Acredita-se que nos humanos, o ligamento vocal se desenvolva após o nascimento. No entanto, não há consenso na literatura sobre qual a faixa etária de seu aparecimento. Muitos estudos indicam que no neonato, a lâmina própria da prega vocal apresenta-se com algumas fibras esparsas sem uma organização particular. O principal objetivo deste estudo foi obter informação a respeito das fibras colagênicas e do sistema elástico (sob a luz dos conhecimentos atuais sobre a matriz extracelular), na lâmina própria de pregas vocais de fetos no período perinatal. Laringes obtidas por autópsia de fetos entre sete a nove meses foram estudadas através de microscopia de luz e eletrônica de transmissão. Fibras contendo colágeno foram identificadas através do Método da Picrossírius-polarização, fibras do sistema elástico foram descritas utilizando-se o método de Resorcina-fucsina de Weigert após oxidação com oxona. Os resultados histoquímicos coincidem com as observações da microscopia eletrônica, evidenciando populações de fibras de colágeno segregadas em diferentes compartimentos na lâmina própria. Assim, em sua região central as fibras de colágeno se mostraram finas, fracamente birrefringentes de coloração esverdeada, enquanto que as regiões superficiais e profundas apresentaram fibras grossas de colágeno com forte birrefringência de cor vermelho-amarelada, quando estudadas através do método da Picrossírius-polarização. Estas características sugerem que as fibras finas da região central são compostas principalmente por colágeno tipo III, enquanto o colágeno tipo I predomina nas regiões superficial e profunda, em concordância com as observações da literatura relacionada com o estudo da prega vocal de adultos. Assim como o componente colagênico, as fibras do sistema elástico mostraram uma distribuição diferencial ao longo da lâmina própria. Em certo sentido, esta distribuição é complementar àquela das fibras de colágeno: a região central, na qual fibras colagênicas eram escassas e finas, apresentou maior densidade de fibras do sistema elástico, em comparação às regiões superficial e profunda. Assim, a presença de um padrão de distribuição diferencial das fibras da matriz extracelular na prega vocal humana fetal equivalente à descrição clássica do ligamento vocal adulto nos permitiu concluir que o ligamento vocal já está presente ao nascimento. As implicações funcionais destes achados foram discutidas. O conceito corrente de que os estímulos externos, como a fonação, são essenciais para a determinação da estrutura em camadas da lâmina própria, faz com que nossos resultados sejam surpreendentes ao evidenciar a presença de uma distribuição complexa e organizada dos componentes do tecido conjuntivo na lâmina própria de pregas vocais de fetos no período perinatal. A idéia de que a contribuição genética poderia desempenhar um papel importante na organização destas camadas, independentemente do estímulo mecânico, poderia explicar melhor a presença das estruturas observadas já ao nascimento, uma vez que o mecanismo genético pode agir antes de qualquer estímulo mecânico externo, como a fonação. / It is currently believed that, in humans, the vocal ligament develops after birth. However, there is no consensus in the literature about the age of its surge. Most papers describe that in the newborn, the lamina propria shows the presence of some sparse fibers without any particular organization. The main purpose of this study was to obtain information regarding collagenous and elastic system fibers (in the light of the current knowledge on extracellular matrix) in the lamina propria of fetal vocal fold. Larynges obtained from autopsy of human fetuses aged seven to nine months were studied by means of light and electron microscopy. Collagen containing fibers were assessed by the Picrosirius-polarization method, elastic system fibers were described using Weigert\'s resorcin-fuchsin with previous oxidation with oxone. The histochemical results coincide with the electron microscope observations in showing collagen populations segregated into different compartments of the lamina propria. Thus, in its central region the collagen shows up as thin, weakly birefringent, greenish fibers while the superficial and deep regions consist of thick collagen fibers which display a strong birefringence of red or yellow color when studied with the aid of the Picrosirius-polarization method. These characteristics strongly suggest that the thin fibers in the central region are composed mainly of type-III collagen, whereas type-I collagen predominates in the superficial and deep regions, in agreement with the observations in the literature pertaining to studies of adult vocal folds. As well as collagen, the elastic system fibers show a differential distribution throughout the lamina propria. This distribution is complementary, in a sense, to that of the collagen fibers: the central region, with thin collagenous fibers, presents the greatest density of elastic system fibers in comparison to the superficial and deep regions. Thus, the presence of a differential distribution of the extracellular matrix fibers in the fetal vocal fold equivalent to the classical description of the adult vocal ligament allowed the conclusion that a vocal ligament is already present in the newborn. The functional implications of the foregoing findings are discussed. Current ideas sustaining that stimuli like phonation are essential to the determination of the layered structure of the lamina propria would make it surprising that a newborn baby could present a complex and organized distribution of connective tissue components as our results show to be the case. The idea that genetic contrivance instead should play a role in the organization of these layers seems to explain better the observed structures once it would act before any mechanical stimulus similar to phonation could take place.

