Effects of the intraperitoneal administration of ebulin f in six-week-old Swiss mice on the evolution of their survival (Kaplan-Meyer plots) (left) and body weight (right). Effects of intraperitoneal administration of ebulin f in six-month-old Swiss mice on the evolution of their survival (Kaplan-Meyer plots) (left) and body weight (right). Effects of intraperitoneal administration of ebulin f in 12-month-old Swiss mice on the evolution of their survival (Kaplan-Meyer plots) (left) and body weight (right). The decrease in cell renewal and resulting atrophy lead to a shortening of both intestines. AcknowledgmentsThis study was supported by UVa-GIR (Recognized Research Group from the Valladolid University) and Complutense University UCM-CAM (Comunidad Autonoma de Madrid-Universidad Complutense de Madrid) research group 950247. I study the evolution of reproductive modes in insects at different levels, from symbiont-induced alterations to the genes underlying these important life-history traits. Genetic basis of sex ratio behaviourIn parasitoid wasps, like in other haplodiploid species, sons develop from unfertilised, haploid eggs, whereas daughters develop from fertilized, diploid eggs. Genomics of parasitoid life-history traitsAn organisma€™s life-history defines the key processes that determine its survival, growth, and reproduction. Symbiont-induced alterations to reproductionIn my PhD project at Leiden University, I characterized the evolutionary consequences of parthenogenesis-inducing Wolbachia bacteria in the Drosophila parasitoid Leptopilina clavipes. IntroductionAging is a multi-factorial process involving changes in the structure and function of organs and tissues; among these, vital organs, such as the lungs and intestines, are of particular interest. Effects on the LungVenous and capillary congestion was a common finding in both the control and experimental animals, but to a greater extent in the latter (Figure 4).
Venous congestion can be observed in the three groups, but whereas the structure of the lung appears normal in the control specimens, pneumonia in an advanced phase of hepatization is present in (B) and (C).
Hemorrhage is present inside a bronchus lumen in a lung with pneumonia in the red hepatization phase.
Venous and capillary congestion can be seen in a pneumonic focus showing a chronic inflammatory infiltrate.
Normal Lieberkuhn’s crypts with mitoses (arrow) can be seen in A; crypts appear atrophic and show apoptosis (arrow) in (B).
This gives females full control of the sex ratio they produce when laying eggs on a host.I am interested in the genetic constraints of adaptive sex allocation in the parasitoid Nasonia.
Evolution fine-tunes life-histories to optimally match an organisms environment by selecting heritable adaptations.
Using a combination of field work, population genetics, cytology, QTL mapping and behavioural analysis, I showed that the Wolbachia infection is irreversible and that it causes reproductive barriers that can be considered as the first steps in speciation between infected and uninfected populations.As a postdoctoral researcher at Lyon University, I studied the mechanisms behind an obligatory Wolbachia-infection in Asobara tabida, another Drosophila parasitoid. We performed experiments to assess whether ebulin f administration to six- and 12-month-old mice would trigger higher toxicity than that displayed in six-week-old mice. The small intestine of mammals has very important functions, such as the digestion of food and the absorption of the resulting nutrients [1,2,3]. In a few experimental specimens, pneumonia appeared in its initial phase, while others displayed hemorrhage during red hepatization (Figure 5), whilst in some animal, lungs appeared in the grey hepatization phase. Erythrocytes have lost their eosinophilia and are being phagocytized by macrophages (arrows).
In contrast, differences between small intestine lengths at 12 months old, as well as between large intestine lengths at six months old were not statistically significant in accordance with one-way ANOVA. My current work is concerned with developing and applying genomic tools for studying parasitoid life-history traits.The life-history of a parasitoid wasp determines for a large part how successful it will be as a pest controller.
In the present report, we present evidence indicating that the toxicological effects of ebulin f after its i.p. The main functions of the colon are the absorption of water and electrolytes and to allow the fermentation of undigested macronutrients by the intestinal microbiota.
The condensation areas, also including a chronic inflammatory infiltrate and thickening of the alveolus-capillary wall, seemed to be independent of the dose of toxin injected, but were more severe in 12-month-old animals, with the appearance of certain atypical nuclei with dysplasia (Figure 6). In this study, we present evidence that adult mice (six-month- and 12-month-old animals) are more sensitive to ebulin f administration than young mice (six-week-old animals).

