Alterações cutâneas do cão no Kala-Azar sul-americano

According to E. Chagas (1938), South-American Kala Azar is a widespread disease from the jungle, several cases being reported from North Brazil (Estado do Pará: Marajó Island, Tocantins and Gurupi river valleys; Estados do Piauí and Ceará: coast and hinterland). Other cases were found in Northeast Brazil (Estados de Pernambuco, Alagôas and Sergipe: coast and hinterland; Estado da Bahia: hinterland). A few cases were described from Estado de Mato-Grosso (Brazil), Provincia de Salta and Território do Chaco (Argentine), and Zona contestada do Chaco (Paraguai-Bolívia). A well defined secondary anemia associated with enlargement of the liver and spleen are the chief symptoms. Death usually occurs in cachexia and with symptoms of heart failure. Half the patients were children aged less than ten years (CHAGAS, CASTRO & FERREIRA, 1937). Quite exhaustive epidemiological researches performed by CHAGAS, FERREIRA, DEANE, DEANE & GUIMARÃES (1938) in Municipio de Abaeté (Estado do Pará, Brazil) gave the incidence of 1.48% for the natural infection in human, 4.49% in dogs, and 2.63% in cats. The infection was arcribed (CUNHA & CHAGAS, 1937) to a new species of Leishmania (L. chagasi). Latter CUNHA (1938) state, that it is identical to L. infantum. ADLER (1940) found that so far it has been impossible to distinguish L. chagasi from L. infantum by any laboratory test but a final judgment must be reserved until further experiments with different species of sandflies have been carried out. Skin changes in canine Kala Azar were signaled by many workers, and their importance as regards the transmission of the disease is recognized by some of them (ADLER & THEODOR, 1931, 2. CUNHA, 1933). Cutaneous ulcers in naturally infected dogs are referred by CRITIEN (1911) in Malta, by CHODUKIN & SCHEVTSCHENKO (1928) in Taschkent, by DONATIEN & LESTOCQUARD (1929) and by LESTOCQUARD & PARROT (1929) in Algeria, and by BLANC & CAMINOPETROS (1931) in Greece. Depilation is signaled by YAKIMOFF & KOHL-YAKIMOFF (1911) in Tunis, by YAKIMOFF (1915) in Turkestan. Eczematous areas or a condition described as "eczema furfurace" is sometimes noted in the areas of depilation (DONATIEN & LESTOCQUARD). The skin changes noticed by ADLER & THEODOR (1932) in dogs naturally infected with Mediterranean Kala Azar can be briefly summarized as a selective infiltration of macrophages around hair follicles including the sebaceous glands and the presence of infected macrophages in normal dermis. The latter phenomenon in the complete absence of secondary infiltration of round cells and plasma cells is the most striking characteristic of canine Kala Azar and differentiates it from L. tropica. In the more advanced stages the dermis is more cellular than that of normal dogs and may even contain a few small dense areas of infiltration with macrophages and some round cells and polymorphs. The external changes, i. e., seborrhea and depilation are roughly proportional to the number of affected hair follicles. In dogs experimentally infected with South-American Kala Azar the parasites were regularly found in blocks of skin removed from the living animal every fortnight (CUNHA, 1938). The changes noticed by CUNHA, besides the presence of Leishmania, were