Objetivo: Este trabalho avaliou o recobrimento radicular obtido por duas técnicas de cirurgia plástica periodontal em pacientes fumantes graves, através de análises clínica, histológica e laboratorial. Materiais e Métodos: Foram selecionados 20 pacientes fumantes graves que apresentavam ao menos duas retrações classes I ou II de Miller em dentes não molares, bilateralmente. Os parâmetros clínicos avaliados foram: profundidade de sondagem (PS), nível clínico de inserção (NCI), altura (AR) e largura das retrações (LR), altura (AM) e espessura da mucosa queratinizada (EM) e altura e base das papilas mesiais e distais à retração. Um lado recebeu retalho posicionado coronalmente, enquanto no outro lado foi utilizada a técnica de retalho estendido. Ambos os procedimentos foram associados ao enxerto de tecido conjuntivo subepitelial. Biópsias foram feitas durante os procedimentos cirúrgicos para quantificação dos vasos sanguíneos utilizando de avaliação imunohistoquímica. Coleta de saliva foi realizada pré tratamento e após 3 e 6 meses, como forma de avaliar, por meio de ensaio imunoenzimático (ELISA), a exposição ao fumo durante o período completo do estudo. Resultados: Os parâmetros clínicos avaliados não apresentaram diferença estatística entre grupos ou intra-grupos. Os pacientes mantiveram a mesma exposição ao fumo durante os 6 meses avaliados. O recobrimento radicular obtido foi baixo para ambas as técnicas estudadas (43,18% para técnica padrão e 44,52% para técnica teste). O retalho apresentou menor quantidade de vasos sanguíneos do que o tecido do enxerto (13,49 ± 5,48 contra 18,69 ± 4,87, p= 0,0001). Conclusão: O fumo prejudicou intensamente os resultados de ambas as técnicas empregadas, reduzindo o recobrimento radicular, a frequência de recobrimento total e a quantidade de vasos sanguíneos nos tecidos. Não houve diferença entre as técnicas testadas. / Background: The aim of this study was to evaluate the root coverage obtained by two different periodontal plastic techniques in severe smokers, analyzing clinical, laboratorial and histological parameters. Methods: Twenty severe smokers were selected. Each patient had bilateral Miller class I or II gingival recessions in non-molar teeth. Clinical measurements of probing pocket depth (PPD), clinical attachment level (CAL), recession height (RH) and recession width (RW), keratinized mucosa height (KMH) and keratinized mucosa thickness of (KMT) and height and width of the adjacent papillae were determined at baseline, 3 and 6 months. One side received coronally positioned flap, while the other side received the extended flap technique, both procedures in conjuction with the subeptelial connective tissue graft (SCTG). Biopsies from SCTGs and flaps were prepared for immunohistochemistry. Blood vessels were identified and counted by coexpression of Factor-VIII related antigen stained endothelial cells. Salivary cotinine samples were taken at baseline and after 3 and 6 months, as an indicator of the level of exposure to nicotine. Results: Between groups or intra groups analysis showed no statistical differences for the evaluated clinical parameters. Patients maintained the same exposure to smoke during the 6 months evaluation periods. Both techniques had low root coverage (standard technique: 43.18% and test technique: 44.52%). Histomorphometry of biopsies revealed blood vessel densities of 13.49 ± 5.48 vessels/200x field for the flaps, and of 18.69 ± 4.87 vessels/200x field for the grafts (p = 0.0001). Conclusion: The extend flap technique did not differ from standart technique for recession treatment of severe smoking patients.

