The retrospective design is a significant bias concerning the dat

The retrospective design is a significant bias concerning the data collected, which can have an impact on results. Randomized studies are difficult to implement, given the small number of patients, acquisition of one specific technique per referral center, and the fact that the patients are often specifically referred for a selected technique of treatment. However, we think that the consistency of treatment Selleck Apitolisib over the whole study and the detailed

long-term clinical outcome allow us to provide data useful to clinical practice. In conclusion, flexible endoscopy has become a new standard for ZD treatment when performed with adequate equipment inspired by rigid diverticuloscopes. This approach has a lower rate of adverse events than open surgery or endoscopic stapling techniques. Moreover, the risk of general anesthesia in elderly patients can be avoided because airways are protected by the diverticuloscope. Recurrence

is about 25% in the long term but is easily amenable to successful repeated endoscopic treatment. “
“Zenker’s diverticulum (ZD) is an acquired disease that is formed by outpouching of hypopharyngeal mucosa between the inferior pharyngeal constrictor and the cricopharyngeus muscle in an area of junctional muscle weakness known as Killian’s triangle.1 Although the need for myotomy in addition to surgical correction of the diverticulum has been well described, it is only in the past 20 years that more clear insight has emerged on the pathophysiology of ZD despite its original description in the 1700s.1 Specifically, Cook et al2 elegantly demonstrated Cabozantinib molecular weight that in patients with ZD, the upper esophageal sphincter is fibrotic, contributing to reduced compliance, incomplete opening, and therefore increased pressure proximal to the cricopharyngeal Phosphoribosylglycinamide formyltransferase outlet. This leads to a “blow out” of the weakest part of the pharyngeal wall and formation of the diverticulum. Other studies have been conflicting in demonstrating

an increased tone within the sphincter.1 Nevertheless, these and other data have reinforced that cricopharyngeal myotomy is essential in relieving symptoms and preventing recurrent diverticula.1 The traditional approach to cricopharyngeal myotomy and to diverticulectomy or -pexy has been open through a lateral neck incision. In the past 2 decades, however, this operation has been shifting to a transoral endoscopic approach using a rigid endoscope because of equal efficacy, shorter hospital stay, faster return to oral intake, and lower morbidity because of a reduction in adverse events compared with open surgery.3 Such adverse events may include injury to the recurrent laryngeal nerve, mediastinitis, and fistula.1 Not all patients with ZD are amenable to transoral endoscopic therapy using a rigid endoscope.

Nevertheless nearly all amino acid residues that compose the basi

Nevertheless nearly all amino acid residues that compose the basic/aromatic and basic/hydroxyl clusters proposed as interaction PD0325901 datasheet surface of APETx2 with ASIC3 [16] and [25],

are conserved in U-AITX-Bg1c (see Fig. 5B). These are R17, R31, F15, Y16, Y32, F33 (basic/aromatic cluster), and S9, K10 (basic/hydroxyl cluster) in APETx2 (see Suppl. Fig. 1B), which are represented by R18, K19, Y15, W16, Y32, F33 (basic/aromatic cluster) and T9, K10 (basic/hydroxyl cluster) in U-AITX-Bg1c. Moreover, although R31 is absent in U-AITX-Bg1c it is worthy of mentioning that R36 is spatially near to R18 and K19; therefore it can be considered as part of the basic/aromatic cluster. Regarding APETx1, it has been proposed an interaction surface comprising the aromatic residues Y5, Y32, and F33, two basic residues, K8 and K18, and three aliphatic amino acids, G7, G31 and L34 [15]. More recently K18 and L34/F33/Y32 have been proposed to be involved in the interaction with hERG channel [86]. Among the new APETx-like peptides, U-AITX-Bg1d

is the closest to APETx1 regarding the conservation of all these amino acid residues, which are represented by W5, Y32, F33, K10, K17, G7, G31, and M34 (see Fig. 5B). Interestingly, as observed also in Fig. 5B, the other peptides U-AITX-Bg1a and 1b do not show positively charged amino acid residues located closely to R17 and R31 positions of APETx2. Those molecules only present a single K8, which is exposed together with F5 and W5 near the N-termini of U-AITX-Bg1a and 1b, respectively. In addition, the electrostatic potentials of such molecules ABT-888 solubility dmso vary a lot, and U-AITX-Bg1a and 1b are the less charged ones. On the contrary, U-AITX-Bg1c and 1e present the most dense positive surfaces. In Suppl. Fig. 1C and D we also depict the electrostatic potentials of APETx1, APETx2, BcIV and the putative new U-AITX-Ael1a.

