Orientador: Maria Regina Sposto / Banca: Cláudia Maria Navarro / Banca: Enilson Antônio Sallum / Resumo: O objetivo deste trabalho foi avaliar a prevalência e severidade do crescimento gengival em pacientes brasileiros sob terapia com nifedipina, por meio do uso do "Novo Índice Clínico para Crescimento Gengival Induzido por Drogas(Índice DIGO)". O estudo foi realizado em 35 pacientes sob terapia com nifedipina (grupo teste ) e em um grupo controle de 35 pacientes. Foram feitos os registros das variáveis demográficas (idade e sexo do paciente), farmacológicas (dose e tempo de uso da nifedipina) e das variáveis periodontais (índice de placa, índice gengival, profundidade de sondagem, nível de inserção clínico, sangramento à sondagem) e do crescimento gengival. O teste Z (nível de significância a 5% e p<0.05) e a correlação de Spearman foram usados para a comparação dos resultados entre os grupos teste e controle. Os resultados mostraram que o crescimento gengival foi observado em 68% dos pacientes e que não houve associação entre o crescimento gengival e as variáveis demográficas e as variáveis farmacológicas. No entanto, houve associação entre o crescimento gengival e as variáveis periodontais, exceto para o índice de placa. Podemos concluir que a inflamação gengival apresentou influência no crescimento gengival associado à nifedipina. / Abstract: The aim of this study was to evaluate the occurrence of nifedipine induced GO in patients and the risk factors associated using a New Clinical Index for Drug Induced Gingival Overgrowth (DIGO) and the relation between The study was carried out with 35 patients under treatment with nifedipine (test group) and 35 patients without treatment (control group). There was assessed the characteristics of demographic (age, gender), pharmacological (dose, time of use), periodontal variables (plaque index, gingival index, probing depth, attachment level, bleeding on probing) and gingival overgrowth of the sample. The test Z (significance level at 5% - p< 0.05) and the Spearman correlation were used to compare data in test and control groups. Gingival overgrowth was noticed in 68% of patients. The statistical analyses showed no association between the gingival overgrowth and demographic and pharmacological variables, except for the plaque index. However, there was an association between the gingival overgrowth and periodontal variables. Further, it is possible to conclude that the presence of gingival inflammation was the main factor of risk to promote nifedipine-induced gingival overgrowth. / Mestre

Orientador: Janete Dias Almeida / Banca: José Benedito Oliveira Amorim / Banca: Emília Ângela Loschiavo Arisawa / Resumo: A hiperplasia fibrosa inflamatória (HFI) é um processo proliferativo não neoplásico, encontrado em mucosa bucal, geralmente decorrente de traumatismos crônicos. A Espectroscopia Raman fornece informações dos tecidos avaliados através de suas propriedades ópticas. Tem sido empregada em estudos biológicos para a caracterização de alterações neoplásicas. No entanto, são escassos os estudos que envolvam processos inflamatórios. Objetivou-se caracterizar através da Espectroscopia FT-Raman HFI com tecidos bucais normais de mucosa jugal, avaliando a especificidade e a sensibilidade deste método, relacionando os espectros obtidos aos achados histopatológicos. Foram utilizados 19 amostras de HFI e 6 amostras de mucosa normal (MN) que localizavam-se em mucosa jugal, obtendo-se, totalizando 92 espectros de HFI e 27 espectros de MN. Os resultados demonstrados pela análise dos componentes principais revelaram que PC3 e PC5 foram os responsáveis por uma melhor classificação, podendo ser observada no gráfico de loading plots como espectros com picos invertidos. Pela análise de discriminantes linear, foi possível observar que 19 espectros de MN e 85 espectros de HFI foram classificados corretamente, correspondendo, respectivamente, a 70,4 % e 92,4 % do total de dados. A curva de Roc revelou um valor preditivo do modelo de 0,87. Concluiu-se que a análise dos espectros Raman permitiu detectar similaridades e diferenças biológicas e bioquímicas entre HFI e MN de mucosa jugal, demonstrando que a Espectroscopia Raman apresenta sensibilidade e especificidade no diagnóstico de processos proliferativos não neoplásicos. Observou-se correlação entre os achados histopatológicos e os obtidos pela