Estudo das superfícies dorsal e ventral da língua e da superfície de interface epitélio-tecido conjuntivo de Bradypus torquatus, foi realizado empregando-se os métodos de microscopia de luz e microscopia eletrônica de varredura. As línguas de Bradypus torquatus foram removidas, dissecadas e seccionadas em 3 partes: rostral, média e caudal e tratadas pelos métodos de microscopia de luz e eletrônica de varredura. Os resultados revelaram que a língua apresenta um ápice arredondado, um corpo e uma raiz. A mucosa dorsal mostra três tipos de papilas linguais: filiformes, fungiformes e vaIadas e uma superfície inferior lisa. Histologicamente, observou-se que a superfície dorsal da língua contém as papilas filiformes, fungiformes e vaIadas, distribuídas nas regiões rostral, média e caudal. A camada epitelial é de espessura variável e possui os estratos basal, espinhosa, granular e córnea, mostrando as células epiteliais. A lâmina própria é caracterizada por um tecido conjuntivo denso formando as projeções papilares longas, curtas e arredondadas. As imagens tridimensionais obtidas pela microscopia eletrônica de varredura, mostraram numerosas papilas filiformes localizadas essencialmente nas regiões rostral e média. Poucas papilas fungiformes estão presentes na superfície dorsal da língua, tendo na maioria localizada nas margens da extremidade rostral da língua, exibindo as formas arredondadas e baixas. Elas são cobertas de células epiteliais queratinizadas. Na região caudal notou-se duas papilas vaIadas, contendo valas profundas e a superfície coberta de epitélio escamoso estratificado onde pode ser identificados poros gustativos. Nas papilas fungiformes identificou-se vários poros gustativos na superfície. As amostras tratadas com a solução de hidróxido de sódio e examinado ao microscópio eletrônico de varredura, revelaram após a remoção total da camada epitelial, o tecido conjuntivo denso em disposição original apresentando papilas conjuntivas de diferentes formas e tamanhos. As regiões rostral e média evidenciaram numerosas papilas correspondentes às filiformes e fungiformes e na região caudal, a papila conjuntiva da papila vaIada apresentou uma forma alongada. Na superfície foram notadas pequenos forames de ductos de glândulas salivares e projeções papilares maiores e menores em aspectos tridimensionais. Na superfície de interface epitélio-tecido conjuntivo, foram notados espessos feixes de fibras colágenas. Cortes corados por Picro-sírius e analisados por meio de microscopia de luz polarizada evidenciaram a coloração de papilas conjuntivas em aspecto vermelho indicando a presença de fibras colágenas do tipo I e fibras colágenas de coloração esverdeada, tipo III. / The study of the dorsal and lingual surfaces and the surface of interface epithelium-connective tissue of Bradypus torquatus\'tongue was performed employing the light and scanning electron microscopy techniques. The tongue of Bradypus torquatus were removed, dissected and sectioned in three parts: rostral, medium and caudal regions and prepared by light and scanning electron microscopy methods. The results revealed that the tongue present a round apex, dorsum and a root. The dorsal mucosae showed three lingual papillae: filiforms, fungiforms and vallate and a ventral smooth surface. Histologically, it was observed that the dorsal surface of tongue contains the filiform, fungiform and vallate papillae located in the rostral, medium and caudal regions.The epitheliallayer is variable in thickness and possess the basal, spinosum, granular and queratinized layers, showing the epithelial cells. The lamina propria is characterized by a dense connective tis sue forming the long, short and rounded papilar projections. The three-dimensional images obtained by scanning electron microscopy, showed numerous filiform papillae located essentially in the rostral and medium regions. Few fungiform papillae are present in the dorsal surface of tongue, usually located in the border of rostral apex of tongue exhibiting the rounded and low in formo They are covered by queratinized epithelial cells. In the caudal region was noted two vallate papillae containing a deep vala and the surface was covered by stratified squamous epithelial cells. The superior third of surface of vallate papillae was covered by stratified squamous epithelial cells where may be identified the gustatory taste pores. In the fungiform papillae, several taste pores are located on the surface. The samples treated with NaOH solution and examined by scanning electron microscopy revealed afier removal of epitheliallayer, the dense connective tissue- in original disposition, presenting different sizes and shapes of conjuntive papillae. The rostral and medium regions envidentiated numerous papillae corresponding to the filiform and fungiform papillae and in the caudal region, the connective tissue papillae of vallate papillae presented a round formo On the surface were noted small foraminae of lingual salivary gland ducts and small and great papillary projections in three-dimensional images. AIso, on the surface of interface epithelium-connective tissue, were noted thick bundles of collagen fibers. The specimens stained with Picrosirius and examined by po larized light microscopy revealed the connective tissue stained in red colour indicating the collagen fibers type I and the type III, those stained in green colour collagen fibers.

