RESUMO
Chlorpromazine (CPZ) is an antipsychotic phenothiazine which is still commonly prescribed though it causes idiosyncratic toxicity such as cholestasis. CPZ toxicity mechanisms involve oxidative stress among others. Cigarette smoke (CS) causes deleterious effects through diverse mechanisms such as oxidative stress. CS alters drug metabolizing enzymes expression and drug transporters expression and activity in animal cell models as well as in Saccharomyces cerevisiae. CS therefore alters pharmacokinetic and pharmacodynamics of many drugs including CPZ and caffeine whose toxicity is promoted by CS condensate (CSC). CSC interaction with CPZ toxicity deserves investigation. In this study, CSC exerted mild toxicity on Saccharomyces cerevisiae which resisted to this chemical stress after several hours. CPZ toxicity on yeast was dose-dependent and the cells resisted to CPZ up to 40 µM after 24 h of treatment. Yeast cells treated simultaneously with CPZ and a nontoxic CSC dose were less sensitive to CPZ. CSC probably triggers cross-resistance to CPZ. Using Sod1 mutant strain, we showed that this gene is potentially involved in the potential cross-resistance. Other genes encoding stress-related transcription factors could be involved in this process. Nicotine and cadmium chloride, which caused a dose-dependent toxicity individually, acted with CPZ in an additive or synergistic manner in terms of toxicity. Although our results cannot be extrapolated to humans, they clearly show that CSC and its components interact with CPZ toxicity.
Assuntos
Clorpromazina , Saccharomyces cerevisiae , Animais , Clorpromazina/toxicidade , Humanos , Estresse Oxidativo , Saccharomyces cerevisiae/genética , Fumaça/efeitos adversos , FumarRESUMO
Impaired adipogenesis is associated with the development of insulin resistance and an increased risk of type 2 diabetes (T2D). GATA Binding Protein 3 (GATA3) is implicated in impaired adipogenesis and the onset of insulin resistance. Therefore, we hypothesize that inhibition of GATA3 could promote adipogenesis, restore healthy fat distribution, and enhance insulin signaling. Primary human preadipocytes were treated with GATA3 inhibitor (DNAzyme hgd40). Cell proliferation, adipogenic capacity, gene expression, and insulin signaling were measured following well-established protocols. BALB/c mice were treated with DNAzyme hgd40 over a period of 2 weeks. Liposomes loaded with DNAzyme hgd40, pioglitazone (positive), or vehicle (negative) controls were administered subcutaneously every 2 days at the right thigh. At the end of the study, adipose tissues were collected and weighed from the site of injection, the opposite side, and the omental depot. Antioxidant enzyme (superoxide dismutase and catalase) activities were assessed in animals' sera, and gene expression was measured using well-established protocols. In vitro GATA3 inhibition induced the adipogenesis of primary human preadipocytes and enhanced insulin signaling through the reduced expression of p70S6K. In vivo GATA3 inhibition promoted adipogenesis at the site of injection and reduced MCP-1 expression. GATA3 inhibition also reduced omental tissue size and PPARγ expression. These findings suggest that modulating GATA3 expression offers a potential therapeutic benefit by correcting impaired adipogenesis, promoting healthy fat distribution, improving insulin sensitivity, and potentially lowering the risk of T2D.
