16-Bromoepiandrosterone, an activator of the mammalian immune system, inhibits glucose 6-phosphate dehydrogenase from Trypanosoma cruzi and is toxic to these parasites grown in culture

Glucose 6-phosphate dehydrogenase (G6PDH) catalyzes the first step of the pentose-phosphate pathway which supplies cells with ribose 5-phosphate (R5P) and NADPH. R5P is the precursor for the biosynthesis of nucleotides while NADPH is the cofactor of several dehydrogenases acting in a broad range of biosynthetic processes and in the maintenance of the cellular redox state. RNA interference-mediated reduction of G6PDH levels in bloodstream-form Trypanosoma brucei validated this enzyme as a drug target against Human African Trypanosomiasis. Dehydroepiandrosterone (DHEA), a human steroidal pro-hormone and its derivative 16 alpha-bromoepiandrosterone (16BrEA) are uncompetitive inhibitors of mammalian G6PDH. Such steroids are also known to enhance the immune response in a broad range of animal infection models. It is noteworthy that the administration of DHEA to rats infected by Trypanosoma cruzi, the causative agent of Human American Trypanosomiasis (also known as Chagas` disease), reduces blood parasite levels at both acute and chronic infection stages. In the present work, we investigated the in vitro effect of DHEA derivatives on the proliferation of T. cruzi epimastigotes and their inhibitory effect on a recombinant form of the parasite`s G6PDH (TcG6PDH). Our results show that DHEA and its derivative epiandrosterone (EA) are uncompetitive inhibitors of TcG6PDH, with K(i) values of 21.5 +/- 0.5 and 4.8 +/- 0.3 mu M, respectively. Results from quantitative inhibition assays indicate 16BrEA as a potent inhibitor of TcG6PDH with an IC(50) of 86 +/- 8 nM and those from in vitro cell viability assays confirm its toxicity for T. cruzi epimastigotes, with a LD(50) of 12 +/- 8 mu M. In summary, we demonstrated that, in addition to host immune response enhancement, 16BrEA has a direct effect on parasite viability, most likely as a consequence of TcG6PDH inhibition. Crown Copyright (C) 2010 Published by Elsevier Ltd. All rights reserved.

The defensive functions of plant inhibitors are not restricted to insect enzyme inhibition

Three plant proteinase inhibitors BbKI (kallikrein inhibitor) and BbCI (cruzipain inhibitor) from Bauhinia bouhinioides, and a BrTI (trypsin inhibitor) from B. rufa, were examined for other effects in Callosobruchus maculatus development; of these only BrTI affected bruchid emergence. BrTI and BbKI share 81% identities in their primary sequences and the major differences between them are the regions comprising the RGD and RGE motifs in BrTI. These sequences were shown to be essential for BrTI insecticidal activity, since a modified BbKI [that is a recombinant form (BbKIm) with some amino acid residues replaced by those found in BrTI sequence] also strongly inhibited insect development. By using synthetic peptides related to the BrTI sequence, YLEAPVARGDGGLA-NH(2) (RGE) and IVYYPDRGETGL-NH(2) (RGE), it was found that the peptide with an RGE sequence was able to block normal development of C. maculatus larvae (ED(50) 0.16% and LD(50) 0.09%), this being even more effective than the native protein. (C) 2009 Elsevier Ltd. All rights reserved.

Immobilization of lutetium bisphthalocyanine in nanostructured biomimetic sensors using the LbL technique for phenol detection

This study describes the development of amperometric sensors based on poly(allylamine hydrochloride) (PAH) and lutetium bisphthalocyanine (LuPc(2)) films assembled using the Layer-by-Layer (LbL) technique. The films have been used as modified electrodes for catechol quantification. Electrochemical measurements have been employed to investigate the catalytic properties of the LuPc(2) immobilized in the LbL films. By chronoamperometry, the sensors present excellent sensitivity (20 nA mu M(-1)) in a wide linear range (R(2) = 0.994) up to 900 mu M and limit of detection (s/n = 3) of 37.5 x 10(-8) M for catechol. The sensors have good reproducibility and can be used at least for ten times. The work potential is +0.3 V vs. saturated calomel electrode (SCE). In voltammetry measurements, the calibration curve shows a good linearity (R(2) = 0.992) in the range of catechol up to 500 mu M with a sensitivity of 90 nA mu M(-1) and LD of 8 mu M. (C) 2011 Elsevier B.V. All rights reserved.