utilização da técnica. / Abstract: Inflamatory fibrous hyperplasia (IFH) is considered a non-neoplasic proliferative process that generally occurs in association to trauma. This pathology has typical hystopathological features in both epithelial and conjunctive tissues. Raman spectroscopy of pathological tissues has become a reality that can contribute to knowledge enhancement about biochemical alteration making possible the differential diagnosis of oral pathologies. The objective of the present study was to use FT-Raman Spectroscopy to identify biological and biochemical alterations that exist between oral IFH in buccal mucosa and normal tissue from the same site. Therefore 92 spectra of inflammatory fibrous hyperplasia from 19 patients were compared to 27 spectra of normal tissue from6 patients. It was observed that the relation between the data of IFH and normal tissue was more evident when the Principal Components Analysis (PCA) of all spectra were calculated and when the Principal Components (PC) PC3 vs. PC5 were analysed. Results of Box Plot show great differences in normal group. This was confirmed by the analysis of the Loading Plot of the PC's that showed great modifications between normal and IFH identified by inverted peaks in Raman band 500 e 1110 cm-1; 1300, 1580 and 1730 cm-1. These bands correspond mainly to molecular vibrations of lipids, collagens (I and III) and proteins. The predictive value of diagnostic model was calculated by linear discriminates analysis that showed 0,87 ROC curve. Biological and biochemical similarities and differences between IFH and normal tissues were confirmed by Raman Spectroscopy when compared to hystopathologycal analysis. / Mestre

Introdução: A reestenose pós tratamento endovascular de lesões ateroscleróticas em artérias periféricas é a principal desvantagem desta técnica minimamente invasiva. A inflamação vascular após angioplastia com balão e/ou implante de stent desempenha um papel importante na proliferação de células do músculo liso vascular e posterior crescimento de uma neoíntima, e vários marcadores inflamatórios têm sido referidos como potenciais preditores dessa complicação, porém os fatores que contribuem para a estenose intra-stent no segmento vascular periférico não foram completamente elucidados. Recentemente, tem-se sugerido que a superfície revestida de stents recobertos possa impedir a reestenose de forma mais eficaz do que os stents convencionais. Objetivo: Avaliar o papel do sistema calicreína-cinina (SCC), do óxido nítrico (NO) e das metaloproteinases (MMP), mediadores inflamatórios importantes e que contribuem ativamente para a reparação de tecidos, no processo de reestenose arterial devido a hiperplasia intimal, pós angioplastia com stent recoberto no segmento fêmoro-poplíteo, com a intenção de contribuir com novas medidas terapêuticas. Método: Foi realizado um estudo prospectivo envolvendo 27 pacientes submetidos à angioplastia com stent revestido no segmento fêmoro-poplíteo, selecionados no Ambulatório de Cirurgia Vascular e Endovascular do HCFMRP/USP. Foram estudados os seguintes marcadores: sistema calicreína-cininas - com quantificação dos substratos (cininogênio de alto e baixo peso molecular - CAPM / CBPM) e da atividade das enzimas (calicreína plasmática e tecidual e cininase II); a determinação dos níveis de nitrito e nitratos para a avaliação de óxido nítrico; dosagem das MMPs 2 e 9 e dos níveis circulantes de seus inibidores (inibidores teciduais das metaloproteinases - TIMPs [1 e 2]). Amostras de sangue foram coletadas antes do implante do stent, 24 horas e seis meses após o procedimento. Foi realizado ultrasson Doppler após seis meses, e, na presença de alterações, realizada angiografia para comprovação da presença de reestenose. Resultados: Quatro (14,8%) dos vinte sete pacientes estudados desenvolveram reestenose (>= 50%) em seis meses. Esses pacientes tiveram níveis significativamente mais baixos de CAPM (24h, P <0,05) e de CBPM (antes - P <0,05; 24 horas P <0,01; 6 meses P <0,05); níveis mais baixos de TIMP 2 ( seis meses P<0,05) comparados ao grupo sem reestenose. As atividades da calicreína plasmática e tecidual, da cininase II, NO e MMPs tiveram comportamento semelhante entre os pacientes com e sem reestenose. Conclusão: As taxas de reestenose foram baixas com o uso de stents revestidos no segmento fêmoro-poplíteo comparativamente aos