Submitted by Irene Nascimento (irene.kessia@ufpe.br) on 2016-10-06T17:36:12Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) PPG CCS Mestrado MARCLEBIO JUL2016.pdf: 4899039 bytes, checksum: 6ddba26db5ccea9cfece89934d95f230 (MD5) / Made available in DSpace on 2016-10-06T17:36:12Z (GMT). No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) PPG CCS Mestrado MARCLEBIO JUL2016.pdf: 4899039 bytes, checksum: 6ddba26db5ccea9cfece89934d95f230 (MD5) Previous issue date: 2016-07-08 / A doença renal crônica (DRC) caracteriza-se por perda irreversível e progressiva da função renal, com redução da taxa de filtração glomerular ou alteração renal estrutural. É bastante prevalente no Brasil e no mundo, principalmente devido às suas principais etiologias: hipertensão arterial sistêmica e diabetes mellitus. A DRC apresenta elevadas taxas de morbimortalidade, principalmente de origem cardiovascular. Os fatores de risco cardiovascular tradicionais como hipertensão arterial sistêmica, diabetes melllitus, tabagismo, dislipidemia e hipertrofia de ventrículo esquerdo são bem estudados nesta população. Os fatores ditos não-tradicionais como hiperfosfatemia, inflamação, microalbuminúria e anemia aumentam cada vez mais sua importância. Dentre estes nãotradicionais, começa a haver um interesse crescente sobre a prolactina (PRL), que têm seus níveis séricos aumentados em pacientes com DRC devido à uma redução de sua excreção renal e ainda um estímulo para sua secreção pela uremia. A partir disso, objetivou-se realizar um estudo de corte transversal em pacientes com DRC avaliando níveis séricos de PRL e sua correlação com marcadores inflamatórios. Foram avaliados 178 pacientes com DRC (estágios 3, 4 e 5), em um único centro de referência em nefrologia, por um período de 6 meses. Foram incluídos todos os pacientes consecutivos atendidos em ambulatório de referência para DRC (151) e pacientes em hemodiálise (27). Destes, 146 tiveram avaliados os seus níveis de PRL e de marcadores inflamatórios [Interleucina-6 (IL-6), proteína C reativa ultrassensível (PCRus), ferritina e albumina]. Foram estudados os níveis médios de PRL em cada estágio da DRC e sua relação com marcadores inflamatórios. Observou-se que a taxa de filtração glomerular teve relação inversa com os níveis séricos de PRL, ocorrendo também um aumento da prevalência de hiperprolactinemia com a perda de função renal (19% no estágio 3; 32,4% no estágio 4; 40% no estágio 5 conservador e 81,5% no estágio 5 em hemodiálise). Foi visto ainda que nos pacientes submetidos à hemodiálise, os níveis de IL-6 foram estatisticamente mais elevados naqueles com prolactina mais elevada em relação àqueles com prolactina normal (p= 0,046). Houve tendência aos níveis de prolactina serem maiores nos pacientes com IL-6 ou PCRus elevados nos demais estágios da DRC. Não houve diferença estatística entre os níveis de ferritina e albumina nos pacientes com PRL elevada ou normal tanto nos pacientes em tratamento conservador quanto nos pacientes em hemodiálise. Portanto, a prevalência de hiperprolactinemia aumentou com a perda de função renal, e em pacientes com DRC em hemodiálise os níveis de IL-6 são significativamente mais elevados naqueles com prolactina elevada. / Chronic kidney disease (CKD) is characterized by irreversible and progressive loss of renal function with decreased glomerular filtration rate or renal structural change. CKD is a common disease in Brazil and worldwide, mainly due to its main causes: hypertension and diabetes mellitus. The CKD has high morbidity and mortality rates, particularly from cardiovascular causes. Traditional cardiovascular risk factors such as hypertension, diabetes mellitus, smoking, dyslipidemia, and left ventricular hypertrophy are well studied in this population. Nonetheless, nontraditional risk factors, such as hyperphosphatemia, anemia, inflammation, and microalbuminuria, have received increasing attention. Hyperprolactinemia is also gaining attention among the nontraditional risk factors. It has increased serum levels in patients with CKD due to a reduction of renal excretion and also a stimulus to secretion by uremia. From that aimed we conduct a cross-sectional study in patients with CKD assessing serum PRL and its correlation with inflammatory markers. 178 patients were evaluated with CKD (stages 3, 4 and 5), in a single reference center in nephrology, for a period of 6 months. They included all consecutive patients seen at referral center for CKD (151) and hemodialysis patients (27). Of these, 146 were assessed their levels of PRL and inflammatory markers [Interleukin-6 (IL-6), high-sensitive C-reactive protein (hsCRP), ferritin and serum albumin]. PRL average levels were studied in each stage of CKD and its relationship to inflammatory markers. It was concluded that the glomerular filtration rate was inversely related to serum levels of PRL, also been an increase in the prevalence of hyperprolactinemia with the loss of renal function (19% in stage 3, 32.4% in stage 4, 40% in stage 5 conservative and 81.5% in stage 5 on hemodialysis). It was also seen that in patients undergoing hemodialysis, the IL-6 levels were significantly higher in those with higher prolactin compared to those with normal prolactin (p = 0.046). There was a tendency to prolactin levels are higher in patients with IL-6 or high CRP in other stages of CKD. There was no statistical difference between ferritin and albumin levels in patients with high or normal PRL in patients undergoing conservative treatment or hemodialysis. Therefore, the prevalence of hyperprolactinemia increased with loss of renal function, and in hemodialysis patients with CKD IL-6 levels were significantly higher in those with elevated prolactin.