perivascular and diffuse infiltration of the cutis with mononuclears sometimes more marked near hair follicles, as well as depilation, seborrhea and ulceration. The parasites were first discovered and very numerous in the paws. Our material was obtained from dogs experimentally infected by Dr. A. MARQUES DA CUNHA< and they were the subject of a previous paper by CUNHA (1938). In this study, however, several animals were discarded as it was found that they did develop a superimposed infection by Demodex canis. This paper deals with the changes found in 88 blocks of skin removed from five dogs, two infected with two different canine strains, and three with two distinct human strains of South-American Kala Azar. CUNHA'S valuable material affords serial observations of the cutaneous changes in Kala Azar as most of the blocks of skin were taken every fortnight. The following conclusions were drawn after a careful microscopic study. (1) Skin changes directly induced in the dog by the parasites of South-American Kala Azar may b described as an infiltration of the corium (pars papillaris and upper portion of the reticular layer) by histocytes. Parasites are scanty, at first, latter becoming very numerous in the cytoplasm of such cells. Sometimes the histocytes either embedding or not leishman bodies appear as distinct nodes of infiltration or cell aggregations (histocytic granuloma, Figs. 8 and 22) having a perivascular distribution. The capillary loops in the papillae, the vessels of the sweat glands, the subpapillary plexus, the vertical twigs connecting the superficial and deep plexuses are the ordinary seats of the histocytic Kala Azar granulomata. (2) Some of the cutaneous changes are transient, and show spontaneous tendency to heal. A gradual transformation of the histocytes either containing or not leishman bodies into fixed connective tissue cells or fibroblasts occut and accounts for the natural regression just mentioned. Figs. 3, 5, 18, 19 and 20 are good illustrations of such fibroblastic transformation of the histocytic Kala Azar granulomata. (3) Skin changes induced by the causative organism of South-American Kala Azar are neither uniform nor simultaneous. The same stage may be found in the same dog in different periods of the disease, and not the same changes take place when pieces from several regions are examined in the same moment. The fibroblastic transformation of the histocytic granulomata marking the beginning of the process of repair, e. g., was recognised in dog C, in the 196th as well as in the 213rd (Fig. 18) and 231st (Fig. 19) days after the inoculation. (4) The connective tissue of the skin in dogs experimentally infected with South-American Kala Azar is overflowed by blood cells (monocytes and lymphocytes) besides the proliferation in situ of undifferentiated mesenchymal cells. A marked increase in the number of cells specially the "ruhende Wanderzellen" (Figs. 4 and 15) is noticed even during the first weeks after inoculation (prodomal stage) when no leishman bodies are yet found in the skin. Latter a massive infiltration by amoeboid wandering cells similar to typical blood monocytes (Fig. 21) associated to a small number of lymphocytes and plasma cells (Figs. 9, 17, 21, and 24) indicates that the emigration of blood cells...

