The use of minds. Ontology for an ecological life policy

El análisis filosófico, politológico y sociológico de las últimas décadas ha hecho hincapié en la búsqueda de la producción de formas de vida políticamente “cualificadas” o “descalificadas”. En este marco, el concepto de biopolítica ha revolucionado la topografía y los instrumentos de las facetas analíticas de lo político, así como de las relaciones y de los sistemas sociales. Sin embargo, a esta riqueza analítica no se ha ajustado otra reflexión tan rica acerca de las praxis de emancipación a partir de y en las relaciones biopolíticas. En este artículo queremos mostrar las potencialidades teoréticas y prácticas del concepto de uso, y su importancia para la definición de una ontología para una vida ecológica, que sea al mismo tiempo ética y política, si y cuando este concepto permite la superación y la desactivación de los dispositivos biopolíticos dominantes. Desde esta perspectiva, muy útiles son las recientes obras de Giorgio Agamben y Ottavio Marzocca. Ambos autores –aunque de forma diferente– consideran el concepto de uso como elemento para pensar y vivir una nueva manera de existir en el mundo, que libere al bíos de las jaulas biopolíticas y bioeconómicas de nuestro presente.The research on the production of political “qualified” or “unqualified” life forms was at the centre of the philosophical and sociological analytic of the last decades. In this frame, the concept of biopolitics revolutionised the topography and the analytical tools of the political as well as of the social relations and systems, allowing thus to make new connections and analogy visible. To such an analytical abundance, however, does not correspond a comparable reflection about praxis of emancipation from and in the biopolitical relations. In this paper I will show the theoretical and practical potential of the concept of use and its importance for the definition of an ontology of the ecological life, that could be ethical and political, in so much as this concept could allow a suspensión and neutralization of the biopolitical dispositifs. In this perspective the recent works of Giorgio Agamben and Ottavio Marzocca are particularly important. Both authors – in different ways – adopt the concept of use in order to think and to live a new way to be in the world, which unlocks the bios from the biopolitical and bioeconomic cages of our time

El uso de las mentes : ontología para una política de la vida ecológica.

The research on the production of political “qualified” or “unqualified” life forms was at the centre of the philosophical and sociological analytic of the last decades. In this frame, the concept of biopolitics revolutionised the topography and the analytical tools of the political as well as of the social relations and systems, allowing thus to make new connections and analogy visible. To such an analytical abundance, however, does not correspond a comparable reflection about praxis of emancipation from and in the biopolitical relations. In this paper I will show the theoretical and practical potential of the concept of use and its importance for the definition of an ontology of the ecological life, that could be ethical and political, in so much as this concept could allow a suspension and neutralization of the biopolitical dispositifs. In this perspective the recent works of Giorgio Agamben and Ottavio Marzocca are particularly important. Both authors – in different ways – adopt the concept of use in order to think and to live a new way to be in the world, which unlocks the bios from the biopolitical and bioeconomic cages of our time.El análisis filosófico, politológico y sociológico de las últimas décadas ha hecho hincapié en la búsqueda de la producción de formas de vida políticamente “cualificadas” o “descalificadas”. En este marco, el concepto de biopolítica ha revolucionado la topografía y los instrumentos de las facetas analíticas de lo político, así como de las relaciones y de los sistemas sociales. Sin embargo, a esta riqueza analítica no se ha ajustado otra reflexión tan rica acerca de las praxis de emancipación a partir de y en las relaciones biopolíticas. En este artículo queremos mostrar las potencialidades teoréticas y prácticas del concepto de uso, y su importancia para la definición de una ontología para una vida ecológica, que sea al mismo tiempo ética y política, si y cuando este concepto permite la superación y la desactivación de los dispositivos biopolíticos dominantes. Desde esta perspectiva, muy útiles son las recientes obras de Giorgio Agamben y Ottavio Marzocca. Ambos autores –aunque de forma diferente– consideran el concepto de uso como elemento para pensar y vivir una nueva manera de existir en el mundo, que libere al bíos de las jaulas biopolíticas y bioeconómicas de nuestro presente