This makes them good alternatives to chemicals to control pests in agriculture and they are frequently used for this purpose in greenhouses and in the field. My current research aims to use the genetic information of life-history traits to help improve the performance of parasitic wasps in biological control. From a histological point of view, the small intestine has a highly specialized and efficient tissue structure that has a very large surface made up of intestinal folds, villi, microvilli and crypts. My research concerns a range of different aspects of reproductive behaviour, from symbiont-induced reproductive alterations to the evolutionary genetics of behaviour.
We hypothesize that the ebulin f apoptosis-promoting action together with the age-dependent high rate of apoptosis result in an increase in the lectin’s toxicity, leading to a higher lethality level.
Epithelial cells in the gastrointestinal tract have a high turnover rate, and the maintenance of normal cellular balance through apoptosis is crucial for the preservation of normal cellular function [4]. It has been reported that the aged small intestine displays hyper proliferation and a high rate of apoptosis to prevent structurally and functionally stable alterations [5].Structural changes and cell turnover in the epithelial lining of the small intestine are regulated by apoptosis, which contributes to maintaining gut function [5,6]. Associated with this are changes in the absorption of nutrients, such us glucose, vitamins, calcium, magnesium, zinc and copper [7,8,9]. This should be a determining factor in drug absorption for low permeable drugs in the different regions of the gastrointestinal tract. To explain this differential effect, we must consider the lower resistance of the lungs to hazards in more aged animals, since a six-month-old mouse is already rather old, more or less comparable to a man in his sixties; besides, the lung damage found in our study was more severe in 12-month-old animals than in six-month-old ones. Dwarf elder has been used since ancient times, and it is known to have pharmacological properties, namely antioxidant, anti-inflammatory, anti-rheumatic, anti-hemorrhoid and anti-infectious activities, among others [12,13,14,15]. Gastric digestion in vivo and in vitro: How the structural aspects of food influence the digestion process.
Journal of Evolutionary Biology 24, 12-22.Werren JH, Richards S, Desjardins CA, Niehuis O, Gadau J, Colbourne JK, Beukeboom LW et al. A few of the molecules responsible for these activities have been identified, but at the same time, different components of the plant may be toxic if consumed in excess, which is probably due to the presence of ebulins [16,17].
Ebulin f, in the same way as nigrin b, goes from endosomes to lysosomes, where it suffers a degradation that may transform it into inactive products ready to be expelled from the cell, whereas ricin seems to follow a retrograde transport from endosomes to the trans-Golgi network and a final transfer to the rough endoplasmic reticulum [30].Concerning the action mechanism, the effects of ebulin f, similar to those of the related RIL nigrin b, do not seem to be of the type expected simply from an arrest of protein synthesis triggered by a typical translation inhibitor, since the concentration of these proteins required to inhibit translation is higher than that needed to affect cell viability [31,32,33]. We obtained similar results following nasal administration of ebulin blo at the same dose [18]. In addition, it has been reported that aging in mice promoted accelerated apoptosis, leading to the breakdown of intestinal mucosa [34].
In our case, we hypothesize that ebulin f apoptosis-promoting action together with the age-dependent high rate of apoptosis result in a higher toxicity level of the lectin, leading to a higher lethality level.The effects exerted by ebulin f on mice reported here support the notion that they might be pleiotropic and probably comprise transfer arrest along with the promotion of other mechanisms that trigger apoptosis. Additionally, it has recently been reported that type-2 RIP cytotoxicity is mediated by the unfolded protein activated in response to endoplasmic reticulum stress [35]. It has been proposed that the RIP in the plant plays a protective role against insects, viruses and fungi [19]. Furthermore, ricin poisoning promoted the differential expression of at least six proteins in the lungs of mice exposed to aerosolized ricin [36].Work is still underway to analyze in mice the behavioral pattern of cytokines, the activity of the caspase system following administration of sub-toxic and toxic concentrations of ebulin f and the potential changes in lung protein expression due to ebulin f toxicity.