Orientadores: Maria Cristina Volpato, Francisco Carlos Groppo / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-24T18:59:54Z (GMT). No. of bitstreams: 1 Muniz_BrunoVilela_M.pdf: 860734 bytes, checksum: 815e06c4724e0ee7ed82f780fce50958 (MD5) Previous issue date: 2014 / Resumo: A articaína não encapsula em proporção significativa em lipossomas sem gradiente de pH transmembranar. O objetivo deste estudo foi preparar e realizar a caracterização inicial de formulação de articaína associada a lipossomas unilamelares (400nm) com gradiente de pH transmembranar, com sulfato de amônia como tampão interno sobre a viabilidade celular em culturas de queratinócitos humanos (HaCaT) e sobre a liberação de uma interleucina pró-inflamatória (IL-6), comparando com formulações de articaína livre. As células foram expostas às formulações de articaína nas concentrações 0,1%, 0,2% e 0,3% na forma de solução e em suspensão lipossomal (lipossomas unilamelares), além dos controles (soro fisiológico, suspensão lipossomal e meio de cultura). A avaliação da viabilidade celular (redução do MTT - espectrofotometria) foi realizada após 10 min e 4h e a quantificação da IL-6 (imunoensaio de ELISA) após 4h da exposição às formulações. Os resultados foram submetidos aos testes de Kruskal-Wallis com post-hoc de Dunn (viabilidade celular) e de Student-Newman-Keuls (IL-6) com significância de 5%. As vesículas lipossomais mantiveram-se integras após a encapsulação de articaína, apresentando 18,95% de eficiência de encapsulação. A polidispersão dos lipossomas sem anestésico foi de 0,2 ± 0,0, enquanto a dos liposomas contendo articaína variou de 0,56 ± 0,03 a 0,66 ± 0,10. O potencial Zeta variou de -10,5 ± 0,9 a -21,2 ± 0,9 mV. O tamanho das vesículas variou de 622 ± 71,5 a 796 ± 111,95 nm. A viabilidade celular foi diminuída após 10 min de exposição às formulações lipossomais (com e sem articaína) em relação aos demais tratamentos (p<0,05); as formulações lipossomais não diferiram entre si (p>0,05). Não houve diferenças entre os demais tratamentos (p>0,05), os quais não alteraram a viabilidade nesse tempo de exposição. Após exposição por 4h houve diminuição na viabilidade em todas as formulações lipossomais e nas formulações de articaína livre 0,2% e 0,3% (p<0,05), à exceção dos grupos controle e articaína 0,1% (p>0,05). A liberação de IL-6 não foi afetada pelas formulações lipossomais (p>0,05); a articaína livre em todas as concentrações testadas aumentou a liberação de IL-6, tanto em relação ao controle, quanto às formulações lipossomais com a mesma concentração de articaína (p<0,05). Conclui-se que a utilização de lipossomas com gradiente de pH transmembranar aumentou a encapsulação de articaína, entretanto, apresentou toxicidade intrínseca no modelo avaliado / Abstract: Articaine is not encapsulated in significant proportion in liposomes without transmembrane pH gradient. The aim of this study was to perform the initial characterization of a formulation of articaine in unilamelar liposome with transmembranarpH gradient and to observe its effects on human keratinocytes (HaCaT) regarding cellular viability and liberation of interleukin 6 (IL-6) in comparison to plain articaine. HaCaT cells were exposed to plain articaine solutions and liposomal suspensions (0.1%, 0.2% and 0.3% articaine concentrations) and to the controls (saline, liposomal suspension and culture medium). Cell viability (MTT reduction - spectrophotometry) was evaluated at 10 min and 4h after exposure to the treatments; IL-6 was determined after 4 h of cell treatments. Cell viability results were submitted to Kruskal-Wallis test, followed by Dunn post-hoc test; IL-6 results were analyzed by Kruskal-Wallis and Student-Newman-Keuls tests. Significance was set at 5%. Liposome vesicles remained intact after articaine loading and presented encapsulation efficiency of 18.95%. Liposome without anesthetic presented polydispersity index of 0.2 ± 0.0, while liposomes with articaine showed values of 0.56 ± 0.03 to 0.66 ± 0.10. Zeta potentials varied from -10.5 ±0.9 to -21.2 ± 0.9 mV and vesicle sizes from 622 ± 71.5 to 796 ± 111.95 nm. Cell viability decreased after 10 min exposure to the liposomal formulations (with and without articaine) in relation to the other treatments (p<0.05); liposomal formulations did not differ from each other (p>0.05). No differences were found among the other treatments (p>0.05), which did not interfere in cell viability after this exposure time. After 4h exposure cell viability was diminished by all liposomal formulations and by 0.2% and 0.3% plain articaine (p<0.05), except for control groups and 0.1% plain articaine (p>0.05). IL-6 release was not affected by liposomal formulations (p>0.05); all concentrations of plain articaine increased IL-6 release in relation to the controls and to each correspondent liposomal formulation (p<0.05). In conclusion, liposome with transmembrane pH gradient increases articaine encapsulation in relation to that described in the literature, however it presented intrinsic in the model evaluated / Mestrado / Farmacologia, Anestesiologia e Terapeutica / Mestre em Odontologia

