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1.
J Cell Sci ; 137(9)2024 May 01.
Artículo en Inglés | MEDLINE | ID: mdl-38661008

RESUMEN

DPF3, along with other subunits, is a well-known component of the BAF chromatin remodeling complex, which plays a key role in regulating chromatin remodeling activity and gene expression. Here, we elucidated a non-canonical localization and role for DPF3. We showed that DPF3 dynamically localizes to the centriolar satellites in interphase and to the centrosome, spindle midzone and bridging fiber area, and midbodies during mitosis. Loss of DPF3 causes kinetochore fiber instability, unstable kinetochore-microtubule attachment and defects in chromosome alignment, resulting in altered mitotic progression, cell death and genomic instability. In addition, we also demonstrated that DPF3 localizes to centriolar satellites at the base of primary cilia and is required for ciliogenesis by regulating axoneme extension. Taken together, these findings uncover a moonlighting dual function for DPF3 during mitosis and ciliogenesis.


Asunto(s)
Cilios , Mitosis , Factores de Transcripción , Animales , Humanos , Ratones , Axonema/metabolismo , Centriolos/metabolismo , Centrosoma/metabolismo , Cilios/metabolismo , Proteínas de Unión al ADN/metabolismo , Proteínas de Unión al ADN/genética , Inestabilidad Genómica , Células HeLa , Cinetocoros/metabolismo , Huso Acromático/metabolismo , Factores de Transcripción/metabolismo , Factores de Transcripción/genética
2.
Arch Toxicol ; 95(3): 1023-1037, 2021 03.
Artículo en Inglés | MEDLINE | ID: mdl-33426622

RESUMEN

Uranium is widely spread in the environment due to its natural and anthropogenic occurrences, hence the importance of understanding its impact on human health. The skeleton is the main site of long-term accumulation of this actinide. However, interactions of this metal with biological processes involving the mineralized extracellular matrix and bone cells are still poorly understood. To get a better insight into these interactions, we developed new biomimetic bone matrices containing low doses of natural uranium (up to 0.85 µg of uranium per cm2). These models were characterized by spectroscopic and microscopic approaches before being used as a support for the culture and differentiation of pre-osteoclastic cells. In doing so, we demonstrate that uranium can exert opposite effects on osteoclast resorption depending on its concentration in the bone microenvironment. Our results also provide evidence for the first time that resorption contributes to the remobilization of bone matrix-bound uranium. In agreement with this, we identified, by HRTEM, uranium phosphate internalized in vesicles of resorbing osteoclasts. Thanks to the biomimetic matrices we developed, this study highlights the complex mutual effects between osteoclasts and uranium. This demonstrates the relevance of these 3D models to further study the cellular mechanisms at play in response to uranium storage in bone tissue, and thus better understand the impact of environmental exposure to uranium on human bone health.


Asunto(s)
Matriz Ósea/efectos de los fármacos , Modelos Biológicos , Osteoclastos/efectos de los fármacos , Uranio/metabolismo , Animales , Biomimética , Matriz Ósea/metabolismo , Resorción Ósea/metabolismo , Línea Celular Tumoral , Humanos , Ratones , Osteoclastos/metabolismo , Células RAW 264.7 , Distribución Tisular , Uranio/administración & dosificación
3.
Nucleic Acids Res ; 47(6): 2739-2756, 2019 04 08.
Artículo en Inglés | MEDLINE | ID: mdl-30759257

RESUMEN

G-quadruplex ligands exert their antiproliferative effects through telomere-dependent and telomere-independent mechanisms, but the inter-relationships among autophagy, cell growth arrest and cell death induced by these ligands remain largely unexplored. Here, we demonstrate that the G-quadruplex ligand 20A causes growth arrest of cancer cells in culture and in a HeLa cell xenografted mouse model. This response is associated with the induction of senescence and apoptosis. Transcriptomic analysis of 20A treated cells reveals a significant functional enrichment of biological pathways related to growth arrest, DNA damage response and the lysosomal pathway. 20A elicits global DNA damage but not telomeric damage and activates the ATM and autophagy pathways. Loss of ATM following 20A treatment inhibits both autophagy and senescence and sensitizes cells to death. Moreover, disruption of autophagy by deletion of two essential autophagy genes ATG5 and ATG7 leads to failure of CHK1 activation by 20A and subsequently increased cell death. Our results, therefore, identify the activation of ATM by 20A as a critical player in the balance between senescence and apoptosis and autophagy as one of the key mediators of such regulation. Thus, targeting the ATM/autophagy pathway might be a promising strategy to achieve the maximal anticancer effect of this compound.