Cellular and molecular mechanisms of intestinal elongation in mammals: The long and short of it. Regarding the molecular action mechanism, ebulins cause the irreversible inactivation of ribosome by arresting its 28S rRNA, while its B-chain selectively bonds to d-galactose [19,22]. As a general consideration regarding the toxicity described and when taking into account the traditional ingestion of dwarf elder fruits, it becomes most important to employ methods to detoxify ebulin, such as boiling the aqueous extracts of the plant to a level safe enough to inactivate the protein and render it sensitive to degradation by acidic pepsin, as has recently been reported [37]. The latter is one thousand-times more cytotoxic than ebulin in mice due to the great ability of its B chain to bind to the polysaccharide chains situated on the surface of plasma membrane proteins [22,23].Previous studies have described the toxicity of ebulin f and ebulin blo in six-week-old mice [11,18].
Early intestinal targets were the transit amplifying cells present in the small intestine crypts, as well as the cells of the large intestine crypts. The presence of these proteins in certain food products, the concern regarding their possible criminal use or as a bioweapon and also their potential application in medicine make the toxicology of these proteins an area of great interest [18].It is well known that the organic characteristics of living beings vary with age, and in toxicology, in particular, the effects occurring throughout the different stages of life are noteworthy. As pointed out above, an increase in apoptosis is associated with aging, and we hypothesized that ebulin f would increase that process even more and that, therefore, it would increase its toxicity.Ebulin f was isolated by affinity chromatography and characterized by mass spectrometry, as reported previously [10,24].

Its toxicity was first studied in six-week-old mice, and it was found that the intestines were the primary targets for ebulin f when administered by i.p. A comparison of recombination frequencies in intraspecific versus interspecific mapping populations of Nasonia. Since the small intestine of aged mice was reported to undergo a higher rate of apoptosis, we performed experiments to assess whether ebulin f administration to six- and 12-month-old mice would trigger higher toxicity than that displayed in six-week-old animals.
This pasty extract was strained through two layers of cheesecloth, and the fluid was centrifuged at 7500? g for 45 min at 4 °C. The supernatant was centrifuged a second time at the same speed for 30 min and subsequently removed and filtered through two layers of common laboratory filter paper to remove the mucilaginous material.
This simple procedure ensures its removal without the quality and activity of the protein preparation being affected. The filtered extract was chromatographed through acid-treated Sepharose 6B (AT-Sepharose 6B) to obtain d-galactose-binding proteins, as previously reported for the lectins of dwarf elder fruits [10].
The column was washed with the same buffer until the A280 reached values close to 0, and the protein fraction not retained by the AT-Sepharose 6B column was discarded. The retained protein fraction was further eluted with the same buffer containing 0.2 M lactose.
Fractions of 10 mL were collected, and those containing proteins were pooled and concentrated with an Amicon system using a Y10 membrane down to 5 mL. The first 70 mL were discarded, and then, fractions of 2.5 mL were taken and their A280 measured. The purity of the ebulin f was assessed by SDS-PAGE and mass spectrometry, as indicated elsewhere [10]. Treatment with Ebulin fWater-dissolved ebulin f adjusted to the required concentrations for the mice’s weight was i.p. Control mice under the same experimental conditions, but with water, were not affected either with respect to survival or body weight. Two weeks after administration of the toxin, all the surviving animals were anesthetized with isoflurane and sacrificed by decapitation. Is symbiosis evolution influenced by the pleiotropic role of programmed cell death in immunity and development?. Geographic variation in host-selection behaviour in the Drosophila parasitoid Leptopilina clavipes. Sexual functionality of Leptopilina clavipes (Hymenoptera: Figitidae) after reversing Wolbachia-induced parthenogenesis. The genetic basis of male fertility in relation to haplodiploid reproduction in Leptopilina clavipes. Genetic diversity and Wolbachia infection of the Drosophila parasitoid Leptopilina clavipes in western Europe.
Cytology of Wolbachia-induced parthenogenesis in Leptopilina clavipes (Hymenoptera: Figitidae). Evolutionary consequences of Wolbachia-induced parthenogenesis in the parasitoid Leptopilina clavipes. Curing thelytoky in the Drosophila parasitoid Leptopilina clavipes (Hymenoptera: Figitidae).
The response of Leptopilina clavipes (Hymenoptera: Figitidae) to cVA, a major component of Drosophila aggregation pheromone.

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