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1.
Exp Dermatol ; 32(6): 787-798, 2023 06.
Artículo en Inglés | MEDLINE | ID: mdl-36789506

RESUMEN

Hailey-Hailey disease (HHD) is a rare autosomal dominantly inherited disorder caused by mutations in the ATP2C1 gene that encodes an adenosine triphosphate (ATP)-powered calcium channel pump. HHD is characterized by impaired epidermal cell-to-cell adhesion and defective keratinocyte growth/differentiation. The mechanism by which mutant ATP2C1 causes HHD is unknown and current treatments for affected individuals do not address the underlying defects and are ineffective. Notch signalling is a direct determinant of keratinocyte growth and differentiation. We found that loss of ATP2C1 leads to impaired Notch1 signalling, thus deregulation of the Notch signalling response is therefore likely to contribute to HHD manifestation. NOTCH1 is a transmembrane receptor and upon ligand binding, the intracellular domain (NICD) translocates to the nucleus activating its target genes. In the context of HHD, we found that loss of ATP2C1 function promotes upregulation of the active NOTCH1 protein (NICD-Val1744). Here, deeply exploring this aspect, we observed that NOTCH1 activation is not associated with the transcriptional enhancement of its targets. Moreover, in agreement with these results, we found a cytoplasmic localization of NICD-Val1744. We have also observed that ATP2C1-loss is associated with the degradation of NICD-Val1744 through the lysosomal/proteasome pathway. These results show that ATP2C1-loss could promote a mechanism by which NOTCH1 is endocytosed and degraded by the cell membrane. The deregulation of this phenomenon, finely regulated in physiological conditions, could in HHD lead to the deregulation of NOTCH1 with alteration of skin homeostasis and disease manifestation.


Asunto(s)
Pénfigo Familiar Benigno , Humanos , Pénfigo Familiar Benigno/genética , Pénfigo Familiar Benigno/metabolismo , Piel/metabolismo , Queratinocitos/metabolismo , Mutación , Epidermis/metabolismo , ATPasas Transportadoras de Calcio/genética , ATPasas Transportadoras de Calcio/metabolismo , Receptor Notch1/genética , Receptor Notch1/metabolismo
2.
Int J Mol Sci ; 24(8)2023 Apr 09.
Artículo en Inglés | MEDLINE | ID: mdl-37108134

RESUMEN

Nanomaterials are gaining increasing attention as innovative materials in medicine. Among nanomaterials, zinc oxide (ZnO) nanostructures are particularly appealing because of their opto-electrical, antimicrobial, and photochemical properties. Although ZnO is recognized as a safe material and the Zn ion (Zn2+) concentration is strictly regulated at a cellular and systemic level, different studies have demonstrated cellular toxicity of ZnO nanoparticles (ZnO-NPs) and ZnO nanorods (ZnO-NRs). Recently, ZnO-NP toxicity has been shown to depend on the intracellular accumulation of ROS, activation of autophagy and mitophagy, as well as stabilization and accumulation of hypoxia-inducible factor-1α (HIF-1α) protein. However, if the same pathway is also activated by ZnO-NRs and how non-cancer cells respond to ZnO-NR treatment, are still unknown. To answer to these questions, we treated epithelial HaCaT and breast cancer MCF-7 cells with different ZnO-NR concentrations. Our results showed that ZnO-NR treatments increased cell death through ROS accumulation, HIF-1α and endothelial PAS domain protein 1 (EPAS1) activation, and induction of autophagy and mitophagy in both cell lines. These results, while on one side, confirmed that ZnO-NRs can be used to reduce cancer growth, on the other side, raised some concerns on the activation of a hypoxic response in normal cells that, in the long run, could induce cellular transformation.


Asunto(s)
Neoplasias , Óxido de Zinc , Humanos , Mitofagia , Óxido de Zinc/farmacología , Óxido de Zinc/química , Especies Reactivas de Oxígeno/metabolismo , Autofagia , Células MCF-7 , Hipoxia , Factor 1 Inducible por Hipoxia
3.
Lupus ; 31(8): 921-926, 2022 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-35477339