índices publicados de stents não revestidos. Os pacientes que desenvolveram reestenose mostraram níveis reduzidos de cininogênios e de TIMP-2 (seis meses após a angioplastia). Por outro lado, não foi possível demonstrar a participação do óxido nítrico e das metaloproteinases no processo de reestenose / Background: Restenosis after endovascular treatment of atherosclerotic lesions in the peripheral circulation is the major drawback of this minimally invasive technique. Vascular inflammation after balloon angioplasty or stent implantation plays an important role in smooth muscle cells proliferation and subsequent neointima growth, and various inflammatory markers have been reported as potential predictors of this complication, but the factors that contribute to the in-stent stenosis in peripheral vascular segment have not been fully elucidated. Recently, it has been suggested that the coated surface of stents grafts can prevent restenosis more effectively than conventional stents. Objective: The aim of this study was to evaluate the role of the kallikrein-kinin system (KKS), nitric oxide (NO) and metalloproteinases (MMPs), wich are important inflammatory mediators and actively contribute to tissue repair, in the process of arterial restenosis due to intimal hyperplasia, with the aim of developing new interventions. Method: Single-center prospective study with 27 patients with peripheral artery disease (PAD) requiring percutaneous transluminal angioplasty (PTA) and stenting, in the femoropopliteal segment, using coated stents grafts, was performed. The following markers were studied: kallikreinkinin system using the quantification of proteins (high and low weight Molecular kininogen HMWK / LMWK), verification of enzyme activity (tissue kallikrein, plasma kallikrein and kininase II), determination of nitrite and nitrates levels for evaluation of nitric oxide, MMPs 2 and 9 circulating levels and their inhibitors (tissue inhibitors of metalloproteinases [TIMPs 1 and 2]). Serum samples were collected before stent implantation, 24 h and six months after the procedure. Doppler ultrasound was performed after six months, and in the presence of any changes, an angiography was performed to prove the presence of restenosis. Results: Four (14,8%) of the treated patients developed restenosis (>50%) within 6 months. These patients had significantly lower levels of HMWK (24 hours, P < .05), LMWK (before - P < .05; 24 hours - P < .01; 6 months - P < .05) and lower levels of TIMP 2 (6 months < .05) compered to no restenosis group. The activities of plasma and tissue kallikrein, kininase II, NO and MMP had similar behavior among patients with and without restenosis. Conclusion: Restenosis rates were low with the use of coated stents in the femoropopliteal segment compared to published bare metal stents results. Patients with restenosis showed reduced levels of kininogens and TIMP-2 (six months after angioplasty) in patients who developed restenosis. Moreover, it was not possible to demonstrate the involvement of nitric oxide and metalloproteinases in the restenosis process

A hiperplasia do processo coronóide (HPC) é uma variação morfológica que causa limitação na abertura de boca interferindo na mastigação. Normalmente não apresenta sintomatologia dolorosa e o paciente só vai procurar tratamento quando houver dor ou problemas funcionais causados pela compressão do processo coronóide hiperplásico no osso zigomático. Muitos clínicos desconhecem esta alteração e adotam tratamento para disfunção da articulação temporomandibular (ATM). Neste trabalho propôs-se mostrar que a hiperplasia do processo coronóide pode ser observada em radiografias panorâmicas de forma a suspeitar da existência dessa alteração e, aliado aos dados clínicos, o cirurgião-dentista generalista possa encaminhar o paciente ao especialista para que sejam providenciados exames mais complexos que permitam a conclusão do diagnóstico. A amostra constituiu-se de 150 radiografias panorâmicas dentre as quais estavam incluídas imagens sugestivas da hiperplasia do processo coronóide (HPC). Estas radiografias foram submetidas à análise de 3 Radiologistas com um mínimo de 5 anos de experiência na especialidade. Ficou concluído que a radiografia panorâmica é um meio auxiliar para o diagnóstico inicial dessa alteração e o clínico deve estar atento aos possíveis indicativos que esta técnica