Drug-eluting beads loaded with antiangiogenic agents for chemoembolization: in vitro sunitinib loading and release and in vivo pharmacokinetics in an animal model.

PURPOSE: The combination of embolic beads with a multitargeted tyrosine kinase inhibitor that inhibits tumor vessel growth is suggested as an alternative and improvement to the current standard doxorubicin-eluting beads for use in transarterial chemoembolization. This study demonstrates the in vitro loading and release kinetics of sunitinib using commercially available embolization microspheres and evaluates the in vitro biologic efficacy on cell cultures and the resulting in vivo pharmacokinetics profiles in an animal model. MATERIALS AND METHODS: DC Bead microspheres, 70-150 µm and 100-300 µm (Biocompatibles Ltd., Farnham, United Kingdom), were loaded by immersion in sunitinib solution. Drug release was measured in saline in a USP-approved flow-through apparatus and quantified by spectrophotometry. Activity after release was confirmed in cell culture. For pharmacokinetics and in vivo toxicity evaluation, New Zealand white rabbits received sunitinib either by intraarterial injection of 100-300 µm sized beads or per os. Plasma and liver tissue drug concentrations were assessed by liquid chromatography-tandem mass spectroscopy. RESULTS: Sunitinib loading on beads was close to complete and homogeneous. A total release of 80% in saline was measured, with similar fast-release profiles for both sphere sizes. After embolization, drug plasma levels remained below the therapeutic threshold (< 50 ng/mL), but high concentrations at 6 hours (14.9 µg/g) and 24 hours (3.4 µg/g) were found in the liver tissue. CONCLUSIONS: DC Bead microspheres of two sizes were efficiently loaded with sunitinib and displayed a fast and almost complete release in saline. High liver drug concentrations and low systemic levels indicated the potential of sunitinib-eluting beads for use in embolization.

A temporal hierarchy for conspecific vocalization discrimination in humans.

The ability to discriminate conspecific vocalizations is observed across species and early during development. However, its neurophysiologic mechanism remains controversial, particularly regarding whether it involves specialized processes with dedicated neural machinery. We identified spatiotemporal brain mechanisms for conspecific vocalization discrimination in humans by applying electrical neuroimaging analyses to auditory evoked potentials (AEPs) in response to acoustically and psychophysically controlled nonverbal human and animal vocalizations as well as sounds of man-made objects. AEP strength modulations in the absence of topographic modulations are suggestive of statistically indistinguishable brain networks. First, responses were significantly stronger, but topographically indistinguishable to human versus animal vocalizations starting at 169-219 ms after stimulus onset and within regions of the right superior temporal sulcus and superior temporal gyrus. This effect correlated with another AEP strength modulation occurring at 291-357 ms that was localized within the left inferior prefrontal and precentral gyri. Temporally segregated and spatially distributed stages of vocalization discrimination are thus functionally coupled and demonstrate how conventional views of functional specialization must incorporate network dynamics. Second, vocalization discrimination is not subject to facilitated processing in time, but instead lags more general categorization by approximately 100 ms, indicative of hierarchical processing during object discrimination. Third, although differences between human and animal vocalizations persisted when analyses were performed at a single-object level or extended to include additional (man-made) sound categories, at no latency were responses to human vocalizations stronger than those to all other categories. Vocalization discrimination transpires at times synchronous with that of face discrimination but is not functionally specialized.

What can animal memory study bring to the assessment of memory and cognitive skills for intellectual Disability?

Three case studies are presented to investigate the possibility of evaluating memory and cognitive capacities of severe intellectual disability with attention given to the ecological environment. Two 22-year-old male patients and a 27-year-old male patient, all three with severe intellectual disability with no verbal communication skills, were evaluated with a new and original paradigm adapted to study cognition in humans from experimental paradigms. We developed a test based on animal models to complement the "home" scale of the Adolescent and Adult Psychoeducational Profile (AAPEP), an assessment instrument designed for adolescents and adults with severe developmental disabilities. Results show that the new instrument is helpful, not only to staff members who can better understand the poor performances of their patients in daily life activities but also in the elaboration of individual acquisition plans. These preliminary results demonstrate the interest in developing a larger controlled study and in publishing our procedure.

Auditory perceptual decision-making based on semantic categorization of environmental sounds.