In vitro basal cytotoxicity assay applied to estimate acute oral systemic toxicity of grandisin and its major metabolite

Preclinical investigations can start with preliminary in vitro studies before using animal models. Following this approach, the number of animals used in preclinical acute toxicity testing can be reduced. In this study, we employed an in-house validated in vitro cytotoxicity test based on the Spielmann approach for toxicity evaluation of the lignan grandisin, a candidate anticancer agent, and its major metabolite. the 4-O-demethylgrandisin, by neutral red uptake (NRU) assay, on mouse fibroblasts Balb/c 3T3 cell line. Using different concentrations of grandisin and its major metabolite (2.31; 1.16; 0.58; 0.29; 0.14; 0.07; 0.04; 0.002 mu M) in Balb/c 3T3-A31 NRU cytotoxicity assay, after incubation for 48 h, we obtained IC(50) values for grandisin and its metabolite of 0.078 and 0.043 mu M, respectively. The computed LD(50) of grandisin and 4-O-demethylgrandisin were 617.72 and 429.95 mg/kg, respectively. Both were classified under the Globally Harmonized System as category 4. Since pharmacological and toxicological data are crucial in the developmental stages of drug discovery, using an in vitro assay we demonstrated that grandisin and its metabolite exhibit distinct toxicity profiles. Furthermore, results presented in this work can contribute to reduce the number of animals required in subsequent pharmacological/toxicological studies. (C) 2010 Elsevier GmbH. All rights reserved.

Multivariate Analysis for the Classification Differentiation of Brazilian Sugarcane Spirits by Analysis of Organic and Inorganic Compounds

Brazilian sugarcane spirits were analyzed to elucidate similarities and dissimilarities by principal component analysis. Nine aldehydes, six alcohols, and six metal cations were identified and quantified. Isobutanol (LD 202.9 mu gL-1), butiraldehyde (0.08-0.5 mu gL-1), ethanol (39-47% v/v), and copper (371-6068 mu gL-1) showed marked similarities, but the concentration levels of n-butanol (1.6-7.3 mu gL-1), sec-butanol (LD 89 mu gL-1), formaldehyde (0.1-0.74 mu gL-1), valeraldehyde (0.04-0.31 mu gL-1), iron (8.6-139.1 mu gL-1), and magnesium (LD 1149 mu gL-1) exhibited differences from samples.

Automated two-dimensional separation flow system with electrochemical preconcentration, stripping, capillary electrophoresis and contactless conductivity detection for trace metal ion analysis

This paper describes the automation of a fully electrochemical system for preconcentration, cleanup, separation and detection, comprising the hyphenation of a thin layer electrochemical flow cell with CE coupled with contactless conductivity detection (CE-C(4)D). Traces of heavy metal ions were extracted from the pulsed-flowing sample and accumulated on a glassy carbon working electrode by electroreduction for some minutes. Anodic stripping of the accumulated metals was synchronized with hydrodynamic injection into the capillary. The effect of the angle of the slant polished tip of the CE capillary and its orientation against the working electrode in the electrochemical preconcentration (EPC) flow cell and of the accumulation time were studied, aiming at maximum CE-C(4)D signal enhancement. After 6 min of EPC, enhancement factors close to 50 times were obtained for thallium, lead, cadmium and copper ions, and about 16 for zinc ions. Limits of detection below 25 nmol/L were estimated for all target analytes but zinc. A second separation dimension was added to the CE separation capabilities by staircase scanning of the potentiostatic deposition and/or stripping potentials of metal ions, as implemented with the EPC-CE-C(4)D flow system. A matrix exchange between the deposition and stripping steps, highly valuable for sample cleanup, can be straightforwardly programmed with the multi-pumping flow management system. The automated simultaneous determination of the traces of five accumulable heavy metals together with four non-accumulated alkaline and alkaline earth metals in a single run was demonstrated, to highlight the potentiality of the system.

