Antifungal Products Research Articles

Disrupting the Dok3-Card9 Interaction with Synthetic Peptides Enhances Antifungal Effector Functions of Human Neutrophils.

Invasive fungal disease is an emerging and serious public health threat globally. The expanding population of susceptible individuals, together with the rapid emergence of multidrug-resistant fungi pathogens, call for the development of novel therapeutic strategies beyond the limited repertoire of licensed antifungal drugs. Card9 is a critical signaling molecule involved in antifungal defense; we have previously identified Dok3 to be a key negative regulator of Card9 activity in neutrophils. In this study, we identified two synthetic peptides derived from the coiled-coil domain of Card9, which can specifically block Dok3-Card9 binding. We showed that these peptides are cell-permeable, non-toxic, and can enhance antifungal cytokine production and the phagocytosis of human neutrophils upon fungal infection. Collectively, these data provide a proof of concept that disrupting the Dok3-Card9 interaction can boost the antifungal effector functions of neutrophils; they further suggest the potential utility of these peptide inhibitors as an immune-based therapeutic to fight fungal infection.

Characterization of a Haloalkaline Nocardiopsis sp. Strain S10 Isolated from Wastewater and Optimization of Culture Medium for Improving Production of Antifungal Compounds

Background: The emerging antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) are a pressing global concern. Furthermore, the limited number of available antifungal agents requires the discovery of new antimicrobials. Introduction: This paper focused on the isolation and characterization of an actinobacterium designated S10, the study of its antimicrobial activity, on the one hand, and the optimization of antifungal production using statistical methods, on the other hand. Methods and Results: A haloalkaliphilic actinobacterium designated S10 was isolated from a wastewater treatment plant in the Souk-El-Tenine region (Bejaia, Algeria). This strain showed a broad spectrum of activity against bacteria (B. subtilis, L. innocua, M. luteus, S. aureus, MRSA, E. coli, P. aeruginosa, S. Typhi, and V. cholerae), molds (A. niger, G. geotricum, and F. oxysporum) and a pathogenic yeast (C. albicans). The extracellular enzymes producer Nocardiopsis sp. S10 was identified based on morphological, chemical, and 16S rRNA gene sequencing analysis. The best antimicrobial production was achieved on Mincer-Sea-Water (MSW) solid medium, and the antibiotics produced were extracted with ethyl acetate. For antifungal production, central composite design (CCD) and response surface methodology (RSM) were used to optimize the culture medium formulation. Conclusion: Optimal antifungal activity was achieved using a mixture of 2 g. L-1 starch, 2 g. L-1 of yeast extract and 2.75 g. L-1 of peptone at pH = 9.2. The application of the statistical approach stimulated a good yield of anti-C. albicans activity, corresponding to an increase of 72.22% compared to the non-optimized conditions.

Naringenin-Capped Silver Nanoparticles Amalgamated Gel for the Treatment of Cutaneous Candidiasis.