Asunto(s)
Apoptosis/efectos de los fármacos , Proteínas de la Ataxia Telangiectasia Mutada , Autofagia/efectos de los fármacos , Senescencia Celular/efectos de los fármacos , G-Cuádruplex , Neoplasias/patología , Células A549 , Animales , Apoptosis/genética , Proteínas de la Ataxia Telangiectasia Mutada/antagonistas & inhibidores , Proteínas de la Ataxia Telangiectasia Mutada/metabolismo , Autofagia/genética , Línea Celular Tumoral , Senescencia Celular/genética , Daño del ADN/efectos de los fármacos , Células HeLa , Humanos , Ligandos , Masculino , Ratones , Ratones Endogámicos NOD , Ratones Noqueados , Ratones SCID , Neoplasias/genética , Transducción de Señal/efectos de los fármacos , Transducción de Señal/genética , Ensayos Antitumor por Modelo de Xenoinjerto
4.
Cell Microbiol ; 21(1): e12963, 2019 01.
Artículo en Inglés | MEDLINE | ID: mdl-30321912

RESUMEN

Candida albicans is an opportunistic fungal pathogen that colonises the skin as well as genital and intestinal mucosa of most healthy individuals. The ability of C. albicans to switch between different morphological states, for example, from an ellipsoid yeast form to a highly polarised, hyphal form, contributes to its success as a pathogen. In highly polarised tip-growing cells such as neurons, pollen tubes, and filamentous fungi, delivery of membrane and cargo to the filament apex is achieved by long-range delivery of secretory vesicles tethered to motors moving along cytoskeletal cables that extend towards the growing tip. To investigate whether such a mechanism is also critical for C. albicans filamentous growth, we studied the dynamics and organisation of the C. albicans secretory pathway using live cell imaging and three-dimensional electron microscopy. We demonstrate that the secretory pathway is organised in distinct domains, including endoplasmic reticulum membrane sheets that extend along the length of the hyphal filament, a sub-apical zone exhibiting distinct membrane structures and dynamics and a Spitzenkörper comprised of uniformly sized secretory vesicles. Our results indicate that the organisation of the secretory pathway in C. albicans likely facilitates short-range "on-site" secretory vesicle delivery, in contrast to filamentous fungi and many highly polarised cells.


Asunto(s)
Candida albicans/crecimiento & desarrollo , Candida albicans/metabolismo , Citoesqueleto/metabolismo , Hifa/crecimiento & desarrollo , Hifa/metabolismo , Vesículas Secretoras/metabolismo , Candida albicans/ultraestructura , Hifa/ultraestructura , Imagenología Tridimensional , Microscopía Intravital , Microscopía Electrónica
5.
Circ Res ; 122(10): 1369-1384, 2018 05 11.
Artículo en Inglés | MEDLINE | ID: mdl-29523554

RESUMEN

RATIONALE: Macrophages face a substantial amount of cholesterol after the ingestion of apoptotic cells, and the LIPA (lysosomal acid lipase) has a major role in hydrolyzing cholesteryl esters in the endocytic compartment. OBJECTIVE: Here, we directly investigated the role of LIPA-mediated clearance of apoptotic cells both in vitro and in vivo. METHODS AND RESULTS: We show that LIPA inhibition causes a defective efferocytic response because of impaired generation of 25-hydroxycholesterol and 27-hydroxycholesterol. Reduced synthesis of 25-hydroxycholesterol after LIPA inhibition contributed to defective mitochondria-associated membrane leading to mitochondrial oxidative stress-induced NLRP3 (NOD-like receptor family, pyrin domain containing) inflammasome activation and caspase-1-dependent Rac1 (Ras-related C3 botulinum toxin substrate 1) degradation. A secondary event consisting of failure to appropriately activate liver X receptor-mediated pathways led to mitigation of cholesterol efflux and apoptotic cell clearance. In mice, LIPA inhibition caused defective clearance of apoptotic lymphocytes and stressed erythrocytes by hepatic and splenic macrophages, culminating in splenomegaly and splenic iron accumulation under hypercholesterolemia. CONCLUSIONS: Our findings position lysosomal cholesterol hydrolysis as a critical process that prevents metabolic inflammation by enabling efficient macrophage apoptotic cell clearance.


