O endereço abaixo não existe na globo.com This website is using a security service to protect itself from online attacks The action you just performed triggered the security solution There are several actions that could trigger this block including submitting a certain word or phrase You can email the site owner to let them know you were blocked Please include what you were doing when this page came up and the Cloudflare Ray ID found at the bottom of this page The report provides evidence of the weakness of industry self-regulation such as flaws in external audits and certification schemes which do not consistently and effectively identify the risks to human rights and harm prevention On the contrary: they end up providing false guarantees of compliance while at the same time preventing the necessary investigation and interventions Dozens of recurring violations are documented in the study such as the absence of employment contracts inadequate hygiene facilities and failure to provide drinking water and meals and it was very cold in the house,” said one of the workers rescued from a farm nearly all the accounts mention the lack of provision of personal protective equipment by the employer an essential and mandatory item for this work.   20 workers – including a 15-year-old girl and three boys under 18 – were rescued from a farm in the supply chain of Nestlé where they did not receive regular wages and had to use their own personal protection items the farm had an international sustainability certification from the Rainforest Alliance.  “Legislation needs to be passed that addresses forced labor in supply chains and that requires strict alignment with the Guidelines of the OECD (Organization for Economic Co-operation and Development) and other international standards,” said Júlia Mello Neiva director of strengthening the human rights movement at Conectas “A law banning the sale of products made with forced labor is one of the possible paths identified in the report as well as greater guarantees of reparations for affected workers and the identification and disclosure of all those involved in the production process,” said Neiva.  Victims of modern slavery seek reparations through voluntary complaint mechanisms such as the National Contact Points (NCPs) of the OECD Guidelines Conectas and Adere-MG (Rural Workers Alliance of the State of Minas Gerais) filed a complaint to the Brazilian NCP of violations involving modern-day slavery on coffee farms in southern Minas Gerais which were part of the supply chain of Nestlé The extrajudicial complaint was based on data from the Dutch organization Danwatch and the local NGO Repórter Brasil which identified that the multinational company purchased coffee from farms where the beans were harvested by people in conditions akin to slavery Following negotiations between Conectas, Adere-MG and Nestlé, the NCP released a report, in October 2023, with tentative recommendations for the company to eradicate human rights violations in its production chain.  Read more details about the case at this link 2025 - Conectas Human Rights - Postal Code 47 - São Paulo (SP) Brazil - ZIP: 01032-970 - Phone: +55 (11) 3884-7440 We are a global communion of churches in the Lutheran tradition country programs and partner organizations to promote justice reconciliation and dignity for individuals and communities LWF membership represents over 78 million Christians in the Lutheran tradition in 99 countries across the globe as well as ten recognized churches and congregations We strive to put our faith into action within and beyond the communion and seek God’s Word and Spirit to guide us Your partnership truly makes a difference.Together a better world is possible.With Passion for the Church and for the World strengthening relations among young people in Brazil and realizing their connectedness in the global Lutheran communion were vital aspects of the recent national youth gathering IECLB National Council of Evangelical Youth coordinator addressing the young people gathered for the CONGRENAJE The 25th Congresso Nacional da Juventude Evangélica (CONGRENAJE), the national synod meeting of youth in the Evangelical Church of the Lutheran Confession in Brazil (IECLB) brought together more than 1,200 participants in Domingos Martins The CONGRENAJE had the theme “What is your essence?” and referred to Ephesians 5:2b: “Christ loved us and gave his life for us a fragrant offering and sacrifice to God.” Worship and many of the event’s Bible studies subsequently took up passages related to fragrances and aromas unfolding their biblical significance and implications for the lives of young people today.  the National Council of Evangelical Youth coordinator emphasized the importance of the CONGRENAJE in bringing together youth from many regions He also referred to the 200th anniversary of Lutheran presence in Brazil in 2024 highlighting the need to reflect on what is essential and lasting in these changing times.  