Indiana University-Purdue University Indianapolis (IUPUI) / Adult hematopoietic stem cells (HSC) are routinely used to reconstitute hematopoiesis after myeloablation; however, transplantation efficacy and multilineage reconstitution can be limited by inadequate HSC number, or poor homing, engraftment or self-renewal. We have demonstrated that mouse and human HSC express prostaglandin E2 (PGE2) receptors, and that short-term ex vivo exposure of HSC to PGE2 enhances their homing, survival and proliferation, resulting in increased long-term repopulating cell and competitive repopulating unit (CRU) frequency. HSC pulsed with PGE2 are more competitive, as determined by head-to-head comparison in a competitive transplantation model. Enhanced HSC frequency and competitive advantage is stable and maintained upon multiple serial transplantations, with full multi-lineage reconstitution. PGE2 increases HSC CXCR4 mRNA and surface expression and enhances their migration to SDF-1α in vitro and homing to bone marrow in vivo and stimulates HSC entry into and progression through cell cycle. In addition, PGE2 enhances HSC survival, associated with an increase in Survivin mRNA and protein expression and reduction in intracellular active caspase-3. While PGE2 pulse of HSC promotes HSC self-renewal, blockade of PGE2 biosynthesis with non-steroidal anti-inflammatory drugs (NSAIDs) results in expansion of bone marrow hematopoietic progenitor cells (HPC). We co-administered NSAIDs along with the mobilizing agent granulocyte-colony stimulating factor (G-CSF) and evaluations of limiting dilution transplants, assays monitoring neutrophil and platelet recoveries, and secondary transplantations, clearly indicate that NSAIDs facilitate mobilization of a hematopoietic graft with superior functional activity compared to the graft mobilized by G-CSF alone. Enhanced mobilization has also been confirmed in baboons mobilized with G-CSF and a NSAID. Increases in mobilization are the result of a reduction of signaling through the PGE2 receptor EP4, which results in marrow expansion and reduction in the osteoblastic HSC niche. We also identify a new role for cannabinoids, an eicosanoid with opposing functions to PGE2, in hematopoietic mobilization. Additionally, we demonstrate increased survival in lethally irradiated mice treated with PGE2, NSAIDs, or the hypoxia mimetic cobalt chloride. Our results define novel mechanisms of action whereby eicosanoids regulate HSC and HPC function, and characterize novel translational strategies for hematopoietic therapies.

This review summarizes the principles of cancer chemoprevention and discusses the evidence from epidemiologic and experimental studies and preclinical and clinical trials of potential colorectal chemopreventive agents. The putative mechanisms of action of the drugs in chemoprevention and their potential to reduce the incidence and mortality rate of colorectal neoplasms are discussed. The future of colorectal chemoprevention will depend on important new insights into molecular carcinogenesis of colorectal cancer, application of molecular markers as surrogate endpoints, and ultimately on therapeutic targets of prevention in clinical trials.

BACKGROUND AND PURPOSE: Pharmacokinetic assessment of drug tissue permeation following iontophoresis is limited. The depth of ketoprofen tissue permeation following cathodic iontophoresis (4 mA, 40 minutes) and the stereoselectivity of drug delivery were examined in this study. SUBJECTS: Ketoprofen (750 mg) was iontophoresed onto one porcine medial thigh, with passive drug permeation conducted on the other thigh. METHODS: Skin, subcutaneous fascia, and muscle biopsies from the drug delivery sites were harvested and stored separately, and the "R" and "S" ketoprofen enantiomers were determined. Results. Iontophoretic and passive applications yielded equivalent total ketoprofen concentrations in the skin and fascia. In contrast, multivariate analysis demonstrated that the ketoprofen concentration in the first centimeter of muscle following iontophoresis was greater than the drug concentration in the deeper underlying muscle layers and greater than that delivered to any muscle layer following passive delivery. No transcutaneous stereoselective delivery) of ketoprofen was detected. CONCLUSION AND DISCUSSION: Compared with passive delivery, iontophoresis enhances nonstereoselective ketoprofen permeation into the fascia-muscle interface. With delivery to deeper tissue sites, however, there is no apparent enhancement over passive application.

No / To demonstrate the benefits of fluoroscopy-guided intra-articular steroid injection in the hip with varying degrees of disease severity, and to investigate the financial aspects of the procedure and impact on waiting time. MATERIALS AND METHODS: A prospective study was undertaken of patients who underwent fluoroscopic intra-articular steroid injection over the 9-month study period. Comparative analysis of the Oxford hip pain score pre- and 6-8 weeks post-intra-articular injection was performed. Hip radiographs of all patients were categorised as normal, mild, moderate, or severe disease (four categories) based on the modified Kellgren-Lawrence severity scale, and improvement on the Oxford hip pain score on each of these four severity categories were assessed. RESULTS: Within the study cohort of 100 patients, the mean increase in post-procedure hip score of 7.32 points confirms statistically significant benefits of the therapy (p<0.001, 95% confidence interval: 5.55-9.09). There was no significant difference in pre-injection hip score or change in score between the four severity categories (p=0.51). Significant improvement in hip score (p<0.05) was demonstrated in each of the four severity categories 6-8 weeks post-injection. No associated complications were observed. CONCLUSION: The present study confirms that fluoroscopy-guided intra-articular steroid injection is a highly effective therapeutic measure for hip osteoarthritis across all grades of disease severity with significant cost savings and the potential to reduce waiting times.

