La neoplasia Blastica di derivazione dagli elementi Dendritici Plasmacitoidi è una rara ed aggressiva neoplasia ematologica con una prognosi sfavorevole. Non esiste un regime terapeutico standard di prima linea per questi pazienti e solo pochi studi hanno finora esplorato la genetica del BPDCN mostrando un cariotipo complesso e alterazioni genetiche sporadiche. Lo scopo del nostro studio è stato quello di indagare, mediante sequenziamento massivo, lo stato mutazionale di tutto il genoma codificante di 15 casi di BPDCN e della linea cellulare CAL-1. Sulla base di questi dati, abbiamo seguito un approccio di target sequencing per validare, mediante la piattaforma Miseq, mutazioni in 36 geni selezionati. Il WES ha rivelato 199 SNV ricorrenti in 89 geni, tutti legati a condizioni patologiche. I geni più ricorrentemente mutati tra i campioni sono stati ASXL1 e TET2, due regolatori epigenetici, che giocano un ruolo cruciale nella metilazione del DNA e degli istoni e il loro danno porta ad alterazione della struttura della cromatina. Abbiamo quindi deciso di usare i nostri dati di sequenziamento per esplorare il rimodellamento della cromatina nel BPDCN, guardando lo stato mutazionale di tutti i geni coinvolti nella regolazione epigenetica del DNA. Abbiamo trovato 26 geni modificatori epigenetici mutati in quasi tutti i campioni, sei di questi colpiti da mutazioni stop-gain che hanno portato, presumibilmente, ad una perdita funzionale di attività. Partendo proprio da questo punto abbiamo deciso di procedere con la sperimentazione in vivo creando dei modelli murini somministrando 5-Azacitdina, Decitabina e Romidepsina da soli e in combinazione. Inoltre basandoci su studi pregressi abbiamo anche deciso di trattare i topi con Bortezomib. Queste sperimentazioni hanno portato a risultati incoraggianti, specialmente per il trattamento combinato dei topi con Decitabina e Azacitidina che ha portato a riduzione della massa tumorale e maggiore sopravvivenza dei topi trattati rispetto ad i controlli non trattati. / Blastic plasmacytoid dendritic cell neoplasm is a rare but aggressive hematologic malignancy with a poor prognosis. There is no first-line standard treatment regimen established for patients with BPDCN and only a few studies explored the genetics of BPDCN showing a complex karyotype and sporadic genetic defects. The aim of our study was to investigate, using deep sequencing, the mutational status of all BPDCN coding genome in 15 BPDCN cases and the CAL-1 cell line. Based on these data, we designed a resequencing approach to identify mutations in 36 selected genes with Miseq platform. WES revealed 199 recurrent SNV (at least 2/16 cases) in 89 genes, all related to pathological conditions. The most recurrent mutated genes, among samples, were ASXL1 and TET2, two epigenetic regulators, that play a crucial role in DNA and histones methylation and their damage alter the chromatin structure. We thus decided to use our exome sequencing data to explore the chromatin remodelling of BPDCN, interrogating the mutational status of all the genes involved into the epigenetic regulation of DNA. We found 26 epigenetic modifier genes mutated in almost all samples and six of them are affected by stop gain mutations presumably leading to a functional loss of chromatin remodelling activity. Starting from this point, we decided to proceed with in vivo testing by creating mouse models by administering 5-Azacitdina, Decitabine and Romidepsine alone and in combination. In addition, we are relying on previous studies we have also decided to treat the mice with Bortezomib. These experiments have led to encouraging results, especially for the combined treatment of mice with Decitabine and Azacitidine, which led to tumor shrinkage and increased survival of treated mice compared to the untreated controls.

The recent finding that MYC-driven cancers are sensitive to inhibition of the DNA damage response (DDR) pathway, prompted us to investigate the role of DDR pathway as therapeutic target in diffuse large B-cell lymphoma (DLBCL), which frequently overexpresses the MYC oncogene. In a preliminary immunohistochemical study conducted on 99 consecutive DLBCL patients, we found that about half of DLBCLs showed constitutive expression of the phosphorylated forms of checkpoint kinases (CHK) and CDC25c, markers of DDR activation, and of phosphorylated histone H2AX (γH2AX), marker of DNA damage and genomic instability. Constitutive γH2AX expression correlated with c-MYC levels and DDR activation, and defined a subset of tumors characterised by poor outcome. Next, we used the CHK inhibitor PF-0477736 as a tool to investigate whether the inhibition of the DDR pathway might represent a novel therapeutic approach in DLBCL. Submicromolar concentrations of PF-0477736 hindered proliferation in DLBCL cell lines with activated DDR pathway. These results were fully recapitulated with a different CHK inhibitor (AZD-7762). Inhibition of checkpoint kinases induced rapid DNA damage accumulation and apoptosis in DLBCL cell lines and primary cells. These data suggest that pharmacologic inhibition of DDR through targeting of CHK kinases may represent a novel therapeutic strategy in DLBCL. The second part of this work is the clinical, molecular and functional description of a paradigmatic case of primary refractory Burkitt lymphoma characterized by spatial intratumor heterogeneity for the TP53 mutational status, high expression levels of genomic instability and DDR activation markers, primary resistance to chemotherapy and exquisite sensitivity to DDR inhibitors.