Abstract The extracellular matrix is not just a scaffold for cells and tissues, but rather a dynamic part of the human body. Characteristics of collagens, the major protein components of the extracellular matrix, are determined already during synthesis and mutations in genes encoding collagens, unbalance of regulators or dysfunction of collagen modifying enzymes, for instance, can lead to severe clinical complications. Certain hydroxylysine residues formed by lysyl hydroxylases (LHs) function in collagens as precursors of collagen cross-links that stabilize collagenous structures and thereby tissues. In humans, a deficiency of LH1, which is known to hydroxylate lysines in the helical regions of collagen polypeptides, causes Ehlers-Danlos syndrome VIA (EDS VIA). It is characterized e.g. by progressive kyphoscoliosis and hypermobile joints. Mutations in LH2, which is known to hydroxylate lysines in the telopeptides of collagen polypeptides, cause Bruck syndrome type 2 (BS2). BS2 patients suffer from fragile bones and congenital joint contractures, for instance, but the syndrome is usually not lethal. In this work we have generated and analyzed genetically modified LH1 and LH2 null mouse lines to study the in vivo functions and roles of these enzymes. Analyses concentrated also on collagen cross-links that were determined from several null or heterozygous mouse tissues. In the present work we also studied the effects of known BS2 mutations on recombinant human LH2 polypeptides to understand the molecular pathology of the syndrome. As an animal model for human EDS VIA, LH1 null mice had certain characteristics typical for EDS VIA, such as muscular hypotonia, but generally the symptoms were milder. Like EDS VIA patients, the mice have an increased risk of arterial ruptures and ultrastructural changes can be seen in the wall of the aorta, explained by inadequate helical lysine hydroxylation accompanied by a changed cross-linking state of tissues. Similarly, analysis of the LH2 null mouse line demonstrated the importance of the enzyme in cross-link formation. We showed that even a reduced amount of LH2 in adult mice changes the cross-linking pattern in tissues and a total lack of the enzyme leads to embryonic lethality. Furthermore, we demonstrated that LH2 is particularly important in tissue structures supporting blood vessels in the developing mouse embryo or in extraembryonic tissues. Finally, our in vitro studies with recombinant human LH2 polypeptides revealed that the known BS2 mutations severely affect the activity of the enzyme thus explaining the clinical symptoms of the patients, but the mutations do not lead to a total inactivation of the enzyme, which may be critical for the survival of patients. / Tiivistelmä Solunulkoinen matriksi ei ole ainoastaan soluja ja kudoksia tukeva rakenne, vaan se on dynaaminen osa ihmiskehoa. Kollageenien, solunulkoisen matriksin yleisimpien proteiinien ominaisuudet määräytyvät jo kollageenien synteesivaiheessa ja mutaatiot kollageeneja koodittavissa geeneissä, säätelytekijöiden epätasapaino tai esimerkiksi kollageeneja muokkaavien entsyymien toimintahäiriöt voivat johtaa vaikeisiin kliinisiin komplikaatioihin. Tietyt lysyylihydroksylaasien (LH) muodostamat hydroksilysiinitähteet toimivat kollageeneissa kollageeniristisidosten esiasteina. Ristisidokset vakauttavat kollageenirakenteita ja siten myös kudoksia. LH1 hydroksyloi lysiinejä kollageenipolypeptidien kolmoiskierteisellä alueella ja ihmisellä entsyymin puutos aiheuttaa tyypin VIA Ehlers-Danlosin syndrooman (EDS VIA), jossa potilailla on esimerkiksi etenevää kyfoskolioosia ja yliliikkuvat nivelet. Mutaatiot LH2-entsyymissä, joka hydroksyloi lysiinejä kollageenipolypeptidien telopeptidialueilla, aiheuttavat tyypin 2 Bruckin syndrooman (BS2). BS2-potilaat kärsivät mm. luiden hauraudesta ja niveljäykkyydestä, mutta syndrooma ei yleensä ole letaali. Tässä työssä loimme ja analysoimme geneettisesti muunnellut LH1 ja LH2 hiirilinjat, joiden kyseinen LH-geeniaktiivisuus on hiljennetty. Linjojen avulla halusimme tutkia näiden entsyymien toimintaa ja merkitystä in vivo. Analyysit keskittyivät myös kollageeniristisidoksiin, joita tutkittiin useista poistogeenisten tai heterotsygoottisten hiirten kudoksista. Ymmärtääksemme BS2:n molekyylipatologiaa, tutkimme tässä työssä myös tunnettujen BS2-mutaatioiden vaikutuksia ihmisen LH2-rekombinanttiproteiinissa. EDS VIA:n eläinmallina LH1 poistogeenisillä hiirillä on joitakin ominaisuuksia, kuten lihashypotonia, jotka ovat tyypillisiä EDS VIA:lle, mutta yleisesti oireet ovat lievempiä. Kuten EDS VIA-potilailla, hiirillä on kohonnut valtimoiden repeytymisriski ja aortan seinämän ultrarakenteessa voidaankin havaita muutoksia. Oireita voidaan selittää riittämättömällä kollageenien kolmoiskierteisen alueen lysiinien hydroksylaatiolla, joka muuttaa kollageenien ristisidostilaa kudoksissa. Myös LH2-hiirilinjan analysointi osoitti kyseisen entsyymin tärkeyden ristisidosten muodostamisessa. Jo alentunut LH2:n määrä aikuisissa hiirissä muuttaa kudosten kollageeniristisidoksia ja täydellinen entsyymin puuttuminen johtaa sikiön kuolemaan. Lisäksi osoitimme, että LH2 on erityisen tärkeä kudosrakenteissa, jotka tukevat kehittyvän hiiren sikiön tai sikiön ulkopuolisten kudosten verisuonia. In vitro-tutkimukset ihmisen LH2-rekombinanttiproteiinilla paljastivat, että tunnetut BS2-mutaatiot vaikuttavat erittäin haitallisesti entsyymin toimintaan, mikä selittää potilaiden kliiniset oireet, mutta mutaatiot eivät kuitenkaan aiheuta entsyymin täydellistä inaktivaatiota, mikä voi olla kriittistä potilaiden selviytymisen kannalta.

