface of the first cervical vertebrae, dorsally it has a robust elastic tissue tether on the otoccipital and supraoccipital bones. The ventral surface of the proatlas parallels the dorsal margin of the foramen magnum and rests on the dorsal surface of the spinal venous sinus. Experimental manipulation of the proatlas demonstrated that displacement of the proatlas causes pressure changes in both the spinal venous sinus and the enclosed spinal CSF. The results of this study represent the first demonstration of an explicit functional role for the proatlas, the circulation of fluids between the cranial and spinal compartments of the central nervous system.

On the spinal venous sinus of Alligator mississippiensis.

The epidural space of the American alligator (Alligator mississippiensis) is largely filled by a continuous venous sinus. This venous sinus extends throughout the trunk and tail of the alligator, and is continuous with the dural sinuses surrounding the brain. Segmental spinal veins (sl) link the spinal venous sinus (vs) to the somatic and visceral venous drainage. Some of these sl, like the caudal head vein along the occipital plate of the skull, are enlarged, suggesting more functional linkage. No evidence of venous valves or external venous sphincters was found associated with the vs; the relative scarcity of smooth muscle in the venous wall of the sinus suggests limited physiological regulation. The proatlas (pr), which develops between the occipital plate and C1 in crocodylians, is shaped like a neural arch and is fused to the dorsal surface of the vs. The present study suggests that the pr may function to propel venous blood around the brain and spinal cord. The vs effectively encloses the spinal dura, creating a tube-within-a-tube system with the (smaller volume) spinal cerebrospinal fluid (CSF). Changes in venous blood pressure, as are likely during locomotion, would impact dural compliance and CSF pressure waves propagating along the spinal cord.

Amniotic fluid embolism: a reappraisal.

OBJECTIVES: Using cases from our own experience and from the published literature on amniotic fluid embolism (AFE), we seek to improve on existing criteria for diagnosis and discern associated risk factors. Additionally, we propose a novel theory of pathophysiology. METHODS: This retrospective case review includes eight cases of AFE from two hospital systems and 21 from the published literature. All cases were evaluated using the modified criteria for research reporting of AFE by Clark et al. in Am J Obstet Gynecol, 2016;215:408-12 as well as our proposed criteria for diagnosis. Additional clinical and demographic characteristics potentially correlated with a risk of AFE were included and analyzed using descriptive analysis. RESULTS: The incidence of AFE was 2.9 per 100,000 births, with five maternal deaths in 29 cases (17.2 %) in our series. None of the cases met Clark's criteria while all met our criteria. 62.1 % of patients were over the age of 32 years and two out of 29 women (6.9 %) conceived through in-vitro fertilization. 6.5 % of cases were complicated by fetal death. Placenta previa occurred in 13.8 %. 86.2 % of women had cesarean sections of which 52.0 % had no acute maternal indication. CONCLUSIONS: Our criteria identify more patients with AFE than others with a low likelihood of false positives. Clinical and demographic associations in our review are consistent with those previously reported. A possible relationship between cesarean birth and risk of AFE was identified using our criteria. Additionally, we propose a new hypothesis of pathophysiology.

The presence of a foramen of Luschka in the American alligator (Alligator mississippiensis) and the continuity of the intraventricular and subdural spaces.

In humans and most mammals, there is a notch-like portal, the foramen of Luschka (or lateral foramen), which connects the lumen of the fourth ventricle with the subdural space. Gross dissection, light and scanning electron microscopy, and µCT analysis revealed the presence of a foramen of Luschka in the American alligator (Alligator mississippiensis). In this species, the foramen of Luschka is a notch in the dorsolateral wall of the pons immediately caudal to the peduncular base of the cerebellum, near the rostral end of the telovelar membrane over the fourth ventricle. At the foramen of Luschka there was a transition from a superficial pia mater lining to a deep ependymal lining. There was continuity between the lumen of the fourth ventricle and the subdural space, via the foramen of Luschka. This anatomical continuity was further demonstrated by injecting Evans blue into the lateral ventricle which led to extravasation through the foramen of Luschka and pooling of the dye on the lateral surface of the brain. Simultaneous subdural and intraventricular recordings of cerebrospinal fluid (CSF) pressures revealed a stable agreement between the two pressures at rest. Perturbation of the system allowed for static and dynamic differences to develop, which could indicate varying flow patterns of CSF through the foramen of Luschka.

