The Role of the Proteinase Inhibitor Ovorubin in Apple Snail Eggs Resembles Plant Embryo Defense against Predation

BACKGROUND: Fieldwork has thoroughly established that most eggs are intensely predated. Among the few exceptions are the aerial egg clutches from the aquatic snail Pomacea canaliculata which have virtually no predators. Its defenses are advertised by the pigmented ovorubin perivitellin providing a conspicuous reddish coloration. The nature of the defense however, was not clear, except for a screening for defenses that identified a neurotoxic perivitellin with lethal effect on rodents. Ovorubin is a proteinase inhibitor (PI) whose role to protect against pathogens was taken for granted, according to the prevailing assumption. Through biochemical, biophysical and feeding experiments we studied the proteinase inhibitor function of ovorubin in egg defenses. METHODOLOGY/PRINCIPAL FINDINGS: Mass spectrometry sequencing indicated ovorubin belongs to the Kunitz-type serine proteinase inhibitor family. It specifically binds trypsin as determined by small angle X-ray scattering (SAXS) and cross-linking studies but, in contrast to the classical assumption, it does not prevent bacterial growth. Ovorubin was found extremely resistant to in vitro gastrointestinal proteolysis. Moreover feeding studies showed that ovorubin ingestion diminishes growth rate in rats indicating that this highly stable PI is capable of surviving passage through the gastrointestinal tract in a biologically active form. CONCLUSIONS: To our knowledge, this is the first direct evidence of the interaction of an egg PI with a digestive protease of potential predators, limiting predator's ability to digest egg nutrients. This role has not been reported in the animal kingdom but it is similar to plant defenses against herbivory. Further, this would be the only defense model with no trade-offs between conspicuousness and noxiousness by encoding into the same molecule both the aposematic warning signal and an antinutritive/antidigestive defense. These defenses, combined with a neurotoxin and probably unpalatable factors would explain the near absence of predators, opening new perspectives in the study of the evolution and ecology of egg defensive strategies

Saturated Fats: A Perspective from Lactation and Milk Composition

For recommendations of specific targets for the absolute amount of saturated fat intake, we need to know what dietary intake is most appropriate? Changing agricultural production and processing to lower the relative quantities of macronutrients requires years to accomplish. Changes can have unintended consequences on diets and the health of subsets of the population. Hence, what are the appropriate absolute amounts of saturated fat in our diets? Is the scientific evidence consistent with an optimal intake of zero? If not, is it also possible that a finite intake of saturated fats is beneficial to overall health, at least to a subset of the population? Conclusive evidence from prospective human trials is not available, hence other sources of information must be considered. One approach is to examine the evolution of lactation, and the composition of milks that developed through millennia of natural selective pressure and natural selection processes. Mammalian milks, including human milk, contain 50% of their total fatty acids as saturated fatty acids. The biochemical formation of a single double bond converting a saturated to a monounsaturated fatty acid is a pathway that exists in all eukaryotic organisms and is active within the mammary gland. In the face of selective pressure, mammary lipid synthesis in all mammals continues to release a significant content of saturated fatty acids into milk. Is it possible that evolution of the mammary gland reveals benefits to saturated fatty acids that current recommendations do not consider

A highly stable, nondigestible lectin from<i>Pomacea diffusa</i>unveils clade-related protection systems in apple snail eggs

AbstractThe acquisition of egg protection is vital for species survival. Poisonous eggs fromPomaceaapple snails have defensive macromolecules for protection. Here we isolated and characterized a novel lectin called PdPV1 that is massively accumulated in the eggs ofPomacea diffusaand seems part of its protective cocktail. The native protein, an oligomer of ca. 256 kDa, has high structural stability, withstanding 15 min boiling and denaturing by sodium dodecyl sulphate. It resistsin vitroproteinase digestion and displays structural stability between pH 2.0–12.0 and up to 85 °C. These properties, as well as its subunit sequences, glycosylation pattern, presence of carotenoids, size, and global shape resemble those of its orthologs from otherPomacea. Further, like members of thecanaliculataclade, PdPV1 is recovered unchanged in faeces of mice ingesting it, supporting an antinutritive defensive function. PdPV1 also displays a strong hemagglutinating activity specifically recognizing selected ganglioside motifs with high affinity. This activity is only shared with PsSC, a perivitelline from the same clade (bridgesiiclade). As a whole, these results indicate that species in the genusPomaceahave diversified their eggs defences: Those from thebridgesiiclade are protected mostly by non-digestible lectins that lower the nutritional value of eggs, in contrast with protection by neurotoxins of otherPomaceaclades, indicating apple snail egg defensive strategies are clade-specific. The harsh gastrointestinal environment of predators would have favoured their appearance, extending by convergent evolution the presence of plant-like highly stable lectins, a strategy not reported in other animals.Summary statementAnalysis of key snail egg proteins shows evolutionary defensive trends associated with the phylogenetic position, extending by convergent evolution the presence of plant-like defensive strategies not reported in other animals</jats:sec