We have to ask ourselves: What is essential “The theme of our gathering and the anniversary of the IECLB relate well,” Schumann said “Looking back at 50 years of CONGRENAJE and 200 years of Lutheran presence in Brazil we have to ask ourselves: What is essential to highlight the importance of living the essence of Christ’s love in everyday life.  “Jesus brought the essence of love into our world,” Genz said you will ask yourselves what your essence is Remember that the death and resurrection of Jesus Christ are the keys to eternal life May we hold on to Jesus and pour out the precious essence of God’s love.” Youth from ten LWF member churches presented a banner they created It represents their “essence” as a group and shows their shared identity symbols to represent the work of their churches and a globe with a flower native to each of their countries underneath - representing both their diversity and their connection Special guests to the CONGRENAJE included 20 young people from The Lutheran World Federation (LWF) member churches and Latin America and the Caribbean regions.  LWF Youth Secretary Savanna Sullivan welcomed this initiative “Every church has something meaningful to offer and international partners also have something to offer,” she said “Inviting youth from other parts of the world and offering hospitality to them shows an important aspect of being church together as young people in Brazil and beyond I hope that young people in other churches take up the idea of being connected to the communion when planning their gatherings.”  the participants enjoyed a varied program that included lectures and moments of communion and fellowship among young people from different regions of Brazil and other countries One of the highlights of the gathering was the “Youth Shout,” a public act in which young people shared their message with society and social values in the central square of Domingos Martins This is the place where celebrations marking two centuries of Lutheran presence in Brazil were also launched initiating festivities throughout the country in the year to come Chemin du Pavillon 2,1218 Le Grand Saconnex [email protected]+41 (0)22 791 60 00 "Truly I tell you, whatever you did for one of the least of these brothers and sisters of mine, you did for me." – Matthew 25.40 Volume 15 - 2024 | https://doi.org/10.3389/fmicb.2024.1373469 This article is part of the Research TopicNew Insights in Microbial Stress Tolerance MechanismsView all 15 articles Aspergilli comprise a diversity of species that have been extensively studied due to their catabolic diversity An impressive level of structural and functional conservation has been shown for aspergilli regardless of many (yet) cryptic genomic elements We have hypothesized the existence of conserved genes responsive to stress in aspergilli To test the hypothesis of such conserved stress regulators in aspergilli a straightforward computational strategy integrating well-established bioinformatic tools was used as the starting point five transcriptome-based datasets on exposure to organic compounds were used covering three distinct Aspergillus species only one gene showed the same response in all conditions This gene encodes a protein containing a phenylcoumaran benzylic ether reductase-like domain and a Nitrogen metabolite repressor regulator domain (NmrA) Deletion of this gene caused significant phenotypic alterations compared to that of the parental strain across diverse conditions the deletion of AN9181 raised the mutant’s metabolic activity in different nitrogen sources The acquired data supports that AN9181 acts by repressing (slowing down) A nidulans growth when exposed to aromatic compounds in a concentration dependent manner The same phenotype was observed for amphotericin B AN9181 underwent differential upregulation under oxidative stress conditions herein assigned as NmrB (Nitrogen Metabolite Repression Regulator B) builds up the genetic machinery of perception of oxidative stress by negatively regulating growth under such conditions we integrated the transcriptome signatures of different aspergilli (three species) to distinct organic compounds gathering original and publicly available datasets To identify genes showing the same response across the different datasets a straightforward computational strategy that allows comparing transcriptomic-based datasets initially collected in distinct Aspergillus species was used assigned as NmrB (Nitrogen Metabolite Repression Regulator B) showed the same response in all the datasets nidulans single deletion-mutant ∆AN9181 was compared to that of the wild type growth rate and susceptibility to distinct chemical stressors The collected data suggest that NmrB negatively regulates the metabolism of A Bromoquinol (BMQ) was purchased from Alfa Aesar; 6-iodoquinoline (IDQ) and resveratrol (RVT) from TCI Europe; hydrogen peroxide (H2O2) from Merck; dimethyl sulfoxide from Fisher Chemical and the remaining