The anatomical basis of amphibious hearing in the American alligator (Alligator mississippiensis).

The different velocities of sound (pressure waves) in air and water make auditory source localization a challenge for amphibious animals. The American alligator (Alligator mississippiensis) has an extracolumellar cartilage that abuts the deep surface of the tympanic membrane, and then expands in size beyond the caudal margin of the tympanum. This extracolumellar expansion is the insertion site for two antagonistic skeletal muscles, the tensor tympani, and the depressor tympani. These muscles function to modulate the tension in the tympanic membrane, presumably as part of the well-developed submergence reflex of Alligator. All crocodilians, including Alligator, have internally coupled ears in which paratympanic sinuses connect the contralateral middle ear cavities. The temporal performance of internally coupled ears is determined, in part, by the tension of the tympanic membrane. Switching between a "tensed" and "relaxed" tympanic membrane may allow Alligator to compensate for the increased velocity of sound underwater and, in this way, use a single auditory map for sound localization in two very different physical environments.

Dynamic asymmetry in cerebrospinal fluid pressure: An indicator of regional differences in compliance.

Background: Dural compliance influences the shape and magnitude of the cerebrospinal fluid (CSF) pulsations. In humans, cranial compliance is approximately 2× greater than spinal compliance; the differential has been attributed to the associated vasculature. In alligators, the spinal cord is surrounded by a large venous sinus, which suggests that the spinal compartment may have higher compliance than is found in mammals. Methods: Pressure catheters were surgically implanted into the cranial and spinal subdural spaces of eight subadult American alligators (Alligator mississippiensis). The CSF was propelled through the subdural space by orthostatic gradients and rapid changes in linear acceleration. Results: CSF pressure recordings taken from the cranial compartment were consistently, and significantly, larger than those taken from the spinal compartment. After the myodural bridge of Alligator was surgically released, the asymmetry in CSF pressure was decreased. Conclusion: Unlike the situation in humans, the spinal compartment of Alligator has greater compliance than the cranial compartment, presumably due to the presence of the large spinal venous sinus surrounding the dura. The change in CSF pressures after myodural surgical release supports the hypothesis that the myodural bridge functions, at least in part, to modulate dural compliance and the exchange of CSF between the cranial and spinal compartments.

Prioritizing the reassessment of data-deficient species on the IUCN Red List.

Despite being central to the implementation of conservation policies, the usefulness of the International Union for Conservation of Nature (IUCN) Red List of Threatened Species is hampered by the 14% of species classified as data-deficient (DD) because information to evaluate these species' extinction risk was lacking when they were last assessed or because assessors did not appropriately account for uncertainty. Robust methods are needed to identify which DD species are more likely to be reclassified in one of the data-sufficient IUCN Red List categories. We devised a reproducible method to help red-list assessors prioritize reassessment of DD species and tested it with 6887 DD species of mammals, reptiles, amphibians, fishes, and Odonata (dragonflies and damselflies). For each DD species in these groups, we calculated its probability of being classified in a data-sufficient category if reassessed today from covariates measuring available knowledge (e.g., number of occurrence records or published articles available), knowledge proxies (e.g., remoteness of the range), and species characteristics (e.g., nocturnality); calculated change in such probability since last assessment from the increase in available knowledge (e.g., new occurrence records); and determined whether the species might qualify as threatened based on recent rate of habitat loss determined from global land-cover maps. We identified 1907 species with a probability of being reassessed in a data-sufficient category of >0.5; 624 species for which this probability increased by >0.25 since last assessment; and 77 species that could be reassessed as near threatened or threatened based on habitat loss. Combining these 3 elements, our results provided a list of species likely to be data-sufficient such that the comprehensiveness and representativeness of the IUCN Red List can be improved.