Sôbre alguns Phlebotomus e Brumptomyia da Colômbia (Diptera, Psychodidae)

The first case of Kala-azar in Colombia was discovered in Soledad, S. Vicente do Chucuri, Dept. Santander, by Gast-Galvis who viscerotomized a three year old girl deceased in December, 1943. In 1944, fifty-three Phlebotominae were collected in the chicken pen of the girl's house, two new species included. Mangabeira helped by A. Gast Galvis, Juan Antonio Montoya and E. Osorno Mesa, collected some Phlebotomus in that country. The geographical distribution of the species of Phlebotomus collected in Colombia (P. abonnenci, P. camposi, P. columbianus, P. dubitans, P. gasti, P. montoyai, P. saulensis, P. serranus, P. triramulus) and two species of Brumptomyia (B. beaupertuyi and b mesari), are included. our description of the male P. columbianus is based on some specimens found in association with females. However, doubts exist about such association of sexes. There is no correspondence between the length of the spicules and the ducts of spermathecae. Besides, the specimens were not obtained by raising. The following new species are described and compared with previously known ones: a) Phlebotomus gasti sp. n. differs from the other species by a protruding tubercle in the gubernaculum. It has also fewer setae in the tuft of the basistyle, a different length of the inferior gonapophyses, and a differently shaped clasper. b) Phlebotomus dubitans sp. n. differs from P. walkeri and P. deanei (according to personal information from O. Theodor, who examined the types, they are identical to P. williamsi and P. sericeus respectively), mainly because these species have the inferior gonapophyses larger than the basistyle and fewer setae in the basistyle. P. evandroi is separated by the shape of the claspers and by the tuft of setae of the basistyle. P. marajoensis is the closest relative to P. dubitans. There is a possibility of their being synonymous. On the other hand, they can be differentiated by the existence of three extra distal spines in P. marajoensis. There is also a difference in their palpal indexes: for marajoensis I - II - IV - III - V, and for dubitans I - IV (III - II) - V. We notice, too, that the inferior gonapophyses in P. marajoensis is a little shorter. P. marajoensis has a long seta in the basistyle (clearly shown in the original drawing), not found in the new species. c) Phlebotomus montoyai sp. n.: The closest relatives are P. noguchii, P. peruensis, P. pescei, P. quinquifer and P. rickardi. They differ from the new species by the number and length of the setae of the basistyle tuft which are more numerous and longer in the new species. The shapes of their claspers are also different. Other differences are: the basal portion of the basistyle in P. noguchii is very wide (in montoyai it is narrower); the intermediate spine of the dististyle is located on a protruding tubercle ( in the new species there is hardly a tubercle); the spicules are long, and the inferior gonapophyses is longer than the basistyle. P. quinquifer and P. rickardi have a shorter dististyle and narrower wings, with different venation. The main difference, however lies, in the M4, which ends almost at the level of the junction of M1 with M2 (in P. montoyai the M4 ends far behind). In P. peruensis and P. pescei the intermediary spine of the dististyle is closer to the distal spine than to the basal one, whereas in the new species it is situated between the two pairs. Their inferior gonapophyses is longer than the basistyle. d) Brumptomyia mesai sp. n. - Closest relatives are: B. hamatus, B. pentacanthus, B. beaupertuyi which are easily separated from the new species because the tufts of their basistyle have thin and differently shaped hairs. Also their claspers are shaped differently. B. avellari is also easily recognized on account of the twisted aspect of its clasper and because the basal tuft of the basistyle has few setae, B. brumpti tuft of setae arise directly from the basistyle; these setae are stronger than those of the new species. It has 8 blade-like setae located on the inner surface of the distal half, whereas the new species has only six setae. In B. brumpti, there are three median and two terminal spines in the dististyle; in the new species, there are two median and two terminal spines and one between them, which is closer to the two median spines. The comparison with B. galindoi is based in a specimen determined by Fairchild and deposited in the entomological collection of the "Faculdade de Higiene e Saúde Pública da Universidade de S. Paulo". The genitalia of the new species is much shorter, in galindoi the inferior gonapophyses is 0,8 mm long whereas in B. mesai it hardly reaches 0,6 mm. The shape of the clasper and the distribution of its setae are different. The sub-median lamellae, besides being longer in B. galindoi are also longer in comparison with the other parts of the genitalia. The gubernaculum of the new species is longer, thinner, and more pointed; in B. galindoi it is shorter and triangular. In the drawing published by Fairchild and Hertig 91947), the basistyle shows 8 blade-like setae on the distal half, whereas in the new species only six are found.

A rewiew of the subgenus Trichopygomyia Barretto, 1962; with description of a new species from the brazilian Amazon Basin (Diptera: Psychodidae, Phlebotominae)

The subgenus Trichopygomyia Barreto, 1962 of the phlebotomine genus Lutzomyia França, 1924 is reviewed. This subgenus corresponds to the informal species group longispina of Theodor (1965). Lutzomyia (Trichopygomya) ratcliffei n.sp. from the Brazilian Amazon Basin is described. Figures, keys, distribution maps and notes on ecology are presented for all the known forms. The better known species are most frequently encountered in armadillo burrows and, therefore, could well be vectors of Leishmania.

Phlebotominae de Venezuela: Lutzomyia amilcari sp.n. del estado Lara

Se describe el macho de Lutzomyia amilcari sp.n. capturado en el Parque Nacional de Yacambú, Distrito Andrés Eloy Blanco, Estado Lara, Venezuela. La especie puede ser identificada por la forma característica del parámetro. Pertenece al grupo Verrucarum, serie Verrucarum de Theodor (1965).