Also, in the same Suppl. Fig. 1B the distribution of positively charged and aromatic residues in U-AITX-Ael1a suggests that such a peptide Farnesyltransferase may represent a “chimera” of contact surfaces of either APETx1 or APETx2. The crab bioassay is a simple test widely used for the detection of sea anemone toxins [6], [7], [8], [10], [35], [37], [38], [54], [73], [74], [75] and [80], mostly acting on sodium channels. Envenomed crabs exhibit a severe paralysis within seconds or few minutes after the injection of a sodium channel toxin. Reactions comprise an initial spastic and tetanic phase, and a later rigid phase followed by death of the crabs [80]. On the other hand, several sea anemone peptides belonging to other classes of toxins have been also discovered, through a careful observation of symptoms provoked on crabs [35], [37], [38] and [75]. In the present work we tested all fractions obtained by reversed-phase chromatography. In total, 23 toxic fractions (6 from S. helianthus and 17 from B. granulifera) were found ( Table 1).

Other analyses also showed that within good navigators there was

Other analyses also showed that within good navigators there was significantly better decoding of permanence in RSC compared with PHC (t15 = 1.82, p = .04), while for poor navigators there was no such regional difference (t15 = .045, p = .33; Fig. 4). We performed similar comparisons between good and poor navigators for size and visual salience. Mean classifier values: for size – RSC: good mean 49.3% SD 4.9; poor mean 49.8% SD 6.3; PHC: good mean 47.8% SD 3.4; poor mean 47.0% SD 2.6, and for visual salience – RSC: good mean 49.7% SD 4.5; poor mean 47.9% SD 4.5; PHC: good mean 48.7% SD 3.1; poor mean 47.7% SD

3.9. There were no differences between the two groups for either feature in RSC or PHC (all t ≤ 1.14, p > .26) or within each group (all t ≤ 1.92; p > .08). In a set of GSK2118436 in vitro control analyses, we also compared males and females for permanence, size and visual salience, in both RSC and PHC, but found no significant differences based upon sex. To summarise, there were no demographic,

cognitive or structural brain differences between the good and poor navigators. Neither were there any differences in decodable information in RSC and PHC about the size or visual salience of items in view. Furthermore, there was no difference in the ability to predict whether a majority or minority of viewed items were permanent based upon patterns of activity across voxels in PHC. The only difference between the two groups concerned the accuracy with which it was possible to predict whether stimuli containing Quizartinib clinical trial a majority or minority of permanent items were in view, with good navigators having significantly more information about the number of permanent items in view in their

RSC. In a previous fMRI study, we found that the RSC responded in a highly selective manner to only the most permanent items when stimuli were presented singly (Auger et al., 2012). Here we found that in a situation that was more akin to real life, with multiple items in view, the RSC coded for the specific number of permanent items contained in a visual array. Moreover, this effect was selective, and was not apparent for other item features such as size and visual salience. This detailed tracking of the amount of permanent items in view was echoed in the PHC, although the two brain structures diverged when participants were divided into good and poor navigators. There was no difference in the responsivity of the PHC between the two groups, while significantly better decoding of the number of permanent items in view was possible from patterns of activity in the RSC of good compared to poor navigators. Within good navigators, the RSC also facilitated significantly better prediction of landmark permanence than the PHC.

Many optimization methods have been proposed to solve inverse pro

Many optimization methods have been proposed to solve inverse problems, based on the estimation of deterministic and stochastic parameters. Among the deterministic methods, the Levenberg–Marquardt method has been used successfully in several

areas (Kanevce et al., 2005, Mejias et al., 2003 and Mendonça et al., 2005). Among the stochastic methods, the Differential click here Evolution method has been applied less frequently to inverse problems, but has been used by Kanevce et al., 2005, Mariani and Coelho, 2009a, Mariani and Coelho, 2009b, Mariani et al., 2008 and da Silva et al., 2009. Optimization methods for estimating parameter are used to estimate the diffusion coefficient as a function of the minimization of J by the sum of squared differences between the experimental moisture content and the moisture content computed with a diffusion model: equation(5) J=∑1n(Xexp−Xcomp)2where Xexp is the experimental moisture content and Xcomp is the computed moisture content. A set of moisture contents was obtained experimentally