Discriminating complex sounds relies on multiple stages of differential brain activity. The specific roles of these stages and their links to perception were the focus of the present study. We presented 250ms duration sounds of living and man-made objects while recording 160-channel electroencephalography (EEG). Subjects categorized each sound as that of a living, man-made or unknown item. We tested whether/when the brain discriminates between sound categories even when not transpiring behaviorally. We applied a single-trial classifier that identified voltage topographies and latencies at which brain responses are most discriminative. For sounds that the subjects could not categorize, we could successfully decode the semantic category based on differences in voltage topographies during the 116-174ms post-stimulus period. Sounds that were correctly categorized as that of a living or man-made item by the same subjects exhibited two periods of differences in voltage topographies at the single-trial level. Subjects exhibited differential activity before the sound ended (starting at 112ms) and on a separate period at ~270ms post-stimulus onset. Because each of these periods could be used to reliably decode semantic categories, we interpreted the first as being related to an implicit tuning for sound representations and the second as being linked to perceptual decision-making processes. Collectively, our results show that the brain discriminates environmental sounds during early stages and independently of behavioral proficiency and that explicit sound categorization requires a subsequent processing stage.

Extensive circumferential endoscopic mucosal resection with a new rigid esophagoscope: an animal study

RÉSUME Les techniques de résection muqueuse endoscopique utilisées actuellement ne permettent pas d'effectuer une résection circonférentielle d'un long segment de muqueuse oesophagienne. Une telle résection permettrait le traitement de foyers multicentriques de dysplasies de haut grade (HGIN) ou d'adénocarcinomes précoces (AC) développés sur oesophage de Barrett. Sachant que la prévalence des métastases ganglionnaires régionales est d'environ 7 % pour les adénocarcinomes intra-muqueux (Tis,T1a), les méthodes de traitement non-chirurgicales et moins invasives sont actuellement préférées à une oesophagectomie, dont la morbidité et la mortalité restent élevées en comparaison avec les mucosectomies endoscopiques. Un oesophagoscope rigide modifié a été développé à Lausanne en collaboration avec la maison Karl Storz GmbH, de façon à permettre des résections muqueuses étendues dans l'oesophage. Cette étude animale pilote a investigué la faisabilité et la fiabilité des résections muqueuses circonférentielles de différentes longueurs dans l'oesophage du mouton. Des résections circonférentielles de 2,2 cm (n=6), 3,3 cm (n=6), 4,4 cm (n=7) et 5,5 cm (n=5) de longueur ont été effectuées dans l'oesophage de 24 moutons. Elles consistaient en 2 mucosectomies hémi-circonférentielles opposées. Les animaux ont été suivis par des examens endoscopiques à une semaine puis chaque mois pendant 6 mois ou jusqu'à réépithélialisation complète sans sténose. Au cours du processus de guérison, les sténoses cicatricielles ont été traitées par une ou plusieurs dilatations avec les bougies de Savary. Des résections circonférentielles de 2,2 à 5,5 cm de longueur ont été réalisées avec succès dans 23/24 des cas. Une seule perforation, secondaire à une erreur de manipulation, est survenue directement après la mucosectomie. A l'analyse histologique, une profondeur de résection précise à travers la sousmuq Tyrp1ueuse a été obtenue dans 85% des specimens. Les sténoses cicatricielles ont été contrôlées dans 95% des cas par dilatations avec les bougies de Savary. L'oesophagoscope rigide modifié a permis de réaliser des résections muqueuses circonférentielles et étendues en une seule séance endoscopique d'une durée de moins d'une demi-heure. L'utilisation de ce résectoscope chez l'homme devrait permettre l'éradication complète de l'oesophage de Barrett avec HGIN et/ou AC précoce dans un avenir très proche.

Animal Toxins: How is Complexity Represented in Databases?

Peptide toxins synthesized by venomous animals have been extensively studied in the last decades. To be useful to the scientific community, this knowledge has been stored, annotated and made easy to retrieve by several databases. The aim of this article is to present what type of information users can access from each database. ArachnoServer and ConoServer focus on spider toxins and cone snail toxins, respectively. UniProtKB, a generalist protein knowledgebase, has an animal toxin-dedicated annotation program that includes toxins from all venomous animals. Finally, the ATDB metadatabase compiles data and annotations from other databases and provides toxin ontology.