Asunto(s)
Colesterol/metabolismo , Inflamación/metabolismo , Lisosomas/metabolismo , Macrófagos/metabolismo , Oxiesteroles/metabolismo , Esterol Esterasa/metabolismo , Animales , Apoptosis , Transporte Biológico , Ésteres del Colesterol/metabolismo , Eritrocitos/metabolismo , Hidrólisis , Hipercolesterolemia/metabolismo , Inflamasomas/metabolismo , Receptores X del Hígado/metabolismo , Linfocitos/metabolismo , Ratones , Ratones Endogámicos C57BL , Mitocondrias/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Neuropéptidos/metabolismo , Receptores de LDL/metabolismo , Esplenomegalia/metabolismo , Esterol Esterasa/antagonistas & inhibidores , Proteína de Unión al GTP rac1/metabolismo
6.
Anal Chem ; 91(3): 1773-1778, 2019 Feb 05.
Artículo en Inglés | MEDLINE | ID: mdl-30580509

RESUMEN

Energy dispersive X-ray spectrometry (EDX or EDS) is a technique often implemented on scanning electron microscopes and a regularly used method for qualitative characterization of solid catalysts. This Technical Note reports a method for the determination of the metal content in a sulfonated polyether ether ketone in the form of an indium(III) salt. The possibility of quantitative determination of the sulfur/indium ratio by EDX was assessed by calibration with two indium salts (sulfide and sulfate) readily available in good purity. The accuracy of the uncorrected instrument response was better than 1% under our conditions. A protocol for investigating the metal content of the solid catalyst is proposed, also providing information about the homogeneity of the metal distribution. Because of the simplicity of the sample preparation, the small quantity of material needed, and the rapidity of the EDX measurements, the method appears to be promising for quantitative characterization of solid catalysts.

7.
Sci Rep ; 14(1): 1021, 2024 01 10.
Artículo en Inglés | MEDLINE | ID: mdl-38200072

RESUMEN

Legacy radioactive waste can be defined as the radioactive waste produced during the infancy of the civil nuclear industry's development in the mid-20th Century, a time when, unfortunately, waste storage and treatment were not well planned. The marine environment is one of the environmental compartments worth studying in this regard because of legacy waste in specific locations of the seabed. Comprising nearly 70% of the earth's service, the oceans are the largest and indeed the final destination for contaminated fresh waters. For this reason, long-term studies of the accumulation biochemical mechanisms of metallic radionuclides in the marine ecosystem are required. In this context the brown algal compartment may be ecologically relevant because of forming large and dense algal beds in coastal areas and potential important biomass for contamination. This report presents the first step in the investigation of uranium (U, an element used in the nuclear cycle) bioaccumulation in the brown alga Ascophyllum nodosum using a multi-scale spectroscopic and imaging approach. Contamination of A. nodosum specimens in closed aquaria at 13 °C was performed with a defined quantity of U(VI) (10-5 M). The living algal uptake was quantified by ICP-MS and a localization study in the various algal compartments was carried out by combining electronic microscopy imaging (SEM), X-ray Absorption spectroscopy (XAS) and micro X-ray Florescence (µ-XRF). Data indicate that the brown alga is able to concentrate U(VI) by an active bioaccumulation mechanism, reaching an equilibrium state after 200 h of daily contamination. A comparison between living organisms and dry biomass confirms a stress-response process in the former, with an average bioaccumulation factor (BAF) of 10 ± 2 for living specimens (90% lower compared to dry biomass, 142 ± 5). Also, these results open new perspectives for a potential use of A. nodosum dry biomass as uranium biosorbent. The different partial BAFs (bioaccumulation factors) range from 3 (for thallus) to 49 (for receptacles) leading to a compartmentalization of uranium within the seaweed. This reveals a higher accumulation capacity in the receptacles, the algal reproductive parts. SEM images highlight the different tissue distributions among the compartments with a superficial absorption in the thallus and lateral branches and several hotspots in the oospheres of the female individuals. A preliminary speciation XAS analysis identified a distinct U speciation in the gametes-containing receptacles as a pseudo-autunite phosphate phase. Similarly, XAS measurements on the lateral branches (XANES) were not conclusive with regards to the occurrence of an alginate-U complex in these tissues. Nonetheless, the hypothesis that alginate may play a role in the speciation of U in the algal thallus tissues is still under consideration.