Mesenchymal stromal cells (MSC) are adherent, clonogenic, fibroblast-like cells endowing with unique multipotent differentiation potential and immunosuppressive properties. They are considered as promising candidates for regenerative medicine and immunotherapy. <p>MSC can be isolated from different tissue sources including bone marrow (BM), adipose tissue (AT) and Wharton’s Jelly (WJ). Although fulfilling the ISCT criteria required to be recognized as MSC, MSC from these different sources could disclose some differences taking into account their different anatomical origin and ontogeny as well.<p>In the present work, we investigated the influence of MSC source on their immunosuppressive as well as differentiation properties. We further extended our study to the role of the microenvironment (infection and inflammation) on these features. <p>We show that BM-MSC express Toll-like receptors (TLR) from TLR1 to TLR6. In an inflammatory environment, TLR2, 3 and 4 are significantly upregulated. By upregulating TLR3 and TLR4 transcription, inflammation increases BM-MSC responsiveness to LPS (TLR4 ligand) and poly(I:C) (TLR3 ligand) leading to a pro-inflammatory shift of their cytokine profile. The effect of TLR ligation on BM-MSC osteogenic potential is donor dependent. Inflammation as well as stimulation with LPS and poly(I:C) result in a decrease of BM-MSC immunosuppressive capabilities. <p>We further observed that BM-, AT- and WJ-MSC do not have the same pattern of TLR expression and consequently do not respond the same way to bacterial or viral infection. WJ-MSC do not express TLR4 and although TLR3 is present at the protein level it is not functional as its ligation do not trigger cytokine expression. Inflammation modulates this TLR pattern expression by upregulating TLR3 in all three MSC types and TLR4 only in BM-MSC. TLR ligation increases the production of inflammatory cytokines in BM- and AT- but not in WJ-MSC and augments anti-inflammatory cytokines in AT-MSC. Although inflammation increases in all MSC types the secretion of inflammatory cytokines, additional TLR triggering does not further affect WJ-MSC. The immunosuppressive potential of WJ-MSC on mixed leucocytes reaction (MLR) is not affected either by inflammation or by TLR triggering.<p>On the differentiation side, WJ-MSC has the lower potential to differentiate into osteoblast as compared to BM- and AT-MSC, as revealed by alkaline-phosphatase (ALP) activity and by measuring extracellular Ca2+ deposits. However, inflammation is able to strongly increase the osteogenic differentiation of WJ-MSC as calcification and ALP activity appears as early as at day 7. However this latter enzymatic activity remains much lower than that disclosed by BM-MSC. TLR3 or TLR4 triggering does not affect the osteogenesis of WJ-MSC while it increases it in AT- and also, although to lesser extent, in BM-MSC.<p><p>Our work establishes that the source from which MSC is derived is of major importance for the design of MSC based immunointervention. WJ-MSC appear to be the most attractive cell type when an immunosuppressive action is required in an inflammatory or infectious context. Although WJ-MSC are poorly osteogenic, a complete osteogenic differentiation can be obtained under inflammatory conditions. Taking into account their easy accessibility as well as their huge proliferative potential, these data open an avenue for using these cells in regenerative medicine particularly in clinical settings where chronic inflammation or infection have to be considered. <p> / Doctorat en Sciences biomédicales et pharmaceutiques / info:eu-repo/semantics/nonPublished

Este trabalho visou estudar a integração de implantes de colágeno aniônico:elastina obtidos a partir de pericárdio bovino acelularizado e com densidade de carga negativa variável. As cargas negativas foram introduzidas na forma de grupos carboxílicos, por hidrólise seletiva e controlada de grupos carboxamidas de asparagina e glutamina contidos na estrutura primária da proteína. Foram estudados materiais hidrolisados por 24 e 48 horas de tratamento correspondendo, respectivamente, a 46 '+ OU -' e 87 '+ OU -' cargas negativas adicionais em relação ao colágeno nativo. Os implantes foram introduzidos no subcutâneo de rato por tempos de 14, 60, 120 e 180 dias. Os materiais foram caracterizados por análise térmica, microscopia eletrônica de varredura e de transmissão, e os explantes avaliados por microscopia de luz (colorações de Hematoxilina: Eosina, tricrômico de Masson e Verhoeff). O objetivo foi avaliar o uso de matrizes com colágeno modificado para a reconstrução de tecidos moles. Diferentemente do tecido nativo (pericárdio bovino), a resposta biológica de matrizes de colágeno:elastina polianiônica após 14 dias do implante foi caracterizada por uma progressiva redução na fibrose, porém mais importante, não