Bioinformatic study of selection in animal genomes

Les gènes orthologues divergent sur plusieurs aspects durant l'évolution. Après une revue de la littérature cherchant à montrer de la divergence entre les orthologues de l'humain et de la souris, j'ai souligné les différentes causes de cette divergence. En comparant les gènes qui divergent en fonction, je n'ai pas trouvé de lien avec la divergence des séquences, pour cette raison je me suis penché sur l'étude de l'expression. Notamment, j'ai étudié le niveau, la spécificité ainsi que la présence/absence d'expression des orthologues humain-souris liés aux maladies Mendéliennes. Malgré les similarités trouvées entre l'humain et la souris, j'ai détecté une différence d'expression spécifique à une des deux espèces liée a un phénotype précis (gène essentiel/non-essentiel). Cela m'a permis de conclure que la différence sur le plan phénotypique entre l'humain et la souris est mieux expliquée par les patrons d'expression plutôt que le niveau d'expression ou la sélection. J'ai été également intéressé par l'évolution des séquences d'ADN codantes pour des protéines, en particulier sur le rôle de la sélection. J'ai commencé par une étude sur la fiabilité de détection de la sélection positive en comparant des séquences divergentes. J'ai trouvé, en utilisant le model de branche-site que la sélection peut être détectée sur des séquences qui ont divergé il y a plus de 500 millions d'années. J'ai analysé le biais de GC entres les séquences sans trouver une influence sur l'estimation de la sélection positive. Finalement, Je crois que ce travail est une première étape dans l'établissement d'un lien entre la sélection et les patrons d'expression des gènes chez les vertébrés.

Rapid cohort generation and analysis of disease spectrum of large animal model of cone dystrophy.

Large animal models are an important resource for the understanding of human disease and for evaluating the applicability of new therapies to human patients. For many diseases, such as cone dystrophy, research effort is hampered by the lack of such models. Lentiviral transgenesis is a methodology broadly applicable to animals from many different species. When conjugated to the expression of a dominant mutant protein, this technology offers an attractive approach to generate new large animal models in a heterogeneous background. We adopted this strategy to mimic the phenotype diversity encounter in humans and generate a cohort of pigs for cone dystrophy by expressing a dominant mutant allele of the guanylate cyclase 2D (GUCY2D) gene. Sixty percent of the piglets were transgenic, with mutant GUCY2D mRNA detected in the retina of all animals tested. Functional impairment of vision was observed among the transgenic pigs at 3 months of age, with a follow-up at 1 year indicating a subsequent slower progression of phenotype. Abnormal retina morphology, notably among the cone photoreceptor cell population, was observed exclusively amongst the transgenic animals. Of particular note, these transgenic animals were characterized by a range in the severity of the phenotype, reflecting the human clinical situation. We demonstrate that a transgenic approach using lentiviral vectors offers a powerful tool for large animal model development. Not only is the efficiency of transgenesis higher than conventional transgenic methodology but this technique also produces a heterogeneous cohort of transgenic animals that mimics the genetic variation encountered in human patients.

Assessment of a sheep animal model to optimize photodynamic therapy in the oesophagus

Background and Objectives: Precursor lesions of oesophagus adenocarcinoma constitute a clinical dilemma. Photodynamic therapy (PDT) is an effective treatment for this indication, but it is difficult to optimise without an appropriate animal model. For this reason, we assessed the sheep model for PDT in the oesophagus with the photosensitiser meta-(tetra-hydroxyphenyl) chlorin (mTHPC). Materials and Methods: Twelve sheep underwent intravenous mTHPC injection, blood sampling and fluorescence measurements. mTHPC's pharmacokinetics was measured in vivo and in plasma by fluorescence spectroscopy. Biopsies of sheep oesophagus were compared to corresponding human tissue, and the mTHPC's biodistribution was studied under fluorescence microscopy. Finally, the sheep oesophageal mucosa was irradiated, 4 days after mTHPC's injection. Results: Histologically, the sheep and human oesophagus were closely comparable, with the exception of additional fatty tissue in the sheep oesophagus. mTHPC's pharmacokinetics in sheep and human plasmas were similar, with a maximum of concentration in the sheep 10 hours after i.v. injection. mTHPC's pharmacokinetics in vivo reached its maximum after 30-50 hours, then decreased to background levels, as in humans under similar conditions. Two days after injection, mTHPC was mainly distributed in the lamina propria, followed by a penetration into the epithelium. The sheep and human tissue sensitivity to mTHPC PDT was similar. Conclusion: In conclusion, this model showed many similarities with humans as to mTHPC's plasma and tissue pharmacokinetics, and for tissue PDT response, making it suitable to optimise oesophagus PDT. Lasers Surg. Med. 41:643-652,2009. (C) 2009Wiley-Liss,Inc.

Animal Toxins: How is Complexity Represented in Databases?

Peptide toxins synthesized by venomous animals have been extensively studied in the last decades. To be useful to the scientific community, this knowledge has been stored, annotated and made easy to retrieve by several databases. The aim of this article is to present what type of information users can access from each database. ArachnoServer and ConoServer focus on spider toxins and cone snail toxins, respectively. UniProtKB, a generalist protein knowledgebase, has an animal toxin-dedicated annotation program that includes toxins from all venomous animals. Finally, the ATDB metadatabase compiles data and annotations from other databases and provides toxin ontology.