Peripheral T-cell lymphomas-not otherwise specified (PTCL/NOS) are the most common T-cell neoplasms. This study sought to reshape the PTCL/NOS sub-classification (including its two main morphological variants, Lennert lymphoma, LL, and Follicular variant, F-PTCL) based on the correspondence between their molecular features and those of different functional T-cell subsets, also assessing the clinical impact of such an approach. We found that PTCLs/NOS could be divided into groups corresponding to T-cell subsets differently reliant on transcription regulators including mTOR and FOXP3, and identified minimal gene sets discriminating among these groups. Notably, by grouping tumors according to their dependency on master regulators of T-lymphocyte fate, we identified three groups (T-cytotoxic, Treg/TFH, and other-T-helper) characterized by specific genetic patterns and significantly different clinical outcomes. Immunohistochemistry partially substituted for the molecular analysis by consistently recognizing only Treg and TFH cases. Finally, targeted inhibition of MTOR in T-helper cases (that were characterized by genetic lesions targeting the pathway) was proved to be effective ex vivo. We conclude that PTCL/NOS can be divided into subgroups corresponding to different cellular counterparts, characterized by different genetic patterns and possibly sensitivity to specific therapeutic approaches. Furthermore, we identified different gene and microRNA signatures for LL capable of differentiating it from other PTCL/NOS and enriched in cytotoxic function. Moreover, PI3K/Akt/mTOR pathway emerged as novel therapeutic targets for LL. Additionally, LL showed some differences with other PTCL/NOS in terms of clinical features, all supporting its recognition as a distinct entity. Besides, we found that F-PTCL has a distinct molecular signature more similar to PTCL/NOS rather than AITL, and therefore cannot be included among AITLs at least based on GEP, although this necessities more genetic studies. Overall, these results may impact on PTCL classification as well as on future studies aimed to define the more appropriate therapeutic strategy for each identified subgroup/entity.

coordenadoria de aperfeiÃoamento de pessoal de ensino superior / O cÃncer gÃstrico à a quarta neoplasia mais comum em todo o mundo e a segunda causa de mortalidade por cÃncer. Apesar do tratamento com cirurgia e quimioterapia, a sobrevida global em cinco anos de pacientes com cÃncer gÃstrico permanece baixa, uma possÃvel explicaÃÃo para menor eficÃcia da terapia à a presenÃa de cÃlulas tronco cancerosas. VÃrios marcadores, incluindo LGR5 e LGR6, tÃm sido relatados como marcadores de cÃlulas- tronco, normais e cancerosas. LGR5 e LGR6 sÃo proteÃnas transmembranas, do grupo da proteÃna G, ricas em resÃduos leucina, que participam nas vias de sinalizaÃÃo das cÃlulas e ativam fatores de transcriÃÃo relacionados com a formaÃÃo de tumores. O objetivo deste trabalho foi avaliar a imunoexpressÃo de LGR5 e LGR6 como possÃveis marcadores de cÃlulas-tronco no cÃncer gÃstrico primÃrio, metÃstases linfonodais e mucosa gÃstrica histologicamente normal, atravÃs das tÃcnicas de microarranjo tissular (tissue microarray) e imuno-histoquÃmica. O estudo, de carÃter transversal e observacional realizou-se a partir de oitenta e oito (88) peÃas de gastrectomias devido a carcinomas gÃstricos, realizadas no Hospital UniversitÃrio Walter CantÃdio, que fazem parte dos Arquivos do ServiÃo de Patologia e Medicina Legal da Universidade Federal do CearÃ. As relaÃÃes entre a expressÃo diferencial de LGR5 e LGR6 e caracterÃsticas clinico-patolÃgicas foram avaliadas pelo teste do qui-quadrado ou teste exato de Fisher. Um valor de p < 0,05 foi considerado estatisticamente significativo. Foramconsiderados positivos (a partir de relatos prÃvios) casos que apresentaram uma ou mais cÃlulas com imunomarcaÃÃo citoplasmÃtica ou membranar. LGR5 mostrou-se, neste estudo, ser um potencial biomarcador, mais caracterÃstico de cÃlulas-tronco tumorais e nÃo tumorais, do que LGR6. Ambos sÃo expressos em tecido tumoral e nÃo tumoral, com predomÃnio em mucosa histologicamente normal, em terÃo basal da espessura epitelial. LGR5 positivo predominou no histotipo difuso e LGR6 no tipo intestinal de carcinomas gÃstricos. Nos casos positivos dos dois biomarcadores, cÃlulas marcadas sÃo pouco frequentes ou raras, no total de cÃlulas da massa tumoral e da mucosa gÃstrica normal. LGR6 esteve presente no tumor gÃstrico primÃrio em nÃmero de cÃlulas muito superior ao encontrado nas respectivas metÃstases linfonodais, diferenÃa nÃo observada em relaÃÃo a LGR5. Com exceÃÃo da relaÃÃo de cada biomarcador com um histotipo especÃfico, jà citada, nÃo houve relaÃÃo entre as demais variÃveis clinico-patolÃgicas e a expressÃo de LGR5 e LGR6. / Gastric cancer is the fourth most common neoplasm in the world and the second leading cause of cancer mortality. Despite the treatment with surgery and chemotherapy, the overall five-year survival of patients with gastric cancer remains low, a