The current research was aimed to synthesize a phytomolecule, naringenin (NRG)-mediated silver nanoparticles (NRG-SNPs) to study their antifungal potential against Candida albicans (C. albicans) and Candida glabrata (C. glabrata). The NRG-SNPs were synthesized by using NRG as a reducing agent. The synthesis of NRG-SNPs was confirmed by a color change and surface plasmon resonance (SPR) peak at 425nm. Furthermore, the NRG-SNPs were analyzed for size, PDI, and zeta potential, which were found to be 35 ± 0.21nm, 0.19 ± 0.03, and 17.73 ± 0.92mV, respectively. In silico results demonstrated that NRG had a strong affinity towards the sterol 14α-demethylase. The docking with ceramide revealed the skin permeation efficiency of the NRG-SNPs. Next, the NRG-SNPs were loaded into the topical dermal dosage form (NRG-SNPs-TDDF) by formulating a gel using Carbopol Ultrez 10 NF. The MIC50 of NRG solution and TSC-SNPs against C. albicans was found to be 50µg/mL and 4.8µg/mL, respectively, significantly (P < 0.05) higher than 0.3625µg/mL of NRG-SNPs-TDDF. Correspondingly, MIC50 results were calculated against C. glabrata and the results of NRG, TSC-SNPs, NRG-SNPs-TDDF, and miconazole nitrate were found to be 50µg/mL, 9.6µg/mL, 0.3625µg/mL, and 3-µg/mL, respectively. Interestingly, MIC50 of NRG-SNPs-TDDF was significantly (P < 0.05) lower than MIC50 of miconazole nitrate against C. glabrata. The FICI (fractional inhibitory concentration index) value against both the C. albicans and C. glabrata was found to be 0.016 and 0.011, respectively, which indicated the synergistic antifungal activity of NRG-SNPs-TDDF. Thus, NRG-SNPs-TDDF warrants further in depth in vivo study under a set of stringent parameters for translating in to a clinically viable antifungal product.

CysB Is a Key Regulator of the Antifungal Activity of Burkholderia pyrrocinia JK-SH007.

Burkholderia pyrrocinia JK-SH007 can effectively control poplar canker caused by pathogenic fungi. Its antifungal mechanism remains to be explored. Here, we characterized the functional role of CysB in B. pyrrocinia JK-SH007. This protein was shown to be responsible for the synthesis of cysteine and the siderophore ornibactin, as well as the antifungal activity of B. pyrrocinia JK-SH007. We found that deletion of the cysB gene reduced the antifungal activity and production of the siderophore ornibactin in B. pyrrocinia JK-SH007. However, supplementation with cysteine largely restored these two abilities in the mutant. Further global transcriptome analysis demonstrated that the amino acid metabolic pathway was significantly affected and that some sRNAs were significantly upregulated and targeted the iron-sulfur metabolic pathway by TargetRNA2 prediction. Therefore, we suggest that, in B. pyrrocinia JK-SH007, CysB can regulate the expression of genes related to Fe-S clusters in the iron-sulfur metabolic pathway to affect the antifungal activity of B. pyrrocinia JK-SH007. These findings provide new insights into the various biological functions regulated by CysB in B. pyrrocinia JK-SH007 and the relationship between iron-sulfur metabolic pathways and fungal inhibitory substances. Additionally, they lay the foundation for further investigation of the main antagonistic substances of B. pyrrocinia JK-SH007.

Biosensor-Enabled Discovery of CaERG6 Inhibitors and Their Antifungal Mode of Action against Candida albicans.

Fungal infections caused by opportunistic pathogens, such as Candida albicans, are generally underappreciated by the public in spite of their high mortality rates. Antifungal arsenals are extremely limited. Herein, based on biosynthetic pathway comparison and functional characterization, CaERG6, a crucial sterol 24-C-methyltransferase involved in the biosynthesis of ubiquitous ergosterol in C. albicans, was set up as an antifungal target. CaERG6 inhibitors were identified from the in-house small-molecule library by a biosensor-based high-throughput screening. The CaERG6 inhibitor NP256 (palustrisoic acid E) is a potential antifungal natural product that acts by inhibiting ergosterol biosynthesis, downregulating the gene expression level in hyphal formation, blocking biofilm formation, and disrupting morphological transition in C. albicans. NP256 enhances C. albicans susceptibility to some known antifungals significantly. The present study demonstrated the CaERG6 inhibitor NP256 as a potential class of antifungal compound for monotherapy or combinatory therapy.

Identification of Fcl-29 as an Effective Antifungal Natural Product against Fusarium graminearum and Combinatorial Engineering Strategy for Improving Its Yield.

