Inibição e reversão da agregação plaquetária de eqüinos in vitro com o uso de ketoprofeno, fenilbutazona e flunixim meglumine

Como são várias as enfermidades e os distúrbios que induzem à hipercoagulabilidade e à pré-ativação de plaquetas em eqüinos. A atividade de medicamentos utilizados para controle dessas enfermidades sobre a agregação de plaquetas pode, não apenas servir para avaliar sua evolução, como também a resposta terapêutica. Com o objetivo de avaliar a prevenção e a reversão da agregação plaquetária de eqüinos in vitro foram utilizados os antiinflamatórios não esteroidais (AINES): ketoprofeno, fenilbutazona e flunixim meglumine. A comparação demonstrou que a fenilbutazona e o ketoprofeno previnem a agregação de plaquetas de eqüinos induzida pelo ADP, de forma mais eficaz do que o flunixim-meglumine e, superior ao fragmento monoclonal de anticorpo Reopro, sendo semelhante a dos bloqueadores de receptores de membrana Ro-438857 e RGDS. Quanto a reverão da agregação plaquetária tanto a fenilbutazona quanto o ketoprofeno demonstraram efeitos dose-dependente.

Componente lento da cinética do VO2: determinantes fisiológicos e implicações para o desempenho em exercícios aeróbios

A cinética do consumo de oxigênio (VO2) e a resposta do lactato sanguíneo durante o exercício de carga constante em diferentes intensidades permitem caracterizar os domínios moderado, pesado e severo do exercício. Em exercício de intensidade constante, o perfil da resposta do VO2, analisada por ajustes exponenciais, apresenta as fases cardiodinâmica, fundamental e lenta. A ocorrência do componente lento (CL) tem sido associada a fatores como recrutamento de fibras do tipo II e acúmulo de metabólitos, como lactato, íons H+, fosfato inorgânico e ADP. O CL expressa uma redução da eficiência muscular e tem sido associado à menor tolerância de exercícios aeróbios de intensidade alta. O presente estudo teve por objetivo detalhar a fundamentação teórica sobre sua ocorrência, a influência na tolerância ao exercício, bem como prover os diferentes procedimentos adotados em sua quantificação.

Síntese e avaliação farmacológica de novos compostos híbridos úteis para tratamento das complicaçôes da anemia falciforme

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Structural and functional characterization of the chaperone Hsp70 from sugarcane. Insights into conformational changes during cycling from cross-linking/mass spectrometry assays

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

CrataBL, a lectin and Factor Xa inhibitor, plays a role in blood coagulation and impairs thrombus formation

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Antitumor activity of irradiated riboflavin on human renal carcinoma cell line 786-O

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Structural and kinetic studies of Schistosoma mansoni adenylate kinases

The human parasite Schistosoma mansoni is totally dependent on the purine salvage pathway in order to supply large quantities of purine precursors for its energy and DNA biosynthetic needs. Adenylate kinase (ADK) is responsible for the conversion of AMP (produced by the adenosine kinase reaction) into ADP, which is subsequently converted into ATP by nucleoside diphosphate kinase (NDPK). ADK and NDPK are the most active enzymes of the pathway, probably reflecting an evolutionary adaptation due to the intense use of the branch in which they participate. However, notwithstanding their importance very little information has been accumulated found regarding these enzymes. In this work two adenylate kinases from S. mansoni were cloned and heterologously expressed in Escherichia coil. The purified products were utilized in activity assays, and displayed kinetic parameters similar to the corresponding human orthologous proteins. The cytosolic S. mansoni ADK was crystallized and its structure solved allowing us to detect a difference in the nucleotide binding site when compared with the human ortholog. (C) 2012 Elsevier B.V. All rights reserved.