Pharmacogenetics of Warfarin: Development of a Dosing Algorithm for Brazilian Patients

A dosing algorithm including genetic (VKORC1 and CYP2C9 genotypes) and nongenetic factors (age, weight, therapeutic indication, and cotreatment with amiodarone or simvastatin) explained 51% of the variance in stable weekly warfarin doses in 390 patients attending an anticoagulant clinic in a Brazilian public hospital. The VKORC1 3673G>A genotype was the most important predictor of warfarin dose, with a partial R(2) value of 23.9%. Replacing the VKORC1 3673G>A genotype with VKORC1 diplotype did not increase the algorithm`s predictive power. We suggest that three other single-nucleotide polymorphisms (SNPs) (5808T>G, 6853G>C, and 9041G>A) that are in strong linkage disequilibrium (LD) with 3673G>A would be equally good predictors of the warfarin dose requirement. The algorithm`s predictive power was similar across the self-identified ""race/color"" subsets. ""Race/color"" was not associated with stable warfarin dose in the multiple regression model, although the required warfarin dose was significantly lower (P = 0.006) in white (29 +/- 13 mg/week, n = 196) than in black patients (35 +/- 15 mg/week, n = 76).

Development of a new stir bar sorptive extraction coating and its application for the determination of six pesticides in sugarcane juice

In this article, a novel polydimethylsiloxane/activated carbon (PDMS-ACB) material is proposed as a new polymeric phase for stir bar sorptive extraction (SBSE). The PDMS-ACB stir bar, assembled using a simple Teflon (R)/glass capillary mold, demonstrated remarkable stability and resistance to organic solvents for more than 150 extractions. The SBSE bar has a diameter of 2.36 mm and a length of 2.2 cm and is prepared to contain 92 mu L of polymer coating. This new PDMS-ACB bar was evaluated for its ability to determine the quantity of pesticides in sugarcane juice samples by performing liquid desorption (LD) in 200 mu L of ethyl acetate and analyzing the solvent through gas chromatography coupled with mass spectrometry (GC-MS). A fractional factorial design was used to evaluate the main parameters involved in the extraction procedure. Then, a central composite design with a star configuration was used to optimize the significant extraction parameters. The method used demonstrated a limit of quantification (LOQ) of 0.5-40 mu g/L, depending on the analyte detected; the amount of recovery varied from 0.18 to 49.50%, and the intraday precision ranged from 0.072 to 8.40%. The method was used in the analysis of real sugarcane juice samples commercially available in local markets.

Experimental Chemotherapy against Trypanosoma cruzi Infection Using Ruthenium Nitric Oxide Donors

The ruthenium NO donors of the group trans-[Ru(NO)(NH(3))(4)L](n+), where the ligand (L) is N-heterocyclic H(2)O, SO(3)(2 -), or triethyl phosphite, are able to lyse Trypanosoma cruzi in vitro and in vivo. Using half-maximal (50%) inhibitory concentrations against bloodstream trypomastigotes (IC(50)(try)) and cytotoxicity data on mammalian V-79 cells (IC(50)(V79)), the in vitro therapeutic indices (TIs) (IC(50)(V79)/IC(50)(try)) for these compounds were calculated. Compounds that exhibited an in vitro TI of >= 10 and trypanocidal activity against both epimastigotes and trypomastigotes with an IC(50)(try/epi) of <= 100 mu M were assayed in a mouse model for acute Chagas` disease, using two different routes (intraperitoneal and oral) for drug administration. A dose-effect relationship was observed, and from that, the ideal dose of 400 nmol/kg of body weight for both trans-[Ru(NO)(NH(3))(4)isn](BF(4))(3) (isn, isonicotinamide) and trans-[Ru(NO)(NH3) 4imN](BF4) 3 (imN, imidazole) and median (50%) effective doses (ED50) of 86 and 190 nmol/kg, respectively, were then calculated. Since the 50% lethal doses (LD(50)) for both compounds are higher than 125 mu mol/kg, the in vivo TIs (LD(50)/ED(50)) of the compounds are 1,453 for trans-[Ru(NO)(NH(3))(4)isn](BF(4))(3) and 658 for trans-[Ru(NO)(NH(3))(4)imN](BF(4))(3). Although these compounds exhibit a marked trypanocidal activity and are able to react with cysteine, they exhibit very low activity in T. cruzi -glycosomal glyceraldehyde-3-phosphate dehydrogenase tests, suggesting that this enzyme is not their target. The trans-[Ru(NO)(NH(3))(4)isn](BF(4))(3) and trans-[Ru(NO)(NH(3))(4)imN](BF(4))(3) compounds are able to eliminate amastigote nests in myocardium tissue at 400-nmol/kg doses and ensure the survival of all infected mice, thus opening a novel set of therapies to try against trypanosomatids.