compounds from Sigma Aldrich miconazole (Mic) and 2,7-dichlorofluorescin diacetate (DCFH-DA) Aspergillus fumigatus CEA17 reference strain was propagated at 37°C in solid complete medium [1% D-glucose, 0.2% Peptone, 0.1% Yeast extract, 0.1% Casamino acids, 50 mL of a 20× salt solution, 0.1% trace elements, 0.1% Vitamin solution, 2% agar, pH 6.5 with NaOH]. The composition of the trace elements, vitamins, and nitrate salts has been described previously (Kafer, 1977) The Minimal Inhibitory Concentration (MIC) for fungal growth inhibition of IDQ, initially acquired from The Pathogen Box (www.pathogenbox.org/), was defined following the standard methodology implemented by the Clinical and Laboratory Standards Institute (CLSI, 2018) The compound’s antifungal activity was analyzed by serial dilutions using MIC assay (0 to 25 μM) in MOPS buffered RPMI 1640 medium (Sigma-Aldrich) Plates were incubated at 37°C without shaking for 48 h Non-inoculated controls were done in parallel The preparation of total RNA samples for RNA-seq Expression Profiling was as follows. Erlenmeyer flasks (125 mL) were used to inoculate 1×107 spores in 30 mL of Vogel’s Minimal Media (Vogel, 1956) and incubated for 16 h at 37°C 0.5x MIC (=0.35 μM) or 2x MIC (=1.4 μM) of IDQ was added and incubated for 4 h at 37°C Six replicates for each condition were prepared the cultures were filtered and frozen immediately in liquid nitrogen Total RNA from six mycelia per condition were extracted using RNeasy Plant Mini Kit (Qiagen) according to the manufacturer’s protocol a TissueLyser LT (Qiagen) for cell disruption and approximately 30 mg of poly(vinylpolypyrrolidone) per sample RNA quality (integrity) was evaluated using a Nucleic Acid QC - Fragment Analyzer we performed a whole-genome protein domain analysis for the protein-translated genome of the “receiver species,” aiming to obtain hints on the putative functions played by the co-expressed genes necessary when a functional annotation is lacking Schematic overview of the COmpregensive CO-expression Analysis (COCOA) strategy developed in this work (A) – the iterative use of the well-established bioinformatics tools OrthoFinder InterProScan and InteractiVenn upon selection of suitable and meaningful transcriptome-based datasets is displayed Laboratory assays implemented to understand the biological relevance of the gene identified by COCOA Results of the orthology analysis performed using the genomes of eight Aspergillus spp (C) – the number of genes of each analyzed genome (D) the percentage of genes in orthogroups and (E) the number (n) of species-specific orthogroups are depicted assays used minimal medium with glucose (10 g·L−1) and nitrate (MMGN) as follows: thiamine (0.01 g·L−1) 5% (v/v) nitrate salts solution [NaNO3 (120.0 g·L−1) MgSO4·7H2O (10.4 g·L−1) and KH2PO4 (30.4 g·L−1)] 0.1% (v/v) trace elements solution [ZnSO4·7H2O (22.0 g·L−1); H3BO3 (11.0 g·L−1); MnCl2·4H2O (5.0 g·L−1); FeSO4·7H2O (5.0 g·L−1); CoCl2·6H2O (1.7 g·L−1); CuSO4·5H2O (1.6 g·L−1); Na2MoO4·2H2O (1.5 g·L−1) and Na4EDTA (50.0 g·L−1)] the media was jellified with 1.5% agar (solid cultivation) The MMG is similar to that described above except that sodium nitrate was removed Most functional assays used MMG supplemented or not with a defined amount of a N source as mentioned in the results section in all comparative assays of mutant versus parental strain the medium was supplemented with the essential nutrients: uracil (1.12 g·L−1) uridine (1.22 g·L−1) riboflavin (2.5 mg·L−1) and pyridoxine (0.05 mg·L−1) EC50 levels of each organic compound were assessed on basis of hyphal radial growth rate for the strain A nidulans FGSC A4 grown in petri dishes (55 mm) containing the MMGN (jellified) and supplemented with either BaP (1.0–3.5 mM); BMQ (0.1–1.0 mM); Sal (75–400 mM); IDQ (0.002–1.0 mM); RVT (0.005–1.5 mM); or sodium benzoate (25–200 mM) Controls without the organic compounds were also made The assay was carried out by inoculating 2×105 conidia into the center of the plate and incubate at 30°C for 120 h Radial growth of mycelia (colony diameter in cm) was measured using a Vernier caliper (error ± 0.05 mm) The plates that did not displayed visible growth were visualized under microscope to confirm growth inhibition The EC50 values were calculated using the XLSTAT tool (Microsoft Excel) with the Gompertz model and 0.2 mM for TCS to match the range used for the other organic compounds For the gene expression analysis of AN9181 in the presence of H2O2 A1145 strain (1×106 spores·mL−1) was pre-grown during 140 h in 12-well plates (2 mL of MMG per well) at 30°C with gentle agitation (100 rpm) H2O2 was added to the cells at a defined concentration (1.0 –4.0 mM) and incubated at 30°C for 30 min Controls without addition of H2O2 were grown as well Mycelia samples were collected from the 12-well plates and immediately frozen using liquid nitrogen until further analysis Statistical analyzes used the XL-STAT (Addinsoft) software