Phlébotomes du Nicaragua: I. Description de la femelle de Lutzomyia zeledoni Young et Murillo, 1984 (Diptera, Psychodidae), et implication épidémiologique de la présence de Lutzomyia longipalpis sur la façade Pacifique

Les auteurs décrivent la femelle de Lutzomyia zeledoni Young et Murillo, 1984 du Groupe Vespertilionis Theodor, 1965. Cette espèce a été capturée en milieu péridomestique en compagnie de Lutzomyia longpalpis Lutz et Neiva, 1912 répertorié pour la première fois du Nicaragua. Il est mis l'accent sur l'importance épidémiologique de cette dernière espèce récoltée en abondance dans plusieurs stations de la façade pacifique.

Descripción del macho y de la hembra de Lutzomyia larensis n.sp. (Diptera: Psychodidae, Phlebotominae) del estado Lara, Venezuela

Es dada la descripción del macho y de la hembra de una nueva espécie de flebotomíneo recolectado en tronco de árboles en el Parque Nacional de Yacambú, Estado Lara, Venezuela. La nueva espécie pertenece al subgénero Helcocyrtomyia, Barrto, 1962 o al grupo vexator, série peruensis de Theodor (1965) y de Young & Fairchild (1974). El macho se identifica por la presséncia de 5 espinos en el dististilo y por el mechón de cerdas largas en el basistilo, implantadas en tubérculos gruesos y queratinizados. La hembra a la vez presenta armadura dl cibário con 4 dientes largos y una espermateca segmentada semejante a las de otras espécies de la série peruensis.

Extracellular matrix components in peripheral nerve repair: how to affect neural cellular response and nerve regeneration?

Peripheral nerve injury is a serious problem affecting significantly patients' life. Autografts are the "gold standard" used to repair the injury gap, however, only 50% of patients fully recover from the trauma. Artificial conduits are a valid alternative to repairing peripheral nerve. They aim at confining the nerve environment throughout the regeneration process, and providing guidance to axon outgrowth. Biocompatible materials have been carefully designed to reduce inflammation and scar tissue formation, but modifications of the inner lumen are still required in order to optimise the scaffolds. Biomicking the native neural tissue with extracellular matrix fillers or coatings showed great promises in repairing longer gaps and extending cell survival. In addition, extracellular matrix molecules provide a platform to further bind growth factors that can be released in the system over time. Alternatively, conduit fillers can be used for cell transplantation at the injury site, reducing the lag time required for endogenous Schwann cells to proliferate and take part in the regeneration process. This review provides an overview on the importance of extracellular matrix molecules in peripheral nerve repair.

Wissen um Stimmung : zu einem vagen Begriff in Robert Musils Frühwerk

Diese Studie untersucht die Poetik und Wirkungsästhetik von Robert Musils Die Verwirrungen des Zöglings Törleß (1906) und Die Vereinigungen (1911) anhand des Schlüsselbegriffs >Stimmung<. Wegen der Vagheit des Begriffs und seiner Relevanz in Psychologie, Philosophie und Ästhetik um 1900 wird das Wissen um >Stimmung< im ersten Teil der Studie diskursgeschichtlich beschrieben. Es kann dabei anhand von Robert Mayer, Gustav Theodor Fechner, Hermann von Helmholtz und Wilhelm Wundt gezeigt werden, dass >Stimmungen< als psychologische Zustände wie ästhetische Phänomene in einem engen Verhältnis mit der thermodynamischen Theorie und der Denkfigur des psychophysischen Parallelismus standen. >Stimmungen< galten zum Ende des 19. Jh. einerseits als gleichermaßen experimentell unzugängliche wie grundlegende psychologische Dispositionen, die Körper, Emotion und Intellekt umfassen können, und andererseits als energetisch konzipiertes Verhältnis des Einzelnen zur Außenwelt, das sich in Schwingungen und Strahlen äußert. Im zweiten und dritten Teil wurden die Wissensübertragungen und die genuin literarischen Ausdifferenzierungen von >Stimmungen< in einer dezidiert textnahen Lektüre von Musils Frühwerk entwickelt. Die Textanalyse zeigte, dass in Musils Frühwerk auf thematischer, metaphorologischer, poetologischer und wirkungsästhetischer Ebene von Stimmungen konstituiert wird. Von herausragender Bedeutung sind dabei immaterielle Phänomene. In Auseinandersetzung mit der Ästhetik des Fin de Siècle und dem >Psychophysischen< formuliert der Roman Die Verwirrungen des Zöglings Törleß eine Poetik des Duftes, mit der das dichtungstheoretische Ideal einer gleichsam emotional wie rational wirksamen Literatur umgesetzt werden soll. Die Novellen Die Vollendung der Liebe und Die Versuchung der stillen Veronika übertragen psychologisches Stimmungswissen in den literarischen Text und differenzieren es zu ästhetischen Strukturen aus. Zwei verschiedene Leitkonzepte konnten in Die Vereinigungen identifiziert werden: Während Die Versuchung der stillen Veronika eine Poetik bzw. eine Wirkungsästhetik der Wellen entwirft, formuliert Die Vollendung der Liebe eine literarische Anthropologie anhand musikalischen Metaphern und Texturen aus.

