Effect of Annatto on Micronuclei Induction by Direct and Indirect Mutagens in HepG2 Cells

Annatto (AN), a natural food colorant rich in carotenoids, has been reported as being an effective antioxidant, but little is known about its potential chemopreventive properties. In this Study, we evaluated the ability of AN to protect human hepatoma cells (HepG2) from micronucleus (MN) induction against three different mutagens: benzo(a)pyrene (B(a)P), doxorubicin (DXR), and methyl methanesulfonate (MMS). In an attempt to clarify the possible mechanism of anti mutagenicity of AN, three protocols of treatment were applied (pretreatment; simultaneous treatment, and post-treatment with AN following treatment with the mutagens). Also, cells exposed only to AN were assayed for cytotoxicity and mutagenicity. A dosage up to 10 mu g/ml of AN was devoid of mutagenic activity. Protective effects were seen on micronuclei induced by B(a)P and DXR using pre and simultaneous treatment, but AN had no significant effect on MN induction by MMS in any of the protocols. Our results also show that exposure of cells to concentrations of AN higher than 10 mu g/ml decreased cell viability. Taken together, our findings indicate that AN presents antimutagenic activity in vitro, but its protective effect is dependent on the mutagen and on type of treatment suggesting its potential use as a chemopreventive agent. Environ. Mol. Mutagen. 50:808-814, 2009. (C) 2009 Wiley-Liss, Inc.

Coding Strategies of Indirect Reported Speech in Typological Perspective

The purpose of this paper is to provide a cross-linguistic survey of the variation of coding strategies that are available for the grammatical distinction between direct and indirect speech representation with a particular focus on the expression of indirect reported speech. Cross-linguistic data from a sample of 42 languages will be provided to illustrate the range of available grammatical coding strategies.

Australian import managers' purchasing decision behaviour: An empirical study

Reports a pilot study of the relative importance of import decision variables as rated by Australian managers. A systematic sample qi 104 Australian managers representing different companies participated in the study. Australian importers rated product quality as the most important variable when importing products from overseas followed by long-term suppliers' dependability, product style/feature, price, and timely delivery. Australian managers who import consumer products find the domestic import duties and tariffs to be more important than did those who import industrial products. Larger volume importers regard the product brand name reputation to be more important than did those who import smaller volumes.

Aspects of the biology of the yabby Trypea australiensis (Dana) (Decapoda : Thalassinidea) and the potential of burrow counts as an indirect measure of population density

The thalassinidean shrimp Trypea australiensis (the yabby) commonly occurs on intertidal sandflats and subtidal regions of sheltered embayments and estuaries along the east coast of Australia and is harvested commercially and recreationally for use as bait by anglers. The potential for counts of burrow openings to provide a reliable indirect estimate of the abundance of yabbies was examined on intertidal sandflats on North Stradbroke Island (Queensland, Australia). The relationship between the number of burrow openings and the abundance of yabbies was generally poor and also varied significantly through time, casting doubt on previous estimates of abundance for this species based on unvalidated hole counts. Spatial and temporal variation in population density, the size at maturity and the reproductive period of the yabby were also assessed. Except for an initial peak in abundance as a result of recruitment, the density of yabbies was constant throughout the study but considerably less than that estimated from a previous study in the same area. Ovigerous females were recorded at 3 mm carapace length (CL) which is smaller than previously recorded for this species and thalassinideans in general. Small ovigerous females were found throughout the study, including the summer months, which is unusual for thalassinideans in the intertidal zone. It was hypothesised that in the intertidal zone, small female yabbies may be able to balance the metabolic demands of reproduction and respiration at higher temperatures than can larger females allowing them to reproduce in the warmer months.

Indirect certainty equivalents for the firm facing price and production uncertainty

Producer decisionmaking under uncertainty is characterized using indirect objective functions. The characterization is for the class of producers with continuous and nondecreasing preferences over stochastic incomes who face both price and production uncertainty. (C) 2002 Elsevier Science B.V. All rights reserved.