Reumatoid artrit (RA) är en kronisk inflammationssjukdom som symmetriskt drabbar kroppens leder och går i skov. En inflammationsprocess uppstår i synovialmembranet (ledhinnan) som börjar förtjockas och utsöndra ett exsudat till synovialvätskan, Exsudatet innehåller mycket erosiva inflammatoriska molekyler, inklusive cytokinerna TNF, IL-1 (interleukin) pch IL-6. De onormala cytokinkoncentrationerna förstör med tiden leden. Detta leder till leddeformationer, smärta och stelhet. Sjukdomen involverar även andra organ än enbart rörelseapparaten. RA drabbar strax under 1% av befolkningen, varav kvinnor är överrepresenterade. Diagnosen av RA kan vara mycket svår att ställa eftersom sjukdomen saknar ett enskilt laboratorietest eller ett standardiserat mätinstrument som korrelerar till sjukdomsaktivitet. Till följd därav används mätinstrument av sammanslagan parametrar för att utvärdera sjukdomen. Methotrexat (MTX) räknas som förstahandspreparat vid RA och kombineras standarsdmässigt med biologiska läkemedel, vid svårare sjukdomsförlopp. Det anses att biologisk kombinationsterapi är nödvändig för maximal behandlingseffekt, emellertid visar kliniska behandlingsregister att 1/3 av RA-patienter förskrivs biologisk monoterapi. Kombinationsterapi kan även orsaka mer biverkningar och försämrad följsamhet. Syftet med studien är att undersöka effekten och säkerheten av receptorantagonisten Tocilizumab (TCZ) med avseende på onoterapi vid RA. Arbetet är en litteraturstudie som genomförts genom sökningar i PubMed. Totalt lästes 7 artiklar i fulltext, varav 5 inkluderades i studieanalysen. Utifrån de primära effektmåtten i artiklarna, visade TCZ-monoterapi en signifikant behandlingsskillnad hos RA patienter med medelhög till hög sjukdomsaktivitet. Resultatet visar ingen bättre behandlingseffekt av att kombinera MTX med TCZ jämfört med monoterapi. TCZ-monoterapi hämmar även strukturella ledskador. TCZ-monoterapi anses som en säker och generellt tolererbar behandling.

M.Tech. (Homoeopathy) / Overweight and obesity is an excessive accumulation of fat and can be classified using the body mass index (BMI). Being overweight has a whole host of health-related risks, which includes metabolic syndrome and diabetes mellitus type II to name a few (Goedecke et al. 2005). Interleukin 6 (IL-6), secreted from the adipose tissue surrounding the waist line is responsible for a state of systemic chronic low level inflammation and plays a crucial role in the development of diseases associated with being overweight (Fantuzzi, 2005). Food sensitivities further contributes to this state of low level chronic inflammation. Symptoms of food sensitivities include, difficulty losing weight as well as over eating, which hinders weight loss in over weight individuals (Joyal, 2010). Anti-inflammatory diets such as the Mediterranean diet focuses on a diet rich in antiinflammatory foods, which is known to decrease chronic low level inflammation caused by being overweight and obese. (Galland, 2010). The aim of this study was to determine the comparative effect of the Mediterranean diet to a modified Mediterranean diet (excludes common food sensitivities) on the circulating levels of the inflammatory cytokine interleukin-6 (IL-6) in overweight females by measuring weight loss and IL-6 levels. The study was an experimental comparative study involving thirty female participants who were between the ages of 20-45 and were overweight (BMI 25-30 kg/m²). Potential participants attended an initial interview where they were screened by means of inclusion and exclusion criteria as well as a physical exam that included vitals, weight (kg), waist measurement (cm), hip-to-waist ratio and calculation of BMI. Individuals who met these criteria were sent for blood tests to measure their Interleukin 6 levels (IL-6). Those individuals with an IL-6 level of greater than 0.96pg/ml were then contacted to set up a first appointment. Participants were allocated to either the modified Mediterranean diet or to the Mediterranean diet using matched pairs. Participants attended a three week as well as a 6 week follow up to tract weight-loss progress.