RESUMEN

BACKGROUND: Several data have demonstrated the occurrence of erosive arthritis in Systemic Lupus Erythematosus (SLE) patients. However, a few studies have focused on the pathogenic mechanisms involved in this feature. The implication of oral pathogens has been proved in Rheumatoid Arthritis: in particular, Porphyromonas gingivalis (Pg), by inducing citrullination, could trigger autoimmune response. Here, we evaluated amount of Pg on the tongue in a cohort of SLE patients with arthritis, focusing on the association with the erosive phenotype. METHODS: SLE patients with arthritis were enrolled. DAS28 was applied to assess activity. Erosive damage was evaluated by ultrasound at level of MCP (metacarpophalangeal) and PIP (proximal interphalangeals) joints. All subjects underwent a tongue cytologic swab in order to quantify the amount of Pg (real-time PCR). The bacterium expression was obtained from the ratio between the patient's DNA amount and that obtained from healthy subjects. RESULTS: 33 patients were enrolled (M/F 3/30; median age 47 years, IQR 17; median disease duration 216 months, IQR 180): 12 of them (36.4%) showed erosive damage, significantly associated with ACPA positivity (p = 0.03) and higher values of DAS28 (p = 0.01). A mean ratio of 19.7 ± 31.1 was found for Pg amount. Therefore, we used Pg mean values as threshold, identifying two groups of patients, namely, highPg and lowPg. Erosive damage was significantly more frequent in highPg patients in comparison with lowPg (60.0% vs 26.0%, p = 0.001). Furthermore, highPg patients showed higher prevalence of skin manifestations, serositis, and neurological involvement (p = 0.005, p = 0.03, p = 0.0001, respectively). CONCLUSION: The possible contribution of oral microbiota in SLE erosive arthritis was here evaluated for the first time, finding a significant association between erosive damage and higher expression of Pg at tongue level.


Asunto(s)
Artritis Reumatoide , Lupus Eritematoso Sistémico , Autoanticuerpos , Biopelículas , Humanos , Porphyromonas gingivalis , Lengua/patología
4.
Environ Res ; 191: 110209, 2020 12.
Artículo en Inglés | MEDLINE | ID: mdl-32937173

RESUMEN

Air pollution is recognized as the world's largest environmental health risk. In this work we evaluated in vivo the effects of three relevant components of atmospheric dusts (brake dust, wood pellet ash and Saharan dust) employing the animal model Caenorhabditis elegans. Main endpoints of C. elegans such as life span, brood size and oxidative stress were addressed by exposing the nematodes to different dust concentrations. Brake dust and pellet ash affected the life span and increased significantly the oxidative stress of exposed nematodes, while Saharan dust showed no effects. Water soluble and insoluble fractions of these dusts were used to investigate the impact of the single fraction on C. elegans. The two fractions of brake dust and pellet ash exerted different effects on C. elegans endpoints in terms of life span and oxidative stress response. These fractions acted in different ways on the worm susceptibility to infection of two human pathogens (Staphylococcus aureus and Pseudomonas aeruginosa) affecting the sek-1 gene expression. In conclusion, our study showed that C. elegans is a valuable tool to investigate in vivo possible effects of atmospheric dusts.


Asunto(s)
Contaminantes Atmosféricos , Contaminantes Ambientales , África del Norte , Contaminantes Atmosféricos/toxicidad , Animales , Caenorhabditis elegans , Polvo , Humanos
5.
Appl Microbiol Biotechnol ; 104(20): 8937-8948, 2020 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-32875367

RESUMEN

The probiotic bacteria are helpful for nutritional and therapeutic purposes, and they are commercially available in various forms, such as capsules or powders. Increasing pieces of evidence indicate that different growth conditions and variability in manufacturing processes can determine the properties of probiotic products. In recent years, the lipidomic approach has become a useful tool to evaluate the impact that probiotics induce in host physiology. In this work, two probiotic formulations with identical species composition, produced in two different sites, the USA and Italy, were utilized to feed Caenorhabditis elegans, strains and alterations in lipid composition in the host and bacteria were investigated. Indeed, the multicellular organism C. elegans is considered a simple model to study the in vivo effects of probiotics. Nematodes fat metabolism was assessed by gene expression analysis and by mass spectrometry-based lipidomics. Lipid droplet analysis revealed a high accumulation of lipid droplets in worms fed US-made products, correlating with an increased expression of genes involved in the fatty acid synthesis. We also evaluated the lifespan of worms defective in genes involved in the insulin/IGF-1-mediated pathway and monitored the nuclear translocation of DAF-16. These data demonstrated the involvement of the signaling in C. elegans responses to the two diets. Lipidomics analysis of the two formulations was also conducted, and the results indicated differences in phosphatidylglycerol (PG) and phosphatidylcholine (PC) contents that, in turn, could influence nematode host physiology. Results demonstrated that different manufacturing processes could influence probiotics and host properties in terms of lipid composition. KEY POINTS: • Probiotic formulations impact on Caenorhabditis elegans lipid metabolism; • Lipidomic analysis highlighted phospholipid abundance in the two products; • Phosphocholines and phosphatidylglycerols were analyzed in worms fed the two probiotic formulations.