Asunto(s)
Ascophyllum , Residuos Radiactivos , Uranio , Humanos , Femenino , Bioacumulación , Ecosistema , Espectroscopía de Absorción de Rayos X , Alginatos
8.
J Neurochem ; 125(6): 921-31, 2013 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-23331044

RESUMEN

EFhd2 is a conserved calcium-binding protein, abundant within the central nervous system. Previous studies identified EFhd2 associated with pathological forms of tau proteins in the tauopathy mouse model JNPL3, which expresses the human tau(P301L) mutant. This association was validated in human tauopathies, such as Alzheimer's disease (AD). However, the role that EFhd2 may play in tauopathies is still unknown. Here, we show that EFhd2 formed amyloid structures in vitro, a capability that is reduced by calcium ions. Electron microscopy (EM) analyses demonstrated that recombinant EFhd2 formed filamentous structures. EM analyses of sarkosyl-insoluble fractions derived from human AD brains also indicated that EFhd2 co-localizes with aggregated tau proteins and formed granular structures. Immunohistological analyses of brain slices demonstrated that EFhd2 co-localizes with pathological tau proteins in AD brains, confirming the co-aggregation of EFhd2 and pathological tau. Furthermore, EFhd2's coiled-coil domain mediated its self-oligomerization in vitro and its association with tau proteins in JNPL3 mouse brain extracts. The results demonstrate that EFhd2 is a novel amyloid protein associated with pathological tau proteins in AD brain and that calcium binding may regulate the formation of EFhd2's amyloid structures. Hence, EFhd2 may play an important role in the pathobiology of tau-mediated neurodegeneration.


Asunto(s)
Enfermedad de Alzheimer/metabolismo , Proteínas Amiloidogénicas/metabolismo , Encéfalo/metabolismo , Proteínas de Unión al Calcio/metabolismo , Proteínas tau/metabolismo , Enfermedad de Alzheimer/patología , Amiloide/metabolismo , Animales , Encéfalo/patología , Humanos , Ratones , Ratones Transgénicos , Ovillos Neurofibrilares/metabolismo , Ovillos Neurofibrilares/patología , Multimerización de Proteína , Estructura Terciaria de Proteína , Proteínas tau/genética
9.
Orig Life Evol Biosph ; 42(6): 587-609, 2012 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-23254852

RESUMEN

Identification of the earliest traces of life is made difficult by the scarcity of the preserved microbial remains and by the alteration and potential contamination of the organic matter (OM) content of rocks. These factors can confuse interpretations of the biogenicity and syngenicity of fossilised structures and organic molecules found in ancient rocks. In order to improve our knowledge of the fossilisation processes and their effects at the molecular level, we made a preliminary study of the fate of OM during experimental fossilisation. Changes in the composition and quantity of amino acids, monosaccharides and fatty acids were followed with HPLC, GC and GC-MS analyses during 1 year of silicification of the hyperthermophilic Archaea Methanocaldococcus jannaschii. Although the cells themselves did not fossilise and the accompanying extracellular polymeric substances (EPS) did, our analyses showed that the OM initially present in both cells and EPS was uniformly preserved in the precipitated silica, with amino acids and fatty acids being the best preserved compounds. This study thus completes previous data obtained by electron microscopy investigations of simulated microbial fossilisation and can help better identification and interpretation of microbial biosignatures in both ancient rocks and in recent hydrothermal formations and sediments.