Le fer est un élément essentiel à de nombreux processus physiologiques fondamentaux. Lors d'une infection, une forte compétition entre l'hôte et l'agent pathogène a lieu ; alors que la bactérie a besoin d'acquérir le fer de l'hôte, pour son développement et sa virulence, l'hôte déploie de nombreux mécanismes pour protéger ses stocks de fer. Il sécrète notamment un peptide capable de moduler l'homéostasie du fer au niveau systémique, l'hepcidine, qui va causer une diminution de la quantité de fer sérique, une rétention intracellulaire du fer et donc une restriction du fer accessible aux pathogènes. Lors d'une inflammation, l'expression de l'hepcidine est décrite dans la littérature pour être principalement induite via l'activation de deux voies de signalisation : la voie STAT3 et la voie BMP/SMAD, l'altération de chacune de ces voies conduisant à un défaut d'induction d'hepcidine. Cependant, nos précédentes observations publiées ont infirmé le rôle de l'IL6, ligand de la voie STAT3, dans la régulation de l'hepcidine en réponse au LPS, et ont suggéré l'implication d'un peptide appartenant à la famille du TGFb, l'activine B, dans la régulation de l'hepcidine via l'activation de la voie BMP/SMAD in vivo. Dans cette étude, nous nous sommes intéressés au rôle de l'activine B dans la régulation de l'hepcidine in vivo, lors d'une infection. Grâce à l'utilisation de souris invalidées pour le gène codant l'activine B (Inhbb-/-), nous avons confirmé que l'activine B était un ligand endogène de la voie BMP/SMAD dans le foie, puisqu'elle induit la phosphorylation des effecteurs SMAD en réponse au LPS. Cependant, nous avons pu clairement démontrer que l'activine B, et donc l'augmentation de la phosphorylation des effecteurs SMAD, ne participaient pas à la régulation de l'hepcidine in vivo, en réponse à l'infection. Nous nous sommes alors intéressés à l'implication de l'IL6 dans la régulation de l'hepcidine. Alors que l'absence d'Il6 n'altère pas l'induction de l'hepcidine in vivo en réponse au LPS, cette cytokine semble jouer un rôle clé pour la réponse de l'hôte lors d'une infection par une bactérie. Dans ce contexte, la littérature décrit l'importance de l'IL6 pour une réponse immunitaire protectrice de l'hôte lors d'une infection. Lors d'une infection, nous proposons donc l'implication d'une nouvelle voie de signalisation dans l'expression de l'hepcidine. De plus, nous suggérons un rôle important de l'IL6, non pas dans la régulation transcriptionnelle de l'hepcidine, mais pour la protection de l'hôte lors d'une infection bactérienne. Enfin, nos résultats montrent qu'un niveau d'activation basal de la voie BMP/SMAD est requis pour une induction d'hepcidine lors d'inflammation, et que l'augmentation de la phosphorylation des effecteurs SMAD observée en réponse à l'inflammation ne participe pas à cette régulation. / Iron is essential for several fundamental metabolic processes. During infection, a strong competition between the host and the pathogen occurs; while the bacteria needs to acquire iron from the host, for its growth and virulence, hosts have developed several mechanisms to protect its iron stores. In particular, the host produces a peptide in order to modulate systemic iron homeostasis, hepcidin. Hepcidin decreases the amount of circulating iron, causes intracellular iron retention and thus a restriction of accessible iron to pathogens. During inflammation, hepcidin expression is described in the literature to be mainly mediated through activation of two signaling pathways: the STAT3 and the BMP/SMAD pathways. Impairment of one of these pathways leads to an impaired hepcidin induction. However, our previous published observations did not support the role of IL6, the major ligand of STAT3 pathway, in the regulation of hepcidin in response to LPS, but suggested the involvement of another protein, that belongs to the TGFb family, activin B, in the regulation of hepcidin via the activation of the BMP/SMAD pathway in vivo. In this study, we investigated the role of activin B in the regulation of hepcidin in vivo, during infection. By using knockout mice for the gene encoding activin B (Inhbb-/-), our results suggest that activin B is not involved in the regulation of hepcidin in vivo in response to infection. We then investigated the function of IL6 in the regulation of hepcidin. Although the absence of IL6 does not affect the induction of hepcidin in vivo in response to LPS, this cytokine appears to play a key role in the host response during bacterial infection. Indeed, the literature describes the importance of IL6 for a protective immune response of the host during infection. During infection, we hypothesize that another signaling pathway regulates hepcidin expression. In addition, we suggest an important role of IL6, not in the transcriptional regulation of hepcidin, but for the host protection during a bacterial infection. Finally, our results also show that basal level of BMP/SMAD pathway is required for an appropriate induction of hepcidin during inflammation.