Differences in gene expression with a p-value below 0.05 were considered statistically significant Cell viability was evaluated using the XTT assay in 96-well plates (200 μL per well) using Malt Extract medium or MMG supplemented with 10 mM N source and an inoculum of 1×106 spores·mL−1 and incubated at 30°C for 24 h (triplicates) 10 μL of a solution containing 4.6 mg·mL−1 of XTT and 0.104 mg·mL−1 of menadione was added to each well and the absorbance measured (460 nm) The strains’ spore germination fitness number of spores that are able to form a colony was evaluated by spreading 100 spores onto solid media then counting the colony forming units (CFUs) daily The radial growth diameter of each strain (inoculum: 1 μL of a suspension of 2×108 spores·mL−1) in solid MMG supplemented or not with specific compounds (viz CSP and ITZ) was measured after five days of incubation (30°C The MIC of each antifungal was determined using the micro-broth dilution method (MMG testing specific concentration ranges for Amph B (250–550 mg·L−1) CSP (60–120 mg·L−1) and ITZ (0.2–0.5 mg·L−1) An inoculum of 1×106 spores·mL−1 was used and the plates were incubated at 30°C for 48 h The lowest concentration that showed no growth under microscopic observation was considered the MIC Intracellular ROS was quantified using DCFH-DA 30°C); then 2.5 μg·mL−1 of DCFH-DA was added and incubated (30°C H2O2 was added to the cells at increasing concentrations The cell suspension was disrupted in a TissueLyzer LT (Qiagen) with a metal bead at a maximum speed (3 cycles of 1 min) The fluorescence intensity of the supernatant (recovered by centrifugation: 12000 g 10 min) was measured using a Tecan Infinite M Nano+ Microplate (Männedorf Switzerland) as follows: excitation length 485/9; emission 528/20; optics The fluorescence intensity (per mycelia dry weight) was normalized against the control (no H2O2 added) as well as a comparable percentage of genes included in orthogroups and number of species-specific orthogroups This gene is part of an orthogroup that contains two genes in A Different expressed genes in the five transcriptomic datasets (A) – upon orthology transformation of the transcriptome-based datasets to the “receiver species” Aspergillus nidulans AN9181 was the only gene found consistently up-regulated in all the analyzed datasets BaP (Benzo[a]pyrene); BMQ (Bromoquinoline); Sal (Sodium Salicylate); IDQ (6-iodoquinoline) and RVT (Resveratrol) (B) – the AN9181 orthogroup comprises 25 genes present in the genomes of the eight orthology-analyzed Aspergillus species which are separated into two distinct clusters in the gene-tree obtained upon OrthoFinder analysis (C,D) – targeted gene expression analysis (assessed by RT-qPCR) is displayed for AN9181 (C) and AN8970 (D) This past evidence raises the hypothesis that AN9181 participates in stress responses the gene expression analysis did not support the idea of a concerted action of the two genes of the AN9181 orthogroup in A we focused the remaining analyzes on the AN9181 This result suggests that AN9181 strongly influences the fitness of conidia germinating in medium having low availability of poor nitrogen sources Phenotype features of A1145 and ΔAN9181 (A,B) – the morphology and pigmentation of A1145 strain (A) and ΔAN9181 strain (B) after cultivation at 30°C for five days in Malt Extract Agar (C–F) – colony forming units of A1145 (black) and ΔAN9181 (gray) strains plated on Malt Extract Agar (C); minimal medium glucose supplemented with 10 mM nitrogen from different sources (NH4)2SO4 (D); 10 mM NaNO3 (E); no nitrogen added (F) (G,H) – cell viability and proliferation measured by XTT assay A1145 (black) and ΔAN9181 (gray) strains were grown in Malt Extract or in minimal medium glucose supplemented with 10 mM nitrogen from the different nitrogen sources (G) including amino acids (H) Values and error bars represent the mean and the standard deviation of triplicates Significant differences (Student’s t-test) are marked with asterisks [*) * p ≤ 0.05; ** p ≤ 0.01 and *** p ≤ 0.001 Collectively the results suggest that AN9181 influences the utilization of N sources (hence also spore germination fitness) in a nitrogen type specific manner the mutant phenotype suggests that AN9181 participates in the regulation of nitrogen catabolism in A resembling NmrA negative regulation of N utilization in many rich N sources Further assays are however needed to better understand AN9181 regulatory network in the context of nitrogen utilization (A–C) relative growth of the A1145 (black) and ΔAN9181 (gray) strains grown in minimal medium glucose (solid) supplemented with the EC50 concentrations of each organic compound compared to the control condition (no chemical addition) 0.45 mM) (A); and with increasing concentrations of sodium salicylate (B) or sodium benzoate (C) *p ≤ 0.05; **p ≤ 0.01 and ***p ≤ 0.001 The absence of a phenotype in three (out of five) of the organic compounds herein tested (Figure 4A) is likely