possible explanation for less effective therapy is the presence of cancer stem cells. Several markers, including LGR5 and LGR6, have been reported as markers of normal and cancerous stem cells. LGR5 and LGR6 are transmembrane proteins of the G protein group, rich in leucine residues, that participate in cell signaling pathways and activate transcription factors related to tumor formation. The objective of this work was to evaluate the immunoexpression of LGR5 and LGR6 as possible markers of stem cells in primary gastric cancer, lymph node metastases and histologically normal gastric mucosa through tissue microarray and immunohistochemistry techniques. The cross-sectional and observational study was carried out from eighty-eight (88) pieces of gastrectomies due to gastric carcinomas performed at the Walter CantÃdio University Hospital, which are part of the Archives of the Service of Pathology and Legal Medicine of the Federal University Of CearÃ. The relationships between LGR5 and LGR6 differential expression and clinical-pathological characteristics were assessed by the chi-square test or Fisher's exact test. A value of p <0.05 was considered statistically significant. Positive (from previous reports) were considered cases that presented one or more cells with cytoplasmic or membrane immunostaining. LGR5 was shown to be a potential biomarker, more characteristic of tumor and non-tumor stem cells, than LGR6 in this study. Both are expressed in tumoral and non-tumoral tissue, predominantly in histologically normal mucosa, in a basal third of epithelial thickness. Positive LGR5 predominated in the diffuse histotype and LGR6 in the intestinal type of gastric carcinomas. In the positive cases of the two biomarkers, marked cells are infrequent or rare, in the total cells of the tumor mass and the normal gastric mucosa.

The present study evaluated the effects of L lysine in intestinal and urothelial epithelia in augmentation of bladder and ureterosigmoidostomy in rats.A total of 66 female rats 9 weeks old were split divided in to 9 experimental groups. The animals from groups: I , II and III were subjected to bladder augmentation with colon segment (AV) and treated with L lysine, celecoxib and H2O, respectively. The groups: IV, V and VI were subjected to ureterosigmoidostomy (US) treated with L lysine, celecoxib and H2O in that order. Groups: VII, VIII and IX (non- operated controls) received L lysine, celecoxib and H2O respectively. The dosage of L lysine was 150mg/ kg/ body weight and the celecoxib was 20mg/kg/ body weight. The effects of L lysine on the colon epithelium of rats subjected to US was initially evaluated by analysis Aberrant Cripts Foci (ACF) at stereostopic microscopy, after fixation with formaldehyde and staining with methylene blue. Followed by histopathological study of ureteral epithelium, colonic and bladder in all groups, stained with hematoxylin and eosin and PAS Alcian Blue. Rare ACF were found in all mice with US and compared between groups. There was no statistically significant difference between groupsOn histopathology with hematoxylin and eosin, mild to moderate hyperplasia was observed in intestinal and urothelial epithelia at the site of anastomosis in all animals submitted to cystoplasty (Groups I and III), but transitional metaplasia of the intestinal glandular epithelium was more accentuated in Group I (p=0.045). There were no inflammatory cells, dysplasia or atypia. In epithelia of the left ureter and colon of mice with US a mild inflammatory infiltrate composed of lymphocytes, moderate to severe intensity, had urothelial hyperplasia of moderate to severe intensity in all the ureters, three polyps in different ureters and inflammatory polyps in colon . There were no dysplasia or atypia. The staining with Alcian Blue, noticed an important decrease of goblet cells and mucin in colon in all operated rats. In the histopathology there is similarity in the quality and quantity of injuries. Thus, we conclude that L-Lysine did not influence the carcinogenesis of intestinal epithelia, and urothelial of rats submitted to US and AV with colon segment, at times, doses and methods evaluated.However, the L-lysine stressed the "urothelial metaplasia" in intestinal segment bladder and ureter transitional hyperplasia in rats subjected to ureterosigmoidostomies. / Avalia os efeitos da L lisina na carcinogÃnese de epitÃlios intestinal e urotelial nasampliaÃÃes vesicais e ureterossigmoidostomias em ratas. O total de 66 ratas com nove semanas de idade foi dividido em nove grupos. Os animais dos grupos I, II e III foram submetidas a ampliaÃÃo vesical com segmento de colo (AV) e tratados com L lisina, celecoxibe e H2O, respectivamente. Os do grupo IV, V e VI foram submetidos a ureterossigmoidostomia (US) e tratados com L lisina, celecoxibe e H2O nesta ordem. Os grupos VII, VIII e IX (controles nÃo operados), receberam L lisina, celecoxibe e H2O, respectivamente. A dose de L lisina foi de 150 mg/kg/peso e o celecoxibe foi de 20 mg/kg/peso. Os efeitos da L lisina