Immature Animals Research Articles

Induced hypothyroidism alters articular cartilage in skeletally immature miniature swine

ABSTRACT Purpose/Aim Thyroid hormone has been implicated in the normal growth and development of articular cartilage; however, its effect on a disease state, such as hypothyroidism, is unknown. The purpose of this investigation was to compare normal articular cartilage from proximal femurs of immature miniature swine to proximal femurs from hypothyroid-induced immature miniature swine. Materials and Methods Two 11-week-old male Sinclair miniature swine were made hypothyroid by administration of 6-propyl-2-thiouracil (PTU) in their drinking water; two control animals did not receive PTU. At 25 weeks of age, the animals were euthanized and their proximal femurs were fixed and decalcified. Samples were sectioned and analyzed by histology to define extracellular matrix (ECM) structure, immunohistochemistry (IHC) to identify types II and X collagen, and histomorphometry to assess articular cartilage mean total and localized height and cell density. Statistics included nested mixed-effects ANOVA with p ≤ 0.05 considered statistically significant. Results Compared to controls, hypothyroid articular cartilage demonstrated statistically significant quantitative differences in mean tissue height, mean cell density and type II collagen localized zone height. Qualitative differences in ECM proteoglycans and overall collagen types were also found. Type X collagen was not detected in either hypothyroid or control articular cartilage specimens. Conclusions Significant changes in articular cartilage structure in hypothyroid compared to control immature miniature swine suggest that thyroid hormone is critical in the growth and development of articular cartilage. Clinical Significance Understanding articular cartilage development in immature animal models may provide insight into healing or repair of degenerative human articular cartilage.

Target detection in healthy 4-week old piglets from a passive two-tone auditory oddball paradigm

BackgroundPassive auditory oddball tests are effort independent assessments that evaluate auditory processing and are suitable for paediatric patient groups. Our goal was to develop a two-tone auditory oddball test protocol and use this clinical assessment in an immature large animal model. Event-related potentials captured middle latency P1, N1, and P2 responses in 4-week old (N = 16, female) piglets using a custom piglet 32- electrode array on 3 non-consecutive days. The effect of target tone frequency (250 Hz and 4000 Hz) on middle latency responses were tested in a subset of animals.ResultsResults show that infrequent target tone pulses elicit greater N1 amplitudes than frequent standard tone pulses. There was no effect of day. Electrodes covering the front of the head tend to elicit greater waveform responses. P2 amplitudes increased for higher frequency target tones (4000 Hz) than the regular 1000 Hz target tones (p < 0.05).ConclusionsTwo-tone auditory oddball tests produced consistent responses day-to-day. This clinical assessment was successful in the immature large animal model.

Perampanel exhibits anticonvulsant action against pentylentetrazol-induced seizures in immature rats

Perampanel is a new antiepileptic drug with unique mechanism of action – antagonism of AMPA receptors. Its action in immature animals is not yet sufficiently known therefore we started to study anticonvulsant action of perampanel pretreatment (1–20 mg/kg i.p.) against seizures elicited by pentylenetetrazol. Three age groups of rats were examined – 12, 18 and 25 days old. Perampanel selectively suppressed the tonic phase of generalized seizures in the two younger groups and whole tonic-clonic seizures in the 25-day-old group. It exhibited also an anticonvulsant action against minimal clonic seizures present in control 18- and 25-day-old rats.Perampanel is an effective anticonvulsant drug even at very early stages of brain development.