Introdução: Os antiinflamatórios não esteróides (AINEs), incluindo a aspirina, são drogas largamente utilizadas para tratamento das doenças inflamatórias e da dor, e que podem causar efeitos colaterais sérios, causando considerável morbidade e mortalidade, relacionadas á doença ulcerosa, duodenal e gástrica, particularmente ao sangramento gastrointestinal. O risco relativo global de complicações gastroduodenais é de três a dez vezes, maior nos usuários de AINEs, quando comparado com indivíduos sadios. Cerca de 25% dos usuários crônicos dos antiinflamatórios não esteroides (AINEs) deverão desenvolver doença ulcerosa, e de 2 a 4% deverão apresentar sangramento ou perfuração. Mais de 17.000.000 de norte americanos utilizam vários tipos de drogas antiinflamatórios não esteróides (AINEs) diariamente e que provocam mais de 100.000 hospitalizações e cerca de 7000 a 10.000 mortes por ano nos Estados Unidos da América do Norte, fazendo desta família de drogas uma das mais comumente usadas em todo planeta. Cerca de 50% das lesões observadas em endoscopias de controle, ocorrem sem que o paciente tenha qualquer tipo de sintoma. Acredita-se que houve recrudescimento da prevalência de lesões digestivas pela substituição dos antiinflamatórios COX-2 pelos antiinflamatórios tradicionais, principalmente pela ausência de cuidados na prevenção deste tipo de ocorrência, em populações consideradas de risco. Objetivos: a) avaliar a prevalência de lesões e complicações digestivas secundárias ao uso de AINEs; b) qual é o perfil clínico deste paciente atendido em razão de queixas digestivas e a relação destas com os achados endoscópicos. Materiais e métodos: estudo aberto, prospectivo, multicêntrico avaliando consecutivamente 1.231 pacientes submetidos a exame de endoscopia digestiva alta em virtude de queixas digestivas, única ou associadas, como: 1) pirose; 2) dor epigástrica; 3) dor abdominal; 4) náusea; 5) vômito. Antes da realização do exame de endoscopia digestiva alta, os pacientes respondiam a questionário cujo objetivo era avaliar o início e o tipo de queixa clínica, o uso de medicamentos e possíveis complicações associadas como sangramento digestivo. Os critérios de inclusão foram: pacientes de ambos os sexos com idade mínima de 18 anos e que tivessem sintomas prévios iniciados, no máximo, há 14 dias antes da realização do exame de endoscopia digestiva alta. Os critérios de exclusão foram os de pacientes que se recusaram a participar do estudo e/ou de assinar o Termo de Consentimento Livre e Esclarecido, os incapazes de responder ao questionário, os com idade inferior aos 18 anos, os pacientes que já haviam realizado cirurgia gástrica e pacientes portadores de insuficiência renal ou hepática. Resultados: Foram avaliados 1.213 pacientes de 18 a 82 anos sendo que 65% destes eram do sexo feminino, 13,1% eram fumantes e 15,6% referiam ingestão de bebidas alcoólicas. A utilização de AINEs foi mais frequente no sexo feminino, porém número de complicações foi maior nos pacientes do sexo masculino (sangramentos foi duas vezes maior; p=0,045 e a ocorrência de úlcera quase 1,5 vezes maior; p=0,041). Os principais sinais e sintomas relatados foram epigastralgia e pirose (67% e 62%, respectivamente). Os 1.213 pacientes foram alocados em dois grupos: Grupo I - AINE composto por 228 (18,8%) e o Grupo II - Não AINEs (NAINEs) por 985 (81,2%) pacientes.. O exame de endoscopia digestiva alta foi normal em 3,9% dos pacientes do grupo I e em 10,7% dos do grupo II (p< 0,001). A probabilidade de um paciente que não utiliza AINE ter endoscopia digestiva alta normal é 2,5 vezes maior quando comparado aos que utilizaram AINEs (p=0,001). As presenças de lesões erosivas ou ulceradas no estômago e duodeno também foram mais frequentes nos pacientes do Grupo I quando comparado aos do Grupo II. Observa-se que é maior a incidência de lesões, tanto erosivas quanto ulceradas no estômago quando comparadas ao duodeno (erosões: 49,12% vs 13,60 respectivamente, p=0,001; úlceras: 14,04% vs 11,84% respectivamente, p= 0,05). O risco de hemorragia digestiva, 12 vezes maior (6,14% vs 0,51%) nos pacientes que fizeram uso de AINEs sendo o estômago o sítio de maior prevalência de sangramento. Não se observou diferença estatística quando analisada a presença de esofagite erosiva nos dois grupos. Conclusões: Evidenciamos frequência maior de úlcera gástrica, úlcera duodenal e sangramento digestivo nos pacientes que utilizaram AINEs. Não foram encontradas relações entre os achados endoscópicos e os sintomas dispépticos. Não observamos influência dos AINEs no aparecimento de esofagite erosiva / Introduction: The non steroidal anti-inflammatory drugs (NSAID), including aspirin, are drugs widely used in the treatment of inflammatory diseases and pain. This use may cause serious side-effects, leading to considerable morbidity and mortality related to ulcer, duodenal and gastric disease, especially gastrointestinal bleeding. The overall relative risk of gastroduodenal complications is three to ten times higher in users of NSAID, compared to healthy individuals. Around 25% of the chronic users of non steroidal