Asunto(s)
Fósiles , Sedimentos Geológicos/química , Methanococcales/química , Compuestos Orgánicos/química , Origen de la Vida , Evolución Biológica , Cromatografía de Gases , Cromatografía Líquida de Alta Presión , Cromatografía de Gases y Espectrometría de Masas
10.
Elife ; 112022 03 28.
Artículo en Inglés | MEDLINE | ID: mdl-35343437

RESUMEN

Phytophthora species cause diseases in a large variety of plants and represent a serious agricultural threat, leading, every year, to multibillion dollar losses. Infection occurs when their biflagellated zoospores move across the soil at their characteristic high speed and reach the roots of a host plant. Despite the relevance of zoospore spreading in the epidemics of plant diseases, individual swimming of zoospores have not been fully investigated. It remains unknown about the characteristics of two opposite beating flagella during translation and turning, and the roles of each flagellum on zoospore swimming. Here, combining experiments and modeling, we show how these two flagella contribute to generate thrust when beating together, and identify the mastigonemes-attached anterior flagellum as the main source of thrust. Furthermore, we find that turning involves a complex active process, in which the posterior flagellum temporarily stops, while the anterior flagellum keeps on beating and changes its gait from sinusoidal waves to power and recovery strokes, similar to Chlamydomonas's breaststroke, to reorient its body to a new direction. Our study is a fundamental step toward a better understanding of the spreading of plant pathogens' motile forms, and shows that the motility pattern of these biflagellated zoospores represents a distinct eukaryotic version of the celebrated 'run-and-tumble' motility class exhibited by peritrichous bacteria.


Microorganisms of the Phytophthora genus are serious agricultural pests. They cause diseases in many crops, including potato, onion, tomato, tobacco, cotton, peppers, and citrus. These diseases cause billions of dollars in losses each year. Learning more about how the tiny creatures disseminate and reach host plants could help scientists develop new ways to prevent such crop damage. The spore cells of Phytophthora, also known as zoospores, have two appendages called flagella on their bodies. A tinsel-shaped flagellum is near the front of the creature and a long smooth filament-like flagellum is near the posterior. Zoospores use their flagella to swim at high speeds through liquid toward potential plant hosts. Their complex swimming patterns change in response to different physical, chemical, and electrical signals in the environment. But exactly how they use their flagella to generate these movements is not clear. Tran et al. reveal new details about zoospore locomotion. In the experiments, Tran et al. recorded the movements of zoospores in a tiny 'swimming pool' of fluid on top of a glass slide and analyzed the movements using statistical and mathematical models. The results uncovered coordinated actions of the flagella when zoospores swim in a straight line and when they turn. The tinsel-like front flagellum provides most of the force that propels the zoospore forward. To do this, it beats with an undulating wave pattern. It shifts the beating to a breast-stroke pattern to change direction. The posterior flagellum provides a smaller forward thrust and temporarily pauses during turns. The study provides new details about zoospore's movements that may help scientists develop new strategies to control these pests. It also offers more information about how flagella coordinate their actions to switch speeds or change directions that may be of interest to other scientists studying organisms that use flagella to move.


Asunto(s)
Phytophthora , Natación , Cilios , Flagelos , Enfermedades de las Plantas , Plantas
11.
J Colloid Interface Sci ; 590: 260-267, 2021 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-33548609

RESUMEN

HYPOTHESIS: The formation of porous nanostructures on surfaces and the control of their size and shape is fundamental for various applications. The creation of nanotubes is particularly difficult to implement without the aid of hard and rigid templates. Recently, methods that form nanotubular structures in a straightforward manner and without direct templating, e.g. soft templating, have been highly sought after. Here we propose the use of "soft templating" via self-assembly of conducting monomers during electropolymerization in organic solvents as a mean to form porous, nanotubular features. EXPERIMENTS: Naphtho[2,3-b]thieno[3,4-e][1,4]dioxine (NaphDOT) is employed as monomer for electropolymerizations conducted in dichloromethane and chloroform containing varying amounts of water. SEM analyses of the resulting surfaces confirms the strong capacity of NaphDOT to form vertically aligned nanotubes. Polymerization solutions analyzed by DLS and TEM reveal the presence of micelles prior to electropolymerization, and the size of the micelles correlates with the inner diameter of the nanotubes formed. FINDINGS: We show that micelles in polymerization solutions are stabilized by both monomers and electrolytes. We propose a mechanism where reverse micelles are forming a soft-template responsible for the formation of porous nanostructures during electropolymerization in organic, non-polar solvents. In this mechanism, the monomer and electrolyte assume the role of surfactant in the reverse micelle system.