Asunto(s)
Caenorhabditis elegans , Probióticos , Animales , Italia , Lipidómica , Longevidad
6.
J Nanobiotechnology ; 17(1): 17, 2019 Jan 25.
Artículo en Inglés | MEDLINE | ID: mdl-30683113

RESUMEN

BACKGROUND: Dental caries is a recognized worldwide public health problem. Despite being one of the most effective strategies against dental caries, the excessive use of fluorine may result in a potential risk of developing dental fluorosis especially in children under age of six. The purpose of this work is to analyze a fluorine-free toothpaste containing Biomimetic Hydroxyapatite to assess enamel re-mineralizing and repairing properties. RESULTS: The study was performed in vitro and in vivo, comparing the hydroxyapatite toothpaste with two others toothpaste containing different fluorine concentrations. The coating effect of the micro-structured Hydroxyapatite nanoparticles reintegrates the enamel with a biomimetic film reproducing the structure and the morphology of the biologic Hydroxyapatite of the enamel. As demonstrated, the coating is due to the deposit of a new layer of apatite, which presents fewer particles than the natural enamel, not based on the chemical-physical changes occurring in fluorinated toothpastes. Moreover, it shows resistance to brushing as a consequence of chemical bonds between the synthetic and natural crystals of the enamel. CONCLUSIONS: The use of Biomimetic Hydroxyapatite toothpastes has proven to be a valuable prevention measure against dental caries in primary dentition since it prevents the risk of fluorosis.


Asunto(s)
Biomimética , Esmalte Dental/efectos de los fármacos , Durapatita/farmacología , Remineralización Dental/métodos , Niño , Materiales Biocompatibles Revestidos/química , Caries Dental/prevención & control , Esmalte Dental/patología , Durapatita/química , Fluoruros/farmacología , Humanos , Streptococcus mutans/efectos de los fármacos , Diente Primario , Pastas de Dientes/química , Pastas de Dientes/farmacología
7.
Int J Mol Sci ; 20(20)2019 Oct 10.
Artículo en Inglés | MEDLINE | ID: mdl-31658751

RESUMEN

Probiotics exert beneficial effects on host health through different mechanisms of action, such as production of antimicrobial substances, competition with pathogens, enhancement of host mucosal barrier integrity and immunomodulation. In the context of ageing, which is characterized by several physiological alterations leading to a low grade inflammatory status called inflammageing, evidences suggest a potential prolongevity role of probiotics. Unraveling the mechanisms underlying anti-ageing effects requires the use of simple model systems. To this respect, the nematode Caenorhabditis elegans represents a suitable model organism for the study of both host-microbe interactions and for ageing studies, because of conserved signaling pathways and host defense mechanisms involved in the regulation of its lifespan. Therefore, this review analyses the impact of probiotics on C. elegans age-related parameters, with particular emphasis on oxidative stress, immunity, inflammation and protection from pathogen infections. The picture emerging from our analysis highlights that several probiotic strains are able to exert anti-ageing effects in nematodes by acting on common molecular pathways, such as insulin/insulin-like growth factor-1 (IIS) and p38 mitogen-activated protein kinase (p38 MAPK). In this perspective, C. elegans appears to be advantageous for shedding light on key mechanisms involved in host prolongevity in response to probiotics supplementation.


Asunto(s)
Caenorhabditis elegans/microbiología , Interacciones Huésped-Parásitos/fisiología , Probióticos/farmacología , Animales , Caenorhabditis elegans/efectos de los fármacos , Senescencia Celular , Inmunidad , Inflamación , Factor I del Crecimiento Similar a la Insulina/metabolismo , Longevidad , Modelos Animales , Estrés Oxidativo , Fragmentos de Péptidos/metabolismo , Transducción de Señal , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismo
8.
Int J Mol Sci ; 19(6)2018 Jun 20.
Artículo en Inglés | MEDLINE | ID: mdl-29925776

RESUMEN

The term orthodisease defines human disorders in which the pathogenic gene has orthologs in model organism genomes. Yeasts have been instrumental for gaining insights into the molecular basis of many human disorders, particularly those resulting from impaired cellular metabolism. We and others have used yeasts as a model system to study the molecular basis of Hailey-Hailey disease (HHD), a human blistering skin disorder caused by haploinsufficiency of the gene ATP2C1 the orthologous of the yeast gene PMR1. We observed that K. lactis cells defective for PMR1 gene share several biological similarities with HHD derived keratinocytes. Based on the conservation of ATP2C1/PMR1 function from yeast to human, here we used a yeast-based assay to screen for molecules able to influence the pleiotropy associated with PMR1 deletion. We identified six compounds, Kaempferol, Indirubin, Lappaconite, Cyclocytidine, Azomycin and Nalidixic Acid that induced different major shape phenotypes in K. lactis. These include mitochondrial and the cell-wall morphology-related phenotypes. Interestingly, a secondary assay in mammalian cells confirmed activity for Kaempferol. Indeed, this compound was also active on human keratinocytes depleted of ATP2C1 function by siRNA-treatment used as an in-vitro model of HHD. We found that Kaempferol was a potent NRF2 regulator, strongly inducing its expression and its downstream target NQO1. In addition, Kaempferol could decrease oxidative stress of ATP2C1 defective keratinocytes, characterized by reduced NRF2-expression. Our results indicated that the activation of these pathways might provide protection to the HHD-skin cells. As oxidative stress plays pivotal roles in promoting the skin lesions of Hailey-Hailey, the NRF2 pathway could be a viable therapeutic target for HHD.