anti-inflammatory drugs (NSAID) will develop ulcer disease, and 2 to 4% will present bleeding or perforation. More than 17,000,000 North Americans use several kinds of non steroidal anti-inflammatory drugs (NSAID) on a daily basis. This causes more than 100,000 hospitalizations and from 7,000 to 10,000 deaths every year in the USA, which makes this drug one of the most commonly used on the planet. About 50% of the lesions observed in endoscopies occur without any kind of symptom. It is believed that there was an increase in the prevalence of digestive lesions due to the replacement of COX-2 anti-inflammatory drugs with traditional anti-inflammatory drugs, especially because of the lack of preventive care of this kind of occurrence in at-risk populations. Goals: a) Evaluate the prevalence of lesions and digestive complications, secondary to the use of NSAID; b) Evaluate the clinical profile of the patient seen for digestive complaints and the relation of these complaints with the endoscopic findings. Materials and Methods: Prospective, multi-centric, open study, evaluating consecutively 1,231 patients who underwent upper gastrointestinal endoscopy exam due to digestive complaints in isolation or associated, such as: 1) pyrosis; 2) epigastric pain; 3) abdominal pain; 4) nausea; 5) vomiting. Before performing the exam of upper gastrointestinal endoscopy, patients answered a questionnaire whose goal was to evaluate the onset and kind of clinical complaint, the use of medication and possible complications associated to digestive bleeding. The inclusion criteria were: Patients of both sexes with the minimum age of 18 and whose symptoms had begun up to 14 days before undergoing the upper gastrointestinal endoscopy. Exclusion criteria: patients who refused to participate in the study and/ or who refused to sign the Informed Consent Term, the ones who were unable to respond to the questionnaire, the ones who were under 18 years old, patients who had undergone a previous gastric surgery and patients with kidney or hepatic failure. Results: 1,213 patients with ages ranging from 18-82 were evaluated, 65% of which were female and 13,1% were smokers, 15,6% mentioned they ingested alcoholic beverages. The use of NSAID was more frequent among females. However, the number of complications was higher among males (bleeding occurred twice as much; p=0,045 and the occurrence of ulcer was almost 1,5 times higher; p=0,041). The main signs and symptoms reported were epigastralgia and pyrosis (67% and 62%). The 1,213 patients were divided into two groups: Group I- NSAID, made up by 228 (18,8%) and Group II- Non NSAID, made up by 985 patients (81,2%). The upper gastrointestinal endoscopy was normal in 3,9% of the patients in Group I and in 10,7% of the patients in Group II (p<0,001). A patient who does not use NSAID will be 2,5 times more likely to have normal upper gastrointestinal endoscopy than the one who used NSAID (p=0,001). The presence of erosive or ulcer lesions in the stomach and duodenum was more frequent in Group I patients when compared to those of Group II. It is observed that the incidence of lesions in the stomach, both erosive and ulcer is higher when compared to the duodenum (erosions: 49,12% vs. 13,60, p=0,001; ulcers: 14,04% vs. 11,84, p= 0,05). The risk of digestive bleeding is 12 times higher (6,14% vs. 0,51%) in patients who used NSAID, and the stomach is the site with higher prevalence of bleeding. No statistic difference was observed when the presence of erosive esophagitis in both groups was analyzed. Conclusions: We observed that the frequency of gastric ulcer, duodenal ulcer and digestive bleeding was higher in patients who used NSAID. Relations between the endoscopic findings and the dyspeptic symptoms were not found. The influence of NSAIDs on the appearance of erosive esophagitis was not observed

Introdução: Os antiinflamatórios não esteróides (AINEs), incluindo a aspirina, são drogas largamente utilizadas para tratamento das doenças inflamatórias e da dor, e que podem causar efeitos colaterais sérios, causando considerável morbidade e mortalidade, relacionadas á doença ulcerosa, duodenal e gástrica, particularmente ao sangramento gastrointestinal. O risco relativo global de complicações gastroduodenais é de três a dez vezes, maior nos usuários de AINEs, quando comparado com indivíduos sadios. Cerca de 25% dos usuários crônicos dos antiinflamatórios não esteroides (AINEs) deverão desenvolver doença ulcerosa, e de 2 a 4% deverão apresentar sangramento ou perfuração. Mais de 17.000.000 de norte americanos utilizam vários tipos de drogas antiinflamatórios não esteróides (AINEs) diariamente e que provocam mais de 100.000 hospitalizações e cerca de 7000 a 10.000 mortes por ano nos Estados Unidos da América do Norte, fazendo desta família de drogas uma das mais comumente usadas em todo planeta. Cerca de 50% das lesões observadas em endoscopias de controle, ocorrem sem que o paciente tenha qualquer tipo de sintoma. Acredita-se que houve recrudescimento da prevalência de lesões digestivas pela substituição dos antiinflamatórios COX-2 pelos