Modified 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (abts) method to measure antioxidant capacity of Selected small fruits and comparison to ferric reducing antioxidant power (FRAP) and 2,2'-diphenyl-1-picrylhydrazyl (DPPH) methods

The measurement of antioxidant capacity in fruits differs from that of other biological samples due to their low pH and very low lipophilic antioxidant capacity. In this report, we present a modified 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) method for fruits and compare its performance with the other commonly used antioxidant methods of 2,2'-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP). The antioxidant capacity and reaction kinetics of four phenolic compounds, two antioxidant standards, and five fruits were also investigated. The modified ABTS method prepared at a pH of 4.5 with sodium acetate buffer is highly stable and easily applied to fruit samples as compared to the standard (pH 7.4) version. The measured antioxidant capacity of samples varied with the assay method used, pH, and time of reaction. Traditional antioxidant standards (trolox, ascorbic acid) displayed stable, simple reaction kinetics, which allowed end point analysis with all of assays. Of the phenolic compounds examined, chlorogenic and caffeic acids exhibited the most complex reaction kinetics and reaction rates that precluded end point analysis while gallic acid and quercetin reached stable end points. All fruit extracts exhibited complex and varied kinetics and required long reaction times to approach an end point. Because the antioxidant capacity of fruit extracts is a function of the array of individual antioxidants present, accurate comparisons among fruit samples require that reaction times be standardized and of sufficient length to reach steady state conditions and that more than one assay be used to describe the total antioxidant activity of fruit samples.     

Effect of bread baking on the bioavailability of hydrogen-reduced iron powder added to unenriched refined wheat flour

Elemental iron powders are widely used to fortify flour and other cereal products. Our objective was to test the hypothesis that baking enhances the bioavailability of elemental iron powders by oxidizing Fe(0) to Fe(2+) or Fe(3+). An in vitro digestion/Caco-2 cell culture model and a piglet model were used to measure bioavailability. Bread flour, either unfortified or fortified with hydrogen-reduced (HR) iron powder or FeSO(4) (300 mg Fe/kg flour), was baked into bread. For the in vitro studies, bread samples were treated with pepsin at pH 2, 3, 4, 5, 6, or 7 and subsequently incubated with pancreatic enzymes at pH 7 in a chamber positioned above monolayers of cultured Caco-2 cells. Ferritin formation in the cells was used as an index of iron bioavailability. Ferritin formation in cells fed HR Fe bread was similar to cells fed FeSO(4) bread when the peptic digestion was conducted at a pH 2 but lower when the peptic phase was conducted at pH 3, 4, 5, 6, or 7 (P < 0.05). Pig diets containing 35% dried bread were prepared and fed to cross-bred (Hampshire x Landrace x Yorkshire) anemic pigs in two studies. The rate of increase in hemoglobin Fe over the feeding period was used to calculate relative biological value (RBV), an index of iron bioavailability. In the first pig study, RBV of HR Fe added to flour prior to baking was 47.9% when compared to FeSO(4) fortified flour (P < 0.05). In the second pig study, a third treatment consisting of unfortified bread with HR iron added during diet mixing (after bread baking) was included. RBVs of the HR Fe diet (Fe added after baking) and HR Fe diet (Fe added before baking) were 40.1% and 53.5%, respectively, compared to the FeSO(4) diet. Differences in RBV between the HR Fe (before and after baking) and FeSO(4) (before baking) treatment groups were significant, but the difference between the before and after HR treatment groups was not significant. We conclude that bread baking does not enhance the bioavailability of elemental iron powders.     

Variable response of open-pollinated seedling progeny of avocado to Phytophthora root rot

Phytophthora root rot, caused byPhytophthora cinnamomi Rands, is the most important disease of avocado (Persea americana Miller). In an attempt to identify root rot-resistant rootstocks that could ultimately be used under conditions in southern Florida, we screened open-pollinated progeny of avocado from the National Germplasm Repository in Miami. From 1996 to 1998, a total of 2,355 seedlings from 51 accessions were examined in potting mix artificially infested withP. cinnamomi. Most seedlings developed severe root rot, but tolerance was observed in some families (i.e., progeny of certain accessions). Although the most susceptible families developed mean disease ratings of up to 97% root necrosis, mean ratings for the most tolerant families were less than 60%. There was also a strong relationship between the racial background of the female parent and the tolerance of seedlings. Seedlings of the West Indian race and hybrids between it and the Guatemalan race were significantly more tolerant than those from other parents (P< 0.05). Individuals in several families developed < 50% root necrosis, the arbitrary standard of tolerance in this study. Twelve families accounted for 82% (188 of 229) of the tolerant seedlings, and only two of these did not have a West Indian or Guatemalan × West Indian pedigree. Broad-sense heritability for PRR tolerance was 0.45. This is the first report on the inheritance of PRR tolerance in avocado and on the influence of genotype and racial pedigree under controlled conditions.     