We did it together : Fathers´ experiences of planned home birth in the Nordic countries

Sammanfattning: Bakgrund: för många pappor kan det vara en av livets största och bästa stund att närvara vid när deras barn föds. Det finns studier som beskriver pappors upplevelse av sjukhusförlossning, däremot finns det få studier som beskriver pappors upplevelse av planerad hemförlossning. Syftet med den här studien är att beskriva pappans upplevelser och erfarenheter av planerad hemförlossning. Metod: i denna studie medverkar 105 pappor från de nordiska länderna som har deltagit i planerad hemförlossning mellan 2009-2011. Materialet från en öppen enkätfråga analyserades med hjälp av deduktiv ansats. Den öppna frågan löd ”beskriv gärna förlossningen med egna ord”. Resultat: papporna upplevde den planerade hemförlossningen som lugn och säker, mycket tack vare den professionella barnmorskan och den välbekanta miljön. Att få vara hemma med sin partner och om så önskades, sin familj var högt skattat. Födelseprocessen hemma beskrevs av papporna som att ”vi gjorde det tillsammans” och ”det var vår egen förlossning”. Papporna uttryckte delaktighet i förlossningsflödet. De kände också att de fick ett barn och var en del av en vacker förlossning full av kärlek. Konklusion: att välkomna ett barn hemma i en lugn miljö där paret känner sig trygga och ostörda, kan underlätta en positiv och meningsfull förlossningsupplevelse.

Parent’s experiences of feeding their preterm infant in Neonatal Units in Sweden. : Ethnographic design

Introduction: Studies have shown that having a preterm infant may cause stress and powerlessness for parents. It is important to support parents around the feeding situation, and that the Neonatal Intensive Care Unit (NICU) has appropriate space and place to help the family to bond to each other. For the healthcare professionals it is important to promote skin-to-skin contact and breastfeeding; particularly for preterm infants. There are many studies on parent’s experiences of NICUs and a few studies on parent’s experiences of feeding their infant in the NICU. Objective: The objective of this study was to explore parents experiences of feeding their infant in the NICU. Design: The study was conducted using an ethnographic design. Results: A global theme of ‘The journey in feeding’ was developed from four organising themes: ‘Ways of infant feeding’; ‘Environmental influences’; ‘Relationships’ and ‘Emotional factors’. These themes illustrate the challenges mothers reported with different methods of feeding. The environment had a big impact on parent’s experiences of infant feeding. Some mothers felt that breastfeeding seemed unnatural because their infant was so tiny but breastfeeding and skin-to-skin contact helped them to bond to their infant. The mothers thought it was difficult to keep up with the milk production by only pumping. Routines were not inviting parents to find their own rhythm. They also felt stressed about the weighing. Healthcare professionals had positive and negative influences on the parents. Conclusions: This study demonstrates that while all parents expressed the wish to breastfeed, their ‘journey in feeding’ was highly influenced by method of feeding, environmental, relational and emotional factors. The general focus upon routines and assessing milk intake generated anxiety and reduced relationality. Midwives and neonatal nurses need to ensure that they emphasise and support the relational aspects of parenting and avoid over-emphasising milk intake and associated progress of the infant

Hydrodynamic Simulations Of Vacuum-Driven Storage Tanks

As a result of urbanization, stormwater runoff flow rates and volumes are significantly increased due to increasing impervious land cover and the decreased availability of depression storage. Storage tanks are the basic devices to efficiently control the flow rate in drainage systems during wet weather. Presented in the paper conception of vacuum-driven detention tanks allows to increase the storage capacity by usage of space above the free surface water elevation at the inlet channel. Partial vacuum storage makes possible to gain cost savings by reduction of both the horizontal area of the detention tank and necessary depth of foundations. Simulation model of vacuum-driven storage tank has been developed to estimate potential profits of its application in urban drainage system. Although SWMM5 has no direct options for vacuum tanks an existing functions (i.e. control rules) have been used to reflect its operation phases. Rainfall data used in simulations were recorded at raingage in Czestochowa during years 2010÷2012 with time interval of 10minutes. Simulation results gives overview to practical operation and maintenance cost (energy demand) of vacuum driven storage tanks depending of the ratio: vacuum-driven volume to total storage capacity. The following conclusion can be drawn from this investigations: vacuum-driven storage tanks are characterized by uncomplicated construction and control systems, thus can be applied in newly developed as well as in the existing urban drainage systems. the application of vacuum in underground detention facilities makes possible to increase of the storage capacity of existing reservoirs by usage the space above the maximum depth. Possible increase of storage capacity can achieve even a few dozen percent at relatively low investment costs. vacuum driven storage tanks can be included in existing simulation software (i.e. SWMM) using options intended for pumping stations (including control and action rules ).