antiinflamatórios tradicionais, principalmente pela ausência de cuidados na prevenção deste tipo de ocorrência, em populações consideradas de risco. Objetivos: a) avaliar a prevalência de lesões e complicações digestivas secundárias ao uso de AINEs; b) qual é o perfil clínico deste paciente atendido em razão de queixas digestivas e a relação destas com os achados endoscópicos. Materiais e métodos: estudo aberto, prospectivo, multicêntrico avaliando consecutivamente 1.231 pacientes submetidos a exame de endoscopia digestiva alta em virtude de queixas digestivas, única ou associadas, como: 1) pirose; 2) dor epigástrica; 3) dor abdominal; 4) náusea; 5) vômito. Antes da realização do exame de endoscopia digestiva alta, os pacientes respondiam a questionário cujo objetivo era avaliar o início e o tipo de queixa clínica, o uso de medicamentos e possíveis complicações associadas como sangramento digestivo. Os critérios de inclusão foram: pacientes de ambos os sexos com idade mínima de 18 anos e que tivessem sintomas prévios iniciados, no máximo, há 14 dias antes da realização do exame de endoscopia digestiva alta. Os critérios de exclusão foram os de pacientes que se recusaram a participar do estudo e/ou de assinar o Termo de Consentimento Livre e Esclarecido, os incapazes de responder ao questionário, os com idade inferior aos 18 anos, os pacientes que já haviam realizado cirurgia gástrica e pacientes portadores de insuficiência renal ou hepática. Resultados: Foram avaliados 1.213 pacientes de 18 a 82 anos sendo que 65% destes eram do sexo feminino, 13,1% eram fumantes e 15,6% referiam ingestão de bebidas alcoólicas. A utilização de AINEs foi mais frequente no sexo feminino, porém número de complicações foi maior nos pacientes do sexo masculino (sangramentos foi duas vezes maior; p=0,045 e a ocorrência de úlcera quase 1,5 vezes maior; p=0,041). Os principais sinais e sintomas relatados foram epigastralgia e pirose (67% e 62%, respectivamente). Os 1.213 pacientes foram alocados em dois grupos: Grupo I - AINE composto por 228 (18,8%) e o Grupo II - Não AINEs (NAINEs) por 985 (81,2%) pacientes.. O exame de endoscopia digestiva alta foi normal em 3,9% dos pacientes do grupo I e em 10,7% dos do grupo II (p< 0,001). A probabilidade de um paciente que não utiliza AINE ter endoscopia digestiva alta normal é 2,5 vezes maior quando comparado aos que utilizaram AINEs (p=0,001). As presenças de lesões erosivas ou ulceradas no estômago e duodeno também foram mais frequentes nos pacientes do Grupo I quando comparado aos do Grupo II. Observa-se que é maior a incidência de lesões, tanto erosivas quanto ulceradas no estômago quando comparadas ao duodeno (erosões: 49,12% vs 13,60 respectivamente, p=0,001; úlceras: 14,04% vs 11,84% respectivamente, p= 0,05). O risco de hemorragia digestiva, 12 vezes maior (6,14% vs 0,51%) nos pacientes que fizeram uso de AINEs sendo o estômago o sítio de maior prevalência de sangramento. Não se observou diferença estatística quando analisada a presença de esofagite erosiva nos dois grupos. Conclusões: Evidenciamos frequência maior de úlcera gástrica, úlcera duodenal e sangramento digestivo nos pacientes que utilizaram AINEs. Não foram encontradas relações entre os achados endoscópicos e os sintomas dispépticos. Não observamos influência dos AINEs no aparecimento de esofagite erosiva / Introduction: The non steroidal anti-inflammatory drugs (NSAID), including aspirin, are drugs widely used in the treatment of inflammatory diseases and pain. This use may cause serious side-effects, leading to considerable morbidity and mortality related to ulcer, duodenal and gastric disease, especially gastrointestinal bleeding. The overall relative risk of gastroduodenal complications is three to ten times higher in users of NSAID, compared to healthy individuals. Around 25% of the chronic users of non steroidal anti-inflammatory drugs (NSAID) will develop ulcer disease, and 2 to 4% will present bleeding or perforation. More than 17,000,000 North Americans use several kinds of non steroidal anti-inflammatory drugs (NSAID) on a daily basis. This causes more than 100,000 hospitalizations and from 7,000 to 10,000 deaths every year in the USA, which makes this drug one of the most commonly used on the planet. About 50% of the lesions observed in endoscopies occur without any kind of symptom. It is believed that there was an increase in the prevalence of digestive lesions due to the replacement of COX-2 anti-inflammatory drugs with traditional anti-inflammatory drugs, especially because of the lack of preventive care of this kind of occurrence in at-risk populations. Goals: a) Evaluate the prevalence of lesions and digestive complications, secondary to the use of NSAID; b) Evaluate the clinical profile of the patient seen for digestive complaints and the relation of these complaints with the endoscopic findings. Materials and Methods: Prospective, multi-centric, open study, evaluating consecutively 1,231 patients who underwent upper gastrointestinal endoscopy exam due to digestive complaints in isolation or associated, such as: 1) pyrosis; 2) epigastric pain; 