Introduction: Salivary glands are exocrine organs that produce and secrete saliva into theoral cavity. Cancer in the salivary glands is a rare but fatal disease with a five- year survivalrate of 75%. Cancer in the parotid glands, the largest salivary glands, affects about 100persons every year in Sweden. Patients are offered scheduled follow- up visits for five yearsto enable detection of relapse. Relapses can also be detected at extra follow-up visits initiatedby the patients themselves. Aim: The primary aim was to determine how relapse of parotid malignancies are detected; ata scheduled follow- up visit or at visits initiated by the patients. The secondary aim was tocompare if the survival after relapse depends on how it was detected.  Methods: A retrospective cohort study including patients registered in the Head and NeckCancer Register of Örebro University Hospital. Patients with relapse were included.Information was collected from the register and the patients’ medical charts.   Results: A total of 40 patients were included. 25 relapses were found at patient-initiatedvisits, 11 at follow-up visits and 4 were detected incidentally. The two-year survival rate was56% in the group of self- initiated visits and 54.5% in the group of routine follow-up visits.Result from Log Rank test was p-value 0.565.  Conclusion: Findings suggest that it is more common to find relapses at visits initiated bypatient than at follow-up visits. There was no statistically significant difference in survivalbetween the groups.

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No description available.

Mucoepidermoid carcinom (MEC) är en malign mucin-producerande tumör som förekommer i både stora och små spottkörtlar. Syftet med denna studie var att undersöka histologiskt uttryck av muciner (MUC1, MUC4, MUC5AC, MUC5B, MUC6) i MEC för att eventuellt hitta en korrelation mellan kvalitativt mucinuttryck och tumörgrad. Tolv låg- och fem höggradiga MEC och nio normala spottkörtlar intill tumörvävnad undersöktes med hjälp av immunohistokemi där proverna utvärderades med avseende på färgningsmönster och positivitet i specifika celltyper. Normala spottkörtelceller uttryckte främst cytoplasmatiskt mucin MUC5B. MUC1 och MUC4 uttrycktes i normala spottkörtelgångsceller i ungefär hälften av proverna medan MUC5AC uttryck var sällsynt i normala spottkörtlar. MEC:ar uttryckte MUC1, MUC4, MUC5AC och MUC5B. Den apikala delen av membranet i de bägarceller som omger cystiska hålrum visade den starkaste färgningen för MUC1 och MUC4. Uttryck av MUC4 i bägarceller minskade med ökad histologisk grad. Bägarcellers uttryck av MUC5B:s i låggradig MEC var mindre intensivt än uttrycket av MUC5AC i samma celler. Högre uttryck av MUC5B jämfört med MUC5AC noterades i höggradiga tumörer. Sammanfattningsvis uttrycker MEC olika mängd av muciner än normala spottkörtlar. MUC5AC:s uttryck i MEC verkar vara en metaplastisk funktion och MUC4 tycks relatera till tumörens differentieringsgrad. Förhållandet mellan MUC5AC och MUC5B uttryck skulle kunna vara ett användbart verktyg vid diagnostisering och prognosutvärdering av MEC. / Mucoepidermoid carcinomas (MECs) are malignant epithelial mucin-producing tumours encountered in both major and minor salivary glands. The aim of this study was to investigate the histological characteristics of the expression of mucins (MUC1, MUC4, MUC5AC, MUC5B, MUC6) in MECs in search for a possible correlation between qualitative mucin expression and tumour grade. Twelve low-grade, five high-grade MECs and nine normal salivary glands adjacent to tumour tissue were investigated for these mucins by immunohistochemistry. The samples were evaluated with respect to staining pattern and positivity of specific cell types. Normal acinar cells mainly expressed the cytoplasmic mucin MUC5B. MUC1 and MUC4 were expressed in normal ductal cells in approximately half of the samples whereas MUC5AC expression was rare in normal salivary glands. MECs expressed MUC1, MUC4, MUC5AC and MUC5B. The apical membrane of mucous cells lining the cystic cavities showed the strongest staining for MUC1 and MUC4. The expression of MUC4 in mucous cells decreased with increasing histological grade. Expression of salivary mucin MUC5B in mucous cells in low-grade MECs was less intense compared to the expression of MUC5AC in the same cells. In high-grade tumours, a higher expression of MUC5B compared to MUC5AC was noted. In conclusion, MECs express different mucin quantity compared to normal salivary glands. MUC5AC expression in salivary tumour tissue seems to be a metaplastic feature and MUC4 appears to be related to tumour differentiation grade. The relationship between MUC5AC and MUC5B expression could be a useful tool in the diagnosis and estimation of prognosis of MECs.