foram observadas células características de resposta inflamatória crônica, particularmente nos materiais tratados quimicamente. Após 180 dias, a maioria dos implantes estavam integrados à região implantada. Tais resultados sugerem que matrizes acelulares de colágeno:elastina preparadas pela desvitalização de tecido natural podem ser suportes bastante úteis para a reconstrução de tecidos moles, devido ao seu elevado grau de biocompatibilidade e integração / This work studied the integration of acellular polyanionic collagen:elastin matrices derived of bovine pericardium (BP) with variable negative charge. Negative charges were introduced in the material by selective hydrolisis of carboxamide side chain groups from Asn and Gln present in the primary structure of the protein. Hydrolized materials after 24 and 48 hours of treatment, respectively with 46 '+ OU -' and 87 '+ OU -' extra negative charges, were studied. Implants were placed in the subcutaneous of rats for periods of 14, 60, 120 and 180 days. Materials were characterized by differential scanning calorimetry, SEM and TEM, and explants analysed by optical microscopy (H.E., Masson tricromic and Verhoeff stains). The purpose of this work was to evaluate the use of modified collagen matrices for soft tissue reconstruction. Differently from native tissue (BP), the biological response of polyanionic collagen:elastin matrices after 14 days from implantation was characterized by a progressive decrease in fibrosis, but most important, no characteristic cells of a chronic inflammatory response were observed. After 180 days, most of the implants were integrated to the implant region. The results suggest that acellular collagen:elastin matrices prepared by devitalization of natural tissue due to their high degree of biocompatibility and integration may be potentially useful as a scaffold for soft tissue reconstruction

As social media has emerged to become a key site for contemporary communications and cultural production, the internet meme has penetrated every level of social networking online. Albeit being a global phenomenon with pervasive discursive power in a number of fields ranging from humour to international politics and cyber warfare, comparatively little research has been made into how internet memes work on the discursive level of identity formation and their influence on the formation of internet-based social movements. Using Reddit stock market anarchists r/WallStreetBets as case study, this thesis will use Critical Discourse Analysis to analyze how internet memes work on the level of socio-political formations and how their function can be understood in relation to entropic social environments online. This thesis investigates how internet memes are used as a tool for creation of motifs for action, identity markers, connective action, and social narrativization within an ambivalent social movement online. Introducing the novel term ”memetic discourse” as a way to understand memes as transferable units of memetically programmed content, this study shows the potential of memes to act as effective yet unstable modes of communication within networked environments.

Vitiligo is an autoimmune disease in which CD8+ T cells selectively destroy melanocytes, leading to a patchy, disfiguring depigmentation of the skin. Our group and others have highlighted the central role of IFN-γ-dependent chemokines in the progression of disease; however, IFN-γ is also reported to have pleiotropic effects on melanocyte biology. We examined whether IFN-γ has a direct role in melanocyte killing. We tested the T-cell effector functions IFN-γ, Fas ligand and perforin by deleting them from autoreactive T cells used to induce vitiligo in mice. We found that disease incidence, disease severity and T cell accumulation in the skin was reduced in mice receiving adoptive transfer of either IFN-γ deficient or Fas ligand deficient gp100-specific T cells; however, perforin was dispensable and led to increased disease scores and T cell accumulation. To determine how melanocytes are affected by IFN-γ signaling during vitiligo, we performed single-cell RNA-sequencing on suction blister biopsies obtained from vitiligo and healthy subjects. We discovered that integrin expression and TGFb2 signaling was decreased only in lesional melanocyte transcriptomes. Moreover, melanocytes appear to participate in their own demise by increasing HLA expression and recruiting effector cells through the chemotactic ligand CCL18. The loss of melanocyte retention factors may explain their clean disappearance from the skin during keratinocyte turnover. Taken together, we believe IFN-γ production by autoreactive T cells in the skin leads to clean loss of melanocytes by downregulation of melanocyte retention factors and by increasing their potential to be detected by effector cells during vitiligo.