Cellular schwannomas of the intracranial and intraspinal compartment: morphological and immunological characteristics compared with classical benign schwannomas.

The concept of cellular schwannoma as an unusual benign tumor is well established for peripheral nerves but has never been tested in neurosurgical series. In order to test the validity of this concept in cranial nerves and spinal roots we performed an analysis of the clinical and morphological characteristics of 12 cellular and 166 classical benign schwannomas. Immunohistochemical detection of antigen expression in Schwann cells including proliferating cell nuclear antigen (PCNA) was also performed. This study shows that cellular schwannomas in neurosurgical series manifest at a lower age than the classical benign variant and occur mainly in the spinal roots. Mitotic activity and sinusoidal vessels appear more frequently in cellular schwannomas and constitute with high cellularity, the most valuable criteria separating both entities. The postoperative course in both types of tumors was free of metastases or sarcomatous changes. Immunoexpression of S-100 protein, vimentin, epithelial membrane antigen and glial fibrillary acidic protein is not statistically different between the two variants. In contrast, PCNA is more highly expressed in cellular schwannomas. These These results confirm the concept that cellular schwannomas are a clinico-pathological variant of benign schwannomas and provide significant support for the introduction of this entity in neurosurgical oncology.

Adipose-derived stem cells enhance peripheral nerve regeneration.

Traumatic injuries resulting in peripheral nerve lesions often require a graft to bridge the gap. Although autologous nerve auto-graft is still the first-choice strategy in reconstructions, it has the severe disadvantage of the sacrifice of a functional nerve. Cell transplantation in a bioartificial conduit is an alternative strategy to create a favourable environment for nerve regeneration. We decided to test new fibrin nerve conduits seeded with various cell types (primary Schwann cells and adult stem cells differentiated to a Schwann cell-like phenotype) for repair of sciatic nerve injury. Two weeks after implantation, the conduits were removed and examined by immunohistochemistry for axonal regeneration (evaluated by PGP 9.5 expression) and Schwann cell presence (detected by S100 expression). The results show a significant increase in axonal regeneration in the group of fibrin seeded with Schwann cells compared with the empty fibrin conduit. Differentiated adipose-derived stem cells also enhanced regeneration distance in a similar manner to differentiated bone marrow mesenchymal stem cells. These observations suggest that adipose-derived stem cells may provide an effective cell population, without the limitations of the donor-site morbidity associated with isolation of Schwann cells, and could be a clinically translatable route towards new methods to enhance peripheral nerve repair.

Role of thyroid hormones and their receptors in peripheral nerve regeneration.