Asunto(s)
Productos Biológicos/farmacología , Quempferoles/farmacología , Kluyveromyces/efectos de los fármacos , Pénfigo Familiar Benigno/terapia , Productos Biológicos/uso terapéutico , Calcio/metabolismo , ATPasas Transportadoras de Calcio/genética , Línea Celular , Pared Celular/efectos de los fármacos , Proteínas Fúngicas/genética , Pleiotropía Genética , Humanos , Quempferoles/uso terapéutico , Queratinocitos/efectos de los fármacos , Kluyveromyces/genética , Mitocondrias/efectos de los fármacos , Factor 2 Relacionado con NF-E2/metabolismo , Estrés Oxidativo/efectos de los fármacos , Pénfigo Familiar Benigno/genética , Cultivo Primario de Células
9.
Anal Bioanal Chem ; 409(6): 1657-1666, 2017 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-27928607

RESUMEN

Airborne bacteria are components of the atmospheric aerosol particles and can be responsible of allergic disease, regardless of their viability. In this paper, we report a method for the determination of total (viable and nonviable) bacterial content in airborne particles, using muramic and dipicolinic acids as biomarkers of bacteria and bacterial spores, respectively. The analytical procedure was optimized with bacteria and spores of Bacillus subtilis. After extraction and purification, the two biomarkers were analyzed by HPLC-ESI-MS/MS and their percentage was evaluated to be used as conversion factor. The present method for the determination of the total bacterial content was then applied to environmental samples, after a proper collection in an urban site. Thanks to the use of a low pressure impactor, capable of fractionating particles into the range of 0.03-10 µm, it was also possible to study the bacterial content in ultrafine, fine, and coarse particulate matter. The results from this study showed that muramic and dipicolinic acids can be determined together in one chromatographic run in reversed phase ion pair chromatography. Bacteria were more abundant than bacterial spores in the urban atmosphere, both showing a higher concentration in the coarse fraction of particles, although bacteria and bacterial spore amounts per unit mass of ultrafine particles were higher than in fine and coarse particles.


Asunto(s)
Bacillus subtilis/aislamiento & purificación , Ácidos Murámicos/aislamiento & purificación , Material Particulado/análisis , Ácidos Picolínicos/aislamiento & purificación , Esporas Bacterianas/aislamiento & purificación , Espectrometría de Masas en Tándem/métodos , Aerosoles/análisis , Microbiología del Aire , Atmósfera/análisis , Cromatografía Líquida de Alta Presión/métodos , Límite de Detección , Ácidos Murámicos/análisis , Ácidos Picolínicos/análisis , Espectrometría de Masa por Ionización de Electrospray/métodos
10.
J Nanobiotechnology ; 15(1): 57, 2017 Aug 01.
Artículo en Inglés | MEDLINE | ID: mdl-28764786

RESUMEN

BACKGROUND: Nanotechnologies are currently revolutionizing the world around us, improving the quality of our lives thanks to a multitude of applications in several areas including the environmental preservation, with the biodeterioration phenomenon representing one of the major concerns. RESULTS: In this study, an innovative nanomaterial consisting of graphene nanoplatelets decorated by zinc oxide nanorods (ZNGs) was tested for the ability to inhibit two different pathogens belonging to bacterial genera frequently associated with nosocomial infections as well as biodeterioration phenomenon: the Gram-positive Staphylococcus aureus and the Gram-negative Pseudomonas aeruginosa. A time- and dose-dependent bactericidal effect in cell viability was highlighted against both bacteria, demonstrating a strong antimicrobial potential of ZNGs. Furthermore, the analysis of bacterial surfaces through Field emission scanning electron microscopy (FESEM) revealed ZNGs mechanical interaction at cell wall level. ZNGs induced in those bacteria deep physical damages not compatible with life as a result of nanoneedle-like action of this nanomaterial together with its nanoblade effect. Cell injuries were confirmed by Fourier transform infrared spectroscopy, revealing that ZNGs antimicrobial effect was related to protein and phospholipid changes as well as a decrease in extracellular polymeric substances; this was also supported by a reduction in biofilm formation of both bacteria. The antibacterial properties of ZNGs applied on building-related materials make them a promising tool for the conservation of indoor/outdoor surfaces. Finally, ZNGs nanotoxicity was assessed in vivo by exploiting the soil free living nematode Caenorhabditis elegans. Notably, no harmful effects of ZNGs on larval development, lifespan, fertility as well as neuromuscular functionality were highlighted in this excellent model for environmental nanotoxicology. CONCLUSIONS: Overall, ZNGs represent a promising candidate for developing biocompatible materials that can be exploitable in antimicrobial applications without releasing toxic compounds, harmful to the environment.