Cardiolipin fingerprinting of leukocytes by MALDI-TOF/MS as a screening tool for Barth syndrome

Barth syndrome (BTHS), an X-linked disease associated with cardioskeletal myopathy, neutropenia, and organic aciduria, is characterized by abnormalities of cardiolipin (CL) species in mitochondria. Diagnosis of the disease is often compromised by lack of rapid and widely available diagnostic laboratory tests. The present study describes a new method for BTHS screening based on MALDI-TOF/MS analysis of leukocyte lipids. This generates a "CL fingerprint" and allows quick and simple assay of the relative levels of CL and monolysocardiolipin species in leukocyte total lipid profiles. To validate the method, we used vector algebra to analyze the difference in lipid composition between controls (24 healthy donors) and patients (8 boys affected by BTHS) in the high-mass phospholipid range. The method of lipid analysis described represents an important additional tool for the diagnosis of BTHS and potentially enables therapeutic monitoring of drug targets, which have been shown to ameliorate abnormal CL profiles in cells

AI is a viable alternative to high throughput screening: a 318-target study

: High throughput screening (HTS) is routinely used to identify bioactive small molecules. This requires physical compounds, which limits coverage of accessible chemical space. Computational approaches combined with vast on-demand chemical libraries can access far greater chemical space, provided that the predictive accuracy is sufficient to identify useful molecules. Through the largest and most diverse virtual HTS campaign reported to date, comprising 318 individual projects, we demonstrate that our AtomNet® convolutional neural network successfully finds novel hits across every major therapeutic area and protein class. We address historical limitations of computational screening by demonstrating success for target proteins without known binders, high-quality X-ray crystal structures, or manual cherry-picking of compounds. We show that the molecules selected by the AtomNet® model are novel drug-like scaffolds rather than minor modifications to known bioactive compounds. Our empirical results suggest that computational methods can substantially replace HTS as the first step of small-molecule drug discovery

Yarrowia lipolytica as a Sustainable Microbial Platform for Circular added value Compounds Production

Yarrowia lipolytica is among the most studied oleaginous yeasts. Out of 600 identified yeast species, only 30 show this feature and Y. lipolytica, due to its non-pathogenicity and its dimorphisms, is the most studied non- conventional yeast.[1] This yeast is naturally able to accumulate lipids to levels exceeding 50% of CDW[2] using different compounds as carbon and energy sources. Waste cooking oils (WCO) refers to oils and fats used for cooking or for the storing of food substances at the commercial, industrial or household levels. In China and Europe their annual production is approximately 500 million tons and 4 million tons, respectively.[3] If not collected, WCO can clog the sewage system creating the so-called “fatbergs”. Due to the environmental issues associated with WCO, its valorization needs to be of utmost importance. WCO have been thoroughly studied for the (bio-)chemical production of fatty acid methyl esters (FAME) and hydrogenated vegetable oil (HVO) which are known as biodiesel. However, this feedstock was shown to be used as ingredient in fermentation media for the production of valuable compounds.[4] In this study, we report the use of WCO as feedstock for the development of a sustainable platform using the non-conventional yeast for the production of different compounds such as lipases, industrially-relevant sugars and intracellular lipids

Glutamine-Derived Aspartate Biosynthesis in Cancer Cells: Role of Mitochondrial Transporters and New Therapeutic Perspectives