Evaluation of local field potentials of MER data with STN DBS electrodes

The field potentials, i.e., the electrical-potentials of the deep brain stimulations (DBS) and microelectrode signal recordings of STN-neurons flanking DBS leads canister bright to proffer functional and empirical retort or titrating therapeutic-surgical-DBSDBS. Nonetheless, predictable square (conservative) DBS leadelectrodes amid four cylindrical macro/electrodes (for macrostimulations) likely under-sample the spatial circulation of sinks and sources in a given area of the brain. This study gives the exploration of the power-spectral-density (PSD) and spatial-feature dimensions of LFP-activity in primate subthalamicnuclei (STN/or s-nuclei) and globus pallidus internus (GPi) by means of unendingly 32embedded-channel directional DBS-array-electrodes. The waveforms of s-nuclei and GPi were acquired as of directional-DBS-micro electrode-arrays in the latent or quiescent state and in a reach-and-retrieval task in twoprimates in adolescent, naïve and immature (raw) animal Parkinson conditions. The potentials of signal acquisitions recordings were evaluated amongst bipolar-pairs of micro electrodes using personage and agglomerative (bunched) electrode configurations, with the latter mimicking the cylindrical macro electrode (for macro stimulations) arrangements/specifications employed in the present - current clinical settings of field-potentials signal acquisition. Signal recordings from these DBS-electrodes shown that, b -oscillatory frequency fluctuations have spatial-finger-prints in s-nuclei and GPi, and that these fluctuations were muted when clustering the electrode contacts together to create cylindrical macro electrodes alike in parallel and in relative dimension to those employed clinically and diagnostically. Furthermore, these atlases depend on parkinsonian condition and whether the subject was quiescent or performing a motor-task. With this research work the development of future adaptive closed-loop (ACL) DBS-therapies to rely on fieldpoten

PECULIARITIES OF MORPHOFUNCTIONAL CHANGES OF MASTICATORY MUSCLES IN IODINE-DEFICIENT CONDITIONS

The problem of iodine deficiency is becoming increasingly important in Ukraine. According to the United Nations Children’s Fund (UNICEF), each and all regions of Ukraine suffer from “hidden hunger”. The World Health Assembly has stated that the elimination of iodine deficiency will be as much a triumph for health care system as the eradication of smallpox and poliomyelitis. The goal of this study was to establish the peculiarities of structural organization of the masticatory muscles of rats in experimentally induced iodine deficiency. The research material involved masticatory muscle of 36 white outbred male rats, which were divided into groups: group І – intact animals; group ІІ – kept in iodine deficiency condition; group III – exposed to iodine deficiency with the addition of goitriferous food. All manipulations with test animals were performed in agreement with the regulations on the protection of vertebrate animals. The methods used in the study included: biochemical, histological, histochemical, submicroscopic, morphometric and statistical research methods. It has been established that under the conditions of iodine deficiency the lumen of arterial vessels in masticatory muscles becomes narrowed, the vascular vessels become thickened due to membrane oedema. The number of hemocapillaries per unit area of muscle fibre goes down in all the studied muscles. The number of muscle fibres with intermediate succinate dehydrogenase activity decreases with the simultaneous increase in muscle fibres with low succinate dehydrogenase activity. Furthermore, the oedematous changes are the most prominent in muscle fibres with low succinate dehydrogenase activity. In conditions of iodine deficiency with the addition of goitriferous products, we can observe the progression of luminal narrowing of the artery and wall thickening in masticatory muscles where oedematous-dystrophic changes are considerably marked. There is a tendency in the growth in haemocapillaries number per 1 μm2. Histostructurally, the loss of cross striation is quite noticeable. The cross-section diameter increases, especially in muscle fibres with low succinate dehydrogenase activity and muscle fibres with high succinate dehydrogenase activity in the masticatory muscle proper of immature animals. Ultrastructurally, the oedematous-dystrophic changes of myofibrils and mitochondria were revealed. Thus, under the conditions of iodine deficient diet the lumen of arterial vessels narrows in the masticatory muscles, their wall thickens due to oedema of the membranes, which progresses in iodine deficient diet with the addition of goitriferous products. The number of hemocapillaries per unit area of muscle fibre decreases in both study groups. Changes in vascular bed lead to the redistribution of the number of muscle fibres and their significant oedema.

