Establishment of a reproducible and minimally invasive ischemic stroke model in swine

The need for advances in the management/treatment options for ischemic stroke patients requires that upcoming preclinical research uses animals with more human-like brain characteristics. The porcine brain is considered appropriate, although the presence of the rete mirabile (RM) prevents direct catheterization of the intracranial arteries to produce focal cerebral ischemia. To develop a reproducible minimally invasive porcine stroke model, a guide catheter and guide wire were introduced through the femoral artery until reaching the left RM. Using the pressure cooker technique, Squid-12 embolization material was deposited to fill, overflow, and occlude the left RM, the left internal carotid artery, and left circle of Willis wing up to the origins of the middle cerebral arteries (MCAs), mimicking the occlusion produced in the filament model in rodents. Longitudinal multimodal cerebral MRI was conducted to assess the brain damage and cerebral blood supply. The technique we describe here occluded up to the origins of the MCAs in 7 of 8 swine, inducing early damage 90 minutes after occlusion that later evolved to a large cerebral infarction and producing no mortality during the intervention. This minimally invasive ischemic stroke model in swine produced reproducible infarcts and shows translational features common to human stroke

The role of artificial intelligence in paediatric cardiovascular magnetic resonance imaging

Artificial intelligence (AI) offers the potential to change many aspects of paediatric cardiac imaging. At present, there are only a few clinically validated examples of AI applications in this field. This review focuses on the use of AI in paediatric cardiovascular MRI, using examples from paediatric cardiovascular MRI, adult cardiovascular MRI and other radiologic experience

Can Long Bone Structural Variability Detect Among-Population Relationships?

Phenotypic traits develop and are maintained by complex interactions between intrinsic (molecular) and extrinsic (environmental) factors. While the influence of intrinsic factors on adult craniomandibular variation has been intensively studied, less is known about limb bones, in part because it is assumed that their plasticity obscures intrinsic signals, especially those fixed early in life. While diaphyseal regions are plastic in response to activity, the extent to which they also reflect (phylo)genetic autocorrelation has not been sufficiently addressed, particularly given the common practice of comparing long bones across populations unevenly dispersed in space and time. Here I investigate the degree to which long bone lengths, joints, and diaphyses vary in their ability to detect intrinsic genetic patterns. I do this by calculating among-population genetic relationships via long bone variation in samples from England, Southern Europe, and South Africa. I then test whether these predictions significantly match those generated via craniofacial variation and, further, whether they are supported by contextual (historical) information. Given the innately plastic nature of diaphyseal regions, I further test whether differences in physical activity can obscure predicted genetic relationships. I do this by adding a temporal component to genetic distance analyses via inclusion of Medieval samples and by partitioning several Southern European samples into “high intensity” and “low intensity” subgroups based on recorded occupational data, using these to generate more genetic predictions. Results show that all three long bone properties reflect among-population genetic structure, with length and joint dimensions doing so at levels comparable to those of the crania. Diaphyses, however, generate lower levels of among-population differentiation, presumably because their plasticity fuels more intrapopulation variation. Despite this, diaphyses still detect key components of population genetic structure, including genetic affinity shared among modern English and descendant South Africans, the close genetic relationships among modern Southern Europeans (even when subdivided by occupation), and the ancestral connection between Medieval and modern English samples. In total, these results suggest that all long bone properties can detect among-population genetic information, and further, that interpretations of behavior from limb bone variation can be strengthened if genetic relationships (or assumed relationships) are controlled for

The function of the TGF-beta and Toll signalling pathways in Tribolium dorsoventral patterning