at discrete times during the unsteady drying process. The literature uses different criteria to evaluate the quality of the fit obtained by the mathematical selleckchem model and optimization methods to simulate the experimental results. In this study, deviations between measured and simulated moisture content were calculated

using the coefficient of determination in successive trials, as follows: equation(6) R2=1−∑(Xexp−Xcomp)2∑(Xexp−X¯exp)2 The Differential Evolution and Levenberg–Marquardt methods were implemented. 3-oxoacyl-(acyl-carrier-protein) reductase Differential Evolution (DE) is an evolutionary algorithm proposed by Storn and Price (1995). Although DE shares similarities with other evolutionary algorithms (EA), it differs significantly in that the search process is guide based on information about the distance and direction of the current population. DE uses the differences between randomly selected vectors (individuals) as the source of random variations for a third vector (individual), referred to as the target vector. Trial solutions are generated by adding weighted difference vectors to the target vector. This process is referred to as the mutation operator in which the target vector is mutated. A crossover step is then applied to produce an offspring, which is only accepted if it improves the fitness of the parent individual. The basic DE algorithm is described in greater detail below with reference to the three evolution operators: mutation, crossover, and selection.

Here we brought together these distinct lines of research by exam

Here we brought together these distinct lines of research by examining properties of the STS in terms of selective response to social stimuli. Normal adult volunteers participated in an ‘audiovisual localiser’ scan during which they were stimulated with auditory, visual, or audiovisual stimuli of people or objects. We proposed, given that face-selective,

voice-selective and integrative regions are found within the STS, that in addition to areas preferring both faces and voices (i.e., ‘people-selective’ regions) there could also be audiovisual regions that are more sensitive to social stimuli, as compared to information from non-social categories, such as objects. We found check details that a restricted portion of the right pSTS was characterised by a conjunction of (1) an ‘integrative’ response, i.e., stronger response to audiovisual stimuli compared to visual and compared C59 wnt to auditory stimuli and (2) ‘people-selectivity’, i.e., preference for social stimuli irrespective of the modality (voice > objects; face > objects). Furthermore, a large region further extending down the trunk of the right STS was observed to be heteromodal: that is, this region was activated

by both faces and voices, but did not necessarily show integrative properties. Forty English-speaking participants (15 males and 25 females; mean age: 25 years ± 5 years) took part in the scan. All had self-reported normal or corrected vision and hearing. The ethical committee from the University of Glasgow approved the study.

All volunteers provided informed written consent before, and received payment for, participation. 24 people (12 males and 12 females) were video-recorded producing a variety of vocal expressions, both speech and non-speech (e.g., saying the word ‘had’, humming, SPTLC1 yawning). Recordings took place in the television studio at the Learning and Teaching Centre, Glasgow University, and participants were paid at the rate of £6 per hour. The participants were filmed under standard studio lighting conditions (standard tungsten light), and sat directly facing the camera, at a distance so that the whole face was in frame. Videos were recorded with 25 frames per second (40 msec per frame) using a Panasonic DVC Pro AJD 610 camera, fitted with a Fujiform A17 × 7.8 BERM-M28 lens, and transferred and edited using Adobe Premier Elements. Within the video recording, vocalisations were recorded with 16-bit resolution at a sampling frequency of 44,100 Hz. Under the same conditions, 24 moving objects producing sound were also filmed (e.g.

Niedobór limfocytów T o fenotypie CD4+ wydaje się być główną przy

Niedobór limfocytów T o fenotypie CD4+ wydaje się być główną przyczyną zaburzonej współpracy między nimi a limfocytami B, czego następstwem jest brak przełączania klas immunoglobulin this website z IgM na IgG, IgA i IgE, prowadzący do głębokiej hipogammaglobu-linemii, z obecną u niektórych

chorych niewielką produkcją IgM [1]. Za SCID przemawia brak cienia grasicy w badaniu RTG klatki piersiowej i mała jej objętość w badaniu ultrasonograficznym [6, 11]. Hipotrofii obwodowych narządów limfatycznych nie stwierdzamy jedynie u chorych z zespołem Omenna (Ryc. 4) (T+B-NK+SCID), który swym odmiennym, na tle reszty SCID, przebiegiem klinicznym przypomina chorobę przeszczep-przeciw-gospodarzowi. Chorzy z tym zespołem wykazują uogólnioną erytrodermię złusz-czającą, hepatosplenomegalię, a obok hipogamma-globulinemii w zakresie IgG, IgA i IgM, podwyższone stężenie IgE i wysoką eozynofilię. Ta niezwykła konstelacja