radiográfica fornece para a sugestão do diagnóstico. / The Coronoid Process Hyperplasia is a morphologic variation, which reduces mouth opening and interferes on mastication. Normally, it doesn\'t have pain symtomatology, and the patient will only look for treatment when there is pain or functional problems caused by compression of the Hyperplasic Coronoid Process on the zygomatic bone. Many practitioners ignore this alteration, and do treatment for termporomandibular joint disorder. This work proposes to show that is possible to suspect the presence of Coronoid Process Hyperplasia on panoramic radiography, and allied with clinical information, general practitioners can forward their patients to a specialist, to make more complex exams which will allow the conclusion of the diagnosis. The sample was composed by 150 panoramic radiographys, where included some suggestive radiographs of Coronoid Process Hyperplasia. These radiographs were analyzed by 3 Radiologists with at least 5 years of experience on this activity. It was concluded that panoramic radiography is an auxiliary way for initial diagnoses of this alteration and practitioners should attent the possible indicatives that this radiograph technique brings for diagnoses suggestion.

Abstract Apoptosis is a genetically programmed mechanism for a multicellular organism to remove cells that are unnecessary, or potentially harmful. The female reproductive system is characterised by a high rate of cellular proliferation. At the same time, apoptosis is also abundant during the normal physiological function of the ovary and endometrium. More than half of the 7 million oocytes that are produced during human ovarian development are deleted before birth and only about 400 oocytes reach the stage of ovulation during the female fertile lifespan. The fate of the non-ovulatory follicles is atresia, occurring through the mechanism of apoptosis. The endometrium goes through radical renewal processes during each menstrual cycle. Apoptosis has been suggested to participate in the regulation of endometrial cellular homeostasis. Errors in this mechanism can result in endometrial diseases such as hyperplasia and cancer. In this work, apoptosis and its regulation were studied in the human fetal and adult ovary, normal endometrium and endometrial pathologies. In fetal ovaries, apoptosis was already abundantly present in oocytes at 13 weeks of gestation. The maximum rate of apoptosis was seen between the 14th and 20th weeks, after which apoptosis decreased towards term. Ovarian Bcl-2 expression was detected in early fetal life during weeks 13 and 14. Bax expression was observed throughout the studied period, from week 13 to 40. The expression of transcription factor GATA-4, which is linked to follicular survival, was localised to the granulosa cells and was high in early fetal life and decreased somewhat towards term. In adult life apoptosis was located in the granulosa cells of the growing follicles. In ovarian biopsies from women homozygous for the inactivating C566T mutation of the FSH receptor, apoptosis or GATA-4 expression was not detected. During corpus luteum regression a peak in apoptosis was detected 10 - 12 days after the LH surge, and was preceded by an increase in 17HSD type 1 and TNF-α expression. During normal menstrual cycles, the highest rate of apoptosis was observed in the menstrual endometrium. This increase in apoptosis was preceded by a decreased Bcl-2/Bax ratio. In endometrial hyperplasia, the rate of apoptosis was similar to that seen during normal proliferation of the endometrium, but an apparent increase was observed in grade II endometrial carcinoma. In grade III carcinoma, the rate of apoptosis was lower than in grade II carcinoma but higher than in hyperplasia. These results indicate that apoptosis is the mechanism behind the substantial oocyte demise during ovarian development. During adult life, apoptosis was mainly localised to the granulosa cells of the growing follicles which do not reach the stage of a dominant follicle. In ovaries where FSH action is abolished, folliculogenesis was impaired and ovarian apoptosis was negligible. Apoptosis is also the underlying mechanism of corpus luteum regression. In the endometrium, apoptosis has a role in rejuvenating the endometrium for growth during the next endometrial cycle and in regulating cellular homeostasis.