Habitual meal frequency and energy intake regulation in partially temporally isolated men

Department of Human Biology, University of Maastricht, Maastricht, The Netherlands. [email protected] OBJECTIVE: Assessment of a possible relationship between habitual as well as manipulated meal frequency, blood glucose pattern, macronutrient- and energy intake (EI), and energy intake regulation in partially temporally isolated men. DESIGN: A partially temporally isolated within-subject design assessing energy intake regulation in spite of intervention. Intervention consisted of manipulating meal frequency by offering iso-energetic (1 MJ) preloads high in fat or carbohydrate (CHO), with the same energy density. We have previously shown that after a high-CHO preload, inter-meal-interval was 1 h, while after a high-fat preload intermeal-interval was 2 h. SUBJECTS: Twenty healthy young (18-31 y) normal weight (body mass index (BMI): 22.8+/-1.9 kg/m(2)) men. MEASUREMENTS: On two separate days, each after a different preload: subsequent subjects' responses to the preload, eg manipulated meal frequency; continuous blood glucose levels and blood glucose patterns: macronutrient composition of food intake; EI; appetite ratings; and taste perception. From controlled 3-day food intake diaries: habitual meal frequency; EI; and macronutrient-intake. RESULTS: Accuracy of energy intake regulation is expressed as minimizing the difference in energy intake, despite intervention. The difference in 24 h EI on the two test days after the preloads (r(2)=0.56; P<0.001) was a function of habitual meal frequency. Variation in energy intake was primarily explained by habitual meal frequency (r(2)=0.76; P<0.0001). Adding macronutrient composition and number of blood glucose declines to this increased the explained variation to 86 and 96%, respectively. Percentage energy from CHO or from fat explained the variation in habitual meal frequency (r(2)=0.84; P<0.0001). Adding the total number of blood-glucose declines to this increased the explained variation to 88%, and adding average baseline blood glucose levels, sweetness perception and hunger suppression during preload consumption increased the explained variation to 91%. Manipulated meal frequency was related to habitual meal frequency (r(2)=0.86; P<0.0001) and was a function of the number of transient and dynamic blood glucose declines (r(2)=0.74; P<0.0001). CONCLUSION: Habitual meal frequency is of greater significance in energy intake regulation in healthy young men than manipulated meal frequency. Healthy young men with a high habitual meal frequency showed lower 24 h EI, and a smaller difference in EI after macronutrient specific preloads, compared to those with a low habitual meal frequency, thus showing a more accurate energy intake regulation. Habitual meal frequency is based upon a cluster of related factors including macronutrient composition of the food, sweetness perception, hunger suppression, blood glucose declines and average baseline blood glucose levels. Publication Types: Clinical Trial Randomized Controlled Tria

Nutrient intake and plate waste from an Australian residential care facility

Objective: To determine the plate waste, energy and selected-nutrient intake, from elderly residents living in a high-level care (HLC) and low-level care (LLC) facility. Design: Three, single, whole day assessments of plate waste, energy, and selected nutrients, using a visual rating plate waste scale. Setting: Long-term residential care establishment. Subjects: One hundred and sixty-nine (93 HLC and 76 LLC) individual daily intakes. Main findings: The mean energy wasted throughout the whole day was 17%. The energy wasted from main meals (16%) was significantly less than the energy wasted at mid-meals (22%, P=0.049). The lowest mean energy wastage occurred at breakfast (8%) compared to lunch (22%) and dinner (25%, P<0.001). The mean (s.d.) daily energy served and consumed was 8.1 (2.0) and 6.6 (2.2) MJ, respectively. There was no difference in energy served or consumed between HLC and LLC residents. On the observation day, 60% of residents consumed less than their estimated energy requirement. The mean calcium intake was 796 (346) mg, and the median (inter-quartile range) vitamin D intake was 1.78 (2.05) mug. Conclusion: On 1 day, more than half the residents surveyed were at risk of consuming an inadequate energy intake, which over-time, may result in body weight loss. Although wastage was not excessive and energy served was adequate, the amount of food eaten was insufficient to meet energy and calcium requirements for a significant number of residents and it is not possible to consume sufficient vitamin D through food sources.JA Grieger and CA Nowso