objawów i zaburzeń laboratoryjnych jest następstwem oligoklonalnej proliferacji limfocytów Th2 [6, 11, 14]. Leczenie chorych z PNO polega głównie na leczeniu występujących zakażeń. Zwykle pacjenci wymagają stosowania wielu różnych antybiotyków, leków przeciwgrzybiczych i przeciwwirusowych. Stosuje się przedłużone leczenie, niejednokrotnie przez wiele tygodni, nawet miesięcy. Wielokrotnie chorzy z PNO wymagają leczenia w warunkach szpitalnych i stosowania leków drogą dożylną. Drugim bardzo ważnym, a może najważniejszym postępowaniem u chorych z PNO jest profilaktyka zakażeń. Przede wszystkim należy pamiętać o przestrzeganiu zasad higieny, toalecie jamy ustnej, odpowiednim odżywieniu i suplementacji witaminami. Każdy pacjent z PNO powinien unikać niepotrzebnego kontaktu ze źródłami zakażenia. Zaleca się unikanie kontaktu z osobami Thiamine-diphosphate kinase chorym. Nie należy pić ze wspólnych

naczyń, wodę – tylko butelkowaną. W niektórych przypadkach wskazane będzie indywidualne nauczanie. W uzasadnionych przypadkach lekarz może zlecić profilaktykę antybiotykową – zwykle stosuje się amoxycylinę w dawce 20 mg/kg m.c./dzień lub azi-tromycynę 1/2 dawki leczniczej 3x w tygodniu [19]. U chorych z deficytem limfocytów T i wysokim ryzykiem zakażenia Pneumocystis jiroveci zaleca się profilaktyczne przyjmowanie kotrimoksazolu w dawce 18-20 mg/kg m.c./dzień. Leczeniem z wyboru u pacjentów z niedoborem przeciwciał jest substytucja immunoglobulinami (Ig). Pierwsze preparaty Ig podawane były domięśniowo, następnie dożylnie, a w ostatnich latach rozpowszechnia się droga podawania podskórnego. Lecze preparatami podskórnymi ma wiele zalet, jest stosowane w domu pacjenta, pozwala na utrzymanie wyższych stężeń IgG przy takiej samej sumarycznej dawce miesięcznej, a koszty takiego leczenia są niż niż preparatów dożylnych. Chorzy z niedoborem przeciwciał IgG wymagają leczenia przez całe życie. Ig wytwarzane są z osocza tysięcy zdrowych dawców, dlatego zawierają IgG skierowane przeciwko różnym typom mikroorganizmów. Preparaty Ig składają się głównie z cząsteczki IgG.

Site specific management actions are also required for controllin

Site specific management actions are also required for controlling specific human impacts and livelihood activities and for adapting to the impacts of broader environmental changes. Also consistent with the literature on good governance and development processes, writings on MPA management emphasize the importance of adopting integrated or nested, integrative,

adaptive, transparent, and participatory management processes. To be effective in achieving their potential, MPAs should not be “islands of protection” but nested within Integrated Coastal Zone Management (ICZM) or Ecoystem-Based Management (EBM) regimes [4], [11], [190], [191] and [192] ABT-199 molecular weight and/or broader networks of MPAs [51], [143] and [193]. Both ICZM and EBM imply the incorporation of social, economic, cultural, political, and environmental considerations or values at the level of the broader land and seascape into management. For PLX3397 example, coral reef MPAs might be more resilient to the impacts of climate change when combined with the reduction of sedimentation and nutrient loading

and land-based and marine sources of pollution [34]. Networks can improve dispersal and connectivity between MPAs as well as spreading risks through replication of habitats and ecosystems [194] and [195]. Horigue et al. [136] also notes that “scaling up MPAs to form networks is a means to improve management of individual MPAs, and coordinate MPA establishment through collective action and sharing of information and experiences”. Additionally, MPAs can be more effective in supporting fisheries if they are nested within a suite of fisheries management actions outside the boundaries of the MPA [45], [48], [73], [196] and [197]. Active implementation of adaptive management – that