Bibliografische Beschreibung Knepper, Robert Darstellung des Verlaufs der Arteria palatina major unter dem Aspekt der Entnahme autologer Bindegewebstransplantate aus dem Gaumen Universität Leipzig, Dissertation 68 Seiten, 43 Literaturangaben, 42 Abbildungen, 12 Tabellen, 4 Anlagen Referat: Die sich stets weiterentwickelnden Verfahren zahnärztlicher Chirurgie bringen auch neue Anforderungen an den Operateur mit sich. Vor allem im Bereich der ästhetischen Korrektur von weichgewebigen Defekten im Bereich des Zahnhalteapparates gilt es, auf dem neuesten Stand zu sein, um moderne Zahnheilkunde anbieten zu können. Profunde Kenntnisse zur Topographie anatomischer Strukturen sind dabei für den Zahnarzt, den Spezialisten für Parodontologie und den Oralchirurgen unabdingbar. Um die dafür notwendige praktische Anleitung zu erhalten, können Mediziner am Institut für Anatomie der Universität Leipzig Kurse absolvieren, so z.B. auch angehende zahnärztliche Implantologen. Hierbei interessiert die Teilnehmer neben dem praktischen Vorgehen auch der Verlauf von Gefäßen und Nerven im Operationsgebiet. Anatomische Atlanten stellen die betreffenden Areale idealisiert und ohne Berücksichtigung der individuellen Variationsbreite dar. Zugänge, wie sie in der Klinik benötigt werden, finden sich selten. Aus diesem Grund wurde in der vorliegenden Arbeit an sechs vollständigen und 21 geteilten humanen Kopfpräparaten (alle Ethanol-Glycerin fixiert) die A. palatina major präpariert und der klinischen Situation gegenübergestellt. Diese Arterie wurde gewählt, da ihr Versorgungsgebiet – der Gaumen – die bevorzugte Spenderregion autologer Bindegewebstransplantate ist. Dadurch wird die Lücke der oft unzureichenden Verknüpfung der klinischen Situation mit dem anatomischen Korrelat geschlossen. Des Weiteren wurde an diesen Präparaten eine in der Literatur als sicher beschriebene Entnahmestelle vermessen und die Werte den bekannten gegenübergestellt. Um diese Messwerte beurteilen zu können, wurde außerdem die Frage gestellt, inwieweit die Fixierung der Präparate deren Beschaffenheit verändert. Dies wurde in einem Versuch mit dentalen Volumentomogrammen von frisch angefertigten und anschließend fixierten Präparaten untersucht. Auf Grund der Tatsache, dass Humanpräparate nur für einen begrenzten Zeitraum zur Verfügung stehen, ist ein wesentlicher Aspekt dieser makroskopisch-anatomischen Arbeit die fotografische Dokumentation der präparierten Gefäßstrecken, bei der ästhetische u.a. Gesichtspunkte beachtet werden müssen. Da dies auch für andere makroskopische Arbeiten am Institut für Anatomie von Interesse ist, wurde dieser Teil so ausführlich beschrieben, dass er als Anleitung dienen kann.:Inhaltsverzeichnis Abkürzungen VI 1. Einleitung 1 1.1. Problemstellung 1 1.2. Anatomische Grundlagen 3 1.2.1. Embryologische Entwicklung 3 1.2.2. Topographie 4 1.2.3. Mikroskopische Anatomie 7 1.3. Entnahme autologer Gewebstransplantate in der zahnärztlichen Chirurgie 7 1.3.1. Physiologie der freien Gewebstransplantation 7 1.3.2. Spenderregionen 9 1.3.3. Fallbeispiel 10 1.3.4. Verletzung der A. palatina major und Blutungskontrolle 11 2. Material und Methode 12 2.1. Untersuchungsmaterial 12 2.1.1. Wesentliche Angaben zum vorliegenden Material 12 2.1.2. Ethanol-Glycerin Fixierung 15 2.2. Präparationstechniken 16 2.3. Messmethoden 20 2.4. Statistische Auswertung 22 2.5. Einfluss der Fixierungsmethode auf das Präparationsmaterials 22 2.5.1. Materialveränderung durch die Fixierung 22 2.5.2. Versuchsaufbau 23 2.5.3. Dentale Volumentomografie 28 2.5.4. Verwendeter Dentaler Volumentomograph 29 2.5.5. DVT-Auswertungssoftware und Messmethode 30 2.6. Fotografische Dokumentation und Nachbearbeitung 32 2.6.1. Verwendete Hardware 32 2.6.2. Verwendete Software 33 2.6.3. Theoretische Grundlagen 35 2.6.4. Durchführung 38 2.6.5. Nachbearbeitung der Bilder 39 3. Ergebnisse 41 3.1. Vergleich zwischen klinischer Situation und anatomischem Präparat 42 3.2. Vermessung der Lage der A. palatina major 50 3.3. Verhalten von Weichgewebe unter dem Einfluss verschiedener Fixantien 52 3.4. Ergebnisse der Fotodokumentationen anatomischer Präparate 56 3.5. Fotografische Darstellung präformierter Körperhöhlen 58 4. Diskussion 