3) abdominal pain; 4) nausea; 5) vomiting. Before performing the exam of upper gastrointestinal endoscopy, patients answered a questionnaire whose goal was to evaluate the onset and kind of clinical complaint, the use of medication and possible complications associated to digestive bleeding. The inclusion criteria were: Patients of both sexes with the minimum age of 18 and whose symptoms had begun up to 14 days before undergoing the upper gastrointestinal endoscopy. Exclusion criteria: patients who refused to participate in the study and/ or who refused to sign the Informed Consent Term, the ones who were unable to respond to the questionnaire, the ones who were under 18 years old, patients who had undergone a previous gastric surgery and patients with kidney or hepatic failure. Results: 1,213 patients with ages ranging from 18-82 were evaluated, 65% of which were female and 13,1% were smokers, 15,6% mentioned they ingested alcoholic beverages. The use of NSAID was more frequent among females. However, the number of complications was higher among males (bleeding occurred twice as much; p=0,045 and the occurrence of ulcer was almost 1,5 times higher; p=0,041). The main signs and symptoms reported were epigastralgia and pyrosis (67% and 62%). The 1,213 patients were divided into two groups: Group I- NSAID, made up by 228 (18,8%) and Group II- Non NSAID, made up by 985 patients (81,2%). The upper gastrointestinal endoscopy was normal in 3,9% of the patients in Group I and in 10,7% of the patients in Group II (p<0,001). A patient who does not use NSAID will be 2,5 times more likely to have normal upper gastrointestinal endoscopy than the one who used NSAID (p=0,001). The presence of erosive or ulcer lesions in the stomach and duodenum was more frequent in Group I patients when compared to those of Group II. It is observed that the incidence of lesions in the stomach, both erosive and ulcer is higher when compared to the duodenum (erosions: 49,12% vs. 13,60, p=0,001; ulcers: 14,04% vs. 11,84, p= 0,05). The risk of digestive bleeding is 12 times higher (6,14% vs. 0,51%) in patients who used NSAID, and the stomach is the site with higher prevalence of bleeding. No statistic difference was observed when the presence of erosive esophagitis in both groups was analyzed. Conclusions: We observed that the frequency of gastric ulcer, duodenal ulcer and digestive bleeding was higher in patients who used NSAID. Relations between the endoscopic findings and the dyspeptic symptoms were not found. The influence of NSAIDs on the appearance of erosive esophagitis was not observed

<p>&Scaron;tavelj (Rumex crispus, Polygonaceae) je vi&scaron;egodi&scaron;nja zeljasta biljka, koja predstavlja bogat izvor fenolnih komponenti. Iako se smatra invazivnim korovom, mlado li&scaron;će &scaron;tavelja je jestivo i često se koristi kao salata. Dalje, upotreba plodova &scaron;tavelja opisana je u srpskoj i turskoj narodnoj medicini u lečenju gastrointestinalnih tegoba. Cilj ovog rada bio je procena in vitro i in vivo antioksidantne/prooksidantne i citotoksične aktivnosti, i određivanje eventualnog in vitro antiinflamatornog efekta vodenog ekstrakta ploda Rumex crispus. Ukupan sadržaj flavonoida određen je spektrofotometrijskom metodom. Kvalifikacija i kvantifikacija flavonoida potvrđena je visokoefikasnom tečnom hromatografijom (HPLC). Antioksidantna aktivnost vodenog ekstrakta ploda &scaron;tavelja procenjena je na osnovu in vitro testova: Ferric-reducing antioxidant power (FRAP), sposobnosti ekstrakta da neutrali&scaron;e slobodne radikale NO&bull;, OH&bull; i DPPH&bull; i uticaja na lipidnu peroksidaciju u lipozomima. Citotoksičnost ispitivanog ekstrakta je određena in vitro na tumorskim ćelijskim linijama: humani karcinom cerviksa (HeLa), adenokarcinom (HT-29) i adenokarcinom dojke (MCF7). Takođe, moguća in vivo hepatoprotektivna i antioksidantna svojstva ekstrakta određena su kod oksidativnog stresa izazvanog CCl4 kod eksperimentalnih životinja. Pored toga, proverena je hipoteza u kojoj testiran ekstrakt pokazuje in vivo antiproliferativnu aktivnost kod Ehrlich-ovih (EAC) i Hepatoma AS30D ćelija, merenjem zapremine ascitesa, procenta vijabilnih ćelija i nivoa nekoliko antioksidantnih enzima. Optimizovan in vitro test za određivanje potencijala inhibicije ciklooksigenaze-1 (COX-1) i 12-lipooksigenaze (12-LOX) preduzet je u svrhu procene antiinflamatornog efekta vodenog ekstrakta ploda R. crispus. HPLC analiza otkrila je da je mikvelianin najdominantniji flavonoidni konstituent ekstrakta. Testirani ekstrakt pokazao je potencijalnu antioksidantnu aktivnost rezultujući velikom moći u neutralizaciji slobodnih radikala, i sposobno&scaron;ću da smanji lipidnu peroksidaciju u lipozomima. Rezultati su ukazali na tkivno-selektivnu citotoksičnost ekstrakta ploda R. crispus in vitro. Najizraženija antitumorska aktivnost primećena je prema HeLa i MCF7 ćelijskim linijama. Podaci sugeri&scaron;u da bi se ispitivani ekstrakt mogao smatrati potencijalnim in vivo hepatoprotektivnim i antioksidantnim agensom, sprečavajući oksidativna o&scaron;tećenja jetre. S druge strane, pomenuti ekstrakt može pokazati in vivo prooksidantna svojstva, uzrokujući oksidativni stres u maligno transformisanim EAC i AS30D ćelijama i smanjujući zapreminu ascitesa i udeo vijabilnih ćelija, u poređenju sa kontrolnom grupom. Promene u aktivnosti antioksidantnih enzima su verovatno posledica indukovanog oksidativnog stresa u EAC i AS30D ćelijama, naročito kod pretretiranih životinja. Vodeni ekstrakt ploda &scaron;tavelja pokazao je COX-1, kao i 12-LOX inhibitornu aktivnost, navodeći da bi ispitivani ekstrakt mogao biti antiinflamatorni agens. Vodeni ekstrakt ploda R. crispus ima potencijalnu antioksidantnu, citotoksičnu i antiinflamatornu aktivnost. Ispoljavanje prooksidantnih svojstava predstavlja mogući mehanizam antiproliferativnog efekta ekstrakta.