Fusarium head blight (FHB), caused by Fusarium graminearum, whose occurrence and prevalence causes 10-70% wheat production loss, is one of the most destructive diseases influencing the production of wheat globally. To identify the potential natural products (NPs) against F. graminearum, we screened 59 Xenorhabdus strains and discovered that the cell-free supernatant (CFS) of X. budapestensis 14 (XBD14) displays the highest bioactivity. Multiple genetic methods coupled with HRMS/MS analysis determined the major antifungal NP to be Fcl-29, a fabclavine derivative. Fcl-29 was found to effectively control FHB of wheat in the field test and demonstrated broad-spectrum antifungal activity against important pathogenic fungi. The production of Fcl-29 was dramatically improved by 33.82-fold with the combinatorial strategy of genetic engineering (1.66-fold) and fermentation engineering (20.39-fold). The exploration of a new biofungicide in global plant protection is now possible.

Antifungal and Antibiofilm Activity of Colombian Essential Oils against Different Candida Strains.

Most Candida species are opportunistic pathogens with the ability to form biofilms, which increases their resistance to antifungal drug therapies and the host immune response. Essential oils (EOs) are an alternative for developing new antimicrobial drugs, due to their broad effect on cellular viability, cell communication, and metabolism. In this work, we evaluated the antifungal and antibiofilm potential of fifty EOs on C. albicans ATCC 10231, C. parapsilosis ATCC 22019, and Candida auris CDC B11903. The EOs' antifungal activity was measured by means of a broth microdilution technique to determine the minimum inhibitory and fungicidal concentrations (MICs/MFCs) against the different Candida spp. strains. The effects on biofilm formation were determined by a crystal violet assay using 96-well round-bottom microplates incubated for 48 h at 35 °C. The EOs from Lippia alba (Verbenaceae family) carvone-limonene chemotype and L. origanoides exhibited the highest antifungal activity against C. auris. The L. origanoides EOs also presented antifungal and antibiofilm activity against all three Candida spp., thus representing a promising alternative for developing new antifungal products focused on yeast infections, especially those related to biofilm formation, virulence factors, and antimicrobial resistance.

Bacillus amyloliquefaciens FG14 as a potential biocontrol strain against rusty root rot of Panax ginseng, and its impact on the rhizosphere microbial community

Rusty root rot is the most destructive disease in ginseng production. The bacterial strain FG14 isolated from the rhizosphere of Panax ginseng was characterised as Bacillus amyloliquefaciens. The inhibition rates of the FG14 bacterial suspension against the mycelial growth, spore production and spore germination of Ilyonectria robusta which causes ginseng rusty root rot were all more than 90% at a concentration of 108 cfu/mL. The sterile fermentation broth of strain FG14 also had significant inhibitory ability. The results of plantgrowth promotion showed that the fresh weight and root length of ginseng seedlings treated with 3 × 108 cfu/mL FG14 bacterial suspension were 4.49 times and 2.12 times greater than the seedlings treated with I. robusta. The soil enzyme activities of urease, phosphatase, invertase, and catalase of strain FG14 and the expression of seven lipopeptide biosynthesis genes were up-regulated, and antifungal metabolite production was stable. The diversity and abundance of bacterial and fungal communities in rhizosphere soil changed during different growth stages of ginseng after inoculation with strain FG14. The relative abundance of the Ilyonectria genus was significantly lower than the water control at the leaf spreading stage of P. ginseng. These results indicate the great potential of B. amyloliquefaciens FG14 for use as a biological agent for the green control of ginseng rusty root rot.

Recent Biomedical Applications of Coupling Nanocomposite Polymeric Materials Reinforced with Variable Carbon Nanofillers.

The hybridization between polymers and carbon materials is one of the most recent and crucial study areas which abstracted more concern from scientists in the past few years. Polymers could be classified into two classes according to the source materials synthetic and natural. Synthetic polymeric materials have been applied over a floppy zone of industrial fields including the field of biomedicine. Carbon nanomaterials including (fullerene, carbon nanotubes, and graphene) classified as one of the most significant sources of hybrid materials. Nanocarbons are improving significantly mechanical properties of polymers in nanocomposites in addition to physical and chemical properties of the new materials. In all varieties of proposed bio-nanocomposites, a considerable improvement in the microbiological performance of the materials has been explored. Various polymeric materials and carbon-course nanofillers were present, along with antibacterial, antifungal, and anticancer products. This review spots the light on the types of synthetic polymers-based carbon materials and presented state-of-art examples on their application in the area of biomedicine.