Priorización de la reevaluación de las especies con datos deficientes en la Lista Roja de la UICN Resumen No obstante que es fundamental para la implementación de políticas de conservación, la utilidad de la Lista Roja de Especies Amenazadas de la Unión Internacional para la Conservación de la Naturaleza (UICN) está limitada por el 14% de especies clasificadas con datos deficientes (DD) debido a que la información para evaluar el riesgo de extinción de estas especies no existía cuando fueron evaluadas la última vez o porque los evaluadores no consideraron la incertidumbre apropiadamente. Se requieren métodos robustos para identificar las especies DD con mayor probabilidad de ser reclasificadas en alguna de las categorías en la Lista Roja UICN con datos suficientes. Diseñamos un método reproducible para ayudar a que los evaluadores de la lista roja prioricen la reevaluación de especies DD y lo probamos con 6,887 especies DD de mamíferos, reptiles, anfibios, peces y Odonata (libélulas y caballitos del diablo). Para cada una de las especies DD en estos grupos, calculamos la probabilidad de ser clasificadas en una categoría con datos suficientes si fuera reevaluada hoy a partir de covariables que miden el conocimiento disponible (e.g., número de registros de ocurrencia o artículos publicados disponibles), sustitutos de conocimiento (e.g., extensión del rango de distribución) y características de la especie ((e.g., nocturnidad); calculamos el cambio en tal probabilidad desde la última reevaluación a partir del incremento en el conocimiento disponible (e.g., registros de ocurrencia nuevos); y determinamos si las especies podrían calificar como amenazadas con base en pérdidas de hábitat recientes a partir de mapas globales de cobertura de suelo recientes. Identificamos 1,907 especies con una probabilidad >0.5 de ser reclasificados en una categoría con datos suficientes; 624 especies cuya probabilidad aumentó en >0.25 desde la última evaluación, y 77 especies que podrían ser reclasificadas como casi en peligro con base en la pérdida de hábitat. Combinando estos 3 elementos, nuestros resultados proporcionaron una lista de especies probablemente con datos suficientes de tal modo que la exhaustividad y la representatividad de la Lista Roja de la UICN pueden ser mejoradas.

Clinical potential of human amniotic fluid stem cells.

OBJECTIVES: To determine whether amniotic fluid derived stem cells maintain their stem cell characteristics (a) after processing by a licensed cell therapy center and (b) after the cells undergo simulated clinical application. METHODS: Amniotic fluid was collected by laparotomy - a small uterine incision was made at proposed site for delivery and a sterile catheter inserted to collect fluid into a sterile bag. After flow stopped the catheter was withdrawn, the cesarean completed and the collected fluid delivered to the cell therapy center for processing and cryostorage. A clinical setting was simulated where amniotic fluid cells received from cell therapy center were thawed at room temperature for a maximum of 3 h and passed through a clinical cell delivery device to monitor cell viability. The cells were examined for viability, stability, growth, differentiation, and markers of stemness. RESULTS: Amniotic fluid stem cells processed from a clinical cell therapy center behave similarly to amniotic fluid stem cells processed in a research laboratory with respects to viability, stability, growth, differentiation and maintain markers of stemness. There were differences due to heterogeneity of samples which were not methodological. Growth in cell culture and differentiation were satisfactory. Simulation of treating the cells in a clinical environment show a general stability in viability of amniotic fluid cells at room temperature for 3 h minimum and when passed through a clinically approved delivery device. CONCLUSIONS: The data indicate human amniotic fluid processed in a clinical facility could be used therapeutically if proven to be safe.

Morphology of the distal tip of the spinal cord in Alligator mississippiensis.