Assuntos
DNA Catalítico , Diabetes Mellitus Tipo 2 , Resistência à Insulina , Adipogenia/genética , Animais , Antioxidantes/uso terapêutico , Catalase , Diabetes Mellitus Tipo 2/metabolismo , Humanos , Insulina/uso terapêutico , Resistência à Insulina/genética , Lipossomos/uso terapêutico , Camundongos , Obesidade/metabolismo , PPAR gama/metabolismo , Pioglitazona/uso terapêutico , Proteínas Quinases S6 Ribossômicas 70-kDa , Superóxido DismutaseRESUMO
Diclofenac (DCF) adverse reactions involve diverse mechanisms in different models. We recently demonstrated that DCF-induced toxicity in HepaRG decreases as they express DCF-metabolizing enzymes. DCF metabolism promotes toxicity in Saccharomyces cerevisiae expressing heterologous cytochromes-P450. N-Acetylcysteine (NAC) is used to treat diverse medical conditions due to its multiple properties (antioxidant, metal chelator, thiol-disulfide disruption). The latter property accounts for its mucolytic effects and broadens its potential molecular targets to signal transduction proteins, ABC transporters and others. Interaction of NAC with DCF effects depends on the experimental model. This study aims to investigate NAC/DCF interaction and the involvement of ABC transporters in wild type and mutant Saccharomyces cerevisiae. DCF inhibited yeast growth in a dose- and time-dependent manner and the cells started adapting to DCF 24-h post-treatment. NAC potentiated DCF-induced toxicity if added prior or parallel to DCF. Pretreatment with NAC increased its potentiation effect and compromised cells adaption to DCF. Post-treatment with NAC potentiated DCF toxicity without compromising adaptation. Moreover, mutant strains in ABC transporters Pdr5, Yor1, Bpt1 or Pdr15, were more sensitive to DCF; while mutant strains in Pdr5, Vmr1 or Pdr12 were more sensitive to NAC/DCF interaction. DCF ± NAC elicited on the mutant strain in Yap1, an oxidative stress-related protein, the same effects as on the wild type. Therefore, oxidative stress does not seem to be key actor in DCF toxicity in our model. Our hypothesis is that NAC potentiation effect is at least due to its ability to disrupt disulfide bridge in proteins required to overcome DCF toxicity in yeast.
Assuntos
Transportadores de Cassetes de Ligação de ATP/metabolismo , Acetilcisteína/toxicidade , Anti-Inflamatórios não Esteroides/toxicidade , Antioxidantes/toxicidade , Diclofenaco/toxicidade , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/efeitos dos fármacos , Transportadores de Cassetes de Ligação de ATP/genética , Anti-Inflamatórios não Esteroides/metabolismo , Diclofenaco/metabolismo , Dissulfetos/metabolismo , Relação Dose-Resposta a Droga , Sinergismo Farmacológico , Genótipo , Mutação , Estresse Oxidativo/efeitos dos fármacos , Fenótipo , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/crescimento & desenvolvimento , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/genética , Fatores de Tempo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismoRESUMO
A phenylethanoid, two steroids, a flavone glucoside and a chalcone have been isolated for the first time from the stems of Calicotome villosa together with a previously isolated flavone glucoside. Their structures were determined by spectroscopic analyses (NMR, HRMS) as basalethanoïd B (1), ß-sitosterol and stigmasterol (2), chrysine-7-O-ß-d-glucopyranoside (3), chrysine 7-((6''-O-acetyl)-O-ß-d-glucopyranoside) (4) and calythropsin (5). The crude extracts and the isolated compounds (except 4), were evaluated for their antioxidant, antimicrobial (against two Gram-positive bacterial strains: Staphylococcus aureus, Bacillus cereus, four Gram-negative bacterial strains: Staphylococcus epidermidis, Klebsiella pneumonia, Acinetobacter baumanii, and three yeasts: Candida albicans, Candida tropicalis, and Candida glabrata), hemolytic, antidiabetic, anti-inflammatory and cytotoxic activity. The crude extracts showed good ability to scavenge the free radical DPPH. Methanol stem extract followed by the dichloromethane stem extract showed moderate antimicrobial potency; furthermore, at 1 mg/mL the methanol extract showed an inhibition of C. albicans growth comparable to nystatin. Dichloromethane, methanol, and aqueous extracts inhibited 98%, 90%, and 80% of HeLa cell proliferation at 2 mg/mL respectively. Weak hypoglycemic and hemolytic effects were exhibited by the crude extracts. Among all the tested compounds, compound 3 showed remarkable hypoglycemic potential (93% at 0.1 mg/mL) followed by compound 5 (90% at 0.3 mg/mL). Compound 5 was the most effective in the DPPH. scavenging assay (100% at 0.1 mg/mL) and cytotoxic assay on HeLa cells (99% and 90% after 24 and 48 h of treatment at 0.1 mg/mL, respectively). No anti-inflammatory effects were displayed by any of the crude extracts or the isolated compounds at any of the tested concentrations.