After peripheral nerve injury in adult mammals, reestablishment of functional connections depends on several parameters including neurotrophic factors, the extracellular matrix, and hormones. However, little is known about the contribution of hormones to peripheral nerve regeneration. Thyroid hormones, which are required for the development and maturation of the central nervous system, are also important for the development of peripheral nerves. The action of triiodothyronine (T3) on responsive cells is mediated through nuclear thyroid hormone receptors (TRs) which modulate the expression of specific genes in target cells. Thus, to study the effect of T3, it is first necessary to know whether the target tissues possess TRs. The fact that sciatic nerve cells possess functional TRs suggests that these cells can respond to T3 and, as a consequence, that thyroid hormone may be involved in peripheral nerve regeneration. The silicone nerve guide model provides an excellent system to study the action of local administration of T3. Evidence from such studies demonstrate that animals treated locally with T3 at the level of transection have more complete regeneration of sciatic nerve and better functional recovery. Among the possible regulatory mechanisms by which T3 enhances peripheral nerve regeneration is rapid action on both axotomized neurons and Schwann cells which, in turn, produce a lasting and stimulatory effect on peripheral nerve regeneration. It is probable that T3 up- or down-regulates gene expression of one or more growth factors, extracellular matrix, or cell adhesion molecules, all of which stimulate peripheral nerve regeneration. This could explain the greater effect of T3 on nerve regeneration compared with the effect of any one growth factor or adhesion molecule.

Exploring Parallels Between Molecular Changes Induced in PNS by Aging and Demyelinating Neuropathies

The peripheral nervous system (PNS) is involved in many age-dependent neurological deficits, including numbness, pain, restless legs, trouble with walking and balance that are commonly found in the elderly. These symptoms generally result from demyelination and/or loss of axonal integrity. However, the precise identity of age-regulated molecular changes in either neuronal or glial compartments of the nerve is unclear. Interestingly, these deficiencies are also present in inherited neuropathies, where the expressivity of the rapid and early onset phenotypes is undeniably more severe than in normal aging. Nevertheless, especially the molecular changes underlying loss of axonal integrity in neuropathy condition are also poorly understood. To unravel molecular mechanisms affected by PNS aging, we used wildtype mice at 17 time-points from day of birth until senescence (28 months-old). For the neuropathy study, we focused on 56 day-old Schwann cell-specific neuropathy-inducing mutants, MPZCre/1/ LpinfE2-3/fE2-3 and MPZCre/1/ScapfE1/fE1 mice, that have, at this age, already developed neuropathic symptoms. Transcriptomes of dissected Schwann cell-containing endoneurium or sensory neuron-containing dorsal root ganglia have been analyzed throughout time or genotypes, using Illumina Bead Chips. Following data validation, we identified groups of differentially expressed genes in the development, aging and in the neuropathic mutants, in both glial and neuronal compartments. We detected substantial differences in the dynamics of changes in gene expression during development and aging between these two compartments. Furthermore, considering the above-mentioned phenotypic similarities, we integrated aging and mutant data. Interestingly, we observed that there are some parallels at the molecular level between processes involved in aging, which leads to less severe and more progressive PNS alterations, and in the rapid onset peripheral neuropathies. Apart from helping the understanding of molecular alterations underlying age-related PNS phenotypes, this data should also contribute to the identification of pathways that could be used as targets for therapeutical approaches to prevent complications associated with both aging and inherited forms of neuropathies.

Identification and characterization of novel molecular mechanisms involved in initiation and progression of myelination