Os macrófagos são células fagocíticas derivadas dos monócitos sanguíneos produzidos pela medula óssea e estão diretamente envolvidos em um conjunto de processos biológicos vitais. Durante o repasto sanguíneo, fêmeas do mosquito Aedes aegypti inoculam saliva na pele de seu hospedeiro vertebrado e, devido sua localização estratégica nos tecidos, os macrófagos estão dentre as primeiras células residentes a ser exposta a saliva. Apesar dessa evidência fisiológica, pouco se sabe sobre os efeitos imunomoduladores da saliva desse mosquito sobre os macrófagos. Assim, o objetivo deste projeto foi avaliar o papel dos componentes salivares de A. aegypti em macrófagos murinos ativados por LPS+IFN-γ e o potencial efeito de uma proteína salivar no desenvolvimento da encefalomielite autoimune experimental (EAE), um modelo experimental para estudo da esclerose múltipla. Em conclusão, o EGS de A. aegypti apresenta efeito imunomodulatório sobre macrófagos peritoneais ativados com LPS e IFN-γ. Além disso, nossos resultados sugerem que a proteína identificada com possível inibidor da IL-6 produzida por macrófagos ativados pode ser capaz de diminuir a polarização de respostas Th1 e Th17, afetando assim o desenvolvimento da EAE. / Macrophages are phagocytic cells derived from blood monocytes produced by the bone marrow and are directly engaged in a set of vital biological processes. During blood feeding, Aedes aegypti female mosquitoes inoculate saliva into the skin of their vertebrate hosts and, due to its strategic location in the tissues, macrophages are possibly among the first resident cells to be exposed to saliva. Despite this physiological evidence, little is known about the immunomodulatory effects of this mosquitos saliva on macrophages. Thus, the aim of this study was to evaluate the role of A. aegypti salivary components in LPS/IFN-γ-activated murine macrophages and the potential effect of a salivary protein on the development of experimental autoimmune encephalomyelitis (EAE), an experimental model for multiple sclerosis studies. In conclusion, A. aegypti SGE presents immunomodulatory effect on peritoneal macrophages activated by LPS/IFN-γ. Furthermore, our results suggest that the protein identified as a putative inhibitor of IL-6 produced by activated macrophages may be able to down modulate the polarization of Th1 and Th17 responses, thus affecting the development of EAE.

Nosso laboratório investiga os mecanismos moleculares que promovem o estabelecimento de padrões de expressão gênica regulados no desenvolvimento em eucariotos superiores. Como modelo, utilizamos o gene de pufe de DNA BhC4-1, que é amplificado e expresso de modo regulado na glândula salivar e na glândula protorácica no final do quarto estadio larval de B. hygida. Estudos funcionais em D. melanogaster resultaram na identificação de módulos cis-reguladores (MCRs) na região promotora do gene BhC4-1. O MCR de glândula anelar de 67 bp (-253/-187), promove a expressão de BhC4-1-lacZ na glândula anelar a partir do final do desenvolvimento embrionário. O MCR de glândula salivar de 129 bp (-186/-58), dirige a expressão do transgene nas glândulas salivares de prépupas. A glândula anelar é o principal órgão endócrino larval e em D. melanogaster é o resultado da fusão das glândulas protorácicas (responsáveis pela síntese de hormônios esteroides), corpus allatum (síntese de hormônio juvenil) e corpus cardiacum (glândula neuroendócrina). Neste trabalho foram obtidas 12 linhagens independentes transformadas com uma construção que contém o fragmento (-253/+40) do promotor do gene de pufe de DNA BhC4-1 clonado à montante do gene repórter GFP. O genótipo destas linhagens foi validado utilizando-se Southern blots. Inicialmente as 12 linhagens obtidas foram analisadas quanto ao padrão de expressão de GFP em larvas de terceiro estadio e em prépupas 2 horas. Em conjunto, esta análise revelou que o padrão de expressão de GFP é bastante variável nestas linhagens. A análise do padrão de expressão da proteína repórter foi estendida em duas linhagens representativas da série (- 253/+40)/GFP. Nestas linhagens a expressão de GFP é inicialmente detectada na glândula salivar durante o estágio de prépupa e na glândula anelar a partir do terceiro estadio larval. Diferentemente do anteriormente observado em linhagens (-253/+40)/lacZ, nestas linhagens não detectamos a expressão de GFP em tempos do desenvolvimento anteriores ao terceiro estadio larval. Experimentos de interação gênica revelaram que na ausência do fator de transcrição br, a expressão de GFP é mantida na glândula anelar e abolida na glândula salivar de larvas de terceiro estadio. Os resultados dos experimentos de interação gênica corroboram dados anteriores que indicavam que o conjunto de fatores de transcrição que regulam a expressão de BhC4-1-lacZ na glândula anelar é distinto daquele que promove a expressão do gene