Assuntos
Anti-Infecciosos/química , Antineoplásicos Fitogênicos/química , Antioxidantes/química , Fabaceae/química , Extratos Vegetais/química , Caules de Planta/química , Anti-Infecciosos/farmacologia , Antineoplásicos Fitogênicos/farmacologia , Antioxidantes/farmacologia , Candida albicans/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Positivas/efeitos dos fármacos , Células HeLa , Humanos , Testes de Sensibilidade Microbiana , Estrutura Molecular , Extratos Vegetais/farmacologiaRESUMO
Paraquat has been shown to be a highly toxic compound for humans and animals, and many cases of acute poisoning and death have been reported over the past few decades. The present study was undertaken to evaluate comprehensively herbicides (Paraquat) and some plant extracts to biochemical aspects of Lymnaea natalensis snails. It was found that the exposure of L. natalensis to Paraquat and plant extracts led to a significant reduction in the infectivity of Fasciola gigantica miracidia to the snail. The glucose level in hemolymph of exposed snails was elevated, while the glycogen showed a decrease in soft tissues when compared with the control group. In addition, the activity level of some enzymes representing glycolytic enzymes as hexokinase (HK), pyruvate kinase (PK), phosphofructokinase (PFK), lactate dehydrogenase (LDH), and glucose phosphate isomerase (GPI) in snail's tissues were reduced in response to the treatment. It was concluded that the pollution of the aquatic environment by herbicide would adversely affect the metabolism of the L. natalensis snails. Snails treated with Agave attenuate, Ammi visnaga, and Canna iridiflora plant had less toxic effect compared to snails treated with Paraquat.
Assuntos
Herbicidas/toxicidade , Lymnaea/efeitos dos fármacos , Paraquat/toxicidade , Extratos Vegetais/toxicidade , Animais , Fasciola/crescimento & desenvolvimento , Glucose-6-Fosfato Isomerase/metabolismo , Hexoquinase/metabolismo , L-Lactato Desidrogenase/metabolismo , Dose Letal Mediana , Lymnaea/metabolismo , Lymnaea/parasitologia , Fosfofrutoquinases/metabolismo , Compostos Fitoquímicos/toxicidade , Piruvato Quinase/metabolismoRESUMO
Global transcriptional regulators are crucial for supporting rapid adaptive responses in changing environments. In Thermococcales, the TrmB sugar-sensing regulator family is well represented but knowledge of the functional role/s of each of its members is limited. In this study, we examined the link between TrmBL4 and the degree of protein secretion in different sugar environments in the hyperthermophilic Archaeon Thermococcus barophilus. Although the absence of TrmBL4 did not induce any growth defects, proteomics analysis revealed different secretomes depending on the sugar and/or genetic contexts. Notably, 33 secreted proteins present in the supernatant were differentially detected. Some of these proteins are involved in sugar assimilation and transport, such as the protein encoded by TERMP_01455 (cyclomaltodextrin glucanotransferase), whereas others have intracellular functions, such as the protein encoded by TERMP_01556 (pyruvate: ferredoxin oxidoreductase Δsubunit). Then, using reverse transcription quantitative polymerase chain reaction experiments, we observed effective transcription regulation by TrmBL4 of the genes encoding at least two ABC-type transporters according to sugar availability.
Assuntos
Proteínas Arqueais , Thermococcus , Thermococcus/genética , Thermococcus/metabolismo , Secretoma , Carboidratos , Açúcares/metabolismo , Proteínas Arqueais/genética , Proteínas Arqueais/metabolismoRESUMO
Carnobacterium maltaromaticum is a non-starter lactic acid bacterium (LAB) of interest in the dairy industry for biopreservation. This study investigated the interference competition network and the specialized metabolites biosynthetic gene clusters (BGCs) content in this LAB in order to explore the relationship between the antimicrobial properties and the genome content. Network analysis revealed that the potency of inhibition tended to increase when the inhibition spectrum broadened, but also that several strains exhibited a high potency and narrow spectrum of inhibition. The C. maltaromaticum strains with potent anti-L. monocytogenes were characterized by high potency and a wide intraspecific spectrum. Genome mining of 29 strains revealed the presence of 12 bacteriocin BGCs: four of class I and eight of class II, among which seven belong to class IIa and one to class IIc. Overall, eight bacteriocins and one nonribosomal peptide synthetase and polyketide synthase (NRPS-PKS) BGCs were newly described. The comparison of the antimicrobial properties resulting from the analysis of the network and the BGC genome content allowed us to delineate candidate BGCs responsible for anti-L. monocytogenes and anti-C. maltaromaticum activity. However, it also highlighted that genome analysis is not suitable in the current state of the databases for the prediction of genes involved in the antimicrobial activity of strains with a narrow anti-C. maltaromaticum activity.