Bioprospecting of desert actinobacteria with special emphases on griseoviridin, mitomycin C and a new bacterial metabolite producing Streptomyces sp. PU-KB10–4

BackgroundBioprospecting of actinobacteria isolated from Kubuqi desert, China for antibacterial, antifungal and cytotoxic metabolites production and their structure elucidation.ResultsA total of 100 actinobacteria strains were selectively isolated from Kubuqi desert, Inner Mongolia, China. The taxonomic characterization revealed Streptomyces as the predominant genus comprising 37 different species, along with the rare actinobacterial genus Lentzea. The methanolic extracts of 60.8% of strains exhibited potent antimicrobial activities against Staphylococcus aureus, Micrococcus luteus, Bacillus subtilis, Escherichia coli, Salmonella enterica, Saccharomyces cerevisiae and high to mild in vitro cytotoxicity against PC3 (prostate cancer) and A549 (lung carcinoma) cell lines. The metabolomics analysis by TLC, HPLC-UV/vis, HPLC-MS and NMR showed the presence of compounds with molecular weights ranging from 100 to 1000 Da. The scale-up fermentation of the prioritized anti-Gram-negative strain PU-KB10–4 (Streptomyces griseoviridis), yielded three pure compounds including; griseoviridin (1; 42.0 mgL− 1) with 20 fold increased production as compared to previous reports and its crystal structure as monohydrate form is herein reported for the first time, mitomycin C (2; 0.3 mgL− 1) and a new bacterial metabolite 4-hydroxycinnamide (3; 0.59 mgL− 1).ConclusionsThis is the first report of the bioprospecting and exploration of actinobacteria from Kubuqi desert and the metabolite 4-hydroxycinnamide (3) is first time isolated from a bacterial source. This study demonstrated that actinobacteria from Kubuqi desert are a potential source of novel bioactive natural products. Underexplored harsh environments like the Kubuqi desert may harbor a wider diversity of actinobacteria, particularly Streptomyces, which produce unique metabolites and are an intriguing source to develop medicinally valuable natural products.

Burkholderia gladioli strain KJ-34 exhibits broad-spectrum antifungal activity.

Plant pathogens are one of the major constraints on worldwide food production. The antibiotic properties of microbes identified as effective in managing plant pathogens are well documented. Here, we used antagonism experiments and untargeted metabolomics to isolate the potentially antifungal molecules produced by KJ-34. KJ-34 is a potential biocontrol bacterium isolated from the rhizosphere soil of rice and can fight multiple fungal pathogens (i.e. Ustilaginoidea virens, Alternaria solani, Fusarium oxysporum, Phytophthora capsica, Corynespora cassiicola). The favoured fermentation conditions are determined and the fermentation broth treatment can significantly inhibit the infection of Magnaporthe oryzae and Botryis cinerea. The fermentation broth suppression ratio is 75% and 82%, respectively. Fermentation broth treatment disrupted the spore germination and led to malformation of hyphae. Additionally, we found that the molecular weight of antifungal products were less than 1000 Da through semipermeable membranes on solid medium assay. To search the potentially antifungal molecules that produce by KJ-34, we used comparative and bioinformatics analyses of fermentation broth before and after optimization by mass spectrometry. Untargeted metabolomics analyses are presumed to have a library of antifungal agents including benzoylstaurosporine, morellin and scopolamine. These results suggest that KJ-34 produced various biological control agents to suppress multiple phytopathogenic fungi and showed a strong potential in the ecological technologies of prevention and protection.