Investigating the gap between citizens` sustainability attitudes and food purchasing behaviour: empirical evidence from Brazilian pork consumers

Consumers worldwide are increasingly concerned with sustainable production and consumption. Recently, a comprehensive study ranked 17 countries in regard to their environmentally friendly behaviour among consumers. Brazil was one of the top countries in the list. Yet, several studies highlight significant differences between consumers` intentions to consume ethically, and their actual purchase behaviour: the so-called `Attitude-Behaviour Gap`. In developing countries, few studies have been conducted on this issue. The objective of this study is therefore to investigate the gap between citizens` sustainability-related attitudes and food purchasing behaviour using empirical data from Brazil. To this end, Brazilian citizens` attitudes towards pig production systems were mapped through conjoint analysis and their coexistence with relevant pork product-related purchasing behaviour of consumers was investigated through cluster analysis. The conjoint experiment was carried Out with empirical data collected from 475 respondents surveyed in the South and Center-West regions of Brazil. The results of the conjoint analysis were used for a subsequent cluster analysis in order to identify clusters of Brazilian citizens with diversified attitudes towards pig production systems, using socio-demographics, attitudes towards sustainability-related themes that are expected to influence the way they evaluate pig production systems, and consumption frequency of various pork products as clusters` background information. Three clusters were identified as `indifferent`, `environmental conscious` and `sustainability-oriented` citizens. Although attitudes towards environment and nature had indeed an influence on citizens` specific attitudes towards pig farming at the cluster level, the relationship between `citizenship` and consumption behaviour was found to be weak. This finding is similar to previous research conducted with European consumers: what people (in their role of citizens) think about pig production systems does not appear to significantly influence their pork consumption choices. Improvements in the integrated management of this chain would better meet consumers` sustainability-related expectations towards pig production systems.

Indirect Emissions from Biofuels: How Important?

A global biofuels program will lead to intense pressures on land supply and can increase greenhouse gas emissions from land-use changes. Using linked economic and terrestrial biogeochemistry models, we examined direct and indirect effects of possible land-use changes from an expanded global cellulosic bioenergy program on greenhouse gas emissions over the 21st century. Our model predicts that indirect land use will be responsible for substantially more carbon loss ( up to twice as much) than direct land use; however, because of predicted increases in fertilizer use, nitrous oxide emissions will be more important than carbon losses themselves in terms of warming potential. A global greenhouse gas emissions policy that protects forests and encourages best practices for nitrogen fertilizer use can dramatically reduce emissions associated with biofuels production.

Parametric change in the grammatical encoding of indirect objects in Brazilian Portuguese

This paper examines the syntax of indirect objects (IO) in Brazilian Portuguese (BP). Adopting a comparative perspective we propose that BP differs from European Portuguese (EP) in the grammatical encoding of IO. In EP ditransitive contexts, IO is found in two configurations - one projected by a (low) applicative head and another one involving a lexical/true preposition. We propose that the former property is contingent upon the presence of dative Case marking: namely, the morpheme `a` that introduces IO (a-DP), whose corresponding clitic pronoun is `lhe/lhes`. In contrast, important changes in the pronominal system, coupled with the increase in the use of the preposition `para` are taken as evidence for the loss of the low applicative construction in BP. Thus only the configuration with the lexical/true preposition is found in (Standard) BP. We argue that the innovative properties of IO in BP are due to the loss of the (3rd person) dative clitic and the preposition `a` as dative Case markers. Under this view, we further account for the realization of IO as a DP/weak pronoun, found in dialects of the central region of Brazil, which points to a similarity with the English Double Object Construction. Finally we show that the connection between the morphological expression of the dative Case and the expression of parameters supports a view of syntactic change according to which parametric variation is determined in the lexicon, in terms of the formal features of functional heads.

Using BCG, MPT-51 and Ag85 as antigens in an indirect ELISA for the diagnosis of bovine tuberculosis

This study evaluated the Mycobacterium tuberculosis protein antigen MPT-51, the trimeric antigen 85 (Ag85) complex, and Bacillus Calmette-Guerin (BCG) in an indirect ELISA to diagnose bovine tuberculosis (TB) from serum samples. Serum was collected from 208 intra-dermal tuberculin test (ITT)-positive and 54 ITT-negative animals from a region where bovine TB is endemic. Using the Ag85 and BCG antigens, the indirect ELISA was able to discriminate ITT-positive from ITT-negative animals. This level of discrimination was not achieved when using the MPT-51 antigen. The highest sensitivity (Se) and specificity (Sp) of the test was found when BCG was used (Se, 82%; Sp, 91%). Further work in different epidemiological settings and with larger numbers of animals will be required to validate these findings. (C) 2009 Elsevier Ltd. All rights reserved.