RESUMO
While competition targeting food-borne pathogens is being widely documented, few studies have focused on competition among non-pathogenic food bacteria. Carnobacterium maltaromaticum is a genetically diverse lactic acid bacterium known for comprising several bacteriocinogenic strains with bioprotective potentialities against the food-borne pathogen Listeria monocytogenes. The aim of our study is to examine the network properties of competition among a collection of 73 strains of C. maltaromaticum and to characterize their individual interaction potential. The performed high-throughput competition assays, investigating 5 329 pairwise interactions, showed that intraspecific competition was major in C. maltaromaticum with approximately 56% of the sender strains antagonizing at least one receiver strain. A high diversity of inhibitory and sensitivity spectra was identified along with a majority of narrow inhibitory as well as sensitivity spectra. Through network analysis approach, we determined the highly nested architecture of C. maltaromaticum competition network, thus showing that competition in this species is determined by both the spectrum width of the inhibitory activity of sender strains and the spectrum width of the sensitivity of receiver strains. This study provides knowledge of the competition network in C. maltaromaticum that could be used in rational assembly of compatible microbial strains for the design of mixed starter cultures.
Assuntos
Antibiose , Carnobacterium/fisiologia , Contaminação de Alimentos , Microbiologia de Alimentos , Listeria monocytogenes/fisiologia , Testes de Sensibilidade Microbiana , Animais , Bacteriocinas , Ligação Competitiva , Produtos Pesqueiros , Peixes/microbiologia , Humanos , Ácido Láctico/metabolismo , Produtos da Carne , Especificidade da EspécieRESUMO
The red microalga Porphyridium cruentum is exploited industrially for its exopolysaccharides (EPS) and pigments production. EPS produced by P. cruentum are partially released and dissolved into the surrounding environment, they can be recovered from the culture medium after removing the cells. This paper presents a parametric study of the ultrafiltration of EPS solutions on organic membrane. The EPS solutions were produced in conditions representative of an industrial production. They were filtered at lab-scale on a flat, PES 50â kDa MWCO membrane in a complete recirculation mode of permeate and retentate. Permeate flux-transmembrane pressure (TMP) curves were established up to the limiting flux for the filtration of solutions with various values of concentration in EPS (0.10-1.06â kg GlcEqâ m-3), fluid tangential velocity (0.3-1.2 mâ s-1) and temperature (20°C and 40°C). The reversible and irreversible parts of fouling were evaluated for each experiment and the critical flux was determined for an intermediate EPS concentration (0.16â kg GlcEqâ m-3). The results showed that EPS solutions had a strong fouling capacity. When filtering the lowest concentrated solution (0.10â kg GlcEqâ m-3) with moderate fouling conditions, the overall fouling resistance was approximately half of the membrane and the share of irreversible/reversible fouling was 88% and 12%. However, the part of reversible fouling becomes predominant when approaching the limiting flux. Permeate fluxes which were obtained allow to estimate that a VRR of approximately 10 could be obtained when concentrating EPS solutions using PES membranes in flat or tubular modules but not in spiral-wound.