Abstract The carbonic anhydrases (CAs) catalyze the reversible hydration of carbon dioxide. In mammals, there are 13 active isoenzymes, which clearly differ in their cell localisation, tissue distributions and functions. CA IX, a unique transmembrane member of the CA gene family, is a tumour-associated protein which is thought to be involved in malignant cell invasion, adhesion and the regulation of cell proliferation. The main focus in the present study was on elucidating the function and expression of CA IX in normal and malignant tissues, especially in the alimentary tract. The functional studies also included CA II, which is regarded as another important CA isoenzyme in the alimentary tract. CA IX immunostaining showed a decrease in the staining intensity of gastric adenomas with increasing dysplasia grade. Well differentiated carcinomas of the intestinal type showed expression comparable to that in the normal mucosa, while expression was decreased in the less differentiated tumours. CA IX deficiency (Car9-/-) genotype and C57/BL6 strain were the main factors which increased the susceptibility of CA IX deficient mice fed on either a normal or high-salt diet to histological abnormalities, including foveolar hyperplasia and glandular atrophy in the gastric body mucosa, while CA II deficiency was associated with only minor histological abnormalities. In a physiological analysis, CA IX played only a minor role in duodenal bicarbonate secretion (DBS), whereas absence of CA II in mice completely abolished the stimulatory effect of E-type prostaglandin 2 (PGE2) on duodenal alkalisation. The results demonstrate that CA IX expression is diminished in most gastric tumours. The variations observed in its expression support the concept that gastric adenomas and carcinomas do not emerge as progressive steps on a single pathway but may instead represent distinct entities with heterogenic genetic backgrounds. In the stomach, CA IX is mainly involved in the regulation of tissue morphogenesis in the body mucosa, while CA II has a major role in maintaining the gastroduodenal acid/base balance.

Magister Scientiae (Medical Bioscience) - MSc(MBS) / The prostate is prone to three pathological processes that include inflammation, benign prostate hyperplasia (BPH) and tumors. According to the center for Disease and Control 1999-2012 report, prostate cancer is the second leading cause of death in the United States. Scientific evidence suggests that up to 30% of men in the general population aged from 50 years and above, irrespective of geographic origin, have foci of prostate neoplastic growth. Unbalanced ROS production and a dysregulated antioxidant defence system have been implicated in prostate cancer development. The transformation of a normal cell into cancer takes a very long period. This observation provides the advantage of using nutraceuticals to prevent, arrest or reverse the cellular and molecular processes of carcinogenesis. Based on scientifically observed positive health roles of green tea (Cameli sinensis) and rooibos (Aspalathus linearis) on major diseases like atherosclerosis, hepatitis and certain types of cancer, this thesis evaluated the effects of these two teas on benign (RPWE 1) and malignant (LNCaP) prostate cells. This was done through the quantification of reactive oxygen species (ROS) using a fluorescence dye 5,6 CM-H2DCFDA, total prostate specific antigen (PSA) levels using a PSA ELISA kit, cell viability using the MTT assay, apoptosis using Tali annexin V stain and cell imaging studies using a Zeiss axiovert 200M inverted fluorescence microscope. Statistical analysis was done using graphpad prism. The findings of this study show that aqueous extracts of green and black tea, fermented and unfermented rooibos and their active compounds epigallocatechin gallate (EGCG) and aspalatin, respectively, are cytotoxic in malignant (LNCaP) prostate cells but exert protective effects in benign (RPWE 1) prostate cells. This thesis implicates the pro-oxidant and anti-oxidant properties of the plant extracts, respectively, for the above mentioned effects. In this regard, tea and rooibos promoted ROS production in malignant (LNCaP) prostate cells, which subsequently promoted cell death of the malignant cells through apoptosis and