Dorsoventral (DV) patterning in Drosophila melanogaster is one of the most well-known gene regulatory networks (GRN) in biology. To investigate if this GRN is conserved during insect evolution, functional analysis of TGF-beta and Toll pathways in the short-germ beetle Tribolium castaneum was performed. In the first part, the function of several BMP/Dpp extracellular modulators, including the products of Tolloid (Tld) and Twisted-gastrulation/Crossveinless (Tsg-Cv), was investigated in Tribolium via parental RNAi (pRNAi). While Tc-tld pRNAi knock-down decreases embryonic BMP activity, Tc-tsg(cv) knock-down completely abolishes it. These observations are strikingly different from those in Drosophila, where tsg is only required for a subset of Dpp activity. These results suggest that Tsg/Cv-like proteins are essential for BMP signalling in Tribolium. Since duplicated copies of tsg(cv)- and tld-related genes are present in the Drosophila melanogaster genome, duplication followed by sub-functionalization of these modulators might have changed the BMP/Dpp gradient during evolution of the dipteran Drosophila lineage. In the second part, a functional analysis of the Toll pathway was performed. This analysis addressed the question of why the Tribolium Dorsal nuclear gradient is not stable, but rapidly shrinks and disappears, in contrast to the stable Drosophila gradient. Negative feedback accounts for this dynamic behavior: Tc-Dorsal and one of its target genes (Tc-Twist) activate transcription of the I-KappaB homolog Tc-cactus, which in turn terminates Dorsal function. Despite its transient role, Tc-Dorsal is strictly required to initiate DV polarity, as in Drosophila. However, unlike Drosophila, embryos lacking Tc-Dorsal display a periodic pattern of DV cell fates along the AP axis, indicating that a self-organizing ectodermal patterning system operates independently of mesoderm or maternal DV polarity cues. The presence of self-organizing patterning systems in short-germ insects like Tribolium is in agreement with a regulative type of embryogenesis proposed by classical fragmentation studies on hemimetabolous insects. These results also elucidate how extraembryonic tissues are organized in short-germ embryos, and how patterning information is transmitted from the early embryo to the growth zone. Altogether, the functional analysis of the TGF-betA and Toll pathways in Tribolium dorsoventral patterning suggests that extensive changes in this GRN have occurred during insect evolution

Axial patterning of the chondrichthyan pharyngeal endoskeleton and the origin of the jaw

Classical comparative anatomical hypotheses propose that the upper and lower jaw evolved through modification of dorsal and ventral gill arch skeletal elements, respectively. These hypotheses were based largely on the skeletal anatomy of chondrichthyans (sharks, skates and holocephalans), but remain largely untested from a developmental perspective. Here, I test 1) whether jaws and gill arches ancestrally shared common developmental patterning mechanisms, and 2) whether the mandibular arch might carry anatomical vestiges of an ancestral gill-arch-like condition. To do this, I have used embryos of a cartilaginous fish, the little skate (Leucoraja erinacea), which has retained an ancestral organisation of the jaw and gill arch endoskeleton, and which possesses a reduced gill-like structure (a “pseudobranch”) on the back of the mandibular arch. Using candidate and RNAseq/differential gene expression analysis and mRNA in situ hybridisation, I find broad conservation of dorsoventral patterning mechanisms within pharyngeal arches – embryonic structures which contain the progenitors of the jaw and gill arch skeleton – as well as unique transcriptional features that may underpin distinct jaw and gill arch morphologies. The latter include unique gene expression features of jaw and gill arch muscle progenitors, and of developing gill lamellae. Finally, it has been historically speculated that the chondrichthyan pseudobranch derives from the hyoid (i.e. 2nd pharyngeal) arch. I demonstrate here, by cell lineage tracing, that the pseudobranch is, in fact, mandibular arch derived, that it shares gene expression features with developing gills, and that its supporting spiracular cartilage develops under the influence of a shh-expressing signalling centre, in a manner that parallels that shh-dependent development of branchial rays (i.e. cartilaginous appendages that support the respiratory lamellae of the gill arches). Taken together, this dissertation presents evidence for serial homology of the jaw and gill arch skeleton, and of an ancestral gill-arch nature of the mandibular arch of vertebrates

Reassessment of the evolutionary history of the late Triassic and early Jurassic sauropodomorph dinosaurs through comparative cladistics and the supermatrix approach

Non-sauropod sauropodomorphs, also known as 'basal sauropodomorphs' or 'prosauropods', have been thoroughly studied in recent years. Several hypotheses on the interrelationships within this group have been proposed, ranging from a complete paraphyly, where the group represents a grade from basal saurischians to Sauropoda, to a group on its own. The grade-like hypothesis is the most accepted; however, the relationships between the different taxa are not consistent amongst the proposed scenarios. These inconsistencies have been attributed to missing data and unstable (i.e., poorly preserved) taxa, nevertheless, an extensive comparative cladistic analysis has found that these inconsistencies instead come from the character coding and character selection, plus the strategies on merging data sets. Furthermore, a detailed character analysis using information theory and mathematical topology as an approach for character delineation is explored here to operationalise characters and reduce the potential impact of missing data. This analysis also produced the largest and most comprehensive matrix after the reassessment and operationalisation of every character applied to this group far. Additionally, partition analyses performed on this data set have found consistencies in the interrelationships within non-sauropod Sauropodomorpha and has found strong support for smaller clades such as Plateosauridae, Riojasauridae, Anchisauridae, Massospondylinae and Lufengosarinae. The results of these analyses also highlight a different scenario on how quadrupedality evolved, independently originating twice within the group, and provide a better framework to understand the palaeo-biogeography and diversification rate of the first herbivore radiation of dinosaurs

