RESUMEN
The role of glycosaminoglycans (GAGs) in modulating bone morphogenetic protein (BMP) signaling represents a recent and underexplored area. Conflicting reports suggest a dual effect: some indicate a positive influence, while others demonstrate a negative impact. This duality suggests that the localization of GAGs (either at the cell surface or within the extracellular matrix) or the specific type of GAG may dictate their signaling role. The precise sulfation patterns of heparan sulfate (HS) responsible for BMP2 binding remain elusive. BMP2 exhibits a preference for binding to HS over other GAGs. Using well-characterized biomaterials mimicking the extracellular matrix, our research reveals that HS promotes BMP2 signaling in the extracellular space, contrary to chondroitin sulfate (CS), which enhances BMP2 bioactivity at the cell surface. Further observations indicate that a central IdoA (2S)-GlcNS (6S) tri-sulfated motif within HS hexasaccharides enhances binding. Nevertheless, BMP2 exhibits a degree of adaptability to various HS sulfation types and sequences. Molecular dynamic simulations attribute this adaptability to the BMP2 N-terminal end flexibility. Our findings illustrate the complex interplay between GAGs and BMP signaling, highlighting the importance of localization and specific sulfation patterns. This understanding has implications for the development of biomaterials with tailored properties for therapeutic applications targeting BMP signaling pathways.
Asunto(s)
Proteína Morfogenética Ósea 2 , Glicosaminoglicanos , Heparitina Sulfato , Transducción de Señal , Proteína Morfogenética Ósea 2/metabolismo , Heparitina Sulfato/metabolismo , Heparitina Sulfato/química , Humanos , Glicosaminoglicanos/metabolismo , Glicosaminoglicanos/química , Sulfatos de Condroitina/química , Sulfatos de Condroitina/metabolismo , Simulación de Dinámica Molecular , Animales , Unión ProteicaRESUMEN
Most of the nitrogen (N) accessible for life is trapped in dinitrogen (N2), the most stable atmospheric molecule. In order to be metabolized by living organisms, N2 has to be converted into biologically assimilable forms, so-called fixed N. Nowadays, nearly all the N-fixation is achieved through biological and anthropogenic processes. However, in early prebiotic environments of the Earth, N-fixation must have occurred via natural abiotic processes. One of the most invoked processes is electrical discharges, including from thunderstorms and lightning associated with volcanic eruptions. Despite the frequent occurrence of volcanic lightning during explosive eruptions and convincing laboratory experimentation, no evidence of substantial N-fixation has been found in any geological archive. Here, we report on the discovery of a significant amount of nitrate in volcanic deposits from Neogene caldera-forming eruptions, which are well correlated with the concentrations of species directly emitted by volcanoes (sulfur, chlorine). The multi-isotopic composition (δ18O, Δ17O) of the nitrates reveals that they originate from the atmospheric oxidation of nitrogen oxides formed by volcanic lightning. According to these first geological volcanic nitrate archive, we estimate that, on average, about 60 Tg of N can be fixed during a large explosive event. Our findings hint at a unique role potentially played by subaerial explosive eruptions in supplying essential ingredients for the emergence of life on Earth.
RESUMEN
A dense system of pre-Hispanic urban centers has been found in the Upano Valley of Amazonian Ecuador, in the eastern foothills of the Andes. Fieldwork and light detection and ranging (LIDAR) analysis have revealed an anthropized landscape with clusters of monumental platforms, plazas, and streets following a specific pattern intertwined with extensive agricultural drainages and terraces as well as wide straight roads running over great distances. Archaeological excavations date the occupation from around 500 BCE to between 300 and 600 CE. The most notable landscape feature is the complex road system extending over tens of kilometers, connecting the different urban centers, thus creating a regional-scale network. Such extensive early development in the Upper Amazon is comparable to similar Maya urban systems recently highlighted in Mexico and Guatemala.
RESUMEN
Glycosaminoglycans (GAGs) play a crucial role in tissue homeostasis by regulating the activity and diffusion of bioactive molecules. Incorporating GAGs into biomaterials has emerged as a widely adopted strategy in medical applications, owing to their biocompatibility and ability to control the release of bioactive molecules. Nevertheless, immobilized GAGs on biomaterials can elicit distinct cellular responses compared to their soluble forms, underscoring the need to understand the interactions between GAG and bioactive molecules within engineered functional biomaterials. By controlling critical parameters such as GAG type, density, and sulfation, it becomes possible to precisely delineate GAG functions within a biomaterial context and to better mimic specific tissue properties, enabling tailored design of GAG-based biomaterials for specific medical applications. However, this requires access to pure and well-characterized GAG compounds, which remains challenging. This review focuses on different strategies for producing well-defined GAGs and explores high-throughput approaches employed to investigate GAG-growth factor interactions and to quantify cellular responses on GAG-based biomaterials. These automated methods hold considerable promise for improving the understanding of the diverse functions of GAGs. In perspective, the scientific community is encouraged to adopt a rational approach in designing GAG-based biomaterials, taking into account the in vivo properties of the targeted tissue for medical applications.