Anemia em idosos está relacionada ao aumento da morbidade e mortalidade desta população. As causas das anemias em idosos podem ser divididas em três grupos: anemia das doenças crônicas (ADC), anemia por deficiência de nutrientes, na qual se inclui a anemia por deficiência de ferro (ADF) e anemias de causas não-identificadas. A hepcidina constitui uma importante ligação entre defesa primária, inflamação e metabolismo do ferro. A hepcidina é induzida, principalmente, pela interleucina-6 (IL-6), atua como regulador negativo da absorção de ferro e é mediadora da retenção de ferro por monócitos e macrófagos durante inflamação ou infecção. Estudos recentes têm demonstrado o papel da produção desse hormônio por monócitos na homeostase do ferro, num modelo autócrino e parácrino. O presente trabalho teve por objetivo geral correlacionar os níveis de IL-6 produzidos por monócitos em cultura e a expressão de hepcidina em monócitos de indivíduos com ADC, com inflamação sem anemia, ADF e com anemias não-identificadas e, por objetivos específicos, verificar a eficiência de parâmetros hematológicos clássicos em avaliar o status férrico de idosos; comparar os níveis de IL-6 produzidos por células monocíticas em cultura, nos diferentes grupos de estudo, e relacioná-los com os parâmetros utilizados para caracterização das anemias; comparar os níveis de expressão de hepcidina em células monocíticas, nos diferentes grupos de estudo, e relacioná-los com o estado inflamatório e com os parâmetros utilizados para caracterização das anemias. Para isso, os pacientes foram avaliados através de parâmetro bioquímicos (glicemia, creatinina sérica, -glutamil transferase, proteínas totais e albumina, por método colorimétrico e proteína C-reativa, por imunoturbidimetria ultra-sensível) e hematológicos (hemograma completo, utilizando o contador de células Micros 45 ABX®, França, e extensão sangüínea corada por Leishman, ferritina sérica, por método imunoquimioluminescente, receptor de transferrina solúvel, por ensaio imunoenzimático e cálculo do índice sTfR/log ferritina). A determinação dos níveis de IL-6 foi feita por imunoensaio enzimático quantitativo em sobrenadante de cultura de monócitos e a dos níveis de expressão do RNAm da hepcidina em monócitos pela Reação em Cadeia da Polimerase em Tempo Real (RT-PCR). Os níveis séricos de ferritina estavam estatisticamente diminuídos na população com ADF, embora sem atingir os valores preconizados para diagnóstico de deficiência de ferro. Os níveis de receptor de transferrina solúvel (sTfR) e o índice sTfR/log ferritina estavam elevados em pacientes com ADF, porém, o índice não aumentou a sensibilidade da medida do receptor para pacientes idosos. Estes resultados obtidos sugerem que valores de normalidade para níveis de ferritina e índice receptor-ferritina devem ser revistos para a população idosa. Houve aumento da concentração de IL-6 em sobrenadante de cultura de monócitos no grupo Inflamação quando comparado com o grupo Anemia. Os níveis de IL-6 correlacionaram-se positivamente com os níveis da proteína C-reativa e número de leucócitos da população em estudo, porém, não houve correlação com os níveis de RNAm de hepcidina expressos por monócitos. A expressão de RNAm de hepcidina em monócitos mostrou correlação positiva com os níveis séricos de ferritina, porém, não foi diferente entre os grupos de estudo. / Anemia in elderly is associated with increased morbidity and mortality in this population. The causes of anemia in elderly can be divided into three groups: anemia of chronic diseases (ACD), anemia of nutrients deficiency, which include iron deficiency anemia (IDA) and unexplained anemias. Hepcidin is an important link between primary defense, inflammation and iron metabolism. The hepcidin is mainly induced by the Interleukin-6 (IL-6), it acts as a negative regulator of iron absorption and it is mediating of iron retention by monocytes and macrophages during inflammation or infection. Recent studies have been demonstrating the role of this hormone production by monocytes in the iron homeostasis, in autocrine and paracrine fashion. The general objective of this study was to correlate the levels of monocyte-derived IL-6 in culture and the monocyte hepcidin mRNA expression in patients with ACD, with inflammation without anemia, IDA and with unexplained anemias. The specific objectives are to verify the efficiency of classic haematological parameters in evaluating the iron status in elderly; to compare the levels of monocyte-derived IL-6 in culture, in different study groups, and to relate them with the parameters used for anemias characterization; to compare the levels of monocyte hepcidin mRNA expression, in different study groups, and to relate them with the inflammatory state and with the parameters used for anemias characterization. For that, the patients were evaluated by biochemical parameter (blood glucose, serum creatinine, -glutamyl transferase, total proteins and albumin, by colorimetric method and high-sensitivity C-reactive protein, measured by immunoturbidimetric assay) and hematological (complete blood count, using the cells accountant Micros 45 ABX®, and peripheral blood film for Leishmans staining morphology, serum ferritin level by immuno-quimioluminescent assay, soluble transferrin receptor, by Enzyme Linked Immuno Sorbent Assay (ELISA) and sTfR/log ferritin index). The determination of IL-6 levels was performed by quantitative ELISA in monocyte culture supernatants and monocyte hepcidin mRNA levels by Real-Time Polymerase Chain Reaction (RT-PCR). Although serum ferritin levels were statistically decreased in IDA population, it did not reach the values recommended for diagnosis of iron deficiency. The soluble transferrin receptor (sTfR) levels and the sTfR/log ferritin index were significantly higher in IDA group, however, the index did not increase the sensibility of the sTfR measure for elderly. These results suggest that normality values for ferritin levels and sTfR/log ferritin index should be reviewed for elderly population. There was increase of levels of monocyte-derived IL-6 in culture in Inflammation group compared with Anemia group. The IL-6 levels were positively correlated with C-reactive protein levels and leukocyte number of the patients, however, there was not correlation with monocyte hepcidin mRNA levels. The monocyte hepcidin mRNA levels showed positive correlation serum ferritin levels, however, it was not different between the study groups.

Die vorliegende Arbeit untersuchte die Verwendbarkeit einer semiquantitativen Auszählung des zellulär sezernierten Zytokins Interleukin-6 (IL-6) und des zerebralen Intermediärfilamentes Saures Gliafaserprotein (GFAP) hinsichtlich ihrer Aussagekraft zur Detektion eines Schädel-Hirn-Traumas (SHT) und der Abschätzung des Überlebenszeitintervalls zwischen Erleiden der Gewalteinwirkung und dem Todeseintritt. Hierzu wurden Hirngewebsproben von 75 Verstorbenen im Rahmen gerichtlich angeordneter Obduktionen verwendet: 54 Fälle wiesen ein zum Tode führendes Schädel-Hirn-Trauma auf (Fallgruppe) und 21 Fälle mit kardiovaskulär bedingtem Todeseintritt wurden als Kontrollgruppe untersucht. Das maximale postmortale Intervall betrug 6 Tage. Gewebsproben definierter Hirnareale (Kortex, Perikontusionszone (PKZ), Hippocampus, Kleinhirn) wurden mit IL-6- und GFAP-Antikörpern immunhistochemisch untersucht und die positiv gefärbten Zellen semiquantitativ erfasst. Die Ergebnisse wurden zu den Überlebenszeiten nach Trauma korreliert und mit der Kontrollgruppe verglichen. Es zeigte sich für IL-6 und GFAP ein statistisch signifikanter Konzentrationsanstieg mit zunehmender Überlebenszeit in der PKZ und in Hypoxie-sensiblen Arealen des Zentralen Nervensystems (ZNS). Die Methode war geeignet, SHT-Fälle von den Kontrollen bei Überschreiten ermittelter Schwellenwerte in Bezug auf die Färbungen zu differenzieren. Die präsentierten Ergebnisse belegen den potentiellen Nutzen der beiden gewählten immunhistochemischen Marker als Ergänzung zur gängigen SHT-Wundalterdiagnostik in der forensischen Praxis.

Kaposi's sarcoma herpesvirus (KSHV) is a cancer-causing virus, primarily affecting AIDS patients. KSHV is found in 3-10% of the U.S. population and can cause a range of cancers in the highly immunosuppressed; these cancers include Kaposi's sarcoma, pleural effusion lymphoma (PEL) and multicentric Castleman's disease (MCD). The current techniques for treating these cancers are relatively ineffective, largely due to their inefficiency at targeting tumors formed by the infection. One protein produced by KSHV, the viral homolog of interleukin-6 (vIL-6), is thought to play a major role in tumor development post-infection. Here a novel animal model is implemented to study the ways vIL-6 affects tumor development through growth factors and other cytokines within infected highly immune-deficient Rag2-/-γc-/- mice. Mice were subcutaneously injected with one of three types of cells: B cells infected with a wild-type (WT) KSHV, B cells infected with mutant KSHV without the gene for viral interleukin 6, and a negative control of uninfected B cells. After allowing time for tumors to develop the mice were sacrificed and the tumors assessed. Analysis of the physical properties of the tumors, as well as markers expressed by the tumors, were used to help determine whether vIL-6 could be an appropriate target when treating these cancers. In this study vIL-6 was seen to influence certain B cell markers (CD30), as well as onset of tumors (with no significant increase in overall tumor mass, but with marginally statistically significant increase in tumor number). This indicates that although vIL-6 could play a small role as a target for cancer, further investigation into the relationship of CD30 in these types of cancers needs to be explored. It was also found that the KSHV viral-infection decreases the development of tumors compared with uninfected immortalized B cells (BJAB). Not only would results from this experiment help develop new treatments, and change the lives of those suffering with cancers induced by KSHV, but they would provide a foundation for future studies with these types of cancers.