is a deliberate cycle of monitoring, evaluation, analysis, planning, and implementation – can serve to continually correct the course of MPA management strategies [24], Mannose-binding protein-associated serine protease [101], [122] and [198]. Adaptive management reflects a shift away from a linear view of the world and recognizes that MPAs are part of a dynamic, non-linear, and complex system [199]. Integrative research stemming from various social and natural science methods and tools in combination with local and traditional knowledge should also inform both broader integration and adaptive management frameworks [40], [45], [53], [73], [79], [122], [143] and [144]. Drew [200], for example, reviews various examples of how folk taxonomy and systematics and local knowledge of populations and ecological relationships can be used to augment western science in MPA management. Finally, there is widespread consensus that meaningful participation in decision-making and inclusion of relevant stakeholders are a necessary pre-cursor to effective management [94] and [122].

The multiple targets of action of quercetin, luteolin and fisetin

The multiple targets of action of quercetin, luteolin and fisetin, make these compounds candidates for drug design against leishmaniasis. Future research could determine whether fisetin, luteolin and quercetin can be used as a lead or prototype drug with multiple targets for the treatment of leishmaniasis. In conclusion, the in vitro and in silico study of these compounds can facilitate rational drug design and the Paclitaxel molecular weight development of new, safer drugs to treat leishmaniasis, using arginase as a

drug target. Moreover, the low IC50 values observed here may lead to the use of flavonoids as dietary supplements for leishmaniasis patients. This research was supported by FAPESP (Fundação de Amparo à Pesquisa do Estado de São Paulo Proc. 2009/08715-3 and Proc. 2012/17059-5). L.C.M. and M.B.G.R. received fellowships from CNPq and FAPESP, respectively. “
“Meat find more consumption from some land-based animals has come under attack due to unclear status regarding many diseases. Colon cancer is among these diseases, and it is one of the major causes of death in western countries (Sesink, Termont, Kleibeuker, & Van der Meer, 1999). It has been recognised that many genetic factors are involved as determinants of colorectal cancer (Fearon

& Jones, 1992), but environmental factors have appeared to contribute to the incidences of colon cancer (MacLennan, 1997). The World Cancer Research Fund panel has judged that the evidence of red meat and processed meat being a cause of colon cancer is convincing (WCRF, 2007), and a western style diet with a high red meat consumption is suggested as a risk factor for colon cancer (Sesink et al., 1999). Increased consumption of meat

can be due to improved efficiency in agriculture, which has then created sufficient amounts of relatively cheap meat products. Animal breeding has so far given most priority to rapid animal growth and cost-effective feeds. But meat should Liothyronine Sodium also have a good oxidative and microbial shelf life. Sufficient oxidative stabilization is paramount for meat flavour. A present understatement is that oxidised food can be consumed as long as the microbiology and sensory quality are acceptable to consumers. Compounds that could increase the genetic instability of colon cells and the appearance of cancer have received much attention (Ferguson, 2010). Lipid or lipid-derived peroxides are a major source of dietary pro-oxidants speculated to be of toxicological importance (Halliwell & Chirico, 1993). An in vitro study on intake of fat and derived peroxides has identified this as one of many important factors in colon cancer ( Angeli et al. 2011). Lipid peroxides are set with an acceptable upper level of 5–10 mmol/kg in oil or fat ( Sattar & Deman, 1976). Peroxide limits are normally not defined for products other than oil/fats. However, it is more common to eat larger amounts of lean meat than of pure oil/fats in a meal.

Therefore, there is great interest in increasing the amounts of i

Therefore, there is great interest in increasing the amounts of isoflavone aglycones in soy products mainly because,

naturally, most of the isoflavones exist in the glycosylated forms (Cheng, Wu, Lin, & Liu, 2013). The enzymatic processing of isoflavone glycosides in soybean products using isolated β-glucosidases has proved to be effective in increasing the concentration of isoflavone aglycones (Horri et al., 2009, Xue et al., 2009 and Yang et al., 2009). It has previously been demonstrated that the use of the immobilised microorganism cells containing enzymes of interest in bioconversion processes is advantageous when compared to the selleck screening library use of the purified enzymes, since the purification step is not necessary and enzymatic stability is higher (Junior et al., 2009). Debaryomyces hansenii is the most common yeast species in protein-rich fermented food, where this