Secondary neurulation is a common feature of vertebrate development, which in non-mammalian and non-anuran vertebrates, results in the formation of a caudal spinal cord. The present study was undertaken to describe the terminal end of the caudal spinal cord in a crocodylian, a group chosen for their unique status of a living-tailed archosaur. The caudal spinal cord of Alligator mississippiensis terminates near the intervertebral joint between the fourth and fifth terminal vertebrae. Prior to this termination, the dorsal root ganglia get proportionately larger, then stop before the termination of the spinal cord; and the gray matter of the spinal cord is lost producing an unusual morphology in which an ependymal-lined central canal is surrounded by only white matter which is not divided into a cauda equina. The inner layer of the meninges (the pia-arachnoid) courses over the distal end of the spinal cord and forms a ventral attachment, reminiscent of a very short Filum terminale; there is no caudal cistern. The dura extends beyond the termination of the spinal cord, continuing for at least the length of the fourth terminal vertebra, forming a structure herein termed the distal meningeal sheath. During its course, the distal meningeal sheath surrounds a mass of mesothelial cells, then terminates as an attachment on the dorsal surface of the vertebra.

The Influence of Movement on the Cerebrospinal Fluid Pressure of the American Alligator (Alligator mississippiensis).

This study was undertaken to document how the cerebrospinal fluid (CSF) pressure varied during movements and physiological activities. Using surgically implanted pressure catheters; the CSF pressure was recorded from sub-adult American alligators (Alligator mississippiensis) under anesthesia and post-recovery. Pressures were recorded during physiological activities (the cardiac cycle; passive and active ventilation); manual manipulation of the anesthetized animals (foot sweeps; tail oscillations; and body bends); as well as voluntary movements post-recovery (changes in body tone; defensive strikes; and locomotion). The CSF pulsations associated with the cardiac cycle had the lowest mean amplitude (3.7 mm Hg); during active ventilation and defensive strikes; the alligators routinely generated CSF pressure spikes in excess of 100 mm Hg. The recorded CSF pressures appear to be caused by a variety of mechanisms including vascular pressure; fluid inertia; and possible physical displacement of the spinal cord. The results of the study suggest that any model of CSF dynamics or perfusion should incorporate the episodic high-pressure CSF pulsations associated with movement.

The acoustical effect of the neck frill of the frill-necked lizard (Chlamydosaurus kingii).

Animals localise sound by making use of acoustical cues resulting from space and frequency dependent filtering of sound by the head and body. Sound arrives at each ear at different times, with different intensities, and with varying spectral content, all of which are affected by the animal's head and the relative sound source position. Location cues in mammals benefit from structures (pinnae) that modify these cues and provide information that helps resolve the cone of confusion and provide cues to sound source elevation. Animals without pinnae must rely on other mechanisms to solve localisation problems. Most non-mammals lack pinna-like structures, but some possess other anatomical features that could influence hearing. One such animal is the frill-necked lizard (Chlamydosaurus kingii). The species' elaborate neck frill has been speculated to act as an aid to hearing, but no acoustical measurements have been reported. In this study, we characterise the frill's influence on the acoustical information available to the animal. Results suggest that the change in binaural cues is not sufficiently large to impact localisation behavior within the species' likely audiometric range; however, the frill does increase gain for sounds directly in front of the animal similar to a directional microphone.

The orbitalauricular chord of Alligator: The unusual functional linkage between the earflap and eyelid of Crocodylians.

One of the distinctive features of the Crocodylia is the presence of a superficial meatal chamber the aperture of which is regulated by two earflaps. The movements of the upper earflap have been detailed by multiple workers, however, the mechanics of the lower earflap remain unresolved. The present study was undertaken to document the mechanics of the lower earflap in the American alligator, Alligator mississippiensis, and to explore the functional bases of coordinated movements between the lower earflap and lower eyelid in this species. This anatomical system was examined using a combination of fresh dissection, histology, and micro-CT analyses applied to post-embryonic specimens. The rostral margin of the lower earflap is tightly bound to a block of dense connective tissue herein termed the orbitalauricular chord. The orbitalauricular chord is anatomically distinct from both a ligament and a tendon. The dorsal surface of the orbitalauricular chord is attached to a slip of the levator palpebra, while the ventral surface is attached to a slip of the depressor palpebra. These attachments produce a simple mechanism for the elevation and depression of the lower earflap, and thus the opening and closing of the meatal aperture. The caudal surface of the orbitalauricular chord has connective tissue links to the rostral margin of the lower earflap. The morphology of the orbitalauricular chord appears to explain both the mechanics of the lower earflap and the functional coupling between the lower eyelid and lower earflap.