</p> / <p>Curly dock (Rumex crispus, Polygonaceae) is a wild perennial herbaceous plant, which products are described as a rich source of phenolic compounds. Apart from being considered a seriously invasive weed, young leaves of curly dock are edible and often used as salad. Furthermore, the use of its fruits has been described in Serbian and Turkish traditional medicine against stomach complaints. The objectives of this study were to evaluate in vitro and in vivo antioxidant/prooxidant and cytotoxic activities, and to determine an eventual in vitro anti-inflammatory effect of the aqueous extract of Rumex crispus fruits. Total flavonoid content was determined by spectrophotometric method. Qualification and quantification of flavonoids were confirmed using High performance liquid chromatography (HPLC). The aqueous extract of curly dock fruits was evaluated for its antioxidant activity by in vitro assays for Ferric-reducing antioxidant power (FRAP), NO&bull;, OH&bull; and DPPH&bull;-free radical scavenging activities and the influence on lipid peroxidation in liposomes. The cytotoxicity of tested extract was examined in vitro in human cervix carcinoma (HeLa), colon adenocarcinoma (HT-29) and breast adenocarcinoma (MCF7). Also, the potential in vivo hepatoprotective and antioxidant properties of investigated extract were determined on CCl4-induced oxidative stress in experimental animals. Furthermore, the hypothesis that the examined extract might show in vivo antiproliferative activity in Ehrlich carcinoma (EAC) and Hepatoma AS30D cells was tested by measuring volume of ascites, percentage of viable cells and level of several antioxidant enzymes. The optimized in vitro test for determination of cyclooxygenase-1 (COX-1) and 12-lipoxygenase (12-LOX) inhibition potency was undertaken in order to estimate an anti-inflammatory effect of aqueous extract of R. crispus fruits. HPLC analysis revealed miquelianin as the most abundant flavonoid constituent of the extract. The tested extract might have an antioxidant activity resulting in scavenging of free radicals and ability to decrease lipid peroxidation in liposomes. The results could indicate tissue-selective cytotoxicity of R. crispus fruit extract in vitro. The most prominent antitumor activity was observed towards HeLa and MCF7 cell lines. The data suggested that investigated extract may be considered as potential in vivo hepatoprotective and antioxidant agent due to prevention of the liver injuries induced by oxidative damage. On the other hand, mentioned extract could exhibit in vivo prooxidant property, causing the oxidative stress in malignant transformed EAC and AS30D cells and reducing volume of ascites and percentage of viable cells, in comparison with control group. Changes in activities of antioxidant enzymes might be the results of induced oxidative stress in EAC and AS30D cells, especially in the pretreated animals. The aqueous extract of curly dock fruits showed COX-1, as well as 12-LOX inhibitory activity, suggesting that tested extract might be an anti-inflammatory agent. It could be concluded that aqueous fruit extract of R. crispus might have antioxidant, cytotoxic and anti-inflammatory activities. The prooxidant properties of examined extract could be the mechanism of potential antiproliferative effect of extract.</p>

Introdução: Atualmente, o diagnóstico das reações de hipersensibilidade a anti-inflamatórios não esteroidais (AINEs) baseia-se na história relatada pelo paciente e, em determinados casos, é realizado o Teste de Provocação. Todavia, este teste pode expor os pacientes a riscos graves, inclusive anafilaxia. Em busca de ferramenta mais segura, tem-se estudado o Teste de Ativação de Basófilos (BAT). Trata-se de um teste in vitro, no qual é possível testar diversos estímulos em uma única amostra de sangue, avaliando a ativação dos basófilos (indicativo de reação de hipersensibilidade), através do aumento da expressão de moléculas na superfície desses leucócitos, como o CD63. Objetivo: Padronizar e validar o BAT para ácido acetilsalicílico (AAS), diclofenaco, dipirona e paracetamol em pacientes com hipersensibilidade a AINEs. Metodologia: Participaram 20 (testados com os quatro AINEs) + 33 (testados somente com AAS) pacientes atendidos no Serviço de Imunologia Clínica e Alergia do HCFMUSP, que apresentaram manifestações cutâneas em até 24 