Aspartate has a central role in cancer cell metabolism. Aspartate cytosolic availability is crucial for protein and nucleotide biosynthesis as well as for redox homeostasis. Since tumor cells display poor aspartate uptake from the external environment, most of the cellular pool of aspartate derives from mitochondrial catabolism of glutamine. At least four transporters are involved in this metabolic pathway: the glutamine (SLC1A5_var), the aspartate/glutamate (AGC), the aspartate/phosphate (uncoupling protein 2, UCP2), and the glutamate (GC) carriers, the last three belonging to the mitochondrial carrier family (MCF). The loss of one of these transporters causes a paucity of cytosolic aspartate and an arrest of cell proliferation in many different cancer types. The aim of this review is to clarify why different cancers have varying dependencies on metabolite transporters to support cytosolic glutamine-derived aspartate availability. Dissecting the precise metabolic routes that glutamine undergoes in specific tumor types is of upmost importance as it promises to unveil the best metabolic target for therapeutic intervention

Mitochondrial carriers of Ustilago maydis and Aspergillus terreus involved in itaconate production: same physiological role but different biochemical features

Itaconic acid (IA) is a naturally occurring dicarboxylic acid with applications in the manufacture of polymers. IA can be produced by fermentation using the fungi Aspergillus terreus or Ustilago maydis as biocatalysts. Indirect evidence has suggested that the mitochondrial carriers U. maydis Um_Mtt1 and A. terreus At_MttA export mitochondrially synthesized cis‐aconitate to the cytosol for IA synthesis using malate as a countersubstrate. Here, by assaying the transport features of recombinant Um_Mtt1 and At_MttA in reconstituted liposomes, we find that both proteins efficiently transport cis‐aconitate, but malate is well transported only by Um_Mtt1 and 2‐oxoglutarate only by At_MttA. Bioinformatic analysis shows that Um_Mtt1 and At_MttA form a distinctive mitochondrial carrier subfamily. Our data show that although fulfilling the same physiological task, Um_Mtt1 and At_MttA have different biochemical features

Mitochondrial citrate carrier deficiency causes neuromuscular transmission impairment

The mitochondrial citrate carrier (CIC), encoded by the SLC25A1 gene, catalyzes the export of citrate from mitochondria to the cytosol where it is broken into into acetyl-CoA and oxaloacetate. We identified for the first time two pathogenic SLC25A1 variants in a patient that suffered from a severe neurodevelopmental syndrome [1]. Recessive mutations in SLC25A1 have been since identified in more than a dozen patients and CIC deficiency has been classified as an inborn disorder of metabolism (OMIM: 615182) whose biochemical hallmark is combined D-2- and L-2-hydroxyglutaric aciduria [2]. More recently we reported a novel homozygous mutation in the SLC25A1 gene in an affected sib pair presenting with myasthenia and impaired neuromuscular junction (NMJ) transmission [3]. Upon functional reconstitution of recombinant proteins into liposomes, we showed that the newly identified mutation caused a milder activity impairment than the previously reported mutations suggesting a fundamental role of CIC in neuromuscular transmission whose defect was previously masked by the harsher phenotypes. Using the CRISPR/CAS9 approach we obtained stable lines of Caenorhabditis elegans knocked-out in the SLC25A1 ortholog that showed resistance to levamisole, a nicotinic acetylcholine receptor agonist, that causes continued stimulation of the worm muscles, leading to paralysis. This phenotype was, at least in part, rescued by the expression of wild-type human SLC25A1 under the control of a neuron-specific promoter, pointing towards an underlying presynaptic defect. Altogether these data demonstrate a conserved role of CIC in neuromuscular transmission and validate the worm C. elegans as a suitable animal model for further investigation of the molecular and cellular underpinnings of the NMJ transmission defect associated to CIC deficiency

FAD/NADH Dependent Oxidoreductases: From Different Amino Acid Sequences to Similar Protein Shapes for Playing an Ancient Function