no epitÃlio do colo de ratos submetidos a US foi inicialmente avaliado pela anÃlise de FCA (focos de criptas aberrantes) a microscopia estereoscÃpica apÃs fixaÃÃo com formol e coloraÃÃo pelo azul de metileno. Seguiu-se estudo histopatolÃgico dos epitÃlios ureterais, cÃlicos e vesicais em todos os grupos, corados pela hematoxilina e eosina e PAS Alcian Blue. Foram encontrados raros FCA, em todas as ratas com US e na comparaÃÃo entre os grupos. NÃo ocorreu diferenÃa estatisticamente significantes entre os grupos. No estudo histopatolÃgico, com hematoxilina e eosina de epitÃlios cÃlicos e vesicais de animais com AV, foram observados hiperplasia urotelial leve a moderada em todos os animais submetidos a cistoplastia, em regiÃo de anastomoses colovesical e maior nÃmero de animais apresentaram âmetaplasia transicionalâ em epitÃlio glandular intestinal no grupo I (p= 0,045). NÃo havia cÃlulas inflamatÃrias, displasias ou atipias. Nos epitÃlios de ureter esquerdo e colo das ratas com US, em meio a infiltrado inflamatÃrio constituÃdo por linfÃcitos, de moderada a acentuada intensidade, havia hiperplasia urotelial de moderada a acentuada, trÃs polipos em distintos ureteres e um polipo inflamatÃrio em colo. NÃo havia displasias ou atipias. Quanto à coloraÃÃo pelo Alcian Blue, notou-se importante diminuiÃÃo de cÃlulas caliciformes e de mucinas em colo, em todas as ratas operadas. Desta forma, conclui-se que a L lisina nÃo influenciou na carcinogÃnese do epitÃlio intestinal das ureterossigmoidostomias em ratas nos tempos e doses, bem como pelo mÃtodo de avaliaÃÃo de criptas aberrantes. A L lisina acentuou a âmetaplasia urotelialâ em segmento intestinal de ampliaÃÃes vesicais e a hiperplasia transicional no ureter das ratas submetidas a ureterossigmoidostomias.

BACKGROUND:Acromegaly is a disorderassociated with increased mortality, mainly due to cardiovascular disease, caused by chronic excessive GH secretion. Regular echocardiographic assessment is recommended to evaluate cardiac morphological disorders, including valvopathy. One of the therapeutic options is the use of cabergoline (CAB). This medication has been associated with lesions to the cardiac valve apparatus when used in patients with ParkinsonÂs disease or Prolactinoma, but data on acromegalic patients are scant.OBJECTIVE:Compare the prevalence of valvopathy among patients with acromegaly who have used cabergoline and those who have not.METHODS:The present study is cross-sectional and observational analytical.Echocardiography was performed for two cardiologists blinded to treatment, and laboratory tests weremade and patientsâ medical records were reviewed for clinical data collection. Patients were divided into two groups according to usage (CAB group) or not (control group)of cabergoline. SPSS v17 was used for statistical analysis. RESULTS:Fifty two patientswere evaluated, 35 in the CAB group and 17 in the control group. The distribution of sex and age, and the prevalence of hypertension, diabetes and obesity were similar in both groups. In the CAB group, the period of drug administration was 36  27 months, and cumulative dose was 262  178mg. The prevalence of regurgitation in any valve was, for CAB group and Control group respectively 42,9 vs 23,5% (P=0,175, both observers). The prevalence of remodeling in any valve was37,1 vs 23,5% (P=0,326, observer 1) and 42,9 vs 23,5% (P=0,175, observer 2). CONCLUSION:In our study, the use of cabergoline was not associatedwith valvulopathy in patients with acromegaly. Our data do not support modification of current guidelines of echocardiographic evaluation in these patients according to the use of cabergoline. / INTRODUÃÃO:A acromegalia à uma doenÃa com elevada mortalidade, principalmente cardiovascular, causada pela secreÃÃocrÃnica excessiva de GH. AvaliaÃÃo periÃdica com ecocardiograma à recomendada de rotina devido ao aumento de alteraÃÃes morfolÃgicasnesses pacientes, incluindo valvopatia. Uma das opÃÃes terapÃuticas para essa patologia à a cabergolina (CAB). Esta medicaÃÃo vem sendo associadaà lesÃo valvar quando utilizada em pacientes com DoenÃa de Parkinson ou Prolactinoma, porÃm hà poucos dados em pacientes portadores de Acromegalia.OBJETIVOS:Comparar a prevalÃncia de alteraÃÃes valvares em portadores de acromegalia que usaram ou nÃo a cabergolina.MÃTODOS:Este estudo foi do tipo transversal, analÃtico observacional. Foi realizadoecocardiograma por dois observadores que desconheciam tratamento da doenÃa, exames laboratoriais e revisÃo de prontuÃrios. Os pacientes foram divididos em dois grupos, de acordo com o uso (grupo CAB) ou nÃo da cabergolina (grupo controle). Para anÃlise dos dados foi utilizado o programa SPSS v23. RESULTADOS:Foram avaliados52 pacientes, sendo 35 do grupo CAB e 17 do grupo controle. A distribuiÃÃo de sexo e idade, e a prevalÃncia de HAS, DM e Obesidade foram semelhantes nos dois grupos. No grupo CAB,o tempo de uso da droga foi 36  27 meses e a dose cumulativa de 262  178mg.Foi observado uma prevalÃncia de refluxo em qualquer valva no grupo CAB e controle respectivamente 42,9 vs 23,5% (P=0,175) por ambos os observadores.A prevalÃncia de alteraÃÃes morfolÃgicas nos grupos CAB e controle foi de 37,1 vs 23,5% (P=0,326, observador 1) e 42,9 vs 23,5% (P=0,175, observador 2). CONCLUSÃO:No nosso estudo o uso de cabergolina nÃo se associou a valvopatia em pacientes portadores de acromegalia. Nossos dados nÃo apoiam a modificaÃÃo do protocolo de acompanhamento ecocardiogrÃfico nos pacientes portadores de acromegalia de acordo com o uso ou nÃo de cabergolina.