horas após exposição a um ou múltiplos AINEs, bem como 13 (quatro AINEs) + 26 (AAS) controles. A técnica consistiu em incubar sangue total com os AINEs já mencionados e, depois, marcar as amostras com anticorpos monoclonais (CD45, anti-IgE e CD63) para posterior leitura por citometria de fluxo. Os resultados obtidos foram comparados com as histórias clínicas e os testes de provocação oral, quando realizados. Resultados: Utilizando os critérios de positividade do BAT empregados na literatura (isto é, porcentagem de CD45+IgE+highCD63+ e índice de estimulação), a sensibilidade e a especificidade variaram de acordo com o AINE: para ácido acetilsalicílico foram 75,0% e 16,7%, respectivamente, diclofenaco, 100% e 0%, dipirona, 23,5% e 66,7%, paracetamol, 40,0% e 42,9%. Após a realização de curvas dose-resposta e tempo-resposta somente com AAS, foi encontrado novo critério de positividade: média de intensidade de fluorescência (MFI) menor do que 6575 representava BAT positivo; com isso, os valores de sensibilidade e especificidade foram: 84,4% e 34,6%, respectivamente. O BAT foi mais sensível em pacientes cuja última reação ocorreu há menos de um ano da data de execução do BAT (93,7%). Conclusão: Devido aos baixos valores de sensibilidade e/ou especificidade, não foi possível padronizar e, por conseguinte, validar o BAT para ácido acetilsalicílico, diclofenaco, dipirona e paracetamol. / Introduction: Currently, the diagnosis of nonsteroidal antiinflammatory drugs (NSAIDs) hypersensivitity is based on patients´ clinical history and drug provocation tests, which are done in selected cases. Nevertheless, this test may expose patients to severe risks, including anaphylaxis. Looking for a safer tool, Basophil Activation Test (BAT) for allergy diagnosis has been studied in the last years. It is an in vitro method where a wide variety of stimuli can be tested, incubating them with the patient\'s blood sample, and observing basophil activation (indication of hypersensitivity) through upregulation of CD63 (or other basophil activation markers) on this leucocyte\'s membrane. Objective: To standardize and validate BAT stimulated with acetylsalicylic acid (ASA), diclophenac, dipyrone and paracetamol in NSAID hypersensitive patients. Methods: Patients which reported immediate reactions (less than 24 hours) after exposure to one or multiple NSAIDs, with cutaneous symptoms were enrolled from Clinical Immunology and Allergy outpatient clinic from HC-FMUSP. BAT with the four NSAIDs was tested on 20 patients and 13 controls and BAT with ASA only, on 33 patients and 26 controls. BAT consisted of incubating whole blood with NSAIDs, then triple-labeled with monoclonal antibodies (CD45, anti-IgE, CD63) for analysis by flow cytometry. BAT results were compared to clinical history and oral provocation tests, when available. Results: According to literature\'s positivity criteria (percentage of CD45+IgE+highCD63+ and stimulation index), sensitivity and specificity varied according to the NSAID tested: for ASA was 75.0% and 16.7% respectively, diclophenac, 100.0% and 0.0%, dipyrone, 23.5% and 66.7%, paracetamol, 40.0% and 42.9%. A new positivity criterion was possible to be defined after further dose-response and time-response curves only for ASA: Mean Fluorescence Intensity lower than 6575 (positive BAT). Accordingly, new sensitivity and specificity for BAT in ASA hypersensitivity were 84,4% and 34,6%. Patients that presented the last reaction in the last year were more likely to present a positive BAT (93.7%). Conclusion: Due to low values for sensitivity and/or specificity, it was not possible to standardize and validate BAT for ASA, diclophenac, dipyrone and paracetamol.

Made available in DSpace on 2016-01-26T12:51:42Z (GMT). No. of bitstreams: 1 rosasoubhia_tese.pdf: 792706 bytes, checksum: bba71696e22270729d09c3130cc047eb (MD5) Previous issue date: 2005-05-25 / Introduction: Tacrolimus (FK 506) is a potent immunosuppressive drug that may cause nephrotoxicity decreasing the renal blood flow (RBF) and glomerular filtration rate (GFR). Conventional non-steroidal anti-inflammatory drugs (NSAIDs) may cause nephrotoxicity, interfering with renal hemodynamics and fluid and eletrolyte homeostasis. Recently, new selective COX-2 inhibitors have been developed producing less side effects (gastric, cardiac and renal) related to COX-1 inhibition. The increasing use of FK 506 and the intensive use of NSAIDs with analgesic or ani-inflammatory purposes, increases the possibility of a drug combination, potentiating the nephrotoxic risk of the two drugs. Objective : Compare the renal function of rats receiving combination therapy with FK and a non-selective COX inhibitor, sodium diclofenac (SD) with those receiving FK and a selective COX-2 inhibitor, rofecoxib (RO). Material and Methods : Male Munich-Wistar rats receiving a low sodium diet (0.06%) for 7 days and gavage treatment for 7 days with FK (2 mg/kg/day), SD (10 mg/kg/day), RO (3 mg/kg/day), FK+SD, FK+RO and vehicle (control) were used. Glomerular filtration rate (GFR, inulin clearance, ml/min/100g); renal blood flow (RBF, Doppler