Prostate cancer (PCa) bone metastases have been reported in ~90% of patients with advanced disease. Bone metastases disrupt tissue homeostasis and weaken the skeleton, resulting in an increased risk of bone fractures and morbidity. Specifically, PCa cells disrupt the crosstalk between critical cells within the tumor/bone microenvironment (osteoblasts, osteoclasts, and immune cells), and utilize this effector-rich environment for cancer survival and growth. Therefore, a key therapeutic objective in malignant skeletal disease management is to eliminate tumors while restoring bone homeostasis. Current treatments include palliative radiotherapy, chemotherapy, or anti-RANK treatments, all of which have considerable side effects such as osteonecrosis of the jaw or enhanced tumor invasion. There remains a critical gap in therapies than can reduce tumor burden and simultaneously restore bone homeostasis. To address this gap, our work explores emerging gene therapy approaches for treating skeletal malignancies by utilizing multifunctional cytokine-based agents that can simultaneously combat tumor growth and promote bone regeneration.<div><br></div><div>We hypothesize that rationally designed cytokine-based gene therapies that can be secreted from skeletal muscle and targeted to the bone/tumor microenvironment, could effectively reduce tumor growth and restore bone cell homeostasis. To test this hypothesis, we adopted two strategies: 1) a second-generation targeted IL-27 cytokine, and 2) a de novodesign of a cytokine-like therapeutic agent (Propeptide) that includes anti-tumorigenic and pro-osteogenic domains. Both strategies share modules with overlapping therapeutic functions, rendering them complementary in their therapeutic application. In this work, we examined the proof of principle for propeptide gene therapy in muscle cells (in vitro models) and assessed the therapeutic efficacy of our cytokine-based biologics in reducing prostate tumor growth and rebalancing bone cell proliferation and differentiation. Our studies resulted in a propeptide construct representative of a cytokine structure comprised of a bundle of helices that we were able to express in cells. Additionally, our work demonstrated the targeting and anti-tumor efficacy of our therapeutic cytokines in cancer and bone cell models. Ultimately, this will provide the framework for innovative peptide and cytokine-based therapeutics that target and treat both the tumor metastases and bone. This approach will facilitate improvement of morbidity and quality of life of prostate cancer patients with bone metastases and could be applicable to other diseases with bone/tumor pathologies. <br><div><br></div></div>

Im klinischen Alltag stellt die Diagnosestellung der Sepsis bei Schwerbrandverletzten eine Herausforderung dar. Ursächlich sind pathophysiologische Besonderheiten der Verbrennungskrankheit, die in ihrer klinischen Präsentation dem Bild einer Sepsis ähnelt und somit die differentialdiagnostische Abgrenzung erschwert. Es existieren aktuell keine Parameter, Scores, oder Sepsiskriterien, die mit ausreichender Kompetenz eine Sepsis bei Brandverletzten feststellen können. Traditionelle Infektionsparameter (WBC, CRP, Fieber) gelten als zu unspezifisch. Das in der allgemeinen Intensivmedizin weithin etablierte Procalcitonin wird bei Brandverletzten ebenfalls kontrovers diskutiert. Für die Sepsis-3-Definition unter Verwendung des SOFA-Scores konnten bislang keine ausreichende Sensitivität und Spezifität nachgewiesen werden. IL-6 ist ein etablierter Marker der Inflammation mit sehr rascher Induktion bei einer Sepsis. Während das Ergebnis von Blutkulturen häufig erst nach 24-48 Stunden vorliegt, ist die Bestimmung des IL-6 innerhalb weniger Stunden möglich. Der diagnostische Wert des IL6 in der Frühphase der Sepsis bei Brandverletzten ist bislang ungeklärt. Das Ziel unserer Studie war es, die mögliche Beziehung zwischen positiven Blutkulturen und Il- 6-Serumspiegel bei schwerbrandverletzten Patienten mit klinischem Verdacht auf eine Sepsis zu überprüfen und