Metabolomic Prediction of Cadmium Nephrotoxicity in the Snail Helix aspersa maxima.

The decontamination of polluted soils is a major socioeconomic issue in many industrialized countries. In situ remediation approaches are nowadays preferred to ex situ techniques, but they require among others the use of bioindicators, which are sensitive to the progressive depollution on health effects. Animal species have been mainly used so far to monitor aquatic and air pollution. Current research focuses on the development of living indicators of soil pollution. In this study, the garden snail Helix aspersa maxima was acutely exposed to cadmium, one major soil contaminant causing severe health effects, including nephrotoxicity. Kidney and hemolymph were sampled and analyzed by a 1H-NMR-based metabonomic approach. Shortly after Cd exposure, numerous metabolic changes occurred in the hemolymph and kidney extracts. Altogether, they were indicative of a switch in energy sources from the Krebs cycle towards b-oxidation and the utilization of stored galactogen polysaccharides. Then, the activation of antioxidant defenses in the renal cells was suggested by the alteration in some precursors of glutathione synthesis, such as glutamate, and by the release of the antioxidant anserin. Cell membrane damage was evidenced by the increased levels of some osmolytes, betaine and putrescine, as well as by a membrane repair mechanism involving choline. Finally, the development of metabolic acidosis was suggested by the elevation in 3-HMG in the hemolymph, and the more pronounced lysine levels were consistent with acute excretion troubles. Cd-induced renal damage was objectified by the increased level of riboflavin, a recognized biomarker of nephrotoxicity.

A population modification gene drive targeting both Saglin and Lipophorin impairs Plasmodium transmission in Anopheles mosquitoes.

Lipophorin is an essential, highly expressed lipid transport protein that is secreted and circulates in insect hemolymph. We hijacked the Anopheles coluzzii Lipophorin gene to make it co-express a single-chain version of antibody 2A10, which binds sporozoites of the malaria parasite Plasmodium falciparum. The resulting transgenic mosquitoes show a markedly decreased ability to transmit Plasmodium berghei expressing the P. falciparum circumsporozoite protein to mice. To force the spread of this antimalarial transgene in a mosquito population, we designed and tested several CRISPR/Cas9-based gene drives. One of these is installed in, and disrupts, the pro-parasitic gene Saglin and also cleaves wild-type Lipophorin, causing the anti-malarial modified Lipophorin version to replace the wild type and hitch-hike together with the Saglin drive. Although generating drive-resistant alleles and showing instability in its gRNA-encoding multiplex array, the Saglin-based gene drive reached high levels in caged mosquito populations and efficiently promoted the simultaneous spread of the antimalarial Lipophorin::Sc2A10 allele. This combination is expected to decrease parasite transmission via two different mechanisms. This work contributes to the design of novel strategies to spread antimalarial transgenes in mosquitoes, and illustrates some expected and unexpected outcomes encountered when establishing a population modification gene drive.

The emergence of the Biodiversity/Health nexus: making biodiversity a health issue.

Before the COVID-19 pandemic, a French epistemic community has forged and promoted a Biodiversity/Health nexus, which legitimizes biodiversity as a health issue. The relationship between biodiversity and health is now part of French local government agendas, after being included in new international programs. Based on observation of this nexus's epistemic community and 35 semi-structured interviews conducted in France between 2017 and 2020, this article aims to show which actors and groups have been forging and promoting this nexus, and to understand how such an emergent environmental nexus challenges the governance of the present biomedical- and technical expertise-based health system. This article discusses environmental nexus from the perspective of building a new cause by reconstituting chains of causality to "demonstrate" the new problem (Barthe, Politix, 23(91), 77-102, 2010), and the growing importance of integration of concepts as a new ideal of policy-making (Cairns & Krzywoszynska, Environmental Science and Policy, 64, 164-170, 2016). As well as a justification (Boltanski & Thevenot, 1991) of their effectiveness in legitimizing the cause of defending biodiversity, environmental nexuses contain a challenge to recognize knowledge, calling for a change in governance methods in a One Health approach.

The salivary protein Saglin facilitates efficient midgut colonization of Anopheles mosquitoes by malaria parasites.