Effects of Vermireactor Modifications on the Welfare of Earthworms Eisenia fetida (Sav.) and Properties of Vermicomposts

Vermicomposting is a method that allows for the environmentally friendly management of plant organic waste through the activity of concentrated earthworm populations. As a result of an appropriately conducted process, it is possible to obtain a valuable organic fertilizer, abundant in nutrients for plants as well as the development of a favorable (qualitatively and quantitatively) earthworm population structure. This paper presents the results of the vermicomposting process of waste plant mass with the use of vermireactors with different designs. In conventional no protective substrate (NPS) vermireactors, plant waste was the only habitat for earthworms, whilst protective substrate (PS) vermireactors also had a medium to shield the earthworm population in the event of the accidental introduction of toxic waste to these invertebrates. A positive effect of the PS vermireactor construction in the protection of an earthworm population against the potential use of stressful waste was found, as populations of Eisenia fetida were monitored and the development was noted during the vermicomposting with both methods. The largest differences between the populations (PS vs. NPS vermireactors) were demonstrated after 40 and 50 days and related to the number of immature animals (PS &gt; NPS by 30.7%; p &lt; 0.05) and cocoons (PS &gt; NPS by 34.8%; p &lt; 0.05). There were no significant differences between the PS and NPS in the biomass of immature E. fetida and cocoons. The vermicomposts obtained were not significantly different form each other but they were characterized by a higher content of C, N, P, K, Ca and Mg compared to the initial waste biomass. Contents of Cu, Mn, Zn, Cd and Pb in vermicomposts did not prevent their use as fertilizers.

Three-Dimensionally-Printed Bioactive Ceramic Scaffolds: Construct Effects on Bone Regeneration.

The utilization of three-dimensionally (3D)-printed bioceramic scaffolds composed of beta-tricalcium phosphate in conjunction with dipyridamole have shown to be effective in the osteogenesis of critical bone defects in both skeletally immature and mature animals. Furthermore, previous studies have proven the dura and pericranium's osteogenic capacity in the presence of 3D-printed scaffolds; however, the effect galea aponeurotica on osteogenesis in the presence of 3D scaffolds remains unclear. Critical-sized (11 mm) bilateral calvarial defects were created in 35-day old rabbits (n = 7). Two different 3D scaffolds were created, with one side of the calvaria being treated with a solid nonporous cap and the other with a fully porous cap. The solid cap feature was designed with the intention of preventing communication of the galea and the ossification site, while the porous cap permitted such communication. The rabbits were euthanized 8 weeks postoperatively. Calvaria were analyzed using microcomputed tomography, 3D reconstruction, and nondecalcified histologic sectioning in order assess differences in bone growth between the two types of scaffolding. Scaffolds with the solid (nonporous) cap yielded greater percent bone volume (P = 0.012) as well as a greater percent potential bone (P = 0.001) compared with the scaffolds with a porous cap. The scaffolds with porous caps also exhibited a greater percent volume of soft tissue (P < 0.001) presence. There were no statistically significant differences detected in scaffold volume. A physical barrier preventing the interaction of the galea aponeurotica with the scaffold leads to significantly increased calvarial bone regeneration in comparison with the scaffolds allowing for this interaction. The galea's interaction also leads to more soft tissue growth hindering the in growth of bone in the porous-cap scaffolds.

Interaction between Mycoplasma hyopneumoniae and host-a review

Mycoplasma hyopneumoniae is the pathogen of porcine enzootic pneumonia (PEP). Due to difficulties in studying the pathogenesis of M. hyopneumoniae for blockage on the establishment of gene operation platform and immature animal model, mycoplasmologists still make progress in understanding the interaction between M. hyopneumoniae and host. In this paper, we review the adhesion and damage of M. hyopneumoniae to host cells, the inflammatory response and immune response of host stimulated by M. hyopneumoniae. Meanwhile, we propose research directions of the pathogenesis of M. hyopneumoniae in the future. This review can provide references for the follow-up study on the interaction between M. hyopneumoniae and host, and provide theoretical basis for effective vaccine and drug development.