60 4.1. Vergleich der Messergebnisse zur Lage der A. palatina major 60 4.2. Schlussfolgerungen aus dem DVT-Versuch 61 4.2.1. Vergleich der Messdaten für Kochen 61 4.2.2. Einfluss von Fixantien auf Weichgewebe 62 4.2.3. Aussagekraft der im DVT-Versuch ermittelten Werte 62 4.3. Kritische Einschätzung des Materials und der Methoden 63 4.3.1. Veränderungen der Anatomie unter Berücksichtigung der Präparation 63 4.3.2. Aussagekraft bezüglich Alter und Geschlecht der Spender 63 4.3.3. Kritische Einschätzung der angewandten Methoden 64 4.4. Weiterverwendung des Materials und der Erkenntnisse 65 4.4.1. Weiterverwendung Bildtafeln 65 4.4.2. Weiterverwendung DVT Versuchsaufbau 66 4.4.3. Weiterverwendung fotografische Anleitung 66 5. Zusammenfassung 67 6. Verzeichnis der Abbildungen 69 7. Verzeichnis der Tabellen 71 8. Literaturverzeichnis 72 9. Danksagung 76 10. Erklärung eigenständige Abfassung 77 11. Lebenslauf 78 Anlagen 79

Les opérations de hernies abdominales sont l’une des chirurgies les plus répandues dans le monde. Pourtant, malgré des progrès considérables en particulier dans le développement des textiles prosthétiques pour consolider la paroi abdominale, le taux de récurrence reste très élevé. Il apparaît donc nécessaire de développer des modèles numériques patient-spécifiques de la paroi abdominale afin de mesurer puis améliorer l’impact des solutions de soins. Les tissus assurant la cohésion et la stabilité de la paroi abdominale sont les gaines rectusiennes antérieure et postérieure et la ligne blanche, qui sont des tissus conjonctifs. Leur structure particulière, composée d’un entremêlement de fibres de collagène et d’élastine sont au cœur de cette étude.Ce mémoire a permis de mettre en évidence la relation entre architecture microscopique et comportement macroscopique des tissus fibreux. Grâce à des campagnes expérimentales combinant essais mécaniques et observations microscopiques, la structure des tissus a pu être identifiée et corrélée aux paramètres mécaniques. Des outils d’analyse d’images tridimensionnelles ont été développés afin d’estimer automatiquement l’anisotropie d’une texture. Les informations recueillies ont alors menées au développement d’un modèle constitutif anisotrope hypo-paramétré. Basé sur une description tridimensionnelle du réseau fibrillaire, le modèle a été écrit de manière à décorréler les paramètres liés à la structure et ceux relatifs à la nature même du matériau. En considérant les paramètres matériau similaires pour tous les individus, le modèle a démontré sa capacité à prédire le comportement mécanique à partir d’informations texturales / Abdominal hernia operations are one of the most common surgeries in the world. However, despite considerable progress, particularly in the development of prosthetic textiles to strengthen the abdominal wall, the recurrence rate remains very high. It therefore appears necessary to develop patient-specific numerical models of the abdominal wall in order to measure and improve the impact of care solutions. The tissues that ensure the cohesion and stability of the abdominal wall are the anterior and posterior rectus sheaths and the linea alba, which are connective tissues. Their particular structure, composed of an intertwining of collagen and elastin fibers, is at the heart of this study. This thesis highlighted the relationship between microscopic architecture and macroscopic behaviour of fibrous tissues. Thanks to experimental campaigns combining mechanical tests and microscopic observations, the structure of tissues has been identified and correlated to mechanical parameters. Three-dimensional image analysis tools have been developed to automatically estimate the anisotropy of a texture. The collected information then led to the development of a hypo-parameterized anisotropic constitutive model. Based on a three-dimensional description of the fibrillary network, the model was written in such a way as to uncorrelate the parameters related to the structure and those related to the nature of the material. By considering similar material parameters for every individual, the model demonstrated its ability to predict mechanical behaviour based on textural information