Assuntos
Microalgas , Porphyridium , Filtração , Membranas Artificiais , Polímeros , UltrafiltraçãoRESUMO
The valorization of a solid carob waste from the Lebanese industry was investigated by optimizing the production of lactic acid using immobilized Lactobacillus rhamnosus in alginate beads and response surface methodology. The results showed that pH and alginate concentration had a significant effect on the production of lactic acid. The fermentation of non-enriched carob waste juice needed an additional nitrogen source to improve lactic acid production and yield. From extracts with 65 g/L sugars, the optimum conditions were found to be 2% for the concentration of alginate, 4% bacteria cells entrapped in beads, 80 rpm agitation speed and pH 6.4. Lactic acid concentration obtained under these conditions was 22 g/L with a yield of 76.9 g/g consumed sugar and a productivity of 1.22 g/L/h. The use of invertase pretreatment increased lactic acid concentration from 22 to 40 g/L, but reduced yield at 66.6%. Finally, cells immobilized in alginate beads could be used for at least five successive cycles.
Assuntos
Indústria Alimentícia , Ácido Láctico/biossíntese , Lacticaseibacillus rhamnosus/metabolismo , Lotus/metabolismo , Eliminação de Resíduos/métodos , Alginatos/química , Células Imobilizadas , Conservação dos Recursos Naturais , Fermentação , Frutas/química , Frutas/metabolismo , Concentração de Íons de Hidrogênio , Cinética , Lacticaseibacillus rhamnosus/citologia , Lotus/química , Nitrogênio/metabolismo , Extratos Vegetais/química , Extratos Vegetais/metabolismoRESUMO
In order to mineralize Metronidazole (MTZ), a process coupling an electro-Fenton pretreatment and a biological degradation was implemented. A mono-compartment batch reactor containing a carbon-felt cathode and a platinum anode was employed to carry out the electro-Fenton pretreatment of MTZ. A total degradation of MTZ (100â¯mgâ¯L-1) was observed at 0.07â¯mA.cm-2 after only 20â¯min of electrolysis. Yet, after 1 and 2â¯h of electrolysis, the mineralization level remained low (16.2% and 32% respectively), guaranteeing a significant residual organic content for further biological treatment. LCMS/MS was used to determine the intermediates by-products and hence to propose a plausible degradation pathway. An increase from 0 to 0.44 and 0.6 for 1 and 2â¯h of electrolysis was observed for the BOD5/COD ratio. Thus, from 1â¯h of electro-Fenton pretreatment, the electrolysis by-products were considered biodegradable. A biological treatment of the electrolysis by-products after 1 and 2â¯h was then realized. The mineralization yields reached very close values, about 84% for 1 and 2â¯h of electrolysis after 504â¯h of biological treatment, namely close to 89% for the overall process, showing the pertinence of the proposed coupled process.
Assuntos
Antibacterianos , Metronidazol , Poluentes Químicos da Água , Antibacterianos/química , Antibacterianos/metabolismo , Antibacterianos/toxicidade , Biodegradação Ambiental , Análise da Demanda Biológica de Oxigênio , Eletrólise , Ferro/química , Lepidium/efeitos dos fármacos , Lepidium/crescimento & desenvolvimento , Metronidazol/química , Metronidazol/metabolismo , Metronidazol/toxicidade , Esgotos , Poluentes Químicos da Água/química , Poluentes Químicos da Água/metabolismo , Poluentes Químicos da Água/toxicidade , Purificação da Água/métodosRESUMO
In this study, the monitoring of reactive oxygen species and the regeneration of the ferrous ions catalyst were performed during electro-Fenton (EF) process to highlight the influence of operating parameters. The removal of metronidazole (MTZ) was implemented in an electrochemical mono-compartment batch reactor under various ranges of current densities, initial MTZ and ferrous ions concentrations, and pH values. It was found that under 0.07â¯mAâ¯cm-2, 0.1â¯mM of ferrous ions and pHâ¯=â¯3, the efficiency of 100â¯mgâ¯L-1 MTZ degradation and mineralization were 100% within 20â¯min and 40% within 135â¯min of electrolysis, respectively. The highest hydrogen peroxide and hydroxyl radical concentrations, 1.4â¯mM and 2.28â¯mM respectively, were obtained at 60â¯min electrolysis at 0.07â¯mAâ¯cm-2. Improvement of the biodegradability was reached from 60â¯min of electrolysis with a BOD5/COD ratio above 0.4, which was reinforced by a respirometric study, that supports the feasibility of coupling electro-Fenton and biological treatment for the metronidazole removal.