Oceanographic Determinants of the Abundance of Common Dolphins (Delphinus delphis) in the South of Portugal

Off mainland Portugal, the common dolphin (Delphinus delphis) is the most sighted cetacean, although information on this species is limited. The Atlantic coast of Southern Portugal is characterized by an intense wind-driven upwelling, creating ideal conditions for common dolphins. Using data collected aboard whale-watching boats (1929 sightings and 4548 h effort during 2010–2014), this study aims to understand the relationships between abundance rates (AR) of dolphins of different age classes (adults, juveniles, calves and newborns) and oceanographic [chlorophyll a (Chl-a) and sea surface temperature (SST)] variables. Over 70% of the groups contained immature animals. The AR of adults was negatively related with Chl-a, but not related to SST values. The AR of juveniles was positively related with SST. For calves and newborns, although the relationship between SST and AR is similar to that observed for juveniles, the effect could not be distinguished from zero. There was no relationship between Chl-a levels and the AR of juveniles, calves and newborns. These results corroborate previous findings that common dolphins tend to occur in highly productive areas demonstrating linkages between their abundance and oceanographic variables, and that this region may be a potential nursery ground.

Coming of age: morphometric variation in the hand skeletons of juvenile and adult Lesser Treeshrews (Scandentia: Tupaiidae: Tupaia minor Günther, 1876)

AbstractMorphometric analyses of the manus skeleton have proven useful in understanding species limits and morphological divergence among tupaiid treeshrews (Scandentia: Tupaiidae). Specimens in these studies are typically limited to mature individuals with fully erupted permanent dentition, which eliminates potentially confounding variation attributable to age, but also can exclude rare taxa and small island populations that are poorly represented in systematic collections. To determine the real limits associated with including immature animals in such studies, we used multivariate analyses to study sexual and age variation of the manus skeleton in two allopatric populations of the Lesser Treeshrew (Tupaia minor Günther, 1876) from the Malay Peninsula and from Borneo that we treated as separate samples. Individuals were aged using dental eruption of the permanent dentition. We also recorded the degree of epiphyseal fusion of the bones of the manus based on x-rays of study skins. We then tested our ability to distinguish the two populations using a series of discriminant function analyses of hand measurements from samples that included varying proportions of immature individuals and adults. We found no evidence of sexual dimorphism in hand proportions, permitting us to combine females and males in our samples. Epiphyseal fusion of the metacarpals and phalanges typically occurs by the time the third molars have completely erupted, and fusion of the distal epiphyses of the radius and ulna typically occurs by the time the permanent fourth premolars are in place. There is occasional asynchrony between dental age and epiphyseal fusion. In both populations, the hands of most infants and subadults provide morphometric values within the range of variation of adults, although they are typically distributed in the lower part of the adult range and have the potential to bias the sample toward lower mean size. The inclusion of infants and subadults when attempting to discriminate between two taxa generally results in lower rates of correct classifications, although the rates increase as the sample of immature individuals is limited to older subadults. As a general rule, we recommend that specimens of infants and subadults continue to be excluded from analyses when exploring taxonomic boundaries among treeshrews. In cases of extremely small sample sizes of adults, however, older subadults—in which the permanent third premolars are erupting or in place—can be used with appropriate caution.

Acute and subchronic toxicity studies of rhein in immature and d-galactose-induced aged mice and its potential hepatotoxicity mechanisms

Rhein is a key ingredient in many herbal remedies and is widely used. However, herbs containing rhein are frequently associated with poisoning incidents, especially in elderly subjects. Acute and subchronic toxicity of rhein in Kunming mice (KM) was investigated in this experiment. Acute toxicity tests showed a 40% lethality at a given rhein dose of 4000 mg/kg, and the LD50 of rhein was calculated by the bliss method to be greater than 2185.6 mg/kg. In subchronic toxicity, d-gal-induced aged and immature animals were randomized into three groups that were exposed to rhein of 0, 175, and 375 mg/kg/d for 75 days, respectively. No mortality was observed in immature mice group, whereas 55.5% (5/9) subjects in aged mice groups died in the high dosage group. AST, ALT, IL-6, TNF-α levels and typical histopathological changes indicate that rhein causes liver injury. In addition, our investigation explored possible hepatotoxic mechanisms of rhein and experimental results showed increased ROS production, NRF-2 and MDA levels and decreased SOD levels, demonstrating that rhein causes oxidative stress. MMP and mitochondrial swelling levels were able to assess the impact of rhein on mitochondrial function. Furthermore, the effect of rhein on apoptosis can be detected by flow cytometry. Our studies suggested that rhein induces oxidative stress leading to mitochondria dysfunction and apoptotic activation. Multidrug resistance protein (MRP) is an efflux transporter protein and is capable of transporting cellular oxidative stress-related substances. To further clarify the role of MRP in rhein induced oxidative stress, we examined MRP expression in the liver. However, the expression of MRP has no statistical significance.