Indirect immune detection of ecdysone receptor (EcR) during the formation of DNA puffs in Bradysia hygida (Diptera, Sciaridae)

Gene amplification occurs in Bradysia hygida salivary glands, at the end of the fourth larval instar. The hormone 20-hydroxyecdysone (20E) triggers this process, which results in DNA puff formation. Amplified genes are activated in two distinct groups. The activity of the first group is dependent on high levels of 20E, while the second group needs low hormone levels. Consequently, the salivary glands of B. hygida constitute an interesting biological model to study how 20E, and its receptors, affect gene amplification and activity. We produced polyclonal antibodies against B. hygida EcR (BhEcR). In western blots a polypeptide of about 66 kDa was detected in salivary gland extracts. The antibodies were also used for indirect immune-localization of BhEcR in polytene chromosomes. RNA-polymerase II was also immune-detected. We did not detect the receptor in chromosome C where the first and second groups of DNA puffs form during DNA puff anlage formation, but it was present during puff expansion. During the active phase of both groups of DNA puffs, RNA polymerase II co-localized with BhEcR. After puff regression, these antigens were not detected. Apparently, EcR plays a direct role in the transcription of amplified genes, but its role in gene amplification remains enigmatic.

Harris-Benedict equation for critically ill patients: Are there differences with indirect calorimetry?

Purpose: The aim of this study was to compare the measured energy expenditure (EE) and the estimated basal EE (BEE) in critically ill patients. Materials and Methods: Seventeen patients from an intensive care unit were randomly evaluated. Indirect calorimetry was performed to calculate patient`s EE, and BEE was estimated by the Harris-Benedict formula. The metabolic state (EE/BEE x 100) was determined according to the following criteria: hypermetabolism, more than 130%; normal metabolism, between 90% and 130%; and hypometabolism, less than 90%. To determine the limits of agreement between EE and BEE, we performed a Bland-Altman analysis. Results: The average EE of patients was 6339 +/- 1119 kJ/d. Two patients were hypermetabolic (11.8%), 4 were hypometabolic (23.5%), and 11 normometabolic (64.7%). Bland-Altman analysis showed a mean of -126 +/- 2135 kJ/d for EE and BEE. Only one patient was outside the limits of agreement between the 2 methods (indirect calorimetry and Harris-Benedict). Conclusions: The calculation of energy needs can be done with the equation of Harris-Benedict associated with lower values of correction factors (approximately 10%) to avoid overfeeding, with constant monitoring of anthropometric and biochemical parameters to assess the nutritional changing and adjust the infusion of energy. (C) 2009 Elsevier Inc. All rights reserved.

Analysis of five polymorphic DNA markers for indirect genetic diagnosis of haemophilia A in the Brazilian population

Hemophilia A is an X-linked, inherited, bleeding disorder caused by the partial or total inactivity of the coagulation factor VIII (FVIII). Due to difficulties in the direct recognition of the disease-associated mutation in the F8 gene, indirect diagnosis using polymorphic markers located inside or close to the gene is used as an alternative for determining the segregation of the mutant gene within families and thus for detecting carrier individuals and/or assisting in prenatal diagnosis. This study characterizes the allelic and haplotype frequencies, genetic diversity, population differentiation and linkage disequilibrium of five microsatellites (F8Int1, F8Int13, F8Int22, F8Int25.3 and IKBKG) in samples of healthy individuals from Sao Paulo, Rio Grande do Sul and Pernambuco and of patients from Sao Paulo with haemophilia A to determine the degree of informativeness of these microsatellites for diagnostic purposes. The interpopulational diversity parameters highlight the differences among the analyzed population samples. Regional differences in allelic frequencies must be taken into account when conducting indirect diagnosis of haemophilia A. With the exception of IKBKG, all of the microsatellites presented high heterozygosity levels. Using the markers described, diagnosis was possible in 10 of 11 families. The F8Int22, F8Int1, F8Int13, F8Int25.3 and IKBKG microsatellites were informative in seven, six, five and two of the cases, respectively, demonstrating the effectiveness of using these microsatellites in prenatal diagnosis and in carrier identification in the Brazilian population.