Optimization Of Groundwater Remediation With New Efficient Parallel Algorithm For Global Optimization

Application of optimization algorithm to PDE modeling groundwater remediation can greatly reduce remediation cost. However, groundwater remediation analysis requires a computational expensive simulation, therefore, effective parallel optimization could potentially greatly reduce computational expense. The optimization algorithm used in this research is Parallel Stochastic radial basis function. This is designed for global optimization of computationally expensive functions with multiple local optima and it does not require derivatives. In each iteration of the algorithm, an RBF is updated based on all the evaluated points in order to approximate expensive function. Then the new RBF surface is used to generate the next set of points, which will be distributed to multiple processors for evaluation. The criteria of selection of next function evaluation points are estimated function value and distance from all the points known. Algorithms created for serial computing are not necessarily efficient in parallel so Parallel Stochastic RBF is different algorithm from its serial ancestor. The application for two Groundwater Superfund Remediation sites, Umatilla Chemical Depot, and Former Blaine Naval Ammunition Depot. In the study, the formulation adopted treats pumping rates as decision variables in order to remove plume of contaminated groundwater. Groundwater flow and contamination transport is simulated with MODFLOW-MT3DMS. For both problems, computation takes a large amount of CPU time, especially for Blaine problem, which requires nearly fifty minutes for a simulation for a single set of decision variables. Thus, efficient algorithm and powerful computing resource are essential in both cases. The results are discussed in terms of parallel computing metrics i.e. speedup and efficiency. We find that with use of up to 24 parallel processors, the results of the parallel Stochastic RBF algorithm are excellent with speed up efficiencies close to or exceeding 100%.

Determinação de elementos traço em amostras ambientais por ICP OES

Neste trabalho é avaliada a aplicação da técnica de ICP OES (Espectrometria de Emissão com Plasma Indutivamente Acoplado) para a determinação de elementos traço em amostras ambientais. Foram investigados os comportamentos de As, Ba, Cd, Co, Cu, Cr, Mn, Ni, Pb, V e Zn, utilizando-se espectrômetro com vista de observação axial/radial do plasma e sistema de detecção baseado em dispositivos de carga acoplada (CCD). No presente estudo, foi avaliado o desempenho dos nebulizadores pneumáticos do tipo concêntrico (Meinhard), “cross flow” e GemCone® acoplados às câmaras de nebulização de duplo passo (Scott) e ciclônica, bem como do nebulizador ultra-sônico para a introdução de amostras no plasma. Investigou-se a robustez do plasma, potência de radiofreqüência (RF), vazão dos gases de nebulização e auxiliar, bem como a altura de observação, para ambas as vistas de observação do plasma. Sob condições otimizadas os limites de detecção (LD), em ng mL-1, para os elementos As, Ba, Cd, Co, Cr, Cu, Mn, Ni, Pb, V e Zn, em solução aquosa de HNO3 5% (v/v), utilizando-se a configuração axial do plasma, foram: 1,1 - 16; 0,002 - 0,32; 0,03 - 1,2; 0,02 - 0,72; 0,03 - 0,82; 0,04 - 3,0; 0,003 - 0,76; 0,08 - 3,8; 0,22 - 8,9; 0,04 - 2,6; e 0,02 - 1,2 respectivamente. Utilizando-se a configuração radial, os LDs (ng mL-1) dos mesmos elementos foram: 10 - 87; 0,01 - 0,91; 0,07 - 3,8; 0,16 - 4,3; 0,13 - 8,1; 0,16 - 4,3; 0,01 - 0,81; 0,43 - 7,6; 1,4 - 37; 0,28 - 6,0 e 0,77 - 9,5 respectivamente. Com relação à nebulização pneumática, os LDs são relativamente mais baixos quando é utilizado o nebulizador concêntrico acoplado à câmara de nebulização ciclônica. LDs ainda melhores são obtidos mediante o uso de nebulização ultra-sônica mas, neste caso, foi observado que o plasma é menos robusto. As metodologias foram desenvolvidas mediante o uso de materiais de referência certificados sendo analisados os seguintes materiais: sedimento marinho (PACS-2/NRCC), folhas de maçã (apple leaves – 1515/NIST) e água (natural water – 1640/NIST), obtendo-se concentrações concordantes com as certificadas, com exceção do Ni em folha de maçã, que não foi detectado utilizando-se a nebulização pneumática e também não pôde ser determinado com exatidão mediante o uso de nebulizador ultra-sônico. Interferências não espectrais observadas na análise de sedimento marinho foram contornadas através da diluição da amostra, ou através da lavagem da câmara de nebulização com solução de HNO3 5% (v/v), por 60 s entre cada ciclo de leitura. Estas interferências não puderam ser contornadas com o uso de padrão interno (PI). A interferência espectral do As sobre o Cd não foi observada fazendo-se a medição do sinal em área de pico, demarcado com somente 3 pontos/pico. Após serem estabelecidas as metodologias de análise, foram analisadas as seguintes amostras não certificadas: água de rio, água subterrânea (poço artesiano), água tratada e de chuva, folhas de eucalipto e bambu, acículas de pinus e infusão de chá preto, sendo possível quantificar baixas concentrações dos elementos investigados, utilizando-se a calibração externa.