Schwann cells synthesize a large amount of membrane that form a specialized structure called myelin that surrounds axons and facilitate the transmission of electrical signal along neurons in peripheral nervous system (PNS). Previous studies demonstrated that both Schwann cell differentiation and de-differentiation (in the situation of a nerve injury or demyelinating disease) are regulated by cell-intrinsic regulators including several transcription factors. In particular, the de-differentiation of mature Schwann cells is driven by the activation of multiple negative regulators of myelination including Sox2, c-Jun, Notch and Pax3, all usually expressed in immature Schwann cells and suppressed at the onset of myelination. In order to identify new regulators of myelination involved in the development of the PNS, we analyzed the gene-expression profiling data from developing PNS and from three models of demyelinating neuropathies. This analysis led to the identification of Sox4, a member of the Sox family of transcription factors, as a potential candidate. To characterize the molecular function of Sox4 in PNS, we generated two transgenic lines of mice, which overexpress Sox4 specifically in Schwann cells. Detailed analysis of these mice showed that the overexpression of Sox4 in Schwann cells causes a delay in progression of myelination between post-natal day 2 (P2) and P5. Our in vitro analysis suggested that Sox4 cDNA can be overexpressed while the protein translation is tightly regulated. Interestingly, we observed that Sox4 protein is stabilized in nerves of the CMT4C mouse, a model of the human neuropathy. We therefore crossed Sox4 transgenic mice with CMT4C mice and we observed that Sox4 overexpression exacerbated the neuropathy phenotype in these mice. While recognized as being crucial for the normal function of both neurons and myelinating glial cells, the processes that regulate the beginning of myelination and the nature of the neuro-glial cross-talk remains mostly unknown. In order to gain insight into the molecular pathways involved in the interactions between neurons and associated glial cells, we developed a neuron-glia co-culture system based on microfluidic chambers and successfully induced myelination in this system by ascorbic acid. Importantly, we observed that in addition to acting on Schwann cells, ascorbic acid also modulate neuronal/axonal NRG1/ErbB2-B3 signalling. The experimental setting used in our study thus allowed us to discover a novel phenomena of propagation for myelination in vitro. The further characterization of this event brought us to identify other compounds able to induce myelination: ADAMs secretases inhibitor GM6001 and cyclic-AMP. The results generated during my thesis project are therefore not only important for the advancement of our understanding of how the PNS works, but may also potentially help to develop new therapies aiming at improvement of PNS myelination under disease conditions. - Les cellules de Schwann synthétisent une grande quantité de membrane formant une structure spécialisée appelée myéline qui entoure les axones et facilite la transmission du signal électrique le long des neurones du système nerveux périphérique (SNP). Des études antérieures ont démontré que la différenciation et la dédifférenciation des cellules de Schwann (dans la situation d'une lésion nerveuse ou d'une maladie démyélinisante) sont régulées par des régulateurs cellulaires intrinsèques, incluant plusieurs facteurs de transcription. En particulier, la dédifférenciation des cellules de Schwann matures est contrôlée par l'activation de plusieurs régulateurs négatifs de la myélinisation dont Sox2, c-Jun, Notch et Pax3, tous habituellement exprimés dans des cellules de Schwann immatures et supprimés au début de la myélinisation. Afin d'identifier de nouveaux régulateurs de myélinisation impliqués dans le développement du SNP, nous avons analysé le profil d'expression génique durant le développement du SNP ainsi que dans trois modèles de neuropathies démyélinisantes. Cette analyse a mené à l'identification de Sox4, un membre de la famille des facteurs de transcription Sox, comme étant un candidat potentiel. Dans le but de caractériser la fonction moléculaire de Sox4 dans le SNP, nous avons généré deux lignées transgéniques de souris qui surexpriment Sox4 spécifiquement dans les cellules de Schwann. L'analyse détaillée de ces souris a montré que la surexpression de Sox4 dans les cellules de Schwann provoque un retard dans la progression de la myélinisation entre le jour postnatal 2 (P2) et P5. Notre analyse in vitro a suggéré que l'ADNc de Sox4 peut être surexprimé alors que la traduction des protéines est quand à elle étroitement régulée. De façon intéressante, nous avons observé que la protéine Sox4 est stabilisée dans les nerfs des souris CMT4C, un modèle de neuropathie humaine. Nous avons donc croisé les souris transgéniques Sox4 avec des souris CMT4C et avons observé que la surexpression de Sox4 exacerbe le phénotype de neuropathie chez ces souris. Bien que reconnus comme étant cruciaux pour le fonctionnement normal des neurones et des cellules gliales myélinisantes, les processus qui régulent le début de la myélinisation ainsi que la nature des interactions neurone-glie restent largement méconnus. Afin de mieux comprendre les mécanismes moléculaires impliqués dans les interactions entre les neurones et les cellules gliales leur étant associés, nous avons développé un système de co-culture neurone-glie basé sur des chambres microfluidiques et y avons induit avec succès la myélinisation avec de l'acide ascorbique. Étonnamment, nous avons remarqué que, en plus d'agir sur les cellules de Schwann, l'acide ascorbique module également la voie de signalisation neuronale/axonale NRG1/ErbB2-B3. Le protocole expérimental utilisé dans notre étude a ainsi permis de découvrir un nouveau phénomène de propagation de la myélinisation in vitro. La caractérisation plus poussée de ce phénomène nous a menés à identifier d'autres composés capables d'induire la myélinisation: L'inhibiteur de sécrétases ADAMs GM6001 et l'AMP cyclique. Les résultats obtenus au cours de mon projet de thèse ne sont donc pas seulement importants pour l'avancement de notre compréhension sur la façon dont le SNP fonctionne, mais peuvent aussi potentiellement aider à développer de nouvelles thérapies visant à l'amélioration de la myélinisation du SNP dans des conditions pathologiques.