12.
Forensic Sci Int ; 324: 110816, 2021 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-34030001

RESUMEN

In forensic anthropology, saw mark analysis plays an important role in cases of criminal dismemberment. Autopsy saw is not used by the perpetrator in cases of dismemberment, but the forensic pathologist may accidentally create false starts with this saw during an autopsy, especially while sampling bones for further analysis, and these autopsy false starts can be confused with false starts produced by the offender. In this study, the characteristics of 20 false starts were compared using stereomicroscopy (SM) and scanning electron microscopy (SEM). These bone false starts were selected at random from a previous study of 100 false starts created by an electrical oscillating autopsy saw on human femoral bones. That study had enabled the categorization of the lesions into two groups ("superficial group" and "deep group") with a 0.52 mm depth cut off, based on the dramatic differences in lesion characteristics between these two groups. In the current study, SEM confirmed the characteristics of the false starts (walls and profile shapes, striae, bone islands and bone debris were studied), and above all explained the mechanism whereby oval bone islands in deep lesions are formed. Bone islands are due to the horizontal and vertical movement of the oscillating autopsy blade.


Asunto(s)
Desmembramiento de Cadáver , Fémur/lesiones , Fémur/ultraestructura , Microscopía Electrónica de Rastreo , Microscopía , Antropología Forense/métodos , Humanos
13.
J Comp Neurol ; 529(6): 1135-1156, 2021 04 15.
Artículo en Inglés | MEDLINE | ID: mdl-32841380

RESUMEN

The echinoderms are a phylum of marine deuterostomes characterized by the pentaradial (five fold) symmetry of their adult bodies. Due to this unusual body plan, adult echinoderms have long been excluded from comparative analyses aimed at understanding the origin and evolution of deuterostome nervous systems. Here, we investigated the neural anatomy of early juveniles of representatives of three of the five echinoderm classes: the echinoid Paracentrotus lividus, the asteroid Patiria miniata, and the holothuroid Parastichopus parvimensis. Using whole mount immunohistochemistry and confocal microscopy, we found that the nervous system of echinoid early juveniles is composed of three main structures: a basiepidermal nerve plexus, five radial nerve cords connected by a circumoral nerve ring, and peripheral nerves innervating the appendages. Our whole mount preparations further allowed us to obtain thorough descriptions of these structures and of several innervation patterns, in particular at the level of the appendages. Detailed comparisons of the echinoid juvenile nervous system with those of asteroid and holothuroid juveniles moreover supported a general conservation of the main neural structures in all three species, including at the level of the appendages. Our results support the previously proposed hypotheses for the existence of two neural units in echinoderms: one consisting of the basiepidermal nerve plexus to process sensory stimuli locally and one composed of the radial nerve cords and the peripheral nerves constituting a centralized control system. This study provides the basis for more in-depth comparisons of the echinoderm adult nervous system with those of other animals, in particular hemichordates and chordates, to address the long-standing controversies about deuterostome nervous system evolution.


Asunto(s)
Evolución Biológica , Sistema Nervioso/anatomía & histología , Paracentrotus/anatomía & histología , Factores de Edad , Animales , Equinodermos , Femenino , Larva , Masculino , Sistema Nervioso/química , Paracentrotus/química
14.
mBio ; 13(1): e0387321, 2021 02 22.
Artículo en Inglés | MEDLINE | ID: mdl-35164565