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / A anemia falciforme (AF) à uma hemoglobinopatia causada por uma mutaÃÃo pontual no gene da &#946;-globina caracterizada por eventos vaso-oclusivos e um estado inflamatÃrio crÃnico. Estudos tem demonstrado novas opÃÃes de tratamento na AF, com a finalidade de potencializar a aÃÃo da HidroxiurÃia (HU) e com isso promover a diminuiÃÃo da dosagem sem comprometer a concentraÃÃo da hemoglobina fetal (HbF), bem como o uso de substÃncias que possam agir no mecanismo inflamatÃrio. O presente estudo teve como objetivo avaliar o efeito da L-arginina e do BAY 73-6691 sobre as concentraÃÃes de TNF-&#945;, IL-8 e Ãxido nÃtrico em neutrÃfilos de pacientes com AF. Participaram do estudo 50 pacientes com diagnÃstico molecular de AF, atendidos no ambulatÃrio de Hematologia do Hospital UniversitÃrio Walter CantÃdio (HUWC) em Fortaleza-Cearà e 30 indivÃduos saudÃveis como grupo controle. Os pacientes foram divididos em dois grupos, de acordo com o uso de HU: SSHU (30 pacientes em uso de HU) e SS (20 pacientes sem uso de HU). Os neutrÃfilos dos pacientes foram extraÃdos do sangue total por diferenÃa de gradiente de densidade e tratados com L-arginina e BAY 73-6691 isoladamente nas concentraÃÃes 0,1, 1, 10 e 100 Âg/mL e com a associaÃÃo L-arginina e BAY 73-6691 na concentraÃÃo de 10 Âg/mL. Um grupo de neutrÃfilos nÃo tratados foi utilizado para comparaÃÃo (HbSS). A citotoxicidade foi avaliada atravÃs da atividade de LDH e ensaio do MTT. Os nÃveis de TNF-&#945; e IL-8 foram determinados por ELISA, e os nÃveis de NO, por ensaio colorimÃtrico. Foi observado que a L-arginina e o BAY 73-6691 isolados causaram toxicidade em neutrÃfilos de ambos os grupos apenas na concentraÃÃo de 100 Âg/mL, enquanto que a associaÃÃo nÃo apresentou efeitos citotÃxicos na concentraÃÃo de 10 Âg/mL. Tanto a L-arginina e o BAY 73-6691 isolados como a associaÃÃo (L-arginina +BAY 73-6691) foram capazes de reduzir significativamente os nÃveis dos marcadores inflamatÃrios TNF-&#945; e IL-8 e de elevar os nÃveis de NO em neutrÃfilos de pacientes do grupo SS, em relaÃÃo ao grupo de neutÃfilos nÃo tratados (HbSS). Os resultados do estudo mostraram que a L-arginina e o BAY 73-6691 sÃo potenciais alternativas terapÃuticas para a AF por serem capazes de aumentar a biodisponibilidade do NO e reduzir o processo inflamatÃrio em neutrÃfilos de pacientes com AF nÃo tratados com HU. / Sickle cell anemia (SCA) is a hemoglobinopathy caused by a point mutation in the &#946;-globin gene characterized by vaso-occlusive events and a chronic inflammatory state. Studies have demonstrated new treatment options in SCA in order to potentiate the action of Hydroxyurea (HU) and thereby promote decreased dosage without compromising fetal hemoglobin concentration, as well as the use of substances that may act in the inflammatory mechanism. The present study aimed to assess the effect of L-arginine and BAY 73-6691 on TNF-&#945; concentration, IL-8 and nitric oxide in neutrophils from patients with sickle cell anemia. The study included 50 patients with a molecular diagnosis of SCA, treated at the hematology clinic at the University Hospital Walter CantÃdio in Fortaleza, CearÃ, and 30 healthy individuals as a control group. The patients were divided into two groups, according to the use of HU: SCAHU (30 patients treated with HU) and SCASS (20 patients not treated with HU). Neutrophils from patients were extracted from whole blood by density gradient difference and treated with L-arginine and BAY 73-6691 alone in the concentrations 0.1, 1, 10 and 100 ug / mL and L-arginine and BAY 73-6691 association at a concentration of 10 ug / ml. A group of untreated cells was used for comparison. Cytotoxicity was assessed by the LDH activity and MTT assay. TNF-&#945; and IL-8 were determined by ELISA and NO levels by colorimetric assay. It has been observed that L-arginine and BAY 73-6691 isolated neutrophils caused toxicity in both groups only at the concentration of 100 ug / ml while the combination had no cytotoxic effects. L-arginine and BAY 73-6691 isolated and the combination were able to reduce levels of inflammatory markers TNF-&#945; and IL-8 and to increase NO levels significantly in the SS group neutrophils in relation to the group untreated cells. The results of the study showed that L-arginine and BAY 73-6691 are potential therapeutic alternatives for SCA to be able to increase the bioavailability of NO and reducing the inflammatory process in neutrophils from patients with SCA not treated with HU.