Papers on Anthropology XX

-M. Toomsalu, A. Arend. Medical Collections in Tartu Old Anatomical Theatre – from August Rauber’s anatomy museum to a multifunctional research and edutainment centre -H. Kaarma. A short overview of the work of anthropologists of the Old Anatomical Theatre -M. Viikmaa. Leiu Heapost – 75 -R. Allmäe. Human bones in Salme I boat-grave, the Island of Saaremaa; Estonia -N. Balciuniene, D. graf von Keyserlingk, A. Valanciute, I. Balnyte,A. Macas, A. Tamasauskas. Changes of the chicken chorioallantoic membrane and the behaviour of the transplanted glioblastoma -A. Barzdina, M. Pilmane, A. Petersons. Grap and NF expression in brain tissue in children and adults after fatal traumatic brain injury -D. Batulevicius, G. Skripkiene, V. Batuleviciene, V. Skripka, A. Dabuzinskiene, D. H. Pauza. Complexity of the frog intracardiac neurons. Intracellular injection study -A. Dabuzinskiene, A. Ratkevicius, D. Batulevicius, G. Skripkiene, V. Skripka, G. Liutkiene, U. A. Bagdonas, S. Razbadauskiene. Gender differences in the human cervicothoracic ganglia -C. Dogan, C. Raschka. Anthropometrical and sport constitutional comparison of German male soccer players and male students of sport sciences -D. Gudiene, I. Balnyte, J. Palubinskiene, A. Valanciute. Age related structural changes in human basilar artery -L. Heapost. Taste sensitivity to PTC and colour blindness in Estonians -P. Hussar, I. Tokin, G. Filimonova, I. Tokin, Ü. Hussar. Dexamethazone-induced T-lymphocyte apoptosis in different lymphoid organs -L. Kalichman, E. Kobyliansky. Hand osteoarthritis and aging: the results of large-scale cross-sectional study -S. Kana, T. Viik. Annual report of the Estonian Naturalists’ Society 2010 -B. Karmakar, E. Kobyliansky. Finger and palmar dermatoglyphics in Muzeina Bedouins from South Sinai: a qualitative traits -D. Kažoka, J. Vētra. Variations in some anthropometrical parameters of the women with the different iris colour in Latvia -J. Kasmel, T. Kasmel. Seventy years of the Anthropology Section of the Estonian Naturalists’ Society (Part III) -R. Kisenauskaite, D. Paskeviciene. The research of physical condition, physical activity and nutrition of teacher education students -R. Kleina, I. Franckevica, M. Sperga, D. Lutinska. The analysis of undiagnosed malignancies -G. Kolesova, J. Vētra. Sexual dimorphism of pelvic morphology variation in live humans -V. Kulvietis, V. Žalgeviečienė, J. Didžiapetriene,D. Bulotienė, R. Rotomskis. Distribution of nanoparticles in the pregnant rat:the morphologic and spectroscopic study -A. Kuzminienė, S. Šalomskaitė-Davalgienė, I. Balnyte, J. Palubinskienė, A. Valančiūte, V. Ulozas. Evaluation of the chicken embryo choriaollantoic membrane model for laryngeal tumor transplantation -J. Limbo. The tooth size in the end of the Estonian iron age -M. Lintsi, H. Kaarma, M. Aunapuu, A. Arend, R. Aule. Correlation between anthropometrical variables and body surface area -M. Matyk, C. Raschka. Body composition and somatotype of European top roller speed skaters -A. Miskova, M. Pilmane, D. Rezeberga. Immunochistochemical distribution of IGF-1, bFGF and their receptors in decidual embryonic and tubal human pregnancy tissue -E. Mozeika, M. Pilmane, J. Kisis. Distribution of human B-defensin 2,TNF-alpha, IL-1alpha, IL-6 and IL-8 in psoriatic skin -P. Männik, R. T. Kibur, A. Arend, M. Aunapuu. Trophinin and integrin β3 expression in the human endometrium. A pilot study -A. Namm, A. Arend, M. Aunapuu. Bone morphogenetic proteins as regulators of neural tube development -E. Pētersone-Gordina, G. Geshards. Dental disease in a 17th–18th century German community in Jelgava, Latvia -L. Pļaviņa. Assessment of the physical activity level for the staff military personnel -V. Russeva. Alternations of cervical vertebrae in two individuals from the Late Antiquty Necropolis from the “Big Mound” near Cabyle, Bulgaria -S. Skuja, V. Groma, R. Kleina. Chronic alcohol abuse is implicated in the oxidative stress and the changes in the neurotrophic factor receptor expression in the human CNS -R. Stamm, M. Stamm, N. Sorgina, S. Koskel. Training programme to develop young volleyballers’ jumping ability -G. Sumeraga, M. Pilmane. Distribution of neuropeptides in nasal and nasopharyngeal mucosa in patients with the post nasal drip syndrome -N. Szoldatits, B. L. Buda, G. A. Tóth. Body proportion changes of nursing home oligophrenics in western Hungary (1991–2011) -A. Valdovska, M. Pilmane. Relation between serum enzymes and liver histopathology in mink with hepatitis -G. Veldre, T. Kums, E. Salum, J. Eha. Relationship between soldiers’ body height-weight category and changes in their spinal column kyphotic curvature during a long-term military missio