species metabolises organic acids and amino acids to regulate the acidity, and also PLX3397 in vitro provides lipolytic and proteolytic activities that contribute to flavor development; the potential of D. hansenii UFV-1 to produce hydrolytic enzymes, specially α-galactosidases, has previously been explored ( Viana et al., 2007). The present study reports the purification and characterisation of an intracellular β-glucosidase from D. hansenii UFV-1, the immobilisation of D. hansenii cells in calcium alginate, and the application of the free and immobilised enzymes for the hydrolysis of isoflavone glucosides in soy molasses. The yeast strain used in this study Mannose-binding protein-associated serine protease was isolated from a dairy environment

in Minas Gerais, Brazil, and maintained in the culture collection of the Laboratory of Microorganism Physiology, BIOAGRO, Federal University of Viçosa (UFV), Brazil. The yeast was identified by the Centraalbureau voor Schimmelcultures, Utrecht, The Netherlands, as D. hansenii (Zopf) Lodder & Kreger-van Rij var fabryi Nakase & Suzuki. In this study it is designated as D. hansenii UFV-1. A stock culture of D. hansenii UFV-1 was maintained at −80 °C in glycerol and YPD medium (1% yeast extract, 2% peptone and 2% glucose). D. hansenii UFV-1 was streaked on an YPD agar surface (1.5% agar) and maintained in an incubation chamber at 30 °C for 36 h. The yeast was then activated in YPD liquid medium and incubated for 12–15 h, 180 rpm at 28 °C. The cells obtained after centrifugation (5000g for 5 min at 4 °C) were inoculated in an YP medium (1% yeast extract, 2% peptone) containing cellobiose, glucose, maltose or cellulose (1%) as the carbon source. After incubation at 28 °C, 180 rpm, for 12, 24, 36 and 48 h, the supernatant was separated by centrifugation (15,000g for 20 min at 4 °C) and the biomass was utilised as a source of the intracellular enzyme.

, located in Atibaia, São Paulo, Brazil The fruits were grown by

, located in Atibaia, São Paulo, Brazil. The fruits were grown by organic and conventional farming in the same geographic region (Atibaia, São Paulo, Brazil) under the same climatic conditions and were collected randomly during the harvest season of each fruit throughout 2007. The organic

fruits had a certificate issued by the Motika Okada Certification (CMO) service. The fruits were harvested in the partially ripe stage (stage of commercialisation) and properly stored MEK inhibitor drugs in cardboard boxes protected against shock. The fruits were transported overland and arrived at the Laboratory of Vitamin Analysis, Department of Nutrition, Federal University of Viçosa, Minas Gerais, Brazil, within 48 h post-harvest. A completely randomised design consisting of two treatments (organic and conventional production system) and six repetitions Dinaciclib chemical structure per treatment was used. The samples were collected randomly during the harvest season of each fruit. The organic and conventional fruits were collected in such

a way to obtain six different repetitions. The production area was divided into six small plots. In each plot, 2 kg of persimmons and 1 kg of acerola and strawberries produced by organic and conventional farming were collected. The six repetitions were sent to the laboratory in a single step, corresponding to 12 kg of persimmons and 6 kg of acerola and strawberries per treatment. After receiving the fruits, each repetition was subdivided into two parts for sample preparation. One half was used for analysis of vitamin C on the same day and was therefore stored

at room temperature. The other half was stored in a refrigerator at approximately 10 °C for sample preparation and analysis of carotenoids on the next day. Persimmons, acerola and strawberries were washed under running water and the non-edible parts (acerola seeds and leaves of persimmon and strawberry) were removed. The fruits were then chopped and homogenised in a multi-purpose food processor for 5 min until complete homogenisation of the sample, thus guaranteeing more reliable sampling. This procedure was performed six times for Edoxaban each treatment (organic and conventional farming). Vitamin C was extracted from the fruits according to the method of Campos et al. (2009). The previously homogenised sample was weighed (about 1 g) and 15 ml extraction solution (3% metaphosphoric acid, 8% acetic acid, 0.3 N sulfuric acid and 1 mM EDTA) was added. Next, the sample was triturated in a micro-homogenizer for 5 min and vacuum filtered through filter paper. The filtrate was diluted in ultrapure water until a volume of 25 ml in a volume balloon and centrifuged at 1789g for 15 min. The supernatant was stored in a refrigerator at about 5 °C until the time for chromatographic analysis. Carotenoids were extracted as described by Rodriguez-Amaya, Raymundo, Lee, Simpson, and Chichester (1976), with some modifications.