HSCT is the intravenous infusion of hematopoietic stem cells with the purpose to recover or renew the production of blood cells and the immune function in patients with malignant hematological disorders. Oral mucositis is one of the most frequent side effects of anticancer therapy. This work aimed to evaluate the hematological and microbiological parameters, therapeutic regimen and incidence of oral mucositis (OM) in patients submitted to HSCT. The study is longitudinal, conducted from December 2014 to October 2015. We evaluated 15 patients with hematologic malignancies indicated for HSCT. Blood samples were collected for accomplishment of the serology for EBV, CMV and HSV patients; we also performed evaluation and odontological adequacy before the conditioning regimen. After performing the transplant, the patients were clinically evaluated for identification of alterations in the oral mucosa, such as mucositis and opportunistic infections and subjective evaluation of pain in the days D+3, D+6, D+9 and D+10. The most widely used drug in protocols of chemotherapy was melphalan, corresponding to 80% (n=12). When evaluating the incidence of oral amendments it was found that, from the total of the sample (n=15), 93.3% of the patients had at least one episode of OM, considering all evaluated days, being the grade I the most prevalent, 48.0% (n=36). The highest number associated with intensity of the pain was 7.0 (seven). Thirteen patients (86.7%) showed no reactive IgM levels of HSV, and in none of them were identified IgM for EBV and CMV. 93.3% (n=14) patients were positive for IgG for both EBV and CMV. Only three patients (20%) had candidiasis. As to the hematological profile, there was a significant increase in the number of patients exhibiting hematocrit below 30.0% (p <0.005), platelets menor que 50.000/mmÂ) (p = 0.011) and leukocytes (p = 0.013). However, only patients who had less than 2,000 leukocytes (p=0.044) showed OM in the D+10 (100%), being the frequency 15.9 higher in these patients. There was no association between OM with the type of basis disease, with the treatment regimen used for conditioning or with other hematological parameters evaluated. We conclude that the patients who underwent hematopoietic stem cell transplantation present high incidence of episodes of OM, but no association with fungal and viral infections. OM showed strong relationship to leukopenia, being grade I the most prevalent, portraying the possible contribution of the buccal adequacy. / O Transplante de cÃlulas tronco hematopoiÃticas (TCTH) consiste na infusÃo intravenosa de cÃlulas progenitoras hematopoiÃticas com o objetivo de recuperar ou renovar a produÃÃo de cÃlulas sanguÃneas e a funÃÃo imune em pacientes com desordens malignas hematolÃgicas. A mucosite oral (MO) à um dos efeitos secundÃrios mais frequentes da terapia antineoplÃsica. Este trabalho teve como objetivo avaliar os parÃmetros hematolÃgicos, microbiolÃgicos, esquema terapÃutico e incidÃncia de MO em pacientes submetidos a TCTH. O estudo à do tipo longitudinal, realizado no perÃodo de dezembro de 2014 a outubro de 2015. Foram avaliados 15 pacientes portadores de neoplasias hematolÃgicas indicados para o TCTH. Foram coletadas amostras de sangue para realizaÃÃo de hemograma completo e sorologia para VÃrus Epstein-Baar (EBV),CitomegalovÃrus (CMV) e VÃrus Herpes Simples (HSV), dos pacientes, como tambÃm realizado avaliaÃÃo odontolÃgica das doenÃas cÃrie e periodontal utilizando os Ãndices de CPO-D e Ãndice de placa visÃvel (IPV) antes do transplante. ApÃs a realizaÃÃo do transplante os pacientes foram avaliados clinicamente para identificaÃÃo de alteraÃÃes da mucosa bucal, tais como mucosite e infecÃÃes oportunistas e avaliaÃÃo subjetiva de dor, nos dias D+3, D+6, D+9 e D+10. O fÃrmaco mais utilizado nos protocolos de quimioterapia foi o melfalano correspondendo a 80% (n=12). Ao se avaliar a incidÃncia das alteraÃÃes orais verificou-se que do total da amostra (n=15), 93,3% dos pacientes apresentaram pelo menos um episÃdio de MO considerando todos os dias avaliados, sendo o grau I o mais prevalente, 48,0%(n=36). Os pacientes apresentaram dor no D+3 e D+10. Treze pacientes (86,7%) nÃo apresentaram nÃveis reativos de IgM para HSV e, em nenhum deles foi identificada IgM para EBV e CMV. 93,3% (n=14) pacientes foram reagentes para IgG tanto para EBV como para CMV. Somente trÃs pacientes (20%) apresentaram candidÃase. Quanto ao perfil hematolÃgico, houve aumento significante do nÃmero de pacientes exibindo hematÃcrito abaixo de 30,0% (p<0,005), plaquetas menor que 50.000/mm (p=0,011) e leucÃcitos menor que 2.000/mm (p=0,145). No entanto, somente os pacientes que se encontravam com menos de 2.000 leucÃcitos (p=0,044) apresentaram MO no D+10 (100%), sendo a frequÃncia 15,9 maior nesses pacientes. NÃo foi encontrada associaÃÃo entre MO com o tipo da doenÃa base, com o esquema terapÃutico utilizado para o condicionamento nem com os demais parÃmetros hematolÃgicos avaliados. Conclui-se que os pacientes submetidos a transplante de cÃlulas tronco hematopoiÃticas apresentam alta incidÃncia de episÃdios de MO, porÃm sem associaÃÃo com infecÃÃes fÃngicas e virais. A MO mostrou forte relaÃÃo com a leucopenia, sendo o grau I mais prevalente, retratando a possÃvel contribuiÃÃo da adequaÃÃo bucal.