RESUMEN

Phosphatidylinositol phosphates are key phospholipids with a range of regulatory roles, including membrane trafficking and cell polarity. Phosphatidylinositol-4-phosphate [PI(4)P] at the Golgi apparatus is required for the budding-to-filamentous-growth transition in the human-pathogenic fungus Candida albicans; however, the role of plasma membrane PI(4)P is unclear. We have investigated the importance of this phospholipid in C. albicans growth, stress response, and virulence by generating mutant strains with decreased levels of plasma membrane PI(4)P, via deletion of components of the PI-4-kinase complex, i.e., Efr3, Ypp1, and Stt4. The amounts of plasma membrane PI(4)P in the efr3Δ/Δ and ypp1Δ/Δ mutants were ∼60% and ∼40%, respectively, of that in the wild-type strain, whereas it was nearly undetectable in the stt4Δ/Δ mutant. All three mutants had reduced plas7ma membrane phosphatidylserine (PS). Although these mutants had normal yeast-phase growth, they were defective in filamentous growth, exhibited defects in cell wall integrity, and had an increased exposure of cell wall ß(1,3)-glucan, yet they induced a range of hyphal-specific genes. In a mouse model of hematogenously disseminated candidiasis, fungal plasma membrane PI(4)P levels directly correlated with virulence; the efr3Δ/Δ mutant had wild-type virulence, the ypp1Δ/Δ mutant had attenuated virulence, and the stt4Δ/Δ mutant caused no lethality. In the mouse model of oropharyngeal candidiasis, only the ypp1Δ/Δ mutant had reduced virulence, indicating that plasma membrane PI(4)P is less important for proliferation in the oropharynx. Collectively, these results demonstrate that plasma membrane PI(4)P levels play a central role in filamentation, cell wall integrity, and virulence in C. albicans. IMPORTANCE While the PI-4-kinases Pik1 and Stt4 both produce PI(4)P, the former generates PI(4)P at the Golgi apparatus and the latter at the plasma membrane, and these two pools are functionally distinct. To address the importance of plasma membrane PI(4)P in Candida albicans, we generated deletion mutants of the three putative plasma membrane PI-4-kinase complex components and quantified the levels of plasma membrane PI(4)P in each of these strains. Our work reveals that this phosphatidylinositol phosphate is specifically critical for the yeast-to-hyphal transition, cell wall integrity, and virulence in a mouse systemic infection model. The significance of this work is in identifying a plasma membrane phospholipid that has an infection-specific role, which is attributed to the loss of plasma membrane PI(4)P resulting in ß(1,3)-glucan unmasking.


Asunto(s)
Candida albicans , Candidiasis , Animales , Ratones , Candida albicans/genética , Candidiasis/microbiología , Membrana Celular/metabolismo , Pared Celular/metabolismo , Modelos Animales de Enfermedad , Proteínas Fúngicas/genética , Hifa , Fosfatos de Fosfatidilinositol/metabolismo
15.
Nat Metab ; 3(10): 1313-1326, 2021 10.
Artículo en Inglés | MEDLINE | ID: mdl-34650273

RESUMEN

Macrophages rely on tightly integrated metabolic rewiring to clear dying neighboring cells by efferocytosis during homeostasis and disease. Here we reveal that glutaminase-1-mediated glutaminolysis is critical to promote apoptotic cell clearance by macrophages during homeostasis in mice. In addition, impaired macrophage glutaminolysis exacerbates atherosclerosis, a condition during which, efficient apoptotic cell debris clearance is critical to limit disease progression. Glutaminase-1 expression strongly correlates with atherosclerotic plaque necrosis in patients with cardiovascular diseases. High-throughput transcriptional and metabolic profiling reveals that macrophage efferocytic capacity relies on a non-canonical transaminase pathway, independent from the traditional requirement of glutamate dehydrogenase to fuel ɑ-ketoglutarate-dependent immunometabolism. This pathway is necessary to meet the unique requirements of efferocytosis for cellular detoxification and high-energy cytoskeletal rearrangements. Thus, we uncover a role for non-canonical glutamine metabolism for efficient clearance of dying cells and maintenance of tissue homeostasis during health and disease in mouse and humans.


Asunto(s)
Aminación , Glutamina/metabolismo , Fosforilación Oxidativa , Animales , Ratones , Fagocitosis
16.
Antibiotics (Basel) ; 9(4)2020 Mar 25.
Artículo en Inglés | MEDLINE | ID: mdl-32218125

RESUMEN

Oral microbiome plays an important part on oral health and endogenous bacteria and fungi should not be eradicated. However, their proliferation must be controlled by oral hygiene care. In vitro, Solidago virgaurea ssp. virgaurea L. (SV) plant extract inhibits the adherence and hyphal formation of a fungus, Candida albicans. It reduces the biomass of Candida-bacterial biofilms but not fungal or bacterial growth. Unlike chemical antiseptics, like triclosan and chlorhexidine for instance, SV is a plant extract easily biodegradable. The purpose of this study was to assess the in vivo effectiveness of SV extract in reducing oral biomass. A randomized, double-blind clinical study, with dental plaque evaluation designed to assess the effectiveness of a fluorinated toothpaste containing SV (Bucovia™, Givaudan, Vernier, Switzerland) was conducted. Sixty-six subjects (SV group n = 33 vs. control n = 33) brushed their teeth twice a day for a 4-week period. Supragingival dental plaque was sampled. Total bacterial load (broad spectral bacterial quantitative Polymerase Chain Reaction (qPCR)), C. albicans and seven bacterial species were quantified by qPCR. In the Intervention group, there was a decrease of Total bacterial load (ΔD0D28 p = 0.005 and ΔD14D28 p = 0.026), Streptococcus mutans (ΔD0D14 p = 0.024) and C. albicans (ΔD0D28 p = 0.022). In the Control group Total bacterial load tended to decrease from baseline to day 28 (ΔD0D28 p = 0.062 and ΔD14D28 p = 0.009). Plaque Index and Gingival Index improved in both groups.