Drilling down the bioequivalence assessment of topical antifungal products: Microstructure and release

In recent years, the regulatory mechanisms for topical generic product bioequivalence (BE) assessment have been subjected to noteworthy changes, with the FDA issuing product specific guidances, and the EMA adopting a more universal approach with the quality and equivalence of topical products draft guideline. The agencies advise on a modular strategy for BE documentation. Nevertheless, their scope, data analysis and criteria are rather distinct.This study aims to tackle bioequivalence assessment issues of complex topical formulations starting by statistical implications of the EMA/FDA approaches concerning the documentation of qualitative (Q1), quantitative (Q2), microstructure (Q3) and performance requirements (Q4). As a model drug product, a bifonazole 10 mg/g cream formulation was selected. For this specific formulation, the commercially available Reference Product (RP) was compared with two comparator products, also commercially available, referred to as comparator product A (CPA) and comparator product B (CPB). The former displays Q1 sameness and Q2 differences, whilst CPB is categorically considered as Q1/Q2 different. Furthermore, intending to establish a regulatory rationale for the submission of a generic product according to the updated regulatory requirements, the RP was likewise compared with a Test Product (TP). This formulation was designed to display equal Q1/Q2 profile to that of the RP. Validated rheology and in vitro release test (IVRT) methods were used to infer on Q3/Q4 characteristics.During rheology studies, statistically significant RP batch to batch differences were observed. Therefore, in an attempt to surpass this heterogeneity, the initial pool of RP batches was expanded to include RP product batches at different lifecycle stages. Despite this effort, it was not possible to classify the RP/TP, RP/CPA or RP/CPB as rheologically equivalent products. Nevertheless, product performance results, retrieved from IVRT, were able to sustain equivalence between the RP and the formulations exhibiting Q1 sameness (TP and CPA). It should however be mentioned, that for some RP batch combinations, the IVRT results failed to demonstrate equivalence according to the EMA requirements. Enlarging the RP batch pool was then a critical step in further understanding an optimum statistical approach for establishing equivalence in product performance.This study highlights the need to that a ‘one-fits-all approach’ may not be an optimum path way for establishing the regulatory strategy and requirements to support generic product bioequivalence.

A Combination Strategy of Multidrug-Sensitive Budding Yeast and Chemical Modifications Enabling to Find a New Overlooked Antifungal Compound, Sakurafusariene, from In-House Fractionated Library.

In this report, we disclose our discovery of a new antifungal natural product, sakurafusariene (1), from an in-house fractionated library of the culture broth of Fusarium sp. FKI-7550 strain by using a combination strategy of multidrug-sensitive yeast and chemical modification. Throughout our investigation, we encountered challenges in the isolation of natural product 1. A chemical modification strategy via alkylation of 1 allowed for removal of the impurities enabling us to elucidate the structure of 1. Furthermore, we synthesized ester derivatives using a method inspired by the isolation study of 1, which gave us valuable information to understand a preliminary structure-activity relationship against Pyricularia oryzae growth inhibitory activity.

Synthesis and characterization of anti-fungal paint production based on bintaro (Cerbera manghas) seed extract as additive