nÃo hà / The preservation of post-extraction alveolar ridge is one of the challenges of dentistry, especially when rehabilitation claim with endosseous implants. As a result of tooth loss, the residual alveoli tends to reabsorb, creating occasions where there is need for grafting surgery to rehabilitation through supported implant prosthesis. This condition can be prevented, among other techniques, by Guided Bone Regeneration. This study aimed to evaluate the effectiveness of a non-absorbable membrane on bone healing after tooth extraction alveoli sites. A study was conducted with 18 patients requiring extraction, a total of 20 sites, prior to the installation of the implant, which sought care in Face Defects Nucleos the Federal University of CearÃ. Was performed prior clinical and radiographic evaluation to surgical procedures, these were divided into two groups: the test group (n = 10) there was installation of non-absorbable membrane, and the control group (n = 10) the membrane was not used. All patients underwent computed tomography cone beam at fifteen and ninety days postoperatively. In both exams vertical and horizontal linear measurements were made through the ImageJ software, post-extraction alveolar center of fifteen and ninety days. It was observed that the retention time in the test group (0.45  0.78) showed if the distance significantly greater than the height of the control group (-2.25  0.97), in which can be seen a significant reduction in bone height (p <0.001). It was noted significant difference in the pattern of variation of the horizontal measured in treated groups with the membrane (0.45  1.92) and control (-1.22  0.49) (p=0,015). The use of non-absorbable membrane did not cause infection, swelling or allergic reaction immunoinflammatory site. The conditions evaluated, clinical and tomographic, noticed a bone maintenance height and widht of fresh alveoli sites can benefit from the installation of endosseous implants. / A preservaÃÃo do rebordo alveolar pÃs-exodontia à um dos desafios da Odontologia, principalmente quando hà pretensÃo de reabilitaÃÃo com implantes endÃsseos. Em consequÃncia da perda dentÃria, o alvÃolo residual tende a reabsorver, criando ocasiÃes em que hà necessidade de cirurgias de enxertia para reabilitaÃÃo atravÃs de prÃtese implanto suportada. Essa condiÃÃo pode ser prevenida, entre outras tÃcnicas, atravÃs da RegeneraÃÃo Ãssea Guiada. O presente estudo objetivou avaliar a eficÃcia de uma membrana nÃo absorvÃvel no reparo Ãsseo de sÃtios de alvÃolos pÃs exodontia. Foi realizado um estudo com 18 pacientes necessitando de exodontia (20 sÃtios cirÃrgicos) prÃvia à instalaÃÃo de implante, que procuraram atendimento no NÃcleo de Defeitos da Face da Universidade Federal do CearÃ. Foi realizada avaliaÃÃo clÃnica e radiogrÃfica prÃvia aos procedimentos cirÃrgicos, estes foram divididos em dois grupos: no grupo teste (n=10) houve instalaÃÃo da membrana nÃo absorvÃvel, e no grupo controle (n=10) a membrana nÃo foi usada. Todos os pacientes realizaram tomografia computadorizada de feixe cÃnico aos quinze e noventa dias de pÃs-operatÃrio. Em ambos os exames foram realizadas mensuraÃÃes lineares verticais e horizontais, atravÃs do software ImageJ, do centro do alvÃolo pÃs-exodontia de quinze e noventa dias. Observou-se que a manutenÃÃo em altura do grupo teste (0,68Â0,57) mostrou-se superior à da distÃncia em altura do grupo controle (-2,25Â0,97), na qual se pÃde perceber reduÃÃo significativa de perda Ãssea (p<0.001). Houve diferenÃa significante no padrÃo de variaÃÃo da medida horizontal nos grupos tratado com a membrana (0,06Â1,20) e controle (-1,22Â0,49) (p=0,015). O uso da membrana nÃo absorvÃvel nÃo gerou infecÃÃo, inchaÃo ou reaÃÃo alÃrgica imunoinflamatÃria local. Nas condiÃÃes avaliadas notou-se de forma clÃnica e tomogrÃfica uma manutenÃÃo Ãssea em altura de sÃtios de alvÃolos frescos, podendo beneficiar a instalaÃÃo de implantes endÃsseos.