Bioelectrical Impedance Analysis and Skinfold Thickness Sum in Assessing Body Fat Mass of Renal Dialysis Patients

Objective: In chronic renal failure patients under hemodialysis (HD) treatment, the availability of simple, safe, and effective tools to assess body composition enables evaluation of body composition accurately, in spite of changes in body fluids that occur in dialysis therapy, thus contributing to planning and monitoring of nutritional treatment. We evaluated the performance of bioelectrical impedance analysis (BIA) and the skinfold thickness sum (SKF) to assess fat mass (FM) in chronic renal failure patients before (BHD) and after (AHD) HD, using air displacement plethysmography (ADP) as the standard method. Design: This single-center cross-sectional trial involved comparing the FM of 60 HD patients estimated BHD and AHD by BIA (multifrequential; 29 women, 31 men) and by SKF with those estimated by the reference method, ADP. Body fat-free mass (FFM) was also obtained by subtracting the total body fat from the individual total weight. Results: Mean estimated FM (kg [%]) observed by ADP BHD was 17.95 +/- 0.99 kg (30.11% +/- 1.30%), with a 95% confidence interval (CI) of 16.00 to 19.90 (27.56 to 32.66); mean estimated FM observed AHD was 17.92 +/- 1.11 kg (30.04% +/- 1.40%), with a 95% CI of 15.74 to 20.10 (27.28 to 32.79). Neither study period showed a difference in FM and FFM (for both kg and %) estimates by the SKF method when compared with ADP; however, the BIA underestimated the FM and overestimated the FFM (for both kg and %) when compared with ADP. Conclusion: The SKF, but not the BIA, method showed results similar to ADP and can be considered adequate for FM evaluation in HD patients. (C) 2012 by the National Kidney Foundation, Inc. All rights reserved.

NGC 2782: a merger remnant with young stars in its gaseous tidal tail

We have searched for young star-forming regions around the merger remnant NGC 2782. By using Galaxy Evolution Explorer far-ultraviolet and near-ultraviolet imaging and H i data we found seven ultraviolet sources, located at distances greater than 26 kpc from the centre of NGC 2782, and coinciding with its western H i tidal tail. These regions were resolved in several smaller systems when Gemini/Gemini multi-object spectrograph (GMOS) r-band images were used. We compared the observed colours to stellar population synthesis models and found that these objects have ages of similar to 1 to 11 Myr and masses ranging from 103.9 to 104.6 M circle dot. By using Gemini/GMOS spectroscopic data we confirm memberships and derive high metallicities for three of the young regions in the tail (12+log(O/H) = 8.74 +/- 0.20, 8.81 +/- 0.20 and 8.78 +/- 0.20). These metallicities are similar to the value presented by the nuclear region of NGC 2782 and also similar to the value presented for an object located close to the main body of NGC 2782. The high metallicities measured for the star-forming regions in the gaseous tidal tail of NGC 2782 could be explained if they were formed out of highly enriched gas which was once expelled from the centre of the merging galaxies when the system collided. An additional possibility is that the tail has been a nursery of a few generations of young stellar systems which ultimately polluted this medium with metals, further enriching the already pre-enriched gas ejected to the tail when the galaxies collided.

Low resolution structural characterization of the Hsp70-interacting protein - Hip - from Leishmania braziliensis emphasizes its high asymmetry

The Hsp70 is an essential molecular chaperone in protein metabolism since it acts as a pivot with other molecular chaperone families. Several co-chaperones act as regulators of the Hsp70 action cycle, as for instance Hip (Hsp70-interacting protein). Hip is a tetratricopeptide repeat protein (TPR) that interacts with the ATPase domain in the Hsp70-ADP state, stabilizing it and preventing substrate dissociation. Molecular chaperones from protozoans, which can cause some neglected diseases, are poorly studied in terms of structure and function. Here, we investigated the structural features of Hip from the protozoa Leishmania braziliensis (LbHip), one of the causative agents of the leishmaniasis disease. LbHip was heterologously expressed and purified in the folded state, as attested by circular dichroism and intrinsic fluorescence emission techniques. LbHip forms an elongated dimer, as observed by analytical gel filtration chromatography, analytical ultracentrifugation and small angle X-ray scattering (SAXS). With the SAXS data a low resolution model was reconstructed, which shed light on the structure of this protein, emphasizing its elongated shape and suggesting its domain organization. We also investigated the chemical-induced unfolding behavior of LbHip and two transitions were observed. The first transition was related to the unfolding of the TPR domain of each protomer and the second transition of the dimer dissociation. Altogether. LbHip presents a similar structure to mammalian Hip, despite their low level of conservation, suggesting that this class of eukaryotic protein may use a similar mechanism of action. (C) 2012 Elsevier Inc. All rights reserved.