na glândula salivar. As linhagens obtidas neste trabalho constituem uma ferramenta a ser utilizada na caracterização de fatores de transcrição tecido-específicos que regulam o gene BhC4-1 na glândula anelar e/ou na glândula salivar a partir do final do desenvolvimento larval. / Our laboratory investigates the molecular mechanisms that promote the establishment of developmentally regulated gene expression patterns in metazoans. As a model, we employ the BhC4-1 DNA puff gene, which is amplified and expressed in a regulated manner in the salivary gland and in the prothoracic gland at the end of the fourth larval instar in B. hygida. Functional studies in D. melanogaster resulted in the identification of cis-regulatory modules (CRMs) in the BhC4-1 promoter region. The 67 bp (-253/-187) ring gland CRM drives BhC4-1-lacZ expression in the ring gland from late embryonic development. The 129 bp (-186/-58) CRM salivary gland drives transgene expression in the prepupal salivary glands. The ring gland is the major endocrine organ, and comprises the prothoracic glands (synthesis of ecdysteroid hormones), corpus allatum (synthesis of juvenile hormone) and corpus cardiacum (neuroendocrine gland). In this work, 12 independent lines transformed with a construct containing the BhC4-1 promoter fragment (- 253/+40) cloned upstream of the reporter gene GFP were obtained. The genotype of each line was validated using Southern blots. Initially, the 12 obtained lines were analyzed to investigate the pattern of GFP expression in the third instar larvae and in the 2 hours prepupae. This initial screening revealed that the pattern of GFP expression is highly variable in these lines. The developmental pattern of GFP expression was extended in two representative (- 253/+40)/GFP lines. In these lines, GFP expression is initially detected in the larval and prepupal salivary glands and in ring gland third instar. Differently from previously observed in (-253/+40)/lacZ lines, in these lines we did not detect GFP expression at developmental times prior to the third larval instar. Gene interaction experiments revealed that in the absence of the br transcription factor, GFP expression is maintained in the ring gland and abolished in the salivary gland of third instar larvae. The results of gene interaction experiments corroborate previous data indicating the set of transcription factors that regulate BhC4-1-lacZ expression in the ring gland is distinct from that which promotes gene expression in salivary glands. The lines obtained in this work constitute a tool to characterize the tissue-specific transcription factors that regulate BhC4-1 gene in the ring gland and/or in the salivary gland from the end of the larval development.

Neste estudo avaliou-se os efeitos do diabetes mellitus (DM) sobre dois sistemas: glândula parótida de ratos e células cultivadas in vitro. Foram avaliados respectivamente a composição da matriz extracelular e a migração de células expostas a elevada concentração de glicose. Na parótida observou-se aumento de colágenos III, IV e V, laminina e fibronectina, mediado por TGFb2. Em células isoladas observou-se que a glicose dificultou a polarização celular, reduziu a velocidade e direcionalidade de migração, reduziu a persistência e estabilidade das protrusões celulares e a maturação de adesões. Estas alterações estão relacionadas à ativação da GTPase Rac1, dependente de estresse oxidativo. Este estudo sugere, pela primeira vez, que: 1) a hipofunção salivar pode envolver um espessamento da lâmina basal de capilares e parênquima por mecanismos previamente observados em outros orgãos-alvo de complicações diabéticas e 2) que a glicose exerce um efeito direto sobre a migração celular, fator que pode contribuir para a cicatrização deficiente em indivíduos diabéticos. / In this study we evaluated the effects of DM on two different systems: the rat parotid gland and in vitro cultured cells. Extracellular matrix composition and the migratory behavior of cells exposed to a high glucose concentration (HG) were evaluated, respectively. In the parotid, DM led to an increase in collagens III, IV and V, laminin and fibronectin, through a TGFb2-dependent mechanism. In cultured cells, HG impaired cell polarization, reduced migration velocity and directionality, reduced the persistence and stability of protrusive cellular processes, as well as adhesion maturation. These effects were related to Rac1 GTPase activation, dependent on the oxidative stress promoted by HG. This study suggests, for the first time, that: 1) salivary hypofunction in DM might involve the thickening of capillary and parenchyma basal lamina, through mechanisms already described in other target organs for diabetic complications and 2) that glucose directly impairs cell migration, and this effect may contribute to the chronic wound healing observed in diabetic patients.