Assuntos
Eletrólise , Peróxido de Hidrogênio/química , Ferro/análise , Metronidazol/química , Oxigênio/análise , Biodegradação Ambiental , Análise da Demanda Biológica de Oxigênio , Radical Hidroxila , Metronidazol/metabolismo , Oxirredução , Poluentes Químicos da Água/química , Poluentes Químicos da Água/metabolismoRESUMO
This article describes a method for high-throughput competition assays using a bioluminescent strain of L. monocytogenes. This method is based on the use of the luminescent indicator strain L. monocytogenes EGDelux. The luminescence of this strain is correlated to growth, which make it suitable to monitor the growth of L. monocytogenes in mixed cultures. To this aim, luminescence kinetics were converted into a single numerical value, called the Luminescence Disturbance Indicator (LDI), which takes into account growth inhibition phenomena resulting in latency increase, decrease in the luminescence rate, or reduction of the maximum luminescence. The LDI allows to automatically and simultaneously handle multiple competition assays which are required for high-throughput screening (HTS) approaches. The method was applied to screen a collection of 1810 strains isolated from raw cow's milk in order to identify non-acidifying strains with anti-L. monocytogenes bioprotection properties. This method was also successfully used to identify anti-L. monocytogenes candidates within a collection of Lactococcus piscium, a species where antagonism was previously described as non-diffusible and requiring cell-to-cell contact. In conclusion, bioluminescent L. monocytogenes can be used in HTS to identify strains with anti-L. monocytogenes bioprotection properties, irrespectively of the inhibition mechanism.
RESUMO
PURPOSE: Superfamily 1 and Superfamily 2 helicases, two of the largest helicase protein families, play vital roles in many biological processes including replication, transcription and translation. Study of helicase proteins in the model microorganisms of archaea have largely contributed to the understanding of their function, architecture and assembly. Based on a large phylogenomics approach, we have identified and classified all SF1 and SF2 protein families in ninety five sequenced archaea genomes. Here we developed an online webserver linked to a specialized protein database named ARCPHdb to provide access for SF1 and SF2 helicase families from archaea. METHODS: ARCPHdb was implemented using MySQL relational database. Web interfaces were developed using Netbeans. Data were stored according to UniProt accession numbers, NCBI Ref Seq ID, PDB IDs and Entrez Databases. RESULTS: A user-friendly interactive web interface has been developed to browse, search and download archaeal helicase protein sequences, their available 3D structure models, and related documentation available in the literature provided by ARCPHdb. The database provides direct links to matching external databases. CONCLUSIONS: The ARCPHdb is the first online database to compile all protein information on SF1 and SF2 helicase from archaea in one platform. This database provides essential resource information for all researchers interested in the field.
Assuntos
Proteínas Arqueais , Biologia Computacional , Bases de Dados de Proteínas , RNA Helicases , Archaea , Sistemas de Gerenciamento de Base de Dados , Interface Usuário-ComputadorRESUMO
The aim of this work was to develop a strategy for second-generation ethanol production from carob solid waste issued from Lebanese food industry. The pros and cons of submerged (SF) and solid-state fermentations (SSF) using S. cerevisiae on ethanol yield and productivity were compared, including the respective roles of upstream and downstream processes, such as the size reduction, or sugar and ethanol recovery processes. The design of experiments methodology was applied. Experimental results demonstrated that SSF applied to cut carob waste from carob syrup preparation was simpler to operate and more cost-effective, maintained yield and productivity (0.458g ethanol/g consumed sugar and 4.3g/(kg waste)/h) in comparison to SF (0.450g ethanol/g consumed sugar and 5.7g/(kg waste)/h), and was able to achieve ethanol production up to 155g/(kg waste) at low water demand, while SF reached only 78g/(kg waste) due to the limitations of the sugar extraction pretreatment.
Assuntos
Fermentação , Saccharomyces cerevisiae , Etanol , FabaceaeRESUMO
Complete impaction of primary teeth is a very rare condition and less seen at the dental office compared with permanent dentition. To report the use of cone-beam computed tomography in the management of a 7-year-old boy with completely impacted maxillary second deciduous molar due to the presence of odontoma and a cystic lesion.