Exploration of the Antiplatelet Activity Profile of Betulinic Acid on Human Platelets

Betulinic acid, a natural pentacyclic triterpene acid, presents a diverse mode of biological actions including antiretroviral, antibacterial, antimalarial, and anti-inflammatory activities. The potency of betulinic acid as an inhibitor of human platelet activation was evaluated, and its antiplatelet profile against in vitro platelet aggregation, induced by several platelet agonists (adenosine diphosphate, thrombin receptor activator peptide-14, and arachidonic acid), was explored. Flow cytometric analysis was performed to examine the effect of betulinic acid on P-selectin membrane expression and PAC-1 binding to activated platelets. Betulinic acid potently inhibits platelet aggregation and also reduced PAC-1 binding and the membrane expression of P-selectin. Principal component analysis was used to screen, on the chemical property space, for potential common pharmacophores of betulinic acid with approved antithrombotic drugs. A common pharmacophore was defined between the NMR-derived structure of betulinic acid and prostacyclin agonists (PGI2), and the importance of its carboxylate group in its antiplatelet activity was determined. The present results indicate that betulinic acid has potential use as an antithrombotic compound and suggest that the mechanism underlying the antiplatelet effects of betulinic acid is similar to that of the PGI2 receptor agonists, a hypothesis that deserves further investigation.

Expression of BAG-1 and PARP-1 in Precursor Lesions and Invasive Cervical Cancer Associated with Human Papillomavirus (HPV)

Cervical cancer remains persistently the second most common malignancies among women worldwide, responsible for 500,000 new cases annually. Only in Brazil, the estimate is for 18,430 new cases in 2011. Several types of molecular markers have been studied in carcinogenesis including proteins associated with apoptosis such as BAG-1 and PARP-1. This study aims to demonstrate the expression of BAG-1 and PARP-1 in patients with low-grade squamous intraepithelial lesions (LSILs), high-grade squamous intraepithelial lesions (HSILs) and invasive squamous cell carcinomas (SCCs) of the uterine cervix and to verify a possible association with HPV infection. Fifty samples of LSILs, 50 samples of HSILs and 50 samples of invasive SCCs of the uterine cervix were analyzed by immunohistochemistry for BAG-1 and PARP-1 expression. PCR was performed to detect and type HPV DNA. BAG-1 expression levels were significantly different between LSILs and HSILs (p = 0,014) and between LSILs and SCCs (p = 0,014). In regards to PARP-1 expression, we found significant differences between the expression levels in HSILs and SCCs (p = 0,022). No association was found between BAG-1 expression and the presence of HPV. However, a significant association was found between PARP-1 expression and HPV positivity in the HSILs group (p = 0,021). In conclusion our research suggests that BAG-1 expression could contribute to the differentiation between LSIL and HSIL/SCC whereas PARP-1 could be useful to the differentiation between HSIL HPV-related and SCC. Further studies are needed to clarify the molecular aspects of the relationship between PARP-1 expression and HPV infection, with potential applications for cervical cancer prediction.