related to the fact that the used concentrations were below the threshold to cause major stress effect under the utilized cultivation conditions the response is also influenced by the regulation of nitrogen catabolism as analyzed above The observation that the growth phenotype of the ΔAN9181 strain is sodium salicylate and sodium benzoate concentration dependent obvious for concentrations above the corresponding EC50 values is consistent with the working hypothesis that AN9181 regulates stress responsive metabolism A significant decrease in the susceptibility of ΔAN9181 mutant strain compared to the parental strain was noticed The MIC for congo red in either strain is higher than 256 mg·L−1 regardless that the upper inhibitory limit could not be precisely determined due to the strong red color of the media All tests were conducted in the same growth media hence the observed differential responses to the antifungal compounds cannot be simply explained by the regulation of nitrogen utilization by AN9181 (A,B) relative growth of the A1145 (black) and ΔAN9181 (gray) strains grown in minimal medium glucose (solid) supplemented with Caspofungin (0.5 mg·L−1) Amphotericin B (150 mg·L−1) Itraconazole (0.01 mg·L−1) hydrogen peroxide (3.5 mM) and menadione sodium bisulfite (0.1 mM) compared to the control condition (no chemical addition) quantified for the A1145 (black) and ΔAN9181 (gray) strains in the presence of different concentrations of H2O2 The fluorescence intensity per biomass dry weight amount of the control was defined as 100 Values and error bars represent the mean and the standard deviation of at least five replicates (D) targeted gene expression analysis (assessed by RT-qPCR) is displayed for AN9181 Fold change relative to the control (without H2O2 addition) and normalized to AN3469 Collectively the results suggest that AN9181 participates indeed in stress responses upon exposure to different chemical agents leading to oxidative stress initially focusing very different biological questions were selected (five datasets in total) The computational strategy herein used allowed to pinpoint AN9181 as the only gene that showed the same differential upregulation in all five datasets The AN9181 orthogroup comprises only two genes in A These genes are located in different clusters of the AN9181 orthogroup gene tree and possess distinct functional domains Only AN9181 showed differential upregulation when the fungus was grown in media containing distinct chemical stressors including two that were used in the previous studies and two additional toxic aromatic compounds subsequent experimental analyzes focused on the gene AN9181 specifically by studying the phenotype of the generated single-deletion mutant compared to the parental strain This included measuring the viability of the conidia (i.e. the metabolic profile in different nitrogen sources the radial growth in the presence of different stress conditions and the minimal inhibitory concentration to different antifungal drugs The acquired data showed that the deletion of the gene AN9181 lead to higher metabolic activities in different N sources and a decreased susceptibility to sodium salicylate congo red and amphotericin B (a clinically relevant antifungal drug) These opening results support the hypothesis that AN9181 is involved in the regulation against different stress responses especially as it is not only conserved in aspergilli but also in several other fungi (e The identification of the AN9181 as a putative regulator of stress in aspergilli could not be anticipated regardless that some of the transcriptome-based datasets used here were generated years ago This strategy can help to identify (yet) cryptic genetic elements by reusing publicly available peer-reviewed transcriptome-based data from a diversity of scientific fields It remains unresolved the nature of its regulation but AN9181 apparently is able to slow down growth and metabolic activity in conditions that would be otherwise harmful for the fungus The absence of orthologs in the human genome (according to OrthoMCL) increases the significance of this inaugural observations especially in the context of invasive aspergillosis The data presented in the study are deposited in the NCBI repository The author(s) declare that financial support was received for the research This work was financially supported by Fundação para a Ciência e a Tecnologia (FCT) by Project MOSTMICRO ITQB with refs UIDB/04612/2020 and UIDP/04612/2020 LS4FUTURE Associated Laboratory (LA/P/0087/2020) and Project FATE with ref JJ is grateful for the working contract PTDC/CTA-AMB/6587/2020 CM and DH are grateful for the fellowships SFRH/BD/118377/2016 and SFRH/BPD/121354/2016 TM is grateful for the working contract financed by national funds under norma transitória D.L The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest The author(s) declared that they were an editorial board member of Frontiers This had no impact on