Asunto(s)
Antibacterianos/química , Grafito/química , Nanotubos/química , Óxido de Zinc/química , Antibacterianos/farmacología , Materiales Biocompatibles/química , Biopelículas/efectos de los fármacos , Grafito/farmacología , Humanos , Pseudomonas aeruginosa/efectos de los fármacos , Staphylococcus aureus/efectos de los fármacos , Óxido de Zinc/farmacología
11.
J Nanobiotechnology ; 15(1): 89, 2017 Dec 12.
Artículo en Inglés | MEDLINE | ID: mdl-29233187

RESUMEN

BACKGROUND: Secondary caries are considered the main cause of dental restoration failure. In this context, anti-biofilm and bactericidal properties are desired in dental materials against pathogens such as Streptococcus mutans. To this purpose, graphene based materials can be used as fillers of polymer dental adhesives. In this work, we investigated the possibility to use as filler of dental adhesives, graphene nanoplatelets (GNP), a non toxic hydrophobic nanomaterial with antimicrobial and anti-biofilm properties. RESULTS: Graphene nanoplatelets have been produced starting from graphite intercalated compounds through a process consisting of thermal expansion and liquid exfoliation. Then, a dental adhesive filled with GNPs at different volume fractions has been produced through a solvent evaporation method. The rheological properties of the new experimental adhesives have been assessed experimentally. The adhesive properties have been tested using microtensile bond strength measurements (µ-TBS). Biocidal activity has been studied using the colony forming units count (CFU) method. The anti-biofilm properties have been demonstrated through FE-SEM imaging of the biofilm development after 3 and 24 h of growth. CONCLUSIONS: A significantly lower vitality of S. mutans cells has been demonstrated when in contact with the GNP filled dental adhesives. Biofilm growth on adhesive-covered dentine tissues demonstrated anti-adhesion properties of the produced materials. µ-TBS results demonstrated no significant difference in µ-TBS between the experimental and the control adhesive. The rheology tests highlighted the necessity to avoid low shear rate regimes during adhesive processing and application in clinical protocol, and confirmed that the adhesive containing the 0.2%wt of GNPs possess mechanical properties comparable with the ones of the control adhesive.


Asunto(s)
Antiinfecciosos/farmacología , Biopelículas/efectos de los fármacos , Cementos Dentales/química , Grafito/química , Streptococcus mutans/fisiología , Antiinfecciosos/química , Cementos Dentales/farmacología , Microscopía Electrónica de Rastreo , Nanoestructuras/química , Estrés Oxidativo/efectos de los fármacos , Reología , Resistencia a la Tracción
12.
Microbiology (Reading) ; 162(11): 1933-1944, 2016 11.
Artículo en Inglés | MEDLINE | ID: mdl-27655407

RESUMEN

In Saccharomyces cerevisiae, Adr1 is a zinc-finger transcription factor involved in the transcriptional activation of ADH2. Deletion of KlADR1, its putative ortholog in Kluyveromyces lactis, led to reduced growth in glycerol, oleate and yeast extract-peptone medium suggesting, as in S. cerevisiae, its requirement for glycerol, fatty acid and nitrogen utilization. Moreover, growth comparison on yeast extract and peptone plates showed in K. lactis a KlAdr1-dependent growth trait not present in S. cerevisiae, indicating different metabolic roles of the two factors in their environmental niches. KlADR1 is required for growth under respiratory and fermentative conditions like KlADH, alcohol dehydrogenase genes necessary for metabolic adaptation during the growth transition. Using in-gel native alcohol dehydrogenase assay, we showed that this factor affected the Adh pattern by altering the balance between these activities. Since the activity most affected by KlAdr1 is KlAdh3, a deletion analysis of the KlADH3 promoter allowed the isolation of a DNA fragment through which KlAdr1 modulated its expression. The expression of the KlADR1-GFP gene allowed the intracellular localization of the factor in K. lactis and S. cerevisiae, suggesting in the two yeasts a common mechanism of KlAdr1 translocation under fermentative and respiratory conditions. Finally, the chimeric Kl/ScADR1 gene encoding the zinc-finger domains of KlAdr1 fused to the transactivating domains of the S. cerevisiae factor activated in Scadr1Δ the transcription of ADH2 in a ScAdr1-dependent fashion.