No presente trabalho, descrevemos na saliva do mosquito Aedes aegypti uma atividade de indução de morte de linfócitos caracterizada pela rápida exposição de fosfatidilserina na superfície celular, incorporação de iodeto de propídio, diminuição do tamanho celular, perda da expressão de CD62L e seletividade para linfócitos naïve. A molécula responsável por essa atividade foi identificada pelo fracionamento do extrato da glândula salivar (EGS) do mosquito por HPLC e espectrometria de massas. O RNA mensageiro que codifica essa molécula está presente abundantemente em fêmeas de A. aegypti mas não em machos, sugerindo sua participação no repasto sanguíneo. RNA de interferência foi utilizado para silenciar o gene responsável por essa atividade em fêmeas de A. aegypti e o EGS desses mosquitos perdeu a capacidade de induzir a morte de linfócitos, confirmando ser essa a molécula responsável pela atividade. Em conclusão, o presente trabalho identifica uma nova molécula presente na saliva de A. aegypti com atividade rápida, potente e seletiva sobre linfócitos. / In the present work, we described in the saliva of Aedes aegypti mosquito an activity of cell death induction for lymphocytes characterized by fast phosphatidylserine exposure in cell surface, incorporation of propidium iodide, cell shrinkage, loss of CD62L expression, and selectivity for naïve lymphocytes. The molecule responsible for this activity was identified by fractionation of mosquito salivary gland extract (SGE) by HPLC and mass spectrometry. The messenger RNA that codifies this molecule is abundantly present in A. aegypti females but not in males, suggesting its role in blood-feeding. RNA interference was employed to silence the gene responsible for this activity in A. aegypti females and the SGE of these mosquitoes lost the ability to induce lymphocyte death, confirming that this is the molecule responsible for the activity. In conclusion, this work identifies a new molecule from A. aegypti saliva with a fast, potent and selective activity on lymphocytes.

Le syndrome sec de Gougerot Sjögren (SGS) est une maladie auto-immune chronique qui présente des dommages progressifs et irréversibles des glandes exocrines lacrymales et salivaires. Cette pathologie affecte entre 0.1 et 3 % de la population et est plus commune chez les femmes avec un ratio de 9 femmes pour 1 homme. Dans la glande salivaire, une infiltration lymphocytaire est observée et est associée avec la destruction de l'épithélium sécrétoire, qui joue un rôle central dans l'initiation et le développement du SGS. Le processus physiopathologique est loin d’être compris et semble dépendant de phénomènes épigénétiques. En effet, des perturbations importantes de la méthylation de l'ADN sont observées dans les cellules épithéliales en lien avec le niveau d’infiltration lymphocytaire des glandes salivaires.L'objectif de ce travail est de mieux comprendre les changements épigénétiques au cours du SGS et en particulier les défauts de méthylation/déméthylation de l’ADN observés dans les Cellules Epithéliales de Glandes Salivaires (SGEC) et leurs rôles dans le développement de la pathologie.Dans ce but, Une étude globale de la méthylation de l'ADN a été réalisée après culture cellulaire destinée à isoler les SGEC de patients SGS. La puce « human methylation 450k » d’Illumina utilisée couvre plus de 485 000 sites CpG du génome. Des analysesbioinformatiques nous ont permis d'obtenir un panel de gènes différentiellement méthylés. Nous avons ainsi mis en évidence l’importance de la voie calcique (déméthylée, connue pour avoir un impact sur la salivation) et de la voie WNT (hyperméthylée). De plus nous avons pu identifier une régulation interféron et montrer l’importance d’un changement du type d'interféron (I à II) lors de l’évolution vers un lymphome de type MALT. En outre, nous montrons qu’il existe une inter-relation forte entre les processus épigénétiques et les facteurs génétiques associés au SGS. Enfin la dernière partie de ce travail met en lumière l’implication d’un environnement inflammatoire dans le contrôle du processus de méthylation/déméthylation de l’ADN dans les cellules épithéliales.Ainsi, les altérations du méthylome des SGECs pourraient contribuer à la pathophysiologie du SGS et à son évolution en un lymphome du MALT, ceci en lien avec son environnement inflammatoire. / Sjögren