Health assessment of harbour porpoises (PHOCOENA PHOCOENA) from Baltic area of Denmark, Germany, Poland and Latvia

Harbour porpoise (Phocoena phocoena), the only resident cetacean species of the Baltic Sea is formed of two subpopulations populations, occurring in the western Baltic, Belt Seas and Kattegat and the Baltic Proper, respectively. Harbour porpoises throughout these areas are exposed to a large number of human activities causing direct and indirect effects on individuals, that might also harm this species on a population level. From Latvia, Poland, Germany and Denmark 385 out of 1769 collected dead harbour porpoises were suitable for extensive necropsy. The animals were collected between 1990 and 2015 and were either by-caught or found dead on the coastline. Following necropsies, histopathological, microbiological, virological and parasitological investigations were conducted. Females and males were equally distributed among the 385 animals. Most animals from the different countries were juveniles between 3 months and 3 years old (varying between 46.5 and 100% of 385 animals per country). The respiratory tract had the highest number of morphological lesions, including lungworms in 25 to 58% and pneumonia in 21 to 58% of the investigated animals. Of those with pneumonia 8 to 33% were moderate or severe. The alimentary, hearing, and haematopoietic systems had inflammatory lesions and parasitic infections with limited health impact. 45.5 to 100% of the animals from the different countries were known by-caught individuals, of which 20 to 100% varying between countries had netmarks. Inflammatory lesions, especially in the respiratory tract were found in higher numbers when compared to control populations in areas with less human activities such as arctic waters. The high number of morphological changes in the respiratory tract and of bycatches especially among immature animals before reaching sexual maturity is of serious concern, as well as the low number of adult animals among the material. Data on health status and the causes of death are valuable for management. A next step in this regard will combine data from health and genetic investigations in order to detect differences between the two populations of the Baltic.

Risk of Attention Deficit Hyper Activity Disorder After Early Exposure to General Anesthesia; A Case Control Study

Background: Over the past decade, following the discovery that developing brain of immature animals was affected by anesthetic agents, the safety of general anesthesia (GA) in early life has been questioned. Objectives: We investigated the association between anesthesia exposure in children and ADHD development. Methods: This case-control study was conducted at pediatric psychology clinic of our institution and a pediatric neurology private clinic during 2019. Firstly the responsible resident of anesthesiology separated new ADHD cases. Then a questionnaire was filled out through an almost 10 minute’s telephone interview. Finally, frequency distribution of GA was compared between ADHD cases and controls. Results: Finally, the data from 210 children were analyzed. Among 105 ADHD cases, 19% had a history of a procedure requiring GA while it was 3.8% in control group. Comparing the two groups a significant difference was observed regarding the age of receiving GA (P = 0.004), gender (P &lt; 0.001), the history of receiving GA (P = 0.001) and the number of anesthesia exposures (P = 0.001). According to logistic regression analysis, male gender (P = 0.001) OR 3.11 (95CI = 1.63 - 5.93) and age (P = 0.003) OR 0.92 (95CI = 0.87 - 0.97) were significant predictors of early exposure to GA and ADHD development. Conclusions: It was revealed that early exposure to GA might be a risk factor for later developing ADHD. Boys might be more sensitive to the long term adverse effects of anesthetic agents than girls. Further prospective well-planned studies are needed to confirm these findings.

Response of the cerebral vasculature to systemic carbon monoxide administration-Regional differences and sexual dimorphism.