Abstrak. Penelitian ini melaporkan sintesis alkaloid dari biji bintaro (Cerbera manghas). Serbuk biji bintaro yang dihasilkan dari sintesis ekstrak biji bintaro (Cerbera manghas) dikarakterisasi menggunakan Spektroskopi Fourier Transform Infrared (FTIR) dan uji fitokimia. Spektrum FTIR hasil pengujian dari sintesis ekstrak biji bintaro menunjukkan puncak kuat pada 3239, 3338, dan 3293 cm-1 dan sudah menunjukkan hasil alkaloid. Hasil pengujian fitokimia juga telah menunjukkan terdapat senyawa alkaloid. Sintesis alkaloid yang telah diperoleh dijadikan zat aditif pada pembuatan cat. Pada penelitian ini telah berhasil dibuat cat yang berbahan dasar cat komersil untuk aplikasi kayu dengan penambahan zat aditif yaitu serbuk biji bintaro (Cerbera manghas). Pembuatan cat diperoleh dengan menggabungkan cat komersil dengan serbuk biji bintaro dengan perbandingan fraksi massa yaitu 100:0, 95:5, 90:10, 85:15. Cat pada penelitian ini diuji karakterisasinya antara lain adalah uji daya rekat, uji densitas, dan uji viskositas. Hasil penelitian menunjukkan bahwa uji daya rekat terbaik dengan fraksi massa cat: serbuk biji bintaro sebanyak 95:5 masuk dalam klasifikasi 5B dengan area terkelupas sebanyak 0,6%, pada uji densitas dan viskositas nilai yang tertinggi terdapat pada fraksi massa 100:0, nilai viskositas dan densitas masih memenuhi kategori cat kayu yaitu standar SNI 8406.6:2017 densitas cat minimal 1,2 g/ dan standar viskositas cat minimal 11,5 poise. Abstract. This study reports on the synthesis of alkaloids from the seeds of bintaro (Cerbera mangos). Bintaro seed powder produced from the synthesis of bintaro seed extract (Cerbera mangos) was characterized using Fourier Transfor Infrared Spectroscopy (FTIR) and phytochemical tests. The FTIR spectrum of the test results from the synthesis of bintaro seed extract showed strong peaks at 3239, 3338, and 3293 cm-1and already showed alkaloid results. Phytochemical test results have also shown that there are alkaloid compounds. The synthesis of alkaloids that have been obtained is used as an additive in the manufacture of paint. In this study, commercial paint-based paints for wood applications have been successfully made with the addition of additives, namely bintaro seed powder (Cerbera mangos). The manufacture of paint is obtained by combining commercial paint with bintaro seed powder with a mass fraction ratio of 100:0, 95:5, 90:10, 85:15. The paint in this study was tested for its characterization, including the adhesion test, density test, and viscosity test. The results showed that the best adhesion test with the mass fraction of paint: bintaro seed powder as much as 95:5 included in the 5B classification with a peeling area of 0.6%, in the density and viscosity test the highest value was found in the mass fraction of 100:0, the value the viscosity and density still meet the wood paint category, namely the standard SNI 8406.6:2017 paint density of at least 1.2 g/m3 and the minimum paint viscosity standard of 11.5 poise.

In vitro evaluation of the herbal cream formulation from methanolic leaf extracts of Tephrosia vogelii Hook.f for topical application

BackgroundTopical infectious are among of global challenges which propelled up with antimicrobial resistances. This scenario compels for drug discovery and development. Herbal formulation is one of important struggle in the drug development with the purpose of overcoming various diseases distressing health care settings. Thus, this study aimed to formulate and evaluate herbal cream from leaf methanolic extracts of Tephrosia vogelii Hook.f for topical therapies.MethodsHerbal cream was prepared by homogeneously mixing up cream base and methanolic leaf extracts of Tephrosia vogelii. Physicochemical properties and in vitro antimicrobial activities of the cream were evaluated. The stability, colour change, washability and texture were parameters used evaluate physicochemical properties of cream. Disc diffusion method was employed to evaluate antimicrobial activities of the cream against Candida albicans (ATCC 90,028), Escherichia coli (ATCC29953) and Staphylococcus aureus (ATCC25923).ResultsFormulated herbal creams; CBTV1, CBTV2 and CBTV3 with methanolic extracts concentrations content of Tephrosia vogelii 0.05%, 0.10% and 0.25%, respectively, were prepared. All cream formulations were stable, washable by water, green in colour and soft. The CBTV3 product exhibited with high prominent antimicrobial activity compared to the rest, henceforth promisingly to be effective for topical skin therapy.ConclusionThe herbal cream formulation of methanolic leaf extract of Tephrosia vogelii at 0.25% appeared to have more antimicrobial performance. Therefore, it is considered as minimal concentration for the herbal cream formulation in future for clinical trials as a potential antifungal and antibacterial agent product for treatment of skin infections.Graphical