Thyroid hormone enhances transected axonal regeneration and muscle reinnervation following rat sciatic nerve injury.

Improvement of nerve regeneration and functional recovery following nerve injury is a challenging problem in clinical research. We have already shown that following rat sciatic nerve transection, the local administration of triiodothyronine (T3) significantly increased the number and the myelination of regenerated axons. Functional recovery is a sum of the number of regenerated axons and reinnervation of denervated peripheral targets. In the present study, we investigated whether the increased number of regenerated axons by T3-treatment is linked to improved reinnervation of hind limb muscles. After transection of rat sciatic nerves, silicone or biodegradable nerve guides were implanted and filled with either T3 or phosphate buffer solution (PBS). Neuromuscular junctions (NMJs) were analyzed on gastrocnemius and plantar muscle sections stained with rhodamine alpha-bungarotoxin and neurofilament antibody. Four weeks after surgery, most end-plates (EPs) of operated limbs were still denervated and no effect of T3 on muscle reinnervation was detected at this stage of nerve repair. In contrast, after 14 weeks of nerve regeneration, T3 clearly enhanced the reinnervation of gastrocnemius and plantar EPs, demonstrated by significantly higher recovery of size and shape complexity of reinnervated EPs and also by increased acetylcholine receptor (AChRs) density on post synaptic membranes compared to PBS-treated EPs. The stimulating effect of T3 on EP reinnervation is confirmed by a higher index of compound muscle action potentials recorded in gastrocnemius muscles. In conclusion, our results provide for the first time strong evidence that T3 enhances the restoration of NMJ structure and improves synaptic transmission.

Numerical Taxonomy of Old World Phlebotominae (Diptera: Psychodidae): 1. Considerations of Morphological Characters in the Genus Phlebotomus Rondani & Berté 1840

Numerical analyses (correspondence analysis, ascending hierarchical classification, cladistic approach) were applied to the morphological characters of the adults of the genus Phlebotomus Rondani & Berté 1840. They confirm the reliability of the classic classifications, and also redefine the taxonomic and phylogenetic position of certain taxa. Thus, Spelaeophlebotomus Theodor 1948, Idiophlebotomus Quate & Fairchild 1961 and Australophlebotomus Theodor 1948 deserve generic rank. Among the vectors of leishmaniasis, the subgenus Phlebotomus Rondani & Berté 1840 is probably ancient. The results attribute an intermediate taxonomic and phylogenetic position to the taxa Euphlebotomus Theodor 1948 and Anaphlebotomus Theodor 1948, and reveal the probable artificial nature of the latter. The comparatively large numbers of species of subgenera Paraphlebotomus Theodor 1948, Synphlebotomus Theodor 1948 and, above all, Larroussius Nitzulescu 1931 and Adlerius Nitzulescu 1931, suggest that they are relatively recent. The development of adult morphological characters, the validity of their use in taxonomy and proposals for further studies are discussed.