Previous studies about the modulation of the vasculature by CO were performed exclusively in male or sexually immature animals. Understanding the sex differences regarding systemic drug processing and pharmacodynamics is an important feature for safety assessment of drug dosing and efficacy. In this work, we used CORM-A1 as source of CO to examine the effects of this gasotransmitter on brain perfusion and the sex-dependent differences. Dynamic contrast-enhanced imaging (DCE)-based analysis was used to characterize the properties of CO in the modulation of cerebral vasculature in vivo, in adult C57BL/6 healthy mice. Perfusion of the temporal muscle, maxillary vein and in hippocampus, cortex and striatum was analysed for 108min following CORM-A1 administration of 3 or 5mg/kg. Under control conditions, brain perfusion was lower in females when compared with males. Under CO treatment, females showed a surprisingly overall reduced perfusion compared with controls (F=3.452, p=.0004), while no major alterations (or even the expected increase) were observed in males. Cortical structures were only modulated in females. A striking female-dominated vasoconstriction effect was observed in the hippocampus and striatum following administration of CO, in this mixed-sex cohort. As these two regions are implicated in episodic and procedural memory formation, CO may have a relevant impact in learning and memory.

Maturity-dependent cartilage cell plasticity and sensitivity to external perturbation

ObjectiveArticular cartilage undergoes biological and morphological changes throughout maturation. The prevalence of osteoarthritis in the aged population suggests that maturation predisposes cartilage to degradation and/or impaired regeneration, but this process is not fully understood. Therefore, the objective of this study was to characterize the cellular and genetic profile of cartilage, as well as biological plasticity in response to mechanical and culture time stimuli, as a function of animal maturity. Methods/designPorcine articular cartilage explants were harvested from stifle joints of immature (2–4 weeks), adolescent (5–6 months), and mature (1–5 years) animals. Half of all samples were subjected to a single compressive mechanical load. Loaded samples were paired with unloaded controls for downstream analyses. Expression of cartilage progenitor cell markers CD105, CD44, and CD29 were determined via flow cytometry. Expression of matrix synthesis genes Col1, Col2, Col10, ACAN, and SOX9 were determined via qPCR. Tissue morphology and matrix content were examined histologically. Post-loading assays were performed immediately and following 7 days in culture. ResultsCD105 and CD29 expression decreased with maturity, while CD44 expression was upregulated in cartilage from mature animals. Expression of matrix synthesis genes were generally upregulated in cartilage from mature animals, and adolescent animals showed the lowest expression of several matrix synthesizing genes. Culture time and mechanical loading analyses revealed greater plasticity to mechanical loading and culture time in cartilage from younger animals. Histology confirmed distinct structural and biochemical profiles across maturity. ConclusionThis study demonstrates differential, nonlinear expression of chondroprogenitor markers and matrix synthesis genes as a function of cartilage maturity, as well as loss of biological plasticity in aged tissue. These findings have likely implications for age-related loss of regeneration and osteoarthritis progression.

IL-1β prevents ILC2 expansion, type 2 cytokine secretion, and mucus metaplasia in response to early-life rhinovirus infection in mice.

Early-life wheezing-associated respiratory infection with human rhinovirus (RV) is associated with asthma development. RV infection of 6-day-old immature mice causes mucous metaplasia and airway hyperresponsiveness which is associated with the expansion of IL-13-producing type 2 innate lymphoid cells (ILC2s) and dependent on IL-25 and IL-33. We examined regulation of this asthma-like phenotype by IL-1β. Six-day-old wild-type or NRLP3-/- mice were inoculated with sham or RV-A1B. Selected mice were treated with IL-1 receptor antagonist (IL-1RA), anti-IL-1β, or recombinant IL-1β. Rhinovirus infection induced Il25, Il33, Il4, Il5, Il13, muc5ac, and gob5 mRNA expression, ILC2 expansion, mucus metaplasia, and airway hyperresponsiveness. RV also induced lung mRNA and protein expression of pro-IL-1β and NLRP3 as well as cleavage of caspase-1 and pro-IL-1β, indicating inflammasome priming and activation. Lung macrophages were a major source of IL-1β. Inhibition of IL-1β signaling with IL-1RA, anti-IL-1β, or NLRP3 KO increased RV-induced type 2 cytokine immune responses, ILC2 number, and mucus metaplasia, while decreasing IL-17 mRNA expression. Treatment with IL-1β had the opposite effect, decreasing IL-25, IL-33, and mucous metaplasia while increasing IL-17 expression. IL-1β and IL-17 each suppressed Il25, Il33, and muc5ac mRNA expression in cultured airway epithelial cells. Finally, RV-infected 6-day-old mice showed reduced IL-1β mRNA and protein expression compared to mature mice. Macrophage IL-1β limits type 2 inflammation and mucous metaplasia following RV infection by suppressing epithelial cell innate cytokine expression. Reduced IL-1β production in immature animals provides a mechanism permitting asthma development after early-life viral infection.