Treadmill locomotion in the American alligator (Alligator mississippiensis) produces dynamic changes in intracranial cerebrospinal fluid pressure.

To examine the influence of movement on cerebrospinal fluid (CSF) dynamics, intracranial subdural pressure recordings were taken from sub-adult alligators (Alligator mississippiensis) locomoting on a treadmill. Pressure recordings documenting the cardiac, ventilatory, and barostatic influences on the CSF were in good agreement with previous studies. During locomotion the CSF exhibits sinusoidal patterns of pressure change that spanned a mean amplitude of 56 mm Hg, some 16 × the amplitude of the cardiac-linked pulsations. These sinusoidal CSF pulsations were closely linked to the locomotor kinematics, particularly the lateral oscillations of the alligator's head. Data recorded from the freely moving alligators suggest that fluid inertia, body cavity pressures, and likely other factors all influence the CSF pressure. The clear relationship between movement and CSF pressure described in this study suggests that the paucity of studies examining human CSF dynamics during movement should be addressed.

A randomized controlled trial of the effects of whole grains versus refined grains diets on the microbiome in pregnancy.

Dietary whole grain consumption has been postulated to have metabolic benefits. The purpose of this study was to compare a pregnancy diet containing 75% of total carbohydrates as refined grains with a diet of 75% of total carbohydrates as whole grains for pregnancy outcomes and effects on the microbiome. Gestational weight gain, glucose tolerance and newborn outcomes were measured on 248 enrolled compliant women from whom a subset of 103 women consented to give 108 vaginal and 109 anal swabs. The data presented here are limited to the patients from whom the vaginal and anal swabs were obtained in order to study the microbiome. A microbiome-16SrRNA survey-was characterized in these samples. Samples and measurements were obtained at the first obstetrical visit, before beginning a prescribed diet (T1-baseline) and after 17-32 weeks on the prescribed diet (T3). Food frequency questionnaires and total plasma alkylresorcinols were used as a measure of whole grain consumption. There were no dietary differences in maternal weight gain, birth weight, or glucose tolerance test. Mothers consuming the whole grains diet showed a trend of gestational decrease in vaginal bacterial alpha diversity, with increasing Lactobacillus-dominance. No significant difference was observed for the anal microbiome. The results suggest that diet modulations of the vaginal microbiome during gestation may have important implications for maternal and neonatal health and in the intergenerational transfer of maternal microbiome. Trial registration: ClinicalTrials.gov Identifier: NCT03232762.

A global reptile assessment highlights shared conservation needs of tetrapods.

Comprehensive assessments of species' extinction risks have documented the extinction crisis1 and underpinned strategies for reducing those risks2. Global assessments reveal that, among tetrapods, 40.7% of amphibians, 25.4% of mammals and 13.6% of birds are threatened with extinction3. Because global assessments have been lacking, reptiles have been omitted from conservation-prioritization analyses that encompass other tetrapods4-7. Reptiles are unusually diverse in arid regions, suggesting that they may have different conservation needs6. Here we provide a comprehensive extinction-risk assessment of reptiles and show that at least 1,829 out of 10,196 species (21.1%) are threatened-confirming a previous extrapolation8 and representing 15.6 billion years of phylogenetic diversity. Reptiles are threatened by the same major factors that threaten other tetrapods-agriculture, logging, urban development and invasive species-although the threat posed by climate change remains uncertain. Reptiles inhabiting forests, where these threats are strongest, are more threatened than those in arid habitats, contrary to our prediction. Birds, mammals and amphibians are unexpectedly good surrogates for the conservation of reptiles, although threatened reptiles with the smallest ranges tend to be isolated from other threatened tetrapods. Although some reptiles-including most species of crocodiles and turtles-require urgent, targeted action to prevent extinctions, efforts to protect other tetrapods, such as habitat preservation and control of trade and invasive species, will probably also benefit many reptiles.