ultrasound, ml/min); mean blood pressure (MBP, intracarotid probe, mmHg); renal vascular resistence ( RVR, mmHg/ml/min); hematocrit (Htc); urinary volume ( UV, &#956;l/min); solute clearance; renal histology; animal weight (g) and FK serum concentration (SCFK, ng/ml) were assessed. Results are presented as a mean±standart deviation and compared by ANOVA followed by Student-Neuman-Keuls test. Results : The GRF of the SD group was 0.98±0.03, RO 1.06±0.04 and FK 0.99±0.06 similar to control values (1.10±0.05). GRF values decreased in the FK+RO (0.86±0.06;p<0.05 vs RO and Control) and FK+SD (0.63±0.06;p<0.001 vs control, FK and SD groups and p<0.01 vs FK+RO) groups. RBF, MBP, RVR and Htc values were similar in all groups. Diuresis was lower in the groups with drug combination, but there was a statistically significant difference only between FK+SD and RO groups (8.38±0.46 vs 12.99±1.22;p<0.05). There were no significant histological chan ges in the treatment groups. The FK+SD group showed statistically significant weigth changes (-18±5) when compared to the Control and RO groups (6±2 and 5±2, respectively; p<0.001) and to the SD an FK+RO groups (0.2±4 and 1±2, respectively; p<0.01). SCFK was significantly decreased (p<0.05) for FK+SD and FK+RO (1.7±0.3 and 1.8±0.4) groups when compared to the FK group (3.2±0.4. Conclusions: The combination of FK and a non-selective COX inhibitor significantly decreased GFR regardless of a RBF decrease or RVR increase, and is probably a result of Kf decrease. The trend to antidiuresis was similar for the combination of FK with both classes of NSAIDs. FK combined to a non-selective COX inhibitor caused a mild systemic toxicity when compared with the COX-2 selective inhibitor. Serum FK concentrations were significantly lower in NSAIDs treated animals. / Introdução: O tacrolimus (FK 506) é uma potente droga imunossupressora, pode causar nefrotoxicidade aguda com diminuição do fluxo sanguíneo renal (FSR) e ritmo de filtração glomerular (RFG). Os antiinflamatórios não-hormonais (AINHs) convencionais podem causar nefrotoxicidade, interferindo na hemodinâmica renal e na homeostase de fluidos e eletrólitos. Recentemente surgiram novas drogas do grupo coxib que são inibidores seletivos da COX-2, e portanto teriam menos efeitos colaterais relacionados à inibição da COX-1 (gástricos, cardíacos e renais). O crescente uso do FK 506 e o intenso uso de AINHs com finalidade analgésica e ou antiinflamatória aumenta a possibilidade de utilização em conjunto, potencializando o risco de nefrotoxicidade das duas drogas. Objetivo: Comparar a função renal de ratos sob os efeitos do uso simultâneo do FK e de um inibidor não-seletivo da COX, o diclofenaco sódico (DS) e do FK e de um inibidor seletivo da COX-2, o rofecoxib (RO). Materiais e Método: Utilizaram-se ratos Munich-Wistar machos em dieta hipossódica (0,06%) por 7 dias e tratamento por gavagem por 7 dias com FK (2 mg/kg/dia), DS (l0mg/kg/dia), RO (3mg/kg/dia), FK+DS, FK+RO e veículo (Contr). Aferidos ritmo de filtração glomerular (RFG, depuração de inulina, ml/min/l00g); o fluxo sanguíneo renal (FSR, ultrasom Doppler, ml/min); a pressão arterial média (PAM, probe intracarotídeo, mmHg); a resistência vascular renal (RVR, mmHg/ml/min); hematócríto (Ht); o volume urinário (VU, pl/min); a depuração de solutos; a histologia renal; o peso dos animais (g) e a concentração sanguínea de FK CSFK, ng/ml). Os resultados são apresentados com médiaserro padrão da média e comparados por ANOVA seguido do teste Student-Neuman-Keuls. Resultados: O RFG do grupo DS foi 0,980,03, do RO foi 1,060,04 e do FK 0,990,06 similares ao controle (1,100,05). Houve queda do RFG nos grupos FK+RO (0,860,06;p<0,Os vs RO e Contr) e FK+DS (0,630,06;p<0,001 vs Contr,DS, RO e FK; p<0,01 vs FK+RO) Nota de Resumo O FSR, a PAM, a RVR e o Ht foram semelhantes em todos os grupos. A diurese foi menor nos grupos com associação de drogas, mas houve diferença estatisticamente significante apenas entre os grupos FK+DS e RO (8,380,46 vs l299l,22;p<0,05). Não ocorreram modificações histológicas significativas nos grupos estudados. O grupo FK+DS apresentou variação de peso (-185) estatisticamente significante em relação aos grupos Contr 62 e RO 52 (p<0,001) e DS 0,24 e FK+RO -12 (p<0,01). A CSFK diminuiu significativamente (p<0,05) para os grupos FK+DS e FK+RO (1,70,3 e 1,80,4) em relação ao grupo FK (3,20,4). Conclusões: A associação do FK com um inibidor não-seletivo da COX causou diminuição mais acentuada do RFG independentemente da diminuição do FSR ou aumento da RVR, sendo provavelmente decorrente da diminuição do Kf. A tendência à antidiurese foi similar para a associação do FK com as duas classes de AINHs. O FK associado com um inibidor não-seletivo da COX causou discreta toxicidade sistêmica quando comparado com inibidor seletivo da COX-2. Nos animais tratados com AINHs, as concentrações sanguíneas do FK foram significativamente menores.

Dissertation (Ph.D.)--University of South Florida, 2007. / Title from PDF of title page. Document formatted into pages; contains 187 pages. Includes vita. Includes bibliographical references.