necrosis. Further to this, tea and rooibos used in this thesis, protected normal prostate cells from the adverse effects of ROS. In this regard, fluorescence microscope photographs showed RPWE 1 cells with low DCF fluorescence compared to the malignant prostate cells. Low magnification light microscope photographs showed RPWE 1 cells with flat polygonal shapes and increased adherence both at low and high concentrations of tea and rooibos. On the contrary, high concentrations of tea and rooibos on malignant (LNCaP) prostate cells induced stress, which made the cells attain irregular shapes and as the stress levels increased, cells became detached and appeared dead. Flow cytometry confirmed the presence of apoptotic and necrotic cell in malignant (LNCaP) prostate cells. In this thesis, EGCG and aspalathin were responsible for the high rates of apoptosis observed whereas green tea and unfermented rooibos induced the highest rate of necrosis. Further to this, tea and rooibos and the main active compounds EGCG and aspalathin, respectively, significantly promoted the reduction of total serum prostate specific antigen (PSA) in malignant prostate cells. In normal prostate cells, these plant extracts maintained the total serum PSA at its basal physiological level. In this thesis, to the best of our knowledge, we report for the first time the cell-specific effects of fermented rooibos, unfermented rooibos and their main active component aspalathin, on prostate cancer cells. We showed that rooibos and aspalathin exert pro-oxidant effects on malignant LNCaP cells and anti-oxidant effects on benign RPWE 1 cells. In conclusion, tea (C. sinensis) and rooibos (A. linearis) and their respective main active compounds, epigallocatechin gallate and aspalathin, are cytotoxic to malignant prostate cells whereas in normal prostate cells, they have protective effects against ROS induced stress. The pro-oxidant and anti-oxidant effects are responsible for the aforementioned effects respectively. The decrease in total serum PSA demonstrate the strong therapeutic effects that tea and rooibos have on malignant (LNCaP) prostate cells. / Malawi Government: Department of Human Resources Development and Management

Purpose: Ultrasonically measured intravesical prostatic protrusion may be a promising noninvasive method of assessing bladder outlet obstruction. Previous investigations of this technique focused on patients with acute urinary retention and symptomatic men identified in urology clinics, which may not reflect the distribution of intravesical prostatic protrusion in community dwelling men. Materials and Methods: In 2006 a total of 322 white men residing in Olmsted County, Minnesota underwent transrectal ultrasound examination which permitted direct measurement of intravesical prostatic protrusion. Cross-sectional associations between lower urinary tract symptoms/benign prostatic enlargement and intravesical prostatic protrusion were measured. Rapid increases in lower urinary tract symptoms/benign prostatic enlargement measures as predictors of severe intravesical prostatic protrusion were also assessed. Results: Overall 10% of these men had an intravesical prostatic protrusion of 10 mm or greater. Greater intravesical prostatic protrusion was weakly correlated with greater prostate volume (rs = 0.28), higher obstructive symptoms (rs = 0.18) and lower peak urinary flow rate (rs = -0.18). Men with the most rapidly growing prostate before intravesical prostatic protrusion measurement were 3 times more likely to have an intravesical prostatic protrusion of 10 mm or greater. Men with an intravesical prostatic protrusion of 10 mm or greater were more likely to use medications for lower urinary tract symptoms/benign prostatic enlargement compared to those with an intravesical prostatic protrusion less than 10 mm (adjusted OR 2.95, 95% CI 1.23-7.06). Conclusions: These population based data provide reference ranges for future studies of intravesical prostatic protrusion as a predictor of adverse urological outcomes. Intravesical prostatic protrusion is significantly correlated with greater prostate volume, higher obstructive symptoms and lower peak urinary flow rate, suggesting that it may have clinical usefulness in predicting the need for treatment.