Asunto(s)
Materiales Biocompatibles , GlicosaminoglicanosRESUMEN
Tungurahua volcano (Ecuador) intermittently emitted ash between 1999 and 2016, enduringly affecting the surrounding rural area and its population, but its health impact remains poorly documented. We aim to assess the respiratory health hazard posed by the 16-17 August 2006 most intense eruptive phase of Tungurahua. We mapped the spatial distribution of the health-relevant ash size fractions produced by the eruption in the area impacted by ash fallout. We quantified the mineralogy, composition, surface texture, and morphology of a respirable ash sample isolated by aerodynamic separation. We then assessed the cytotoxicity and pro-inflammatory potential of this respirable ash toward lung tissues in-vitro using A549 alveolar epithelial cells, by electron microscopy and biochemical assays. The eruption produced a high amount of inhalable and respirable ash (12.0-0.04 kg/m2 of sub-10 µm and 5.3-0.02 kg/m2 of sub-4 µm ash deposited). Their abundance and proportion vary greatly across the deposit within the first 20 km from the volcano. The respirable ash is characteristic of an andesitic magma and no crystalline silica is detected. Morphological features and surface textures are complex and highly variable, with few fibers observed. In-vitro experiments show that respirable volcanic ash is internalized by A549 cells and processed in the endosomal pathway, causing little cell damage, but resulting in changes in cell morphology and membrane texture. The ash triggers a weak pro-inflammatory response. These data provide the first understanding of the respirable ash hazard near Tungurahua and the extent to which it varies spatially in a fallout deposit.
RESUMEN
The automation of liquid-handling routines offers great potential for fast, reproducible, and labor-reduced biomaterial fabrication but also requires the development of special protocols. Competitive systems demand for a high degree in miniaturization and parallelization while maintaining flexibility regarding the experimental design. Today, there are only a few possibilities for automated fabrication of biomaterials inside multiwell plates. We have previously demonstrated that streptavidin-based biomimetic platforms can be employed to study cellular behaviors on biomimetic surfaces. So far, these self-assembled materials were made by stepwise assembly of the components using manual pipetting. In this work, we introduce for the first time a fully automated and adaptable workflow to functionalize glass-bottom multiwell plates with customized biomimetic platforms deposited in single wells using a liquid-handling robot. We then characterize the cell response using automated image acquisition and subsequent analysis. Furthermore, the molecular surface density of the biomimetic platforms was characterized in situ using fluorescence-based image correlation spectroscopy. These measurements were in agreement with standard ex situ spectroscopic ellipsometry measurements. Due to automation, we could do a proof of concept to study the effect of heparan sulfate on the bioactivity of bone morphogenetic proteins on myoblast cells, using four different bone morphogenetic proteins (BMPs) (2, 4, 6, and 7) in parallel, at five increasing concentrations. Using such an automated self-assembly of biomimetic materials, it may be envisioned to further investigate the role of a large variety of extracellular matrix (ECM) components and growth factors on cell signaling.
RESUMEN
Given the growing demand for studies dealing with natural disasters, the research fields of emotion and social cognition require validated picture stimuli of natural hazards. Such material is essential for studying perceptual processes and behaviors of exposed individuals, and it could find practical applications, such as the improvement of communication strategies during crises. We present the Natural Disasters Picture System (NDPS), a database of pictures of natural hazards, with an emphasis on volcanic threats, and their impact on the environment and humans. We first describe in detail the picture selection and database creation. We then report the validation procedure. One hundred twenty participants rated the pictures on the basis of four dimensions: valence, arousal, dominance and certainty. For each picture, we ultimately determined the best-fitting emotion on the basis of its dimensional pattern. The Hierarchical Ascendant Classification, which yielded 4 clusters subdivided into 9 classes, indicated a highly consistent and distinctive classification of the pictures. Overall, 90% of the pictures elicited negative emotions (fear or sadness), and the other 10% induced neutral to positive emotions (e.g., aesthetic emotions). The NDPS offers a new tool for studying natural events and disasters in the field of affective and cognitive sciences, which will benefit from scientific research and its practical applications. The NDPS is unrestrictedly accessible for researchers.