the peer review process and the final decision All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher The Supplementary material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fmicb.2024.1373469/full#supplementary-material bio-transformation and bio-degradation in mycoendophytes General stress response or adaptation to rapid growth in aspergillus nidulans Crossref Full Text | Google Scholar Citric acid from aspergillus Niger: a comprehensive overview PubMed Abstract | Crossref Full Text | Google Scholar The quinoline bromoquinol exhibits broad-spectrum antifungal activity and induces oxidative stress and apoptosis in Aspergillus fumigatus Performance standards for antimicrobial susceptibility testing: Approved 28th (2018) Google Scholar Sources and transformations of chlorophenols in the natural environment PubMed Abstract | Crossref Full Text | Google Scholar OrthoFinder: solving fundamental biases in whole genome comparisons dramatically improves orthogroup inference accuracy STRIDE: species tree root inference from gene duplication events PubMed Abstract | Crossref Full Text | Google Scholar OrthoFinder: phylogenetic orthology inference for comparative genomics Crossref Full Text | Google Scholar Gabaldón Functional and evolutionary implications of gene orthology Crossref Full Text | Google Scholar Sequencing of aspergillus nidulans and comparative analysis with a Aspergillus fumigatus versus genus aspergillus: conservation adaptive evolution and specific virulence genes Lovastatin biosynthesis by aspergillus terreus in a chemically defined medium Functional analysis of the nitrogen metabolite repression regulator gene nmrA in aspergillus flavus Untapped potential: exploiting fungi in bioremediation of hazardous chemicals InteractiVenn: a web-based tool for the analysis of sets through Venn diagrams The role of the Aspergillus fumigatus areA gene in invasive pulmonary aspergillosis Crossref Full Text | Google Scholar Meiotic and mitotic recombination in aspergillus and its chromosomal aberrations PubMed Abstract | Google Scholar HISAT: a fast spliced aligner with low memory requirements Cytochrome P450 monooxygenase-mediated metabolic utilization of benzo[a]pyrene by aspergillus species Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2 Genomics of aspergillus oryzae: learning from the history of Koji mold and exploration of its future The old 3-oxoadipate pathway revisited: new insights in the catabolism of aromatics in the saprophytic fungus aspergillus nidulans Securing a furan-based biorefinery: disclosing the genetic basis of the degradation of hydroxymethylfurfural and its derivatives in the model fungus aspergillus nidulans Twists and turns in the salicylate catabolism of aspergillus terreus revealing new roles of the 3-Hydroxyanthranilate pathway Multiple degrees of separation in the central pathways of the catabolism of aromatic compounds in fungi belonging to the Dikarya sub-kingdom airborne fungal spores upon exposure of a soil Mycobiota to chlorinated aromatic hydrocarbon pollutants Specialisation events of fungal metacommunities exposed to a persistent organic pollutant are suggestive of augmented pathogenic potential Short-term variability and predictors of urinary pentachlorophenol levels in Ohio preschool children Núñez-Corcuera A new protein carrying an NmrA-like domain is required for cell differentiation and development in Dictyostelium discoideum Review of new and future developments in microbial biotechnology and bioengineering: aspergillus system properties and applications Crossref Full Text | Google Scholar Evolution of the human pathogenic lifestyle in fungi PubMed Abstract | Crossref Full Text | Google Scholar FungiDB: an integrated functional genomics database for fungi Variation among biosynthetic gene clusters and cards of virulence across aspergillus species Understanding fungal functional biodiversity during the mitigation of environmentally dispersed pentachlorophenol in cork oak forest soils A three-act play: pentachlorophenol threats to the cork oak forest soils mycobiome A convenient growth medium for Neurospora (medium N) Google Scholar Cap1p is involved in multiple pathways of oxidative stress response in Candida albicans Deep sequencing analysis of transcriptomes in aspergillus flavus in response to resveratrol World Health Organization WHO fungal priority pathogens list to guide research Google Scholar Identification of the Candida albicans Cap1p regulon Crossref Full Text | Google Scholar phenylcoumaran benzylic ether reductase-like domain (IPR045312) Goldman GH and Silva Pereira C (2024) NmrB (AN9181) expression is activated under oxidative stress conditions acting as a metabolic repressor of Aspergillus nidulans Received: 19 January 2024; Accepted: 29 March 2024; Published: 18 April 2024 Copyright © 2024 Jorge, Martins, Domingos, Martins, Hartmann, Goldman and Silva Pereira. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY) distribution or reproduction in other forums is permitted provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited in accordance with accepted academic practice distribution or reproduction is permitted which does not comply with these terms *Correspondence: Celso Martins, Y2Vsc28ubWFydGluc0B1bmlsLmNo; Cristina Silva Pereira, c3BlcmVpcmFAaXRxYi51bmwucHQ= ‡Present addresses: Celso Martins, Center for Integrative Genomics, Faculty of Biology and Medicine, University of Lausanne, Lausanne, SwitzerlandDiego O. Hartmann, Sea4Us – Biotecnologia e Recursos Marinhos S.A., Porto de Pesca da Baleeira, Armazém 8, Sagres, Portugal †These authors have contributed equally to this work Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher. 94% of researchers rate our articles as excellent or goodLearn more about the work of our research integrity team to safeguard the quality of each article we publish. In Brazil, the proverbial goose that lays the golden egg is in reality something closer to a pheasant that excretes coffee beans. At the Camocim coffee farm, deep in the bucolic hills of Espirito Santo state in Brazil’s southeast, jacus — a type of pheasant native to tropical forests there — are considered some of the most astute pickers — or rather, eaters — of coffee cherries. “He chooses the best fruits, the ripest,” said worker Agnael Costa, 23, delicately scooping up droppings left behind by one of the birds between two tree trunks. What goes in as ripe cherries comes out as beans, which can go on to be sold as some of the most delicious — and expensive — coffee in the world. The coffee at Camocim grows in the middle of the lush forest, and the jacus there are wild. “It was this agroforestry system that created the necessary conditions for this exotic coffee to exist here,” farm owner Henrique Sloper said. Domestically, jacu coffee can sell for 1,118 reals (US$224) per kilogram — a price that can rise significantly upon export. Foreign distributors include British department store Harrods, among others. The jacu, with its black feathers and red throat, was not always welcome at the farm. It was originally seen as a hungry pest feeding off precious crops. However, when Sloper discovered “kopi luwak,” an expensive Indonesian coffee made with beans from civet excrement, he figured jacus might make good allies in the quest to make a better brew. Kopi luwak sometimes has a bad reputation, with farmers accused of pressing civets — small mammals similar to weasels — into captivity to boost production. The wild jacus at Camocim roam, eat and poop at their leisure. “It’s 100 percent natural,” farm production supervisor Rogerio Lemke said. “The jacu is within its own natural habitat,” in Brazil’s Atlantic Forest, he added. Linking production to the whims of a jacu’s appetite is just one of the reasons it fetches such a high price. Jacu droppings look a bit like a cereal bar, with beige grains protruding from a blackish, pasty mess. Once collected and dried, the beans are carefully sorted and hulled, and put into a chilled room. “It is naturally expensive. There is no way to make a jacu coffee at a low cost,” Sloper said. All told, jacu coffee represents less than 2 percent of the estate’s production. The bird also carries other advantages, for farmers and those seeking a cheaper — if less intestinally intimate — cup of Joe. “It serves not only as a selector [of the best beans], but also as a harvest alarm” Sloper said. “Where it eats, the coffee is ripe.” While jacus help select the best coffee, there is nothing in particular about their digestive system that imparts a different taste on the beans. “The birds have an extremely short intestinal transit time,” coffee specialist Ensei Neto said. “So there is not exactly any kind of biochemical process, there is no time.” Their digestive process is much faster than civets or elephants — another animal whose dung is used, in Thailand, to produce specialty coffee. Some think the fermentation process stemming from those animals’ longer digestion times imparts a better flavor on the resulting beans. The jacu “only goes for the ripe fruits,” producing a coffee with “sweet notes, with good acidity,” Neto said. “It doesn’t add anything else, but the story is good.” The jacus have found a fan in Poliana Cristiana Prego, 37, a Brazilian tourist who has come to visit Camocim. “It’s a delicious coffee and the story behind its production is very original. It’s a new experience for us,” she said. “Our customers are lovers of exotic products, but also those who value the idea of sustainable development,” Sloper said. “The future of coffee will come from Brazil,” he said. The country is already the world’s top producer. Whether it comes from jacus or otherwise, Brazilian coffee “is showing the world that we really are able to do what nobody else can,” Sloper said. According to the country’s Civil Defence at least 10 people have died and over 17,500 have been displaced As reported on 02 December heavy