The recombination regulator RecX negatively regulates heat-stable antifungal factor (HSAF) biosynthesis in Lysobacter enzymogenes

Bacteria often use multiple transcription factors to regulate specific biological processes. Biosynthesis of heat-stable antifungal factor (HSAF) is regulated by multiple factors in Lysobacter enzymogenes. However, the mechanism of HSAF biosynthesis regulation remains largely unknown. In this study, we screened a potential HSAF biosynthesis regulator, RecX, by a DNA pull-down assay. Deletion of recX resulted in a significant increase in the production of HSAF, and overexpression of recX significantly suppressed HSAF production. Importantly, our results showe that RecX directly binds to the promoter region of the lafB gene to inhibit its transcription and thus decreases HSAF production in L. enzymogenes. These findings reveal the novel mechanism of RecX regulation of antifungal antibiotic production in L. enzymogenes.

Influence of menthol on biofilm formation, ergosterol content, and cell surface hydrophobicity of Candida glabrata.

Resistance to synthetic antifungals has become one of the leading public health challenges around the world. Accordingly, novel antifungal products like naturally occurring molecules can be one of the potential ways to reach efficient curative approaches to control candidiasis. This work evaluated the effect of menthol on cell surface hydrophobicity (CSH), biofilm formation, growth and ergosterol content of Candida glabrata (C. glabrata), a yeast with a high resistance against antifungal agents. Disc diffusion method (susceptibility to synthetic antifungals), broth micro-dilution method (Susceptibility to menthol), MTT reduction assay (biofilm formation), HPLC technique (ergosterol content) and adherence to n-hexadecane (CSH) were employed to determine the influence of menthol against C. glabrata isolates. The MIC range of menthol versus C. glabrata was 1250-5000µg/mL (mean±SD: 3375 ± 1375µg/mL). The mean rate of C. glabrata biofilm formation was decreased up to 97.67, 81.15, 71.21, 63.72, 47.53, 26.31 and 0.051% at 625, 1250, 2500, 5000, 10000, 20000 and 40000µg/mL concentrations, respectively. The percentages of CSH were significantly in groups treated with MIC/2 (17.51±5.52%) and MIC/4 (26±5.87%) concentrations of menthol. Also, the percentage changes in membrane ergosterol were 15.97, 45.34 and 73.40% at 0.125, 0.25 and 0.5mg/mL concentrations of menthol, respectively, in comparison with untreated control. The results showed the menthol impact versus sessile and planktonic C. glabrata cells, and the interference with ergosterol content, CSH, and biofilm formation, which made it a potent natural antifungal.

Life cycle assessment on calcium zincate production methods for rechargeable batteries

The world's energy transition from fossil to renewable energy is unthinkable without further research in energy storage. Decreasing the environmental impacts from the production of energy storage technologies is essential for achieving a green energy transition. Calcium Zincate (CAZN) is used as active material in rechargeable zinc-based batteries (and other products, such as heterogeneous catalysts for biodiesel or antifungal products). They present a low-cost, safer, alternative to Lithium based batteries and are targeted as replacement solutions for lead-acid batteries.We propose a novelty in the synthesis of CAZN, the hydro-micro-mechanical process (HMMS). The residence time of this new route is about 20 times lower than the traditional processes, so its production needs less infrastructure and can deliver quicker at an industrial scale. In addition, laboratory tests indicate that HMMS CAZN has more reaction surface area and the activation of the battery is 1.77 times faster.Using the life cycle assessment (LCA) method, we compare this new process with the current best option, hydro-thermal synthesis (HTS). The cradle-to-gate results per kg of CAZN already indicates that HMMS is an environmentally better alternative for all indicators; especially when considering the normalization of the results with the residence time and the surface area, HMMS delivers better results, with improvements of 97 % in global warming, for instance. With this, we demonstrate that, outside of the cradle-to-gate, variables that make the final products better service units or give more function should be considered as valuable additional information when deciding among alternatives. This also highlights the importance of life cycle thinking when working with chemical processes and substances.In the sensitivity analysis, we developed 7 scenarios related to the energy demand of the processes, and we incorporated the projection in the European electricity mix for 2030 and 2050.