This study aimed to evaluate the new immunohistochemical markers related to neuroendocrine differentiation induction and stem cells with prognostic factors and biochemical recurrence in patients submitted to radical prostatectomy. Therefore, patients operated at the Hospital Walter CantÃdio, Federal University of CearÃ, in the period of 2008-2013, underwent clinical and outpatient follow-up, between the years 2008-2016. Biochemical recurrence was evaluated and was correlated with pathological characteristics and immunohistochemical reactions. Chromogranin (neuroendocrine differentiation), Aurora Kinase A (AURKA), N-MYC, C-MYC and CD44s were performad in paraffined material. From 74 patients underwent surgery was obtained the followup of 69, in a median period of 41 (2-89) months. Neoplastic neuroendocrine differentiation was associated with seminal vesicles infiltration (p = 0.032) and stage (p = 0.030). C-MYC was associated with Gleason score (p = 0.001) and seminal vesicles infiltration (p = 0.014). AURKA was expressed in rare cases. N-MYC protein was negative in all patients. CD44s was associated with lower preoperative PSA levels and lower Gleason scores. Biochemical recurrence was observed in 27.0% of patients. Recurrence was associated with serum preoperative PSA, Gleason score, seminal vesicle invasion and staging at least in one form of analysis. There was no significant association between recurrence and neuroendocrine differentiation, C-MYC and CD44s expression. Therefore, immunohistochemical detection of neuroendocrine differentiation, expression of C-MYC and loss of CD44s were related to more aggressive carcinomas (PSA, Gleason, seminal vesical invasion and/or stage), but no association with biochemical recurrence. Classic prognostic factors were affirmed like biochemical recurrence predictores. / Este estudo objetivou avaliar novos marcadores imuno-histoquÃmicos relacionados à induÃÃo da diferenciaÃÃo neuroendÃcrina e cÃlulas-tronco com fatores de prognÃstico e recorrÃncia bioquÃmica, em pacientes submetidos à prostatectomia radical. Para tanto, pacientes operados no Hospital Walter CantÃdio, Universidade Federal do CearÃ, no perÃodo de 2008 a 2013, foram submetidos a acompanhamento clÃnico-ambulatorial, entre os anos de 2008 a 2016. Avaliou-se a proporÃÃo daqueles que apresentaram recorrÃncia bioquÃmica, bem como as caracterÃsticas clÃnico-patolÃgicas e a marcaÃÃo em reaÃÃes de imuno-histoquÃmica para cromogranina (diferenciaÃÃo neuroendÃcrina), Aurora quinase A (AURKA), N-MYC, C-MYC e CD44s, em material parafinado. De 74 pacientes submetidos à cirurgia, obteve-se acompanhamento de 69, com tempo de seguimento de 41 (2-89) meses; diferenciaÃÃo neuroendÃcrina na neoplasia se associou com infiltraÃÃo de vesÃculas seminais (p=0,032) e estadiamento (p=0,030). C-MYC associou-se com escore de Gleason (p=0,001) e infiltraÃÃo de vesÃculas seminais (p=0,014). AURKA expressou-se em raros casos. N-MYC foi negativo em todos os pacientes. CD44s se associou com menores nÃveis de PSA prÃ-operatÃrio e menores escores de Gleason. Observou-se recorrÃncia bioquÃmica em 27,0% dos pacientes. RecorrÃncia se associou, em pelo menos uma das formas de anÃlise, com nÃveis sÃricos de PSA prÃ-operatÃrio, escore de Gleason, invasÃo de vesÃculas seminais e estadiamento. NÃo houve associaÃÃo significativa entre recorrÃncia e diferenciaÃÃo neuroendÃcrina, C-MYC e CD44s. Dessa forma, nesse estudo, a detecÃÃo imuno-histoquÃmica da diferenciaÃÃo neuroendÃcrina; a expressÃo de C-MYC e a perda da expressÃo de CD44s relacionaram-se com carcinomas mais agressivos (PSA, Gleason, infiltraÃÃo de vesÃcula seminal e/ou estadiamento), porÃm sem associaÃÃo com a recorrÃncia bioquÃmica; bem como confirma a importÃncia de fatores prognÃsticos considerados como clÃssicos em sÃrie regional de pacientes com cÃncer de prÃstata.