A method for constructing demographic profiles in Susscrofa using Logarithm Size Index scaling

Demographic profiling of archaeological assemblages offers a window into the shifting exploitation strategies practiced by ancient societies. High resolution sex-specific age profiles, in particular, have yielded new insights into the evolution of early management practices taken during the initial stages of animal domestication (Zeder, 2001). This paper is the third in a series seeking to develop a method for constructing demographic profiles for Sus scrofa — one of the earliest major livestock species noteworthy for being independently domesticated in both Southwest and East Asia. The first paper in this series (Lemoine et al., 2014) presented a new system for constructing high resolution age profiles using patterns of dental eruption and wear. The second (Zeder et al., 2015) presented a new method for computing high resolution age profiles based on long bone fusion. Both papers used a reference assemblage of modern Eurasian wild boar for developing these new aging schemes, which were then applied to the large assemblage of S. scrofa remains from the Early Holocene age site of Hallan Çemi (c. 11,600 cal. BP) in southeastern Anatolia — an assemblage that previous researchers claimed provided evidence for early steps toward management (Redding, 2005). Here we focus on a subset of this modern reference collection consisting of 34 wild boar skeletons from Turkey, Iran, and Iraq. We employ a Logarithm Size Index scaling (LSI) technique to examine the effect of factors such as age, locality, and, sex on body size in modern wild boar. We find that even though both age and locality have an impact on body size, LSI values derived from select measurements of various different post-cranial elements can be used to classify individual osteological specimens as belonging to immature animals, and, for animals older than about eight months of age, as female or male. We test the reliability of these classifications by comparing the known demographic profiles of the modern reference population against those derived from the LSI classifications. Finally, we apply this new method to the Hallan Çemi assemblage and propose a method for adjusting LSI values to account for temporal size differences encountered between the modern and ancient populations. We also present methods for assessing the reliability of these classifications in the ancient population. We conclude by offering guidelines for the application of this new method to other archaeological assemblages of S. scrofa and other ungulate species that demonstrate strong degrees of sexual dimorphism.

Methods for Monitoring for the Population Consequences of Disturbance in Marine Mammals: A Review

Assessing the nonlethal effects of disturbance and their population-level consequences is a significant ecological and conservation challenge, because it requires extensive baseline knowledge of behavioral patterns, life-history and demography. However, for many marine mammal populations, this knowledge is currently lacking and it may take decades to fill the gaps. During this time, undetected population declines may occur. In this study we identify methods that can be used to monitor populations subject to disturbance and provide insights into the processes through which disturbance may affect them. To identify and address the knowledge gaps highlighted above, we reviewed the literature to identify suitable response variables and methods for monitoring these variables. We also used existing models of the population consequences of disturbance (PCoD) to identify demographic characteristics (e.g. the proportion of immature animals in the population, or the ratio of calves/pups to mature females) that may be strongly correlated with population status and therefore provide early warnings of future changes in abundance. These demographic characteristics can be monitored using established methods such as visual surveys combined with photogrammetry, and capture-recapture analysis. Individual health and physiological variables can also inform PCoD assessment and can be monitored using photogrammetry, remote tissue sampling, hands-on assessment and individual tracking. We then conducted a workshop to establish the relative utility and feasibility of all these approaches for different groups of marine mammal species. We describe how future marine mammal monitoring programs can be designed to inform population-level analysis.