Evaluation of the Antioxidant and Antifungal Actions of the Optimized Crude Extract and Fractions from the Aerial Parts of Acanthospermum hispidum.

This study investigated the phytochemical characteristics of the aerial parts of Acanthospermumhispidum, by chromatographic and spectrophotometric methods, and evaluated the antioxidant and antifungal activities of the crude extract and polyphenol-enriched fractions of the species. The phytochemical prospection showed the presence of polyphenols from the groups of hydroxycinnamic derivatives and flavonoids in the crude extract (CE) and fractions of the aerial parts of A. hispidum . In the chromatographic analysis, it was possible to observe that the fractionation process of the CE with hexane and ethyl acetate was efficient in enriching the fractions in phenolic compounds. This enrichment provided an increase in antioxidant activity by the DPPH and ABTS methods, in which it was observed a higher antioxidant activity for EAF in the DPPH test and higher activity against the ABTS radical by the fractions AqF and RAqF. The extract and fractions were effective against Candida non- Candida albicans strains, mainly against C. glabrata , C. parapsilosis and C. krusei , acting predominantly fungicidal. The results indicate that the aerial parts of A. hispidum can serve as a basis for the development of new antioxidant and antifungal products. Moreover, the fractionation process can contribute to increasing the biological potential of the species.

Cyclic di-GMP regulates bacterial colonization and further biocontrol efficacy of Bacillus velezensis against apple ring rot disease via its potential receptor YdaK.

Bacillus species are among the most investigated beneficial bacteria and widely used in agricultural systems as biological control agents. Its biocontrol efficacy is controlled by diverse regulators. Cyclic diguanylate (c-di-GMP) is a nearly universal second messenger in bacteria and modulates various important physiological processes, including motility, biofilm formation, antifungal antibiotic production and host colonization. However, the impact of c-di-GMP on biocontrol efficacy of beneficial bacteria is unknown. Bacillus velezensis PG12 is an effective biocontrol strain against apple ring rot disease caused by Botryosphaeria dothidea. In this study, the contribution of c-di-GMP to biocontrol efficacy of B. velezensis PG12 was investigated. Deletion of single gene encoding diguanylate cyclase or phosphodiesterase did not affect its biocontrol efficacy against apple ring rot. However, artificial modulation of c-di-GMP level in the cells leads to a significant change of biocontrol efficacy, suggesting that c-di-GMP positively regulates biocontrol efficacy of B. velezensis PG12 against apple ring rot disease. More evidences indicate that c-di-GMP does not affect the antagonistic activity of B. velezensis PG12 against B. dothidea in vitro and in vivo, but positively regulates biofilm formation of B. velezensis PG12 and its colonization on apple fruits. Importantly, deletion of ydaK could rescue the inhibition of biofilm formation, bacterial colonization and biocontrol efficacy caused by low c-di-GMP level, indicating that YdaK is the potential c-di-GMP receptor to regulate biofilm formation, colonization and effective biological control. However, YdaK did not affect the antagonistic activity of B. velezensis PG12 against B. dothidea. Based on these findings, we propose that c-di-GMP regulates biofilm formation, subsequently the bacterial colonization on apple fruits and thus biocontrol efficacy of B. velezensis through its receptor YdaK. This is the first report showing that c-di-GMP plays a role in biocontrol efficacy of beneficial bacteria.