Análise de diagramas cor-magnitude de 5 aglomerados ricos da LMC

Apresentamos a análise de diagramas cor-magnitude (CMDs) profundos para 5 aglomerados ricos da Grande Nuvem de Magalhães (LMC). Os dados fazem parte do projeto HST G07307, entitulado "Formação e Evolução de Aglomerados Estelares Ricos da LMC", e foram obtidos através do HST/WFPC2 nos filtros F555W (~ V) e F814W (~ I), alcançando V555~25. A amostra de aglomerados é composta por NGC 1805 e NGC 1818, os mais jovens (r < 100 Myr), NGC 1831e NGC 1868,de idades intermediárias (400 < r < 1000Myr), e Hodge 14, o mais velho (r > 1200Myr). Discutimos e apresentamos um método de correção dos CMDs para incompleteza da amostra e contaminação por estrelas de campo. O uso eficiente das informações contidas nos CMDs foi feito por meio de comparações entre os CMDs observados e CMDs modelados. O processo de modelamento de CMDs gera uma Seqüência Principal (MS) sintética, que utiliza como entrada do modelo a informação sobre idade (r), metalicidade (Z), Função de Massa do Presente (PDMF), fração de binárias não resolvidas, distância e extinção da luz. As incertezas fotométricas foram empiricamente determinadas a partir dos dados e incorporadas no modelamento. Técnicas estatísticas de comparação entre CMDs em 1D e 2D são apresentadas e aplicadas como métodos objetivos capazes de determinar a compatibilidade entre modelo e observação. Modelando os CMDs da região central dos aglomerados podemos inferir a metalicidade (Z), o módulo de distância intrínseco ((V - Mv)o) e o avermelhamento (E(B-V)) para cada aglomerado. Também determinamos as idades para os 3 aglomerados mais velhos de nossa amostra. Continuação) Através da comparação em 2D os valores encontrados foram os seguintes: para NGC 1805, Z = 0.007:1+-0.003, (V - Mv)o = 18.50:1+- 0.10, E(B - V) = 0.03:1+-0.01; para NGC 1818,Z = 0.005:1+-0.002, (V - Mv)o = 18.50:1+-0.15, E(B - V)~0.00; para NGC 1831, Z = 0.012:1+-0.002, log(r /yr) = 8.70 :I+-0.05, (V - Mv)o = 18.70:1+- 0.05, E(B - V)~ 0.00; para NGC 1868, Z = 0.008:1+-0.002,log(r/yr) =8.95:1+-0.05, (V - Mv)o = 18.70:1+- 0.05, E(B - V) ~0.00; para Hodge 14, Z = 0.008+-0.004, log(r/yr) = 9.23+-0.10, (V - Mv)o = 18.50+-0.15, E(B - V) = 0.02+- 0.02. Estes valores estão de acordo, dentro das incertezas, com os valores obtidos através da análise de linhas fiduciais (lD), o que agrega confiabilidade a estas determinações. Através da análise de CMDs em regiões concêntricas determinamos a dependência espacial da inclinação da PDMF (a) nos aglomerados mais ricos. Para tanto empregamos duas abordagens distintas para a determinação de a: 1) método tradicional em 1D via conversão direta de V555em massa; 2) método em 2D via modelmento de CMDs. Independente do método de análise, todos os aglomerados analisados demonstram claramente o efeito de segregação em massa. Além disso, NGC 1818, NGC 1831 e NGC 1868 apresentam uma possível perda seletiva de estrelas de baixa massa nas suas regiões mais externas.