Asunto(s)
Bases de Datos Factuales , Desastres Naturales , Vigilancia en Salud Pública , Emociones , Miedo , Femenino , Humanos , Masculino , Reconocimiento Visual de Modelos , Estimulación LuminosaRESUMEN
In cases of impending natural disasters, most events are uncertain and emotionally relevant, both critical factors for decision-making. Moreover, for exposed individuals, the sensitivity to the framing of the consequences (gain or loss) and the moral judgments they have to perform (e.g., evacuate or help an injured person) constitute two central effects that have never been examined in the same context of decision-making. In a framed decision-making task with moral dilemma, we investigated whether uncertainty (i.e., unpredictably of events) and a threatening context would influence the framing effect (actions framed in loss are avoided in comparison to the ones framed in gain) and the personal intention effect (unintentional actions are more morally acceptable in comparison to intentional actions) on the perceived moral acceptability of taking action. Considering the impact of uncertainty and fear on the processes underlying these effects, we assumed that these emotions would lead to the negation of the two effects. Our results indicate that the exposure to uncertain events leads to the negation of the framing effect, but does not influence the moral acceptability and the effect of personal intention. We discuss our results in the light of dual-process models (i.e. systematic vs. heuristic), appraisal theories, and neurocognitive aspects. These elements highlight the importance of providing solutions to cope with uncertainty, both for scientists and local populations exposed to natural hazards.
Asunto(s)
Toma de Decisiones/ética , Principios Morales , Incertidumbre , Femenino , Humanos , Masculino , Modelos Estadísticos , Adulto JovenRESUMEN
Lantibiotics are antimicrobial peptides from the bacteriocin family, secreted by Gram-positive bacteria. These peptides differ from other bacteriocins by the presence of (methyl)lanthionine residues, which result from enzymatic modification of precursor peptides encoded by structural genes. Several groups of lantibiotics have been distinguished, the largest of which is the lacticin 481 group. This group consists of at least 16 members, including lacticin 481, streptococcin A-FF22, mutacin II, nukacin ISK-1, and salivaricins. We present the first review devoted to this lantibiotic group, knowledge of which has increased significantly within the last few years. After updating the group composition and defining the common properties of these lantibiotics, we highlight the most recent developments. The latter concern: transcriptional regulation of the lantibiotic genes; understanding the biosynthetic machinery, in particular the ability to perform in vitro prepeptide maturation; characterization of a novel type of immunity protein; and broad application possibilities. This group differs in many aspects from the best known lantibiotic group (nisin group), but shares properties with less-studied groups such as the mersacidin, cytolysin and lactocin S groups.
Asunto(s)
Antibacterianos , Bacteriocinas , Ácidos/metabolismo , Secuencia de Aminoácidos , Antibacterianos/química , Antibacterianos/metabolismo , Antibacterianos/farmacología , Proteínas Bacterianas/genética , Bacteriocinas/química , Bacteriocinas/genética , Bacteriocinas/metabolismo , Bacteriocinas/farmacología , Regulación Bacteriana de la Expresión Génica , Bacterias Grampositivas/genética , Bacterias Grampositivas/metabolismo , Datos de Secuencia Molecular , Familia de Multigenes , Probióticos , Ingeniería de Proteínas , Percepción de Quorum , Relación Estructura-Actividad , Transcripción GenéticaRESUMEN
We previously characterized three Lactococcus lactis promoters, P170, P1 and P3, which are induced by low pH. Here, we identified a novel 14 bp regulatory DNA region centred at around -41.5 and composed of three tetranucleotide sequences, boxes A, C and D. Boxes A and C contribute to P1 activity, whereas box D and the position of boxes ACD (renamed ACiD-box) are essential to P1 activity and acid response. We also identified a trans -acting protein, RcfB, which is involved in P170 and P1 basal activity and is essential for their pH induction. The regulator belongs to the Crp-Fnr family of transcription regulators. Overexpression of rcfB resulted in increased beta-galactosidase activities and lantibiotic lacticin 481 production from P170- and P1-controlled genes, respectively, in acid condition. RcfB is thus probably activated when cells encounter an acid environment. rcfB is co-transcribed with genes encoding an universal stress-like protein and a multidrug transporter. RcfB plays a role in acid adaptation, as the survival rate of an rcfB mutant after a lethal acid challenge was 130-fold lower than that of the wild-type strain, when the bacteria were first grown in acidic medium. The groESL promoter includes a sequence resembling an ACiD-box and the chaperone GroEL production is partly RcfB dependent in acid condition. Our results suggest that the ACiD-box could be the DNA target site of RcfB.