Malaria is caused by the unicellular parasite Plasmodium which is transmitted to humans through the bite of infected female Anopheles mosquitoes. To initiate sexual reproduction and to infect the midgut of the mosquito, Plasmodium gametocytes are able to recognize the intestinal environment after being ingested during blood feeding. A shift in temperature, pH change and the presence of the insect-specific compound xanthurenic acid have been shown to be important stimuli perceived by gametocytes to become activated and proceed to sexual reproduction. Here we report that the salivary protein Saglin, previously proposed to be a receptor for the recognition of salivary glands by sporozoites, facilitates Plasmodium colonization of the mosquito midgut, but does not contribute to salivary gland invasion. In mosquito mutants lacking Saglin, Plasmodium infection of Anopheles females is reduced, resulting in impaired transmission of sporozoites at low infection densities. Interestingly, Saglin can be detected in high amounts in the midgut of mosquitoes after blood ingestion, possibly indicating a previously unknown host-pathogen interaction between Saglin and midgut stages of Plasmodium. Furthermore, we were able to show that saglin deletion has no fitness cost in laboratory conditions, suggesting this gene would be an interesting target for gene drive approaches.

Induction of control genes in intestinal gluconeogenesis is sequential during fasting and maximal in diabetes.

We studied in rats the expression of genes involved in gluconeogenesis from glutamine and glycerol in the small intestine (SI) during fasting and diabetes. From Northern blot and enzymatic studies, we report that only phosphoenolpyruvate carboxykinase (PEPCK) activity is induced at 24 h of fasting, whereas glucose-6-phosphatase (G-6-Pase) activity is induced only from 48 h. Both genes then plateau, whereas glutaminase and glycerokinase strikingly rebound between 48 and 72 h. The two latter genes are fully expressed in streptozotocin-diabetic rats. From arteriovenous balance and isotopic techniques, we show that the SI does not release glucose at 24 h of fasting and that SI gluconeogenesis contributes to 35% of total glucose production in 72-h-fasted rats. The new findings are that 1) the SI can quantitatively account for up to one-third of glucose production in prolonged fasting; 2) the induction of PEPCK is not sufficient by itself to trigger SI gluconeogenesis; 3) G-6-Pase likely plays a crucial role in this process; and 4) glutaminase and glycerokinase may play a key potentiating role in the latest times of fasting and in diabetes.

Polyunsaturated fatty acyl coenzyme A suppress the glucose-6-phosphatase promoter activity by modulating the DNA binding of hepatocyte nuclear factor 4 alpha.

Glucose-6-phosphatase confers on gluconeogenic tissues the capacity to release endogenous glucose in blood. The expression of its gene is modulated by nutritional mechanisms dependent on dietary fatty acids, with specific inhibitory effects of polyunsaturated fatty acids (PUFA). The presence of consensus binding sites of hepatocyte nuclear factor 4 (HNF4) in the -1640/+60 bp region of the rat glucose-6-phosphatase gene has led us to consider the hypothesis that HNF4 alpha could be involved in the regulation of glucose-6-phosphatase gene transcription by long chain fatty acid (LCFA). Our results have shown that the glucose-6-phosphatase promoter activity is specifically inhibited in the presence of PUFA in HepG2 hepatoma cells, whereas saturated LCFA have no effect. In HeLa cells, the glucose-6-phosphatase promoter activity is induced by the co-expression of HNF4 alpha or HNF1 alpha. PUFA repress the promoter activity only in HNF4 alpha-cotransfected HeLa cells, whereas they have no effects on the promoter activity in HNF1 alpha-cotransfected HeLa cells. From gel shift mobility assays, deletion, and mutagenesis experiments, two specific binding sequences have been identified that appear able to account for both transactivation by HNF4 alpha and regulation by LCFA in cells. The binding of HNF4 alpha to its cognate sites is specifically inhibited by polyunsaturated fatty acyl coenzyme A in vitro. These data strongly suggest that the mechanism by which PUFA suppress the glucose-6-phosphatase gene transcription involves an inhibition of the binding of HNF4 alpha to its cognate sites in the presence of polyunsaturated fatty acyl-CoA thioesters.