Optimization of a Staphylococcus aureus adhesion assay for equine corneocytes

Meticillin-resistant Staphylococcus aureus (MRSA) causes serious skin and soft-tissue infections of humans and animals. Multiple strains of MRSA have been characterized, and one in particular, designated as strain USA 500, causes infections predominantly of horses and the people who work with them. The purpose of this study was to optimize an assay which could subsequently be used to compare the relative avidity of different S. aureus strains for equine corneocytes. Corneocytes were collected from the perineal skin of 10 healthy horses onto adhesive discs. The discs were then incubated at 37°C with an S. aureus field strain at each of three concentrations [10(7), 10(8) and 10(9) colony forming units (CFU)/mL] and for each of three incubation periods (45, 90 and 180 min). After standardized rinsing and staining procedures, discs were examined at ×1,000 magnification and areas containing confluent corneocytes photographed. The percentage of surface area occupied by adherent bacteria was analysed using image processing and analysis software. Significant colour space image processing was required to distinguish bacteria from the ubiquitous melanin granules present within equine corneocytes. Objective and subjective methods were used to determine optimal conditions for specific adherence without introducing confounding factors. A bacterial concentration of 10(8) CFU/mL incubated with corneocytes for 45 min produced maximal bacterial adhesion with the least amount of interbacterial clumping. Future studies should utilize these conditions for optimal assay performance.     

Lipoprotein patterns in acrylamide gel electrophoresis

Lipoproteins of serum stained before electrophoresis gave distinct patterns in acrylamide gel of 3-percent concentration. Samples compared in parallel on gel slabs showed qualitative differences in migration rates of beta-lipoprotein bands.     

Peptides isolated from in vitro digests of milk enhance iron uptake by caco-2 cells

Milk proteins, during digestion, produce a range of biologically active peptides. Among those are peptides that may enhance iron absorption. The objective of this project was to investigate the effect of isolated milk peptides on iron uptake. Cow's milk, 0% fat, was subjected to a modified in vitro digestion process. The milk digest was further fractionated by gel filtration. All eluted fractions as well as beta-casein synthetic peptides (a tripeptide and a hexapeptide) were subsequently tested for effects on iron uptake with Caco-2 cell monolayers. Fractions of milk digests obtained through Sephadex G-25 gel filtration had a significant enhancing effect on iron uptake in Caco-2 cells compared to nonfractionated milk digests. Two fractions (P = 0) and the hexapeptide (P < 0.0001) enhanced iron uptake by up to 3-fold, whereas others and the tripeptide had no effect. These results suggest that selected peptides produced during the in vitro digestion of milk may enhance iron absorption; however, it remains to be demonstrated whether this effect may be nutritionally significant.     

Molecular identification and functional expression of porcine Toll-like receptor (TLR) 3 and TLR7

To investigate porcine Toll-like receptors (TLR) responding to viral pathogen associated molecular patterns, the full-length cDNA of porcine TLR3 and TLR7 were identified and characterized. Porcine TLR3 and TLR7 cDNA encode 904- and 1050-amnio-acid polypeptides, respectively. Both porcine TLR3 and TLR7 contain typical functional TLR domains and share about 80% sequence identity to other mammalian orthologues. Tissue expression profiles showed that TLR3 was highly expressed in kidney, duodenum, spleen and liver, and moderately expressed in bone marrow, lung, and skin. Conversely, TLR7 was moderately and constitutively expressed in all tissues evaluated. Stimulation of mammalian cells transfected with porcine TLR3 and TLR7 constructs with TLR3 and TLR7 agonists [poly (I:C) and imiquimod (R837), respectively], and adenovirus elicited activation of interferon regulatory factors (IRFs). These data provide molecular and functional information for porcine TLR3 and TLR7, and implicate their role in mediating immune protection against porcine viral diseases.     

How farmers matter in shaping agricultural technologies: social and structural characteristics of wheat growers and wheat varieties

Science and technology studies (STS) research challenges the concept of technological determinism by investigating how the end users of a technology influence that technology’s trajectory. STS critiques of determinism are needed in studies of agricultural technology. However, we contend that focusing on the agency of end users may mask the role of political-economic factors which influence technology developments and applications. This paper seeks to mesh STS insights with political-economic perspectives by accounting for relationships between availability of diverse technologies, variations in political-economic structures, and farmer interests and characteristics. We present the results of an analysis on the recent development of three wheat varieties: (a) a wheat variety that was modified genetically to tolerate the herbicide glyphosate, (b) wheat varieties with characteristics selected to serve specific markets, (c) and emerging research and development of perennial wheat varieties. Using data obtained through a survey of wheat growers in Washington State, we analyzed whether farmer interest in these three clusters of wheat varieties was associated with distinct individual characteristics and attitudes and whether those characteristics and attitudes are consistent with political economic structures. Although our analysis did not allow us to assess the degree of direct influence that farmers have on the technological development trajectory for these types of wheat, we were able to document variation in technological alternatives and farmer characteristics related to different political-economic trends.     

Anterior cingulate conflict monitoring and adjustments in control

Conflict monitoring by the anterior cingulate cortex (ACC) has been posited to signal a need for greater cognitive control, producing neural and behavioral adjustments. However, the very occurrence of behavioral adjustments after conflict has been questioned, along with suggestions that there is no direct evidence of ACC conflict-related activity predicting subsequent neural or behavioral adjustments in control. Using the Stroop color-naming task and controlling for repetition effects, we demonstrate that ACC conflict-related activity predicts both greater prefrontal cortex activity and adjustments in behavior, supporting a role of ACC conflict monitoring in the engagement of cognitive control.