Electrographic seizures induced by activation of ETA and ETB receptors following intrahippocampal infusion of endothelin-1 in immature rats occur by different mechanisms

We have demonstrated previously that activation of either the ETA or ETB receptor can induce acute electrographic seizures following the intrahippocampal infusion of endothelin-1 (ET-1) in immature (P12) rats. We also demonstrated that activation of the ETA receptor is associated with marked focal ischemia, while activation of the ETB receptor is not.Exploring the mechanisms underlying seizures induced by these two ET-1 receptor interactions can potentially provide insight into how focal ischemia in immature animals produces seizures and whether ischemiarelated seizures differ from seizures not associated with ischemia.To explore these seizure mechanisms we used microdialysis to determine biomarkers associated with seizures in P12 rats following the intrahippocampal infusion of two different agents: (1) ET-1, which activates both the ETA and ETB receptors and causes focal ischemia and (2) Ala-ET-1, which selectively activates only the ETB receptor and does not cause ischemia.Our results show that seizures associated with combined ETA and ETB receptor activation (and ischemia) have a different temporal distribution and microdialysis profile from seizures associated with ETB activation alone (and without ischemia). Seizures with combined activation peak within the first hour after infusion and the microdialysis profile is characterized by a significant increase in the ratio of glutamic acid to GABA. By contrast, seizures with activation of only the ETB receptor peak in the second hour after infusion and microdialysis shows a significant increase in the ratio of leukotriene B4 to prostaglandin E2.These findings suggest that ischemia-related seizures in immature animals involve an imbalance of excitation and inhibition, while non-ischemiarelated seizures involve an inflammatory process resulting from an excess of leukotrienes.

Non-additive effects of adjunct erythropoietin therapy with therapeutic hypothermia after global cerebral ischaemia in near-term fetal sheep.

Recombinant human erythropoietin (rEpo) is neuroprotective in immature animals, but it is unclear whether the combination of high-dose rEpo therapy with therapeutic hypothermia can further improve outcomes. Hypothermia and rEpo independently improved neuronal survival, with greater improvement with hypothermia, and similarly reduced numbers of caspase-3 positive cells and reactive microglia after 7days recovery. Hypothermia, but not rEpo, was associated with markedly improved EEG power, whereas both interventions improved recovery of EEG frequency. There was no significant improvement in any outcome after combined rEpo and hypothermia compared with hypothermia alone, and of concern, the combination was associated with increased numbers of cortical caspase-3-positive cells compared with ischaemia-hypothermia. These data suggest that the mechanisms of neuroprotection with hypothermia and rEpo overlap and, thus, high-dose rEpo infusion does not appear to be an effective adjunct therapy for therapeutic hypothermia. Therapeutic hypothermia for hypoxic-ischaemic encephalopathy (HIE) provides incomplete neuroprotection. Recombinant human erythropoietin (rEpo) is neuroprotective in immature animals, but it is unclear whether adjunct rEpo therapy with therapeutic hypothermia can further improve outcomes. Near-term fetal sheep received sham-ischaemia (n=9) or global cerebral ischaemia for 30min (ischaemia-vehicle, n=8), followed by intravenous infusion of rEpo (ischaemia-Epo, n=8; 5000U/kg loading dose, then 833.3U/kg/h), cerebral hypothermia (ischaemia-hypothermia, n=8), or rEpo plus hypothermia (ischaemia-Epo-hypothermia, n=8), from 3 to 72h post ischaemia. Fetal brains were collected 7days after cerebral ischaemia. Cerebral ischaemia was associated with severe neuronal loss and microglial induction in the parasagittal cortex and subcortical regions. Hypothermia reduced overall neuronal loss, cortical caspase-3 and reactive microglia in the striatum and cortex, with greater recovery of electroencephalographic (EEG) power and spectral edge (SEF) from 48h onwards. rEpo independently improved neuronal survival in the parasagittal cortex, hippocampal CA4 and thalamus, and reduced cortical caspase-3 and activated microglia in striatal and cortical areas, with greater SEF from 120h onwards. However, ischaemia-Epo-hypothermia did not further improve outcomes compared with ischaemia-hypothermia and was associated with increased numbers of cortical caspase-3-positive cells. These findings suggest that although delayed, prolonged treatment with both hypothermia and rEpo are independently neuroprotective, they have overlapping anti-inflammatory and anti-apoptotic mechanisms, such that the delayed, high-dose rEpo infusion for 3days did not materially augment neuroprotection with therapeutic hypothermia.