Morphometrics of the Spinal Cord and Surrounding Structures in Alligator mississippiensis.

Understanding the fluid dynamics of the cerebrospinal fluid requires a quantitative description of the spaces in which it flows, including the spinal cord and surrounding meninges. The morphometrics of the spinal cord and surrounding tissues were studied in specimens of the American alligator (Alligator mississippiensis) ranging from hatchlings through adults. Within any size class of alligators (i.e., hatchlings), along the axial length there are significant differences in the size of the spinal cord, meninges, and vertebral canal; these differences can be used to define discrete cervical, thoracic, lumbar and caudal regions. When compared across the range of body sizes in Alligator, every structure in each spinal region had a distinctive growth rate; thus, the physical arrangements between the structures changed as the alligator grew. The combination of regional differentiation and differential growth rates was particularly apparent in the lumbar meninges where a unique form of lumbar cistern could be identified and shown to decrease in relative size as the alligator ages. This analysis of the spinal cord and surrounding tissues was undertaken to develop a data set that could be used for computational flow dynamics of the crocodilian cerebrospinal fluid, and also to assist in the analysis of fossil archosaurs.

Increasing taxonomic diversity and spatial resolution clarifies opportunities for protecting US imperiled species.

Continental- and regional-scale assessments of gaps in protected area networks typically use relatively coarse range maps for well documented species groups, creating uncertainty about the fate of unexamined biodiversity and providing insufficient guidance for land managers. By building habitat suitability models for a taxonomically diverse group of 2216 imperiled plants and animals, we revealed comprehensive and detailed protection opportunities in the conterminous United States. Summing protection-weighted range-size rarity (PWRSR, the product of the percent of modeled habitat outside of protected areas and the inverse of modeled habitat extent) uncovered novel patterns of biodiversity importance. Concentrations of unprotected imperiled species in places such as the northern Sierra Nevada, central and northern Arizona, the Rocky Mountains of Utah and Colorado, southeastern Texas, southwestern Arkansas, and Florida's Lake Wales Ridge have rarely if ever been featured in continental- and regional-scale analyses. Inclusion of diverse taxa (vertebrates, freshwater mussels, crayfishes, bumble bees, butterflies, skippers, and vascular plants) partially drove these new patterns. When analyses were restricted to groups typically included in previous studies (birds, mammals, and amphibians), up to 53% of imperiled species in other groups were left out. The finer resolution of modeled inputs (990 m) also resulted in a more geographically dispersed pattern. For example, 90% of the human population of the conterminous United States lives within 50 km of modeled habitat for one or more species with high PWRSR scores. Over one-half of the habitat for 818 species occurs within federally lands managed for biodiversity protection; an additional 360 species have over one-half of their modeled habitat on federal multiple use land. Freshwater animals occur in places with poorer landscape condition but with less exposure to climate change than other groups, suggesting that habitat restoration is an important conservation strategy for these species. The results provide fine-scale, taxonomically diverse inputs for local and regional priority-setting and show that although protection efforts are still widely needed on private lands, notable gains can be achieved by increasing protection status on selected federal lands.

Bridging the research-implementation gap in IUCN Red List assessments.

The International Union for Conservation of Nature (IUCN) Red List of Threatened Species is central in biodiversity conservation, but insufficient resources hamper its long-term growth, updating, and consistency. Models or automated calculations can alleviate those challenges by providing standardised estimates required for assessments, or prioritising species for (re-)assessments. However, while numerous scientific papers have proposed such methods, few have been integrated into assessment practice, highlighting a critical research-implementation gap. We believe this gap can be bridged by fostering communication and collaboration between academic researchers and Red List practitioners, and by developing and maintaining user-friendly platforms to automate application of the methods. We propose that developing methods better encompassing Red List criteria, systems, and drivers is the next priority to support the Red List.