17.
Microorganisms ; 8(7)2020 Jul 07.
Artículo en Inglés | MEDLINE | ID: mdl-32645882

RESUMEN

Most pathogenic oomycetes of the genus Phytophthora spread in water films as flagellated zoospores. Zoospores perceive and produce signals attracting other zoospores, resulting in autoaggregation in vitro or biofilm formation on plant surface. The mechanisms underlying intercellular communication and consequent attraction, adhesion and aggregation are largely unknown. In Phytophthora parasitica, the perception of a K+ gradient induces coordinated motion and aggregation. To define cellular and molecular events associated with oomycete aggregation, we combined transcriptomic and ultrastructural analyses. Results indicate involvement of electroception in K+ sensing. They establish that the transcriptome repertoire required for swimming and aggregation is already fully functional at zoospore release. At the time points analyzed, aggregates are mainly constituted of zoospores. They produce vesicular and fibrillary material discharged at cell-to-cell contacts. Consistently, the signature of transcriptome dynamics during transition to aggregates is an upregulation of genes potentially related to vesicular trafficking. Moreover, transcriptomic and functional analyses show a strong enhancement of carbonic anhydrase activity, indicating that pH homeostasis may contribute to aggregation by acting on both zoospore movement and adhesion. This study poses the molecular and cellular bases of aggregative behavior within oomycetes and expands the current knowledge of ion perception-mediated dissemination of propagules in the rhizosphere.

18.
Science ; 369(6505): 863-866, 2020 08 14.
Artículo en Inglés | MEDLINE | ID: mdl-32792402

RESUMEN

Early plant use is seldom described in the archaeological record because of poor preservation. We report the discovery of grass bedding used to create comfortable areas for sleeping and working by people who lived in Border Cave at least 200,000 years ago. Sheaves of grass belonging to the broad-leafed Panicoideae subfamily were placed near the back of the cave on ash layers that were often remnants of bedding burned for site maintenance. This strategy is one forerunner of more-complex behavior that is archaeologically discernible from ~100,000 years ago.


Asunto(s)
Cuevas , Incendios/historia , Horticultura/historia , Poaceae , Antropología , Arqueología , Historia Antigua , Humanos , Sudáfrica
20.
Arthropod Struct Dev ; 49: 10-18, 2019 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-30721756

RESUMEN

In the animal kingdom, macroscopic variations in size, color, and even hairiness are frequently observed between male and female, making the sex of various species easy to discern. In the case of insects, similar variances also exist. While direct observation is a quick and efficient way to differentiate between sexes, there are also variations which are unseen to the naked eye and occur on a micro- or nanoscopic scale. Sometimes, these micro/nanoscopic variations can lead to significant variations in surface properties as a function of sex. Such is the case for the Mecynorhina polyphemus confluens (Kraatz, 1890). In this work, we characterize these micro- and nanoscale differences, and describe their impact on the surface properties (e.g. wettability). It is found that water interacts quite differently with the surface of the cuticle of Mecynorhina polyphenus confluens, depending on the specimen sex. On a female, water spreads readily across the elytra indicating hydrophilic behavior. However, on the surface of the male elytra, strong hydrophobicity is observed. Microscopic observations reveal differences in microscale surface morphology across the male and female cuticle. These observations contribute to a better, global understanding of the wettability behavior observed on M. polyphemus confluens.


Asunto(s)
Escarabajos/anatomía & histología , Escarabajos/química , Animales , Escarabajos/ultraestructura , Femenino , Masculino , Microscopía Electrónica de Rastreo , Caracteres Sexuales , Propiedades de Superficie , Humectabilidad
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