Flavoprotein oxidoreductases are members of a large protein family of specialized dehydrogenases, which include type II NADH dehydrogenase, pyridine nucleotide-disulphide oxidoreductases, ferredoxin-NAD+ reductases, NADH oxidases, and NADH peroxidases, playing a crucial role in the metabolism of several prokaryotes and eukaryotes. Although several studies have been performed on single members or protein subgroups of flavoprotein oxidoreductases, a comprehensive analysis on structure-function relationships among the different members and subgroups of this great dehydrogenase family is still missing. Here, we present a structural comparative analysis showing that the investigated flavoprotein oxidoreductases have a highly similar overall structure, although the investigated dehydrogenases are quite different in functional annotations and global amino acid composition. The different functional annotation is ascribed to their participation in species-specific metabolic pathways based on the same biochemical reaction, i.e., the oxidation of specific cofactors, like NADH and FADH2. Notably, the performed comparative analysis sheds light on conserved sequence features that reflect very similar oxidation mechanisms, conserved among flavoprotein oxidoreductases belonging to phylogenetically distant species, as the bacterial type II NADH dehydrogenases and the mammalian apoptosis-inducing factor protein, until now retained as unique protein entities in Bacteria/Fungi or Animals, respectively. Furthermore, the presented computational analyses will allow consideration of FAD/NADH oxidoreductases as a possible target of new small molecules to be used as modulators of mitochondrial respiration for patients affected by rare diseases or cancer showing mitochondrial dysfunction, or antibiotics for treating bacterial/fungal/protista infections

Molecular and cellular underpinnings of the neurological phenotypes associated to mitochondrial citrate carrier (SLC25A1) deficiency