Asunto(s)
Proteínas Fúngicas/metabolismo , Kluyveromyces/metabolismo , Saccharomyces cerevisiae/genética , Factores de Transcripción/metabolismo , Alcohol Deshidrogenasa/genética , Alcohol Deshidrogenasa/metabolismo , Medios de Cultivo/metabolismo , Proteínas Fúngicas/genética , Eliminación de Gen , Expresión Génica , Regulación Fúngica de la Expresión Génica , Glicerol/metabolismo , Kluyveromyces/genética , Kluyveromyces/crecimiento & desarrollo , Regiones Promotoras Genéticas , Saccharomyces cerevisiae/metabolismo , Factores de Transcripción/genética
13.
Cell Mol Life Sci ; 71(13): 2535-46, 2014 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-24221134

RESUMEN

Candida albicans represents one of the most prevalent species causing life-threatening fungal infections. Current treatments to defeat Candida albicans have become quite difficult, due to their toxic side effects and the emergence of resistant strains. Antimicrobial peptides (AMPs) are fascinating molecules with a potential role as novel anti-infective agents. However, only a few studies have been performed on their efficacy towards the most virulent hyphal phenotype of this pathogen. The purpose of this work is to evaluate the anti-Candida activity of the N-terminal 1-18 fragment of the frog skin AMP esculentin-1b, Esc(1-18), under both in vitro and in vivo conditions using Caenorhabditis elegans as a simple host model for microbial infections. Our results demonstrate that Esc(1-18) caused a rapid reduction in the number of viable yeast cells and killing of the hyphal population. Esc(1-18) revealed a membrane perturbing effect which is likely the basis of its mode of action. To the best of our knowledge, this is the first report showing the ability of a frog skin AMP-derived peptide (1) to kill both growing stages of Candida; (2) to promote survival of Candida-infected living organisms and (3) to inhibit transition of these fungal cells from the roundish yeast shape to the more dangerous hyphal form at sub-inhibitory concentrations.


Asunto(s)
Proteínas Anfibias/farmacología , Péptidos Catiónicos Antimicrobianos/farmacología , Caenorhabditis elegans/efectos de los fármacos , Candida albicans/efectos de los fármacos , Animales , Antiinfecciosos/farmacología , Anuros/metabolismo , Caenorhabditis elegans/microbiología , Candida albicans/patogenicidad , Piel/metabolismo
14.
Gels ; 10(2)2024 Feb 18.
Artículo en Inglés | MEDLINE | ID: mdl-38391480

RESUMEN

Cultural heritage stone materials frequently experience significant discoloration induced by copper corrosion products, especially calcareous stones associated with bronze or copper statues and architectural elements. This alteration originates from the corrosion of unprotected copper, resulting in the formation of various Cu minerals and the migration of soluble ions to adjacent stone materials. Traditional cleaning methods involve mechanical, chemical, and laser techniques, which are generally time-consuming, costly, not ecological, or can possibly damage original materials. The loading of highly effective chelating agents, such as ethylenediaminetetraacetic acid (EDTA), into hydrogels has recently been exploited. However, the preference for synthetic hydrogels has been prominent until now, although they lack renewability and biodegradability and require high costs. This study explores for the first time the potential to clean copper corrosion with bacterial nanocellulose (BC) loaded with EDTA as a biologically based, sustainable, and biodegradable hydrogel. The BC hydrogel was characterised by field emission-scanning electron microscopy (FE-SEM), X-ray diffraction analysis (XRD), attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), simultaneous thermal analysis (TG-DSC), and tensile testing. It revealed a nano-fibrous structure with high crystallinity and purity and mechanical properties suitable for cultural heritage applications. The EDTA-loaded hydrogel effectively removed copper stains from marble after 120 min of application. Micro-Raman and colorimetric analyses assessed the cleaning efficacy. The study introduces bacterial nanocellulose as a green and effective alternative for heritage conservation, aligning with sustainable methodologies in stone conservation.

15.
Microb Cell Fact ; 12: 34, 2013 Apr 15.
Artículo en Inglés | MEDLINE | ID: mdl-23587421

RESUMEN

BACKGROUND: Kluyveromyces marxianus combines the ease of genetic manipulation and fermentation with the ability to efficiently secrete high molecular weight proteins, performing eukaryotic post-translational modifications. It is able to grow efficiently in a wide range of temperatures. The secretion performances were analyzed in the host K. marxianus L3 in the range between 5°C and 40°C by means of 3 different reporter proteins, since temperature appears a key parameter for production and secretion of recombinant proteins. RESULTS: The recombinant strains were able to grow up to 40°C and, along the tested temperature interval (5-40°C), the specific growth rates (µ) were generally lower as compared to those of the untransformed strain. Biomass yields were slightly affected by temperature, with the highest values reached at 15°C and 30°C. The secretion of the endogenous ß-fructofuranosidase, used as an internal control, was efficient in the range of the tested temperature, as evaluated by assaying the enzyme activity in the culture supernatants. The endogenous ß-fructofuranosidase production was temperature dependent, with the highest yield at 30°C. The heterologous proteins HSA, GAA and Sod1p were all successfully produced and secreted between 5°C and 40°C, albeit each one presented a different optimal production temperature (15, 40, 5-30°C for HSA, GAA and Sod1p, respectively). CONCLUSIONS: K. marxianus L3 has been identified as a promising and flexible cell factory. In a sole host, the optimization of growth temperatures for the efficient secretion of each individual protein can be carried out over a wide range of temperatures.