Asunto(s)
Proteínas Bacterianas/genética , Lactococcus lactis/genética , Regiones Promotoras Genéticas/genética , Factores de Transcripción/genética , Ácidos/farmacología , Proteínas Bacterianas/metabolismo , Bacteriocinas/biosíntesis , Secuencia de Bases , Proteínas Portadoras/genética , Proteínas Portadoras/metabolismo , Chaperonina 60/genética , Chaperonina 60/metabolismo , Elementos Transponibles de ADN , Regulación Bacteriana de la Expresión Génica/efectos de los fármacos , Silenciador del Gen , Genes Reguladores , Lactococcus lactis/efectos de los fármacos , Lactococcus lactis/metabolismo , Datos de Secuencia Molecular , Mutación , Operón , Regiones Promotoras Genéticas/efectos de los fármacos , Secuencias Reguladoras de Ácidos Nucleicos , Temperatura , Factores de Transcripción/metabolismo , Transcripción GenéticaRESUMEN
In lantibiotic lacticin 481 biosynthesis, LctT cleaves the precursor peptide and exports mature lantibiotic. Matrix-assisted laser desorption ionization-time of flight mass spectrometry revealed that a truncated form of lacticin 481 is produced in the absence of LctT or after cleavage site inactivation. Production of truncated lacticin 481 is 4-fold less efficient, and its specific activity is about 10-fold lower.
Asunto(s)
Proteínas Bacterianas/metabolismo , Bacteriocinas/biosíntesis , Regulación Bacteriana de la Expresión Génica , Lactococcus lactis/metabolismo , Secuencia de Aminoácidos , Proteínas Bacterianas/genética , Proteínas Bacterianas/farmacología , Enzimas/genética , Enzimas/metabolismo , Lactococcus lactis/efectos de los fármacos , Lactococcus lactis/genética , Datos de Secuencia Molecular , Mutación , Operón , Precursores de Proteínas/genética , Precursores de Proteínas/metabolismo , Espectrometría de Masa por Láser de Matriz Asistida de Ionización DesorciónRESUMEN
The lantibiotic lacticin 481 operon (lctAMTFEG) is mainly transcribed from P1 and P3, two promoters lying upstream of lctA. A weak additional promoter allows independent expression of the immunity genes (lctFEG). Lacticin 481 production by Lactococcus lactis is stimulated by the acidification due to lactic acid production, and by artificially lowering the pH of the medium. This regulation occurs at the transcriptional level, since P1 and P3 are both acid-induced. P1 is weaker but more tightly regulated than P3. As no specific regulator is encoded by the lacticin 481 operon, P1 and P3 are likely controlled by a general regulator.
Asunto(s)
Ácidos/metabolismo , Proteínas Bacterianas/genética , Bacteriocinas/genética , Regulación Bacteriana de la Expresión Génica , Lactococcus lactis/genética , ADN Recombinante , Concentración de Iones de Hidrógeno , Lactococcus lactis/metabolismo , Operón , Regiones Promotoras Genéticas , Transcripción GenéticaRESUMEN
Class I bacteriocins (lantibiotics) and class II bacteriocins are antimicrobial peptides secreted by gram-positive bacteria. Using two lantibiotics, lacticin 481 and nisin, and the class II bacteriocin coagulin, we showed that bacteriocins can be detected without any purification from whole producer bacteria grown on plates by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS). When we compared the results of MALDI-TOF-MS performed with samples of whole cells and with samples of crude supernatants of liquid cultures, the former samples led to more efficient bacteriocin detection and required less handling. Nisin and lacticin 481 were both detected from a mixture of their producer strains, but such a mixture can yield additional signals. We used this method to determine the masses of two lacticin 481 variants, which confirmed at the peptide level the effect of mutations in the corresponding structural gene.
Asunto(s)
Bacillus/metabolismo , Bacteriocinas/metabolismo , Lactococcus lactis/metabolismo , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Bacillus/crecimiento & desarrollo , Proteínas Bacterianas/metabolismo , Medios de Cultivo , Lactococcus lactis/crecimiento & desarrollo , Nisina/metabolismo , Tromboplastina/metabolismoRESUMEN
Galectins are animal lectins that can specifically bind beta-galactosides. Thirteen galectins have already been described. This review focuses on a specific member of this family: galectin-8. This galectin was discovered in prostate cancer cells eight years ago and has been studied extensively in the last few years. The galectin-8 gene ( LGALS8) encodes numerous mRNAs by alternate splicing and the presence of three unusual polyadenylation signals. These mRNAs encode six different isoforms of galectin-8: three belong to the tandem-repeat galectin group (with two CRDs linked by a hinge peptide) and three to the prototype group (with one CRD). Various studies showed that galectin-8 is widely expressed in tumor tissues as well as in normal tissues. The level of galectin-8 expression may correlate with the malignancy of human colon cancers and the degree of differentiation of lung squamous cell carcinomas and neuro-endocrine tumors. Recently, the differences in galectin-8 expression levels between normal and tumor tissues have been used as a guide for the selection of strategies for the prevention and treatment of lung squamous cell carcinoma. These experiments are still under investigation, but demonstrate the potential of galectin-8 research to enhance our understanding of, and possibly prevent, the process of neoplastic transformation.