The appendicular skeleton variability of the Sauropoda Titanosauria from the Upper Cretaceous of Lo Hueco (Cuenca, Spain)

Tesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento de Biología. Fecha de lectura: 24-01-2020Esta tesis tiene embargado el acceso al texto completo hasta el 24-07-2021En este volumen se presentan nuevos datos acerca del esqueleto apendicular de los titanosaurios del yacimiento Campano-Maastrichtiense de Lo Hueco (Cuenca, España). En este yacimiento se ha recuperado una muestra abundante de restos referidos a saurópodos titanosaurios, con varios ejemplares en conexión y decenas de ejemplares aislados. En esta muestra se identifica una elevada variabilidad morfológica en cada tipo de elemento apendicular y la presencia de ejemplares de pequeño tamaño. Hasta ahora solo se ha descrito en el yacimiento una forma exclusiva de titanosaurio, Lohuecotitan pandafilandi. No obstante, los estudios de los abundantes restos encontrados en el yacimiento habían permitido identificar dos morfotipos principales de dientes, dos tipos de basicraneos de titanosaurio, tres posibles morfotipos identificados en el esqueleto axial correspondiente a las vértebras dorsales, y cuatro morfotipos en el estudio de las vértebras caudales. En el presente estudio se explora la elevada variabilidad encontrada en la muestra de restos apendiculares. Para ello se utilizan una serie de técnicas analíticas relacionadas con el machine learning y la morfometría geométrica en 3D con el objetivo de identificar posibles morfotipos que ayuden a explicar esta variabilidad. Se desarrolla un flujo de trabajo de digitalización del ejemplar en 3D, proceso de restauración virtual en caso de ser ejemplares fragmentarios, y su posterior análisis estadístico. Mediante estas técnicas se determina la presencia de dos morfotipos principales. A partir de esta identificación, se procede a la cuantificación de la variabilidad intraespecífica en cada uno de ellos, así como la determinación de posibles secuencias ontogenéticas y la variabilidad debida a cambios durante el crecimiento del esqueleto apendicular de los titanosaurios. Algunos indicios apuntan a la que los dos morfotipos identificados en el yacimiento pertenecerían a dos gremios distintos que tendrían dos estrategias tróficas distintas. En el presente trabajo se discuten las posibles implicaciones en las diferencias morfológicas observadas entre ambos morfotipos principales. Se realiza un modelo aproximado con el que relacionar la morfología general de las extremidades en neosaurópodos con estos dos tipos de gremios y se relacionan los dos morfotipos principales con dos estrategias tróficas congruentes con los datos del estudio de material craneal. La variabilidad intraespecífica observada en cada morfotipo permite determinar sus implicaciones en la codificación de caracteres morfológicos apendiculares. En este trabajo se han identificado varias secuencias ontogenéticas relativas a cada tipo de elemento analizado. Se describe en detalle por primera vez las secuencias de transformaciones ontogenéticas en estos titanosaurios, así como el estadio y tiempo relativo en que se producen dichos cambios y sus implicaciones en las codificaciones de caracteres morfológicosIn the current dissertation a revision of new data of the appendicular skeleton of the Campanian-Maastrichtian fossil site of Lo Hueco (Cuenca, Spain) is presented. This fossil site have yielded an abundant sample of specimens referable to titanosaur sauropods, with several individuals partially articulated and tens of isolated specimens. There has been identified a high morphological variability in each appendicular element and the presence of several small-sized specimens in this sample. Until now, a single titanosaur exclusive form have been described, Lohuecotitan pandafilandi. However, the study of abundant isolated specimens from the fossil site have allowed to identify two main teeth morphotypes, two types of braincase, three morphotypes identified in the axial skeleton of the dorsal region, and four morphotypes among the caudal vertebrae. The current study explores the high variability found in the sample of appendicular elements. For this matter, a series of analytical techniques related with modern machine learning and 3D geometric morphometrics are used with the objective of identifying the probable morphotypes that help explain the morphological variance. A 3D digitizing workflow of the specimens of study is herein proposed, with a new proposal for virtual restoration of fragmentary elements and its incorporation to statistical analyses. Using these techniques it has been identified two main appendicular morphotypes. Based on this morphotypes, the intraspecific variability has been quantified in each of them, the ontogenetic sequences have been identified and the variability related to transformations during titanosaur ontogenetic development. Previous studies indicates that two titanosaur morphotype from Lo Hueco could have been pertain to two different guilds with two different types of feeding niche exploitation. In the current study, the implications of several morphological differences between both main morphotypes are discussed under the hypothesis of differences in the ecomorphological specialization. A statistical proxy model was created to test the relationships between main appendicular morphology with ecomorphological specialization related with the height of the feeding envelope among neosauropods. The results allow relating the two main morphotypes with two different feeding niche exploitation strategies congruent with previous analyses in the cranial material. The observed intraspecific variability in each morphotype allows determining its impact on morphological character scoring. In the current dissertation it has been identified the presence of several ontogenetic sequences in each morphotype. The ontogenetic sequences have been comprehensively described for first time in this group, as well as the ontogenetic stage and relative time estimation of the morphological character changes with implications for character scoringsEsta tesis fue realizada gracias a la Ayuda para Contratos Predoctorales para la Formación de Doctores BES-2013-065509 - Ministerio de Economía y Competitividad. Esta beca doctoral está asociada al Proyecto de Investigación CGL2012-35199 - Ministerio de Economía y Competitivida