Il carrier mitocondriale del citrato (CIC), codificato dal gene SLC25A1, catalizza il trasporto del citrato dalla matrice mitocondriale al citoplasma dove il citrato viene scisso in acetil-CoA e ossalacetato. Mediante sequenziamento dell’esoma, abbiamo identificato per la prima volta due varianti patogeniche del gene SLC25A1 in un paziente affetto da una grave sindrome del neurosviluppo (Edvardson et al. 2013 J Med Genet 50:240-5). In seguito, mutazioni recessive dello stesso gene sono state identificate in numerosi pazienti e la deficienza del carrier del citrato è stata classificata come un disturbo congenito del metabolism (OMIM: 615182) caratterizzato da aciduria dovuta all’accumulo dell’acido D-2- e L-2-idrossiglutarico (Nota et al. 2013 Am J Hum Genet 92:627-31). Più recentemente, abbiamo identificato una nuova mutazione omozigote del gene SLC25A1 in una coppia di fratelli che mostravano sintomi clinici di miastenia e difetti nella trasmissione nervosa a livello delle giunzioni neuromuscolari ma senza apparenti segni di disturbi del neurosviluppo (Chaouch et al. 2014 J Neuromuscul Dis 1:75-90). Abbiamo quindi analizzato in vitro l’effetto delle mutazioni patogeniche sulla funzione del trasportatore del citrato a seguito di espressione ricombinante e ricostituzione in vescicole lipidiche artificiali. Abbiamo così dimostrato che la nuova mutazione causava un effetto meno drastico sull’attività del trasportatore suggerendo che CIC giochi un ruolo fondamentale nella trasmissione muscolare il cui difetto non era stato precedentemente notato negli altri pazienti a causa della maggiore gravità del fenotipo. Allo scopo di indagare sulle conseguenze neurologiche della deficienza di CIC, in esperimenti preliminari abbiamo ridotto l’espressione dei geni ortologhi di SLC25A1 in zebrafish iniettando nell’embrione oligonucleotidi antisenso del tipo “morfolino”. In questo modo abbiamo potuto osservare nell’animale adulto una alterata morfologia degli assoni a livello delle giunzioni neuromuscolari che ci hanno portato a proporre che i difetti della trasmissione neuromuscolare osservati nei pazienti potessero avere origine in anomalie delle terminazioni nervose pre-sinaptiche. Facendo uso della tecnologia CRISPR/CAS9, abbiamo generato linee stabili di Caenorhabditis elegans in cui il gene ortologo di SLC25A1 è stato inattivato. I vermi geneticamente modificati risultano essere particolarmente resitestenti ad un farmaco, il levamisolo, che agendo come un agonista del recettore di tipo “nicotinico” dell’acetilcolina causa una continua stimolazione della fibra muscolare del verme fino a determinarne la paralisi. Questo fenotipo può essere, almeno in parte, corretto dall’espressione del gene SLC25A1 “wild-type” (cioè senza mutazioni) sotto il controllo di un elemento di regolazione trascrizionale che ne permette l’espressione solamente nei neuroni, Questa osservazione fornisce un ulteriore evidenza della compromissione della trasmissione neuromuscolare a livello pre-sinaptico associati alla disfunzione di CIC. Nel loro complesso, i dati ottenuti dimostrano che CIC ha un ruolo nella trasmissione muscolare conservato nel corso dell’evoluzione. A questo proposito, giova ricordare che la trasmissione dei segnali nervosi a livello delle giunzioni neuromuscolari necessita di una continua e massiccia sintesi di acetilcolina nei motoneuroni a partire da colina e acetil-CoA. Mentre il primo composto può essere riciclato dopo il rilascio e l’idrolisi del neurotrasmettitore, il secondo deve essere continuamente generato dal metabolismo ossidativo nei mitocondri dai quali esce sotto forma di citrato grazie a CIC. I nostri risultati, inoltre, validano il verme C. elegans come un modello animale utile per ulteriori indagini sui meccanismi molecolari e cellulari alla base dei difetti della trasmissione nervosa a livello delle giunzioni neuromuscolari associati alla deficienza di CIC.The mitochondrial citrate carrier (CIC) encoded by the SLC25A1 gene, catalyzes the export of citrate from mitochondria to the cytosol where it is broken into into acetyl-CoA and oxaloacetate. By exome sequencing we identified for the first time two pathogenic SLC25A1 variants in a patient that suffered from a severe neurodevelopmental syndrome (Edvardson et al. 2013 J Med Genet 50:240-5). Recessive mutations in SLC25A1 have been since identified in more than a dozen patients and CIC deficiency has been classified as an inborn disorder of metabolism (OMIM: 615182) whose hallmark is combined D-2- and L-2-hydroxyglutaric aciduria (Nota et al. 2013 Am J Hum Genet 92:627-31). More recently we reported a novel homozygous mutation in the SLC25A1 gene in an affected sib pair. Both patients presented with myasthenia and impaired neuromuscular junction (NMJ) transmission whilst no neurodevelopment disorder has been observed (Chaouch et al. 2014 J Neuromuscul Dis 1:75-90). Functional analysis of the pathogenic mutations has been performed upon reconstitution of recombinant proteins into liposomes. Interestingly, the newly identified mutation caused a milder activity impairment than the previously reported mutations suggesting a fundamental role of CIC in neuromuscular transmission whose defect was previously masked by the harsher phenotypes. In preliminary experiments, knocking down the SLC25A1 orthologues by injection of antisense morpholino oligonucleotides in zebrafish embryos we observed short and erratic outgrowth of motor axons toward muscle fiber at NMJ suggesting that transmission impairment could be related to pre-synaptic nerve terminal abnormalities. Using the CRISPR/CAS9 approach we obtained stable lines of Caenorhabditis elegans knocked-out in the SLC25A1 ortholog that showed resistance to levamisole, a nicotinic acetylcholine receptor agonist, that causes continued stimulation of the worm muscles, leading to paralysis. This phenotype was, at least in part, rescued by the expression of wild-type SLC25A1 under the control of a neuron-specific promoter, strongly pointing towards a underlying pre-synaptic defect. Altogether these data demonstrate a conserved role of CIC in neuromuscular transmission. Of note, signal transmission at NMJ relies on massive synthesis in motor neurons of acetylcholine from choline and acetylCoA. While the former can be recycled after neurotransmitter release and breakdown, the latter must be continuously generated by oxidative metabolism in mitochondria from which it is exported in the form of citrate by CIC. Furthermore, our results validate the worm C. elegans as an animal model suitable for further study of the molecular and cellular underpinnings of the NMJ transmission defect associated to CIC deficiency