Asunto(s)
Kluyveromyces/metabolismo , Proteínas Recombinantes/biosíntesis , Biomasa , Glucano 1,4-alfa-Glucosidasa/genética , Glucano 1,4-alfa-Glucosidasa/metabolismo , Humanos , Kluyveromyces/crecimiento & desarrollo , Proteínas Recombinantes/genética , Albúmina Sérica/genética , Albúmina Sérica/metabolismo , Superóxido Dismutasa/genética , Superóxido Dismutasa/metabolismo , Superóxido Dismutasa-1 , Temperatura , beta-Fructofuranosidasa/genética , beta-Fructofuranosidasa/metabolismo
16.
Nano Lett ; 12(6): 2740-4, 2012 Jun 13.
Artículo en Inglés | MEDLINE | ID: mdl-22612766

RESUMEN

We evaluated the toxicity of graphite nanoplatelets (GNPs) in the model organism Caenorhabditis elegans. The GNPs resulted nontoxic by measuring longevity as well as reproductive capability end points. An imaging technique based on Fourier transform infrared spectroscopy (FT-IR) mapping was also developed to analyze the GNPs spatial distribution inside the nematodes. Conflicting reports on the in vitro antimicrobial properties of graphene-based nanomaterials prompted us to challenge the host-pathogen system C. elegans-Pseudomonas aeruginosa to assess these findings through an in vivo model.


Asunto(s)
Caenorhabditis elegans/efectos de los fármacos , Caenorhabditis elegans/fisiología , Grafito/toxicidad , Nanopartículas/toxicidad , Animales , Relación Dosis-Respuesta a Droga , Tasa de Supervivencia
17.
Nanomaterials (Basel) ; 13(6)2023 Mar 16.
Artículo en Inglés | MEDLINE | ID: mdl-36985974

RESUMEN

The emergence of multidrug-resistant bacteria represents a growing threat to public health, and it calls for the development of alternative antibacterial approaches not based on antibiotics. Here, we propose vertically aligned carbon nanotubes (VA-CNTs), with a properly designed nanomorphology, as effective platforms to kill bacteria. We show, via a combination of microscopic and spectroscopic techniques, the ability to tailor the topography of VA-CNTs, in a controlled and time-efficient manner, by means of plasma etching processes. Three different varieties of VA-CNTs were investigated, in terms of antibacterial and antibiofilm activity, against Pseudomonas aeruginosa and Staphylococcus aureus: one as-grown variety and two varieties receiving different etching treatments. The highest reduction in cell viability (100% and 97% for P. aeruginosa and S. aureus, respectively) was observed for the VA-CNTs modified using Ar and O2 as an etching gas, thus identifying the best configuration for a VA-CNT-based surface to inactivate both planktonic and biofilm infections. Additionally, we demonstrate that the powerful antibacterial activity of VA-CNTs is determined by a synergistic effect of both mechanical injuries and ROS production. The possibility of achieving a bacterial inactivation close to 100%, by modulating the physico-chemical features of VA-CNTs, opens up new opportunities for the design of self-cleaning surfaces, preventing the formation of microbial colonies.

18.
Biochim Biophys Acta Gen Subj ; 1867(1): 130255, 2023 01.
Artículo en Inglés | MEDLINE | ID: mdl-36265765

RESUMEN

The mitochondrial translation machinery allows the synthesis of the mitochondrial-encoded subunits of the electron transport chain. Defects in this process lead to mitochondrial physiology failure; in humans, they are associated with early-onset, extremely variable and often fatal disorder. The use of a simple model to study the mitoribosomal defects is mandatory to overcome the difficulty to analyze the impact of pathological mutations in humans. In this paper we study in nematode Caenorhabditis elegans the silencing effect of the mrpl-24 gene, coding for the mitochondrial ribosomal protein L-24 (MRPL-24). This is a structural protein of the large subunit 39S of the mitoribosome and its effective physiological function is not completely elucidated. We have evaluated the nematode's fitness fault and investigated the mitochondrial defects associated with MRPL-24 depletion. The oxidative stress response activation due to the mitochondrial alteration has been also investigated as a compensatory physiological mechanism. For the first time, we demonstrated that MRPL-24 reduction increases the expression of detoxifying enzymes such as SOD-3 and GST-4 through the involvement of transcription factor SKN-1. BACKGROUND: In humans, mutations in genes encoding mitochondrial ribosomal proteins (MRPs) often cause early-onset, severe, fatal and extremely variable clinical defects. Mitochondrial ribosomal protein L-24 (MRPL24) is a structural protein of the large subunit 39S of the mitoribosome. It is highly conserved in different species and its effective physiological function is not completely elucidated. METHODS: We characterized the MRPL24 functionality using the animal model Caenorhabditis elegans. We performed the RNA mediated interference (RNAi) by exposing the nematodes' embryos to double-stranded RNA (dsRNA) specific for the MRPL-24 coding sequence. We investigated for the first time in C. elegans, the involvement of the MRPL-24 on the nematode's fitness and its mitochondrial physiology. RESULTS: Mrpl-24 silencing in C. elegans negatively affected the larval development, progeny production and body bending. The analysis of mitochondrial functionality revealed loss of mitochondrial network and impairment of mitochondrial functionality, as the decrease of oxygen consumption rate and the ROS production, as well as reduction of mitochondrial protein synthesis. Finally, the MRPL-24 depletion activated the oxidative stress response, increasing the expression levels of two detoxifying enzymes, SOD-3 and GST-4. CONCLUSIONS: In C. elegans the MRPL-24 depletion activated the oxidative stress response. This appears as a compensatory mechanism to the alteration of the mitochondrial functionality and requires the involvement of transcription factor SKN-1. GENERAL SIGNIFICANCE: C. elegans resulted in a good model for the study of mitochondrial disorders and its use as a simple and pluricellular organism could open interesting perspectives to better investigate the pathologic mechanisms underlying these devastating diseases.