Microengineering The Neural Tube

Early embryonic development is a complex and highly regulated orchestra of instructive cues that collectively guide naïve stem cells towards progressively more specialized fates. In the neural tube, the precursor structure to the brain and spinal cord, these signals emanate from ‘organizing centers’ surrounding the neural tube. These organizing centers send out soluble cues or morphogens that diffuse tens to hundreds of microns to recipient cells residing in the neural tube. Re-creating this dynamic landscape of cues in vitro is impossible using standard cell culture tools and techniques. However, microfluidics is perfectly suited to fill this gap, allowing precise control over the microenvironment on the same length scale as the developing embryo. A microfluidic device is presented that is able to re-create some of the spatial patterning events that occur during the early development of the neural tube. This platform enables developmental biologists to reverse engineer development from the ground up, enabling researchers to pose radically new experiments to help answer some of the most relevant questions regarding fate specification in the developing neural tube. Here the device is used to guide mouse embryonic stem cells into motor neurons. Importantly, these motor neurons are able to be directed to differentiate in a defined region of the microdevice, a spatial patterning event that is the hallmark of the developing neural tube. For the first time it is now possible to study the effect of development cues on live populations of stem cells. The characterization of these fundamental developmental processes will prove invaluable in understanding how humans acquire both form and function. One day, it may allow researchers to harness these developmental techniques, which have been refined over thousands of years of evolution, to guide patient derived cells into any user defined cell fate

Treatment options for large acetabular defects in hip revision surgery:current options and a custom-made solution

Total hip arthroplasty is referred to as the operation of the century. In this thesis the current treatment options for large acetabular defects are evaluated and a new acetabular implant to treat large acetabular defects is introduced and evaluated. A review of the literature on the current treatment options for large acetabular defects is presented. It showed that Trabecular MetalTM (Zimmer) augment and shells gave the most promising results. Reconstruction with an antiprotrusio cage was the most frequently reported technique, with good results in a physically low demand elderly population. Bone impaction grafting seems not appropriate for pelvic discontinuity and Paprosky type-3B defects. In those cases, a custom-made triflange implant or a cup-cage reconstruction might be the best alternative, but few reports of sufficient quality are available yet. Next a new custom-made acetabular implant for large acetabular defects is introduced including its surgical technique. The accuracy of the placement of the implant is evaluated in a group of 16 patients. This first group is then compared to a second group of 16 patients. The results are promising and a trend towards better surgical accuracy and less complications in the second group is found, despite a higher complexity in cases. Finally, the clinical and radiological follow-up at two years is described using a prospective case series of 50 hips. It shows that this custom-made acetabular implant is a viable option for uncontained acetabular defects and pelvic discontinuity