INTRODUCTION: Specialized pro-resolving lipid mediators (SPM) such as resolvin-D1 (RvD1) act to resolve vascular inflammation and may guard against the progression of restenosis following cardiovascular interventions. Stimulating synthesis of these mediators directly in vascular cells may increase their local availability, and thus, protect against restenotic injury. However, the ability of endothelial (EC) and vascular smooth muscle cells (VSMC) to produce SPMs from their polyunsaturated fatty acid precursor decosahexaenoic acid (DHA) via lipoxygenase (LO) enzymatic transformation remains unknown. We sought to determine whether vascular cells produce SPMs from DHA and, if they do, how inflammation and mechanical injury of the vasculature alter biosynthesis. METHODS: Primary cultures of human saphenous vein endothelial and smooth muscle cells were treated with DHA in cell culture media (+ 10% serum) for 4h-24h. Freshly dissected rabbit aorta was incubated intact or following gentle endothelial denudation in cell culture media (+10% serum) with or without DHA for 48h. SPM levels in media were quantified by LC-MS/MS and ELISA and lipoxygenase expression and localization were assessed by western blotting and immunofluorescence staining, respectively. RESULTS: EC and SMC receiving media without DHA did not synthesize SPMs within the detection limits of the assay, whereas DHA treatment produced 17-HDHA, 14-HDHA, Mar1, RvD5, RvD2, and a dose and time-dependent increase in RvD1 production in EC (10.1 ±1.0 pg for 1000nM at 24h) and SMC (7.4 ± 0.2 pg for 1000nM at 24h). Intact rabbit aorta incubated in DHA+ media produced 0.24 ± 0.05 pg RvD1/mg tissue whereas aorta incubated in DHA− media produced 0.13 ± 0.007 pg RvD1/mg tissue. Moreover, EC-denuded aortas produced less RvD1/mg tissue than intact aortas. 5-LO was expressed in both cell types, however DHA induced 5-LO expression in EC (1.3 fold -DHA) but not in SMC. DHA promoted a nuclear to cytoplasmic shift of 5-LO in both EC and SMC. Finally, TNF-α stimulated an increase in RvD1 production in EC. CONCLUSIONS: Human vascular cells and rabbit vascular tissue can biosynthesize SPMs de novo from their precursor DHA, signifying a new source of SPMs in the vasculature.

Vascular access dysfunction is a cause of morbidity and mortality in chronic kidney disease (CKD) patients that require hemodialysis. The major cause of vascular access failure is venous stenosis due to neointimal hyperplasia (NH). Vascular smooth muscle cells (VSMC) are critical for the development of NH lesions, as they have the ability to modulate their phenotype from a "contractile" to a "synthetic" phenotype in the presence of uremia, through the regulation of sensor genes for uremia danger signals and VSMC-specific differentiation genes. Recent research indicates that Caspase-1 (casp-1) activation plays an essential role in sensing metabolic danger signal-associated molecular patterns and initiating vascular inflammation. Carbamylated LDL, a uremic toxin that has been shown to be found in higher levels in patients with CKD and in CKD murine models when compared to controls, and could play a role in casp-1 activation. Therefore, the goal of this project is to examine the role of cLDL/CKD-driven casp-1 activation in VSMC and CKD-related NH. We have established a CKD mouse model and published on CKD-associated vascular remodeling. We exposed wild type and caspase-1 knockout mice to our CKD model, analyzed and quantified the NH lesion formed. We also examined in vitro and ex-vivo changes in VSMC-specific differentiation genes when exposed to uremic serum and cLDL, in the presence or absence of caspase-1 inhibitor. We found that CKD serum induces with casp-1 activation and phenotypic changes in VSMCs from a "contractile" to a "synthetic" phenotype, which are reversed with casp-1 inhibition. In an ex-vivo model using relative quantification we found that VSMC contractile markers &#945; -Actin, Calponin, SM-22, and Smoothelin gene expression of CKD mouse carotid VSMC were higher in casp-1 knockout mice when compared to wild-type (1.40, 1.28, 1.22, 1.41 respectively). Also using an in-vivo model, relative quantification of &#945;-actin decreased from 1.0 to 0.329 when VSMCs were exposed to uremic serum and but increased back to 0.588 when Caspase-1 inhibitor is added. The relative quantification of Calponin also decreased from 1.0 to 0.394 when exposed to uremic serum and increased back to 0.601 with caspase-1 inhibitor. We also found that caspase-1 deficiency significantly reversed CKD-related vascular remodeling in casp-1 knockout mice and reduced NH volume by 50% from 1,440,023in wild-type mice to 71,069 µm2 in casp-1 knockouts (p-value 0.002). This evidence provides evidence that casp-1 plays a critical role in NH formation. Furthermore our results provide a novel insight over the therapeutic potential of casp-1 inhibitors for CKD induced NH and other inflammation induced vascular remodeling. / Public Health