den prädiktiven Wert des IL-6 mit dem anderer Parameter (PCT, WBC, Tmax und SOFA Score) zu vergleichen. In einer retrospektiven Studie wurden über einen 7-Jahreszeitraum alle Patienten eingeschlossen, bei denen bei klinischem Verdacht auf eine Sepsis Blutkulturen abgenommen sowie IL6, PCT und WBC bestimmt wurden. Die Patienten mit vollständigen Daten wurden gemäß Blutkultur in zwei Gruppen eingeteilt: Blutkultur-positive (BSI) und Blutkultur-negative Gruppe (Non-BSI). Die Gruppen wurden für alle gemessenen Biomarker, SOFA-Score und die maximale Körpertemperatur (T max) verglichen. Insgesamt wurden 101 Patienten eingeschlossen (BSI= 39, non-BSI= 62). Beide Gruppen waren hinsichtlich demographischer Daten und der Verletzungsschwere vergleichbar. IL-6 war bei den Patienten der BSI-Gruppe signifikant höher [1047 (339,9; 9000,5) vs. 198,5 (112,43; 702,52) ng/ l; p = 0,001]. In einer ROC-Analyse ergab sich eine AUC von 0,7 (59; 80,8 %). Somit ist IL-6 ein Biomarker mit moderater Aussagekraft. Als optimaler Cut-Off Wert erwies sich ein IL-6-Serumspiegel von 312,8 ng/ l (Sensitivität 79,5 %, Spezifität 56,5 %). Hinsichtlich der PCT, WBC und der Tmax zeigten die Gruppen keine signifikanten Unterschiede. Der SOFA-Score in der BSI-Gruppe war signifikant höher, allerdings wurde ein Anstieg um mindestens 2 Punkte im Vergleich zum Vortag nicht erreicht. Unsere Ergebnisse deuten darauf hin, daß IL-6 bei klinischem Verdacht auf eine Sepsis bereits frühzeitig eine positive Blutkultur vorhersagen kann. In diesem Kontext sind andere getestete Parameter (WBC, T max, PCT, SOFA-Score) weniger geeignet. Der frühe Anstieg des IL-6 könnte im klinischen Alltag hilfreich sein, da die Ergebnisse der IL-6-Bestimmung im Gegensatz zur Kultivierung von Blutkulturen innerhalb weniger Stunden vorliegen. Insbesondere in unklaren Fällen und Situationen, bei denen ein Zuwarten bezüglich der Einleitung einer Antibiotikatherapie möglich erscheint (z.B. fehlende Organdysfunktion) kann IL-6 einen Beitrag bei der Diagnosestellung leisten und einen Hinweis geben, ob differentialdiagnostische Überlegungen weiter verfolgt werden müssen. Die Bestimmung des IL-6 könnte somit zur Vermeidung eines zu liberalen, unkritischen Antibiotikaeinsatzes und den damit assoziierten Folgen beitragen. Das retrospektive Design und die Fallzahl sind relevante Limitationen unserer Studie. Eine Aussage zu möglichen Unterschieden zwischen grampositiven und gramnegativen Keimen kann daher nicht sicher getroffen werden. Weitere prospektive Untersuchungen mit einer höheren Patientenzahl sind daher unbedingt zu empfehlen.:Inhaltsverzeichnis 1. Einleitung…………………………………………………………………………..2 1.1 Epidemiologie der Schwerbrandverletzungen in Deutschland…...……..2 1.2 Pathophysiologie………………..……..…………………………….……….2 1.2.1 Verbrennungskrankheit………………………………….....…………….3 1.2.2 Schockphase……………....………………………………………..…….3 1.2.3 Phase der Ödemrückresorption…………………….………………..….4 1.2.4 Phase der Inflammation/ Infektion und des Hypermetabolismus…....4 1.2.5 Spätphase………………………………………………………......…..…4 1.3 Sepsis bei schwerbrandverletzten Patienten…………...…………………5 1.4 Stellenwert von Infektionsparametern und Scores in der Sepsis bei schwerbrandverletzten brandverletzten Patienten………………………..….6 1.5 Interleukin 6…………………………………..……………………………….7 1.5.1 Zelluläre Mechanismen und die Rolle von Interleukin 6 in der Verbrennungskrankheit……………..……………………………...………….7 1.5.2 Interleukin 6 in der Sepsis……………………...……………………….8 1.6 Material und Methoden………………...…………………...………………..9 1.7 Fragestellung und Zielsetzung des Forschungsvorhabens…………..…12 2. Publikation…………………………………………………………… ………….13 3. Zusammenfassung der Arbeit..…………………………..……………………..22 4. Literaturverzeichnis…………...……………….…………………………………25 5. Anlagen……………………………………………….…………………………...29 5.1 Erklärung über den wissenschaftlichen Beitrag des Promovenden zur Publikation………………………………………..……………………………….29 5.2 Erklärung über die eigenständige Abfassung der Arbeit…….……………….30 5.3 Lebenslauf…………………………….......………………………………...……31 5.4 Danksagung………………………………………......………………………….32