Asunto(s)
Proteínas de Caenorhabditis elegans , Caenorhabditis elegans , Animales , Humanos , Caenorhabditis elegans/genética , Caenorhabditis elegans/metabolismo , Proteínas de Caenorhabditis elegans/genética , Proteínas de Unión al ADN/metabolismo , Factores de Transcripción/metabolismo , Proteínas Ribosómicas/genética , Proteínas Ribosómicas/metabolismo , Estrés Oxidativo/genética , Superóxido Dismutasa/metabolismo
19.
Microorganisms ; 11(4)2023 Apr 21.
Artículo en Inglés | MEDLINE | ID: mdl-37110510

RESUMEN

Lactic acid bacteria (LAB) share and provide several beneficial effects on human health, such as the release of bioactive metabolites, pathogen competition, and immune stimulation. The two major reservoirs of probiotic microorganisms are the human gastro-intestinal tract and fermented dairy products. However, other sources, such as plant-based foods, represent important alternatives thanks to their large distribution and nutritive value. Here, the probiotic potential of autochthonous Lactiplantibacillus plantarum PFA2018AU, isolated from carrots harvested in Fucino highland, Abruzzo (Italy), was investigated through in vitro and in vivo approaches. The strain was sent to the biobank of Istituto Zooprofilattico Sperimentale della Lombardia ed Emilia Romagna in Italy for the purpose of patent procedures under the Budapest Treaty. The isolate showed high survival capability under in vitro simulated gastro-intestinal conditions, antibiotic susceptibility, hydrophobicity, aggregation, and the ability to inhibit the in vitro growth of Salmonella enterica serovar Typhimurium, Listeria monocytogenes, Pseudomonas aeruginosa, and Staphylococcus aureus pathogens. Caenorhabditis elegans was used as the in vivo model in order to analyse prolongevity and anti-ageing effects. L. plantarum PFA2018AU significantly colonised the gut of the worms, extended their lifespan, and stimulated their innate immunity. Overall, these results showed that autochthonous LAB from vegetables, such as carrots, have functional features that can be considered novel probiotic candidates.

20.
Microbiology (Reading) ; 158(Pt 7): 1694-1701, 2012 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-22504438

RESUMEN

The interplay between calcium metabolism and glycosylation in yeast is largely unknown. In order to clarify this relationship, the effect of a mutation in the KlOCH1 gene, encoding the Golgi α-1,6-mannosyltransferase, on calcium homeostasis was studied in the yeast Kluyveromyces lactis. In particular, the role of the KlMID1 gene, encoding one of the components of the plasma membrane calcium channel (Cch1-Mid1), was investigated. Almost complete suppression of the phenotypes occurring in the mutant strain, ranging from oxidative stress to cell wall alteration, was observed by increased dosage of KlMID1. In addition, the N-glycosylation mutant showed increased calcium accumulation and decreased transcription of KlMID1 and KlCCH1. Moreover, the calcium alterations included an increased expressional profile for the KlPMC1 gene, encoding the vacuolar calcium ion pump. Furthermore, perturbation of endoplasmic reticulum (ER) homeostasis was observed in Kloch1-1 cells. Similarly, down-modulation of calcium signalling genes as well as altered mitochondrial functionality were induced in wild-type cells after treatment with DTT. However, no mitochondrial alteration occurred in the treated cells when KlMID1 was overexpressed. Our results suggest that the ER stress taking place in Kloch1-1 cells appears to be the primary cause of the KlMID1 down-modulation and its resulting effects on the expression of calcium homeostasis genes.


Asunto(s)
Canales de Calcio/metabolismo , Retículo Endoplásmico/metabolismo , Regulación Fúngica de la Expresión Génica , Kluyveromyces/genética , Kluyveromyces/metabolismo , Mitocondrias/metabolismo , Eliminación de Gen , Dosificación de Gen , Expresión Génica , Homeostasis
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