Syndrome (SjS) is a chronic autoimmune disease characterized by a progressive and irreversible damage of exocrine glands, particularly salivary and lacrymal glands. This pathology affects between 0.1 and 3% of the population and is more common in women with a ratio of 9 women for 1 man. In salivary glands, a lymphocytic infiltration is observed and associated with the destruction of the secretory epithelium, which plays a central role in initiation and development of the SjS. The process remains incompletely clarified and contains a strong epigenetic component. Indeed, important disturbances of the process of DNA methylation are observed in epithelial cells and they are linked with the level of lymphocyte infiltration in salivary glands.The objective of this work is to better understand epigenetic changes during the SjS and in particular the DNA methylation/demethylation defects observed in Salivary Gland Epithelial Cells Epithelial (SGEC) and their roles in the pathology development.For that purpose, a global study of DNA methylation was done on 12 patients SGECs with the Infinium HumanMethylation450 BeadChip (Illumina). The 450k HM allows to cover more than 485,000 CpG sites on the whole genome. Bioinformatic analysis allowed us to obtain genes panels which are differentially methylated during this pathological phenomenon. In an interesting way, we identified the potential involvement of the calcic (hypomethylated, known to impact salivation) and WNT pathways (hypermethylated). Besides we were able to identify interferon regulation and the shift from interferon type I to type II with mucosa associated lymphoid tissue (MALT) lymphoma evolution. Furthermore, the simultaneous genomic–epigenomic analysis revealed significant associations between SjS-associated genetic risk factors and epigenetic modifications. Finally, the last part of this work highlights inflammatory environment involvement to control DNA methylation/demethylation in salivary gland epithelial cells.Altogether, alterations of DNA methylation in SGEC may contribute to SjS pathophysiology and evolution to MALT and this in link with the inflammatory environment.

Aedes aegypti é importante vetor de patógeno causador de doença como dengue, febre amarela, febre Chikungunya e Zika. A fêmea realiza repasto sanguíneo a fim de adquirir nutriente para o desenvolvimento dos ovos. Neste contexto, a saliva possui papel fundamental, representando o elo entre o artrópode hematófago, seu hospedeiro vertebrado e o potencial patógeno a ser transmitido. Nessa saliva encontra-se um coquetel farmacológico com diversas atividades biológicas, como a presença de peptídeos antimicrobianos e moléculas com funções imunomoduladoras sobre células do hospedeiro vertebrado, com especial atenção aos macrófagos. Como os macrófagos também estão envolvidos nos mecanismos efetores da resposta contra protozoários do gênero Leishmania e a leishmaniose apresenta-se como uma doença de caráter zoonótico de grande relevância em saúde pública, o objetivo deste trabalho foi de avaliar o efeito do extrato de glândula salivar (EGS) de A. aegypti em infecções por Leishmania. Nossos resultados mostraram aumento da infecção in vivo e in vitro na presença do EGS, sugerindo fortemente que o EGS de A. aegypti é capaz de aumentar a infecção por Leishmania. / Aedes aegypti is an important vector of disease-causing pathogens such as dengue, yellow fever, Chikungunya and Zika fever. The mosquito female takes a blood meal in order to develop the eggs. In this context, a saliva plays a key role, representing the link between the hematophagous arthropod, its vertebrate host and the potential pathogen to be transmitted. During the evolutionary process, these insects developed a salivary cocktail with an arsenal of molecules presenting several immunomodulatory properties in host cells, such as macrophages. As macrophages are also involved in the mechanisms of response against protozoa of the genus Leishmania and leishmaniasis is a zoonotic disease of great relevance in public health, the aim of this work is to evaluate the effect of salivary gland extract (SGE) of A. aegypti in Leishmania infections. Our results showed increased infection, in vivo and in vitro, in the presence of SGE, strongly suggesting that A. aegypti EGS is able to increase infection by Leishmania.