Repercussões pulmonares após isquemia hepática parcial e reperfusão: modelo experimental

OBJETIVO: Descrever um modelo experimental de lesão de isquemia/reperfusão hepática com manifestações sistêmicas, representadas pelo envolvimento pulmonar, que possa ser utilizado por aqueles que pretendem compreender esse fenômeno. MÉTODOS: Ratos Wistar machos (200-250g) foram usados. Quatorze foram alocados em dois grupos, sendo G1 com oito submetidos somente à laparotomia e G2, seis à isquemia e reperfusão hepática. As funções hepática (aminotransferases séricas, respiração mitocondrial, histologia) e pulmonar (teste do azul de Evans) foram analisadas. RESULTADOS: houve diferença estatística significativa entre G1 e G2 ao se comparar valores de AST (24,3 ± 108 e 5406 ± 2263), ALT (88,5 ± 28,5 e 5169 ± 2690), razão de controle respiratório (3,41 ± 0,17 e 1,91 ± 0,55) e relação ADP/O (1,93 ± 0,03 e 1,45 ± 0,27), lesões histológicas (necrose, células inflamatórias, hemorragia, microesteatose) e teste do azul de Evans (194,31 ± 53 e 491,8 ± 141). CONCLUSÃO: O modelo mostrou-se útil para o estudo de lesão de isquemia/reperfusão hepática.

Sanguineous normothermic, intermittent cardioplegia, effects on hypertrophic myocardium: morphometric, metabolic and ultrastructural studies in rabbits hearts

OBJECTIVES: The present investigation aimed to study the protective effect of intermittent normothermic cardioplegia in rabbit's hypertrophic hearts. METHODS: The parameters chosen were 1) the ratio heart weight / body weight, 2) the myocardial glycogen levels, 3) ultrastructural changes of light and electron microscopy, and 4) mitochondrial respiration. RESULTS: 1) The experimental model, coarctation of the aorta induced left ventricular hypertrophy; 2) the temporal evolution of the glycogen levels in hypertrophic myocardium demonstrates that there is a significant decrease; 3) It was observed a time-dependent trend of higher oxygen consumption values in the hypertrophic group; 4) there was a significant time-dependent decrease in the respiratory coefficient rate in the hypertrophic group; 5) the stoichiometries values of the ADP: O2 revealed the downward trend of the values of the hypertrophic group; 6) It was possible to observe damaged mitochondria from hypertrophic myocardium emphasizing the large heterogeneity of data. CONCLUSION: The acquisition of biochemical data, especially the increase in speed of glycogen breakdown, when anatomical changes are not detected, represents an important result even when considering all the difficulties inherent in the process of translating experimental results into clinical practice. With regard to the adopted methods, it is clear that morphometric methods are less specific. Otherwise, the biochemical data allow detecting alterations of glycogen concentrations and mitochondria respiration before the morphometric alterations should be detected

The lactate paradox revisited in lowlanders during acclimatization to 4100 m and in high-altitude natives

[EN] Chronic hypoxia has been proposed to induce a closer coupling in human skeletal muscle between ATP utilization and production in both lowlanders (LN) acclimatizing to high altitude and high-altitude natives (HAN), linked with an improved match between pyruvate availability and its use in mitochondrial respiration. This should result in less lactate being formed during exercise in spite of the hypoxaemia. To test this hypothesis six LN (22-31 years old) were studied during 15 min warm up followed by an incremental bicycle exercise to exhaustion at sea level, during acute hypoxia and after 2 and 8 weeks at 4100 m above sea level (El Alto, Bolivia). In addition, eight HAN (26-37 years old) were studied with a similar exercise protocol at altitude. The leg net lactate release, and the arterial and muscle lactate concentrations were elevated during the exercise in LN in acute hypoxia and remained at this higher level during the acclimatization period. HAN had similar high values; however, at the moment of exhaustion their muscle lactate, ADP and IMP content and Cr/PCr ratio were higher than in LN. In conclusion, sea-level residents in the course of acclimatization to high altitude did not exhibit a reduced capacity for the active muscle to produce lactate. Thus, the lactate paradox concept could not be demonstrated. High-altitude natives from the Andes actually exhibit a higher anaerobic energy production than lowlanders after 8 weeks of acclimatization reflected by an increased muscle lactate accumulation and enhanced adenine nucleotide breakdown.