rain caused a massive landslide along a highway near Guaratuba in Paranà state Emergency teams are still working at the site Twenty-one vehicles were buried in the slide and it was thought at least 30 people could be missing Six others were able to escape the vehicles without needing assistance Heavy rain has affected other areas of the state The number of displaced has fallen slightly over the last few days from 1,152 to 1,079 More heavy rain has fallen and the municipality of Inácio Martins recorded 88.6 mm in 24 hours to 04 December 2022 Heavy rain since late November continues to cause flooding and landslides in Santa Catarina including 10,000 in Massaranduba and 4,000 in Campo Alegre A total of 1,077 people were displaced including 688 in Joinville and 220 in São Bento do Sul The state government reported fatalities in Palhoça A firefighter is still missing Navegantes and another person was reported missing after being swept away by an overflowing river in Brusque Municipalities in a state of emergency are: Araquari Heavy rain has been falling for the last week in the state of Espírito Santo where Civil Defence reported 18 municipalities affected several of which have declared a state of emergency including Cariacica there were a total of 4,051 displaced in the state as a result of the severe weather One fatality was reported in the municipality of Viana where a total of 1,676 people were displaced Santa Teresa municipality recorded 258 mm of rain in 3 days from 01 to 03 December The Doce river at Linhares is above the orange alert level (3 metres) Authorities reported severe weather in the Norte Fluminense region of Rio de Janeiro State Affected municipalities included Carapebus One person died from a lightning strike in Carapebus another was seriously injured and around 85 people were displaced after a landslide in Conceição de Macabu where 2 people died and around 1,000 families have been displaced Macaé recorded 196.6 mm of rain in 48 hours to 01 December 2022 The national Civil Defence reported a total of 16,900 people affected across 5 municipalities in the state Heavy rain has also affected areas further north At least 16 municipalities in the northeastern state of Bahia have declared a state of emergency causing flooding in Prado City where 3,000 people were displaced on 28 November The heavy rain has continued to cause problems since then and as of 04 December a total of 51 municipalities have been affected 9,281 people were displaced and 65,515 were affected according to the state’s Civil Defence There were no reports of missing persons or deaths The worst affected municipalities and those that have declared a state of emergency are: Prado On 29 November the city of Ibotirama recorded 133.4 mm of rain and Barreiras recorded 104.4 mm The following day 75 mm of rain fell in Ribeira do Amparo Amargosa recorded 84.6 mm on 01 December; Porto Seguro 90.4 mm on 02 December and Lençóis 49.5 mm on 03 December Richard Davies is the founder of floodlist.com and reports on flooding news Cookies | Privacy | Contacts © Copyright 2025 FloodList CYFIRMA’s researchers have tracked Maze as early as Jun last year, and on 10 Jun, Maze group have released a long list of companies who have fallen victim to their cyberattacks. Affected companies span from the US and South America to the UAE. The modus operandi of Maze hackers would entail releasing threats of data dump on public forum if ransom is not paid. When such data is made available, other hackers and cybercriminals will have access to the sensitive data, and would continue their separate exploits, including reselling stolen data in dark web marketplaces. Maze operators have targeted a number of high-profile large enterprises, and government-linked corporations, across wide-ranging industries. Their recent breaches include Cognizant, a global IT services powerhouse with 290,000 employees and annual revenue turnover of almost US$17B in 2019, and ST Engineering, a Singapore government-linked company involved in defence and military projects. Here is the latest list of companies who have been compromised by Maze operators and shared in Maze website: Domingos Martins – Brazilian government website containing information related to Domingos Martins municipal. Breached data could include citizens’ information such contact details, taxes, e-government services rendered. Daily Thermetrics – A US firm providing process engineering industries with precise temperature measurement instrumentation. John Christner Trucking – A family trucking business based out of Oklahoma. FERSPED Inc. – A shipping and freight-forwarding company based in Macedonia. Mead O’Brien, Inc. – The company provides valves & valve automation, steam & hot water products and systems, instrumentation products, skid designs, field services, surveys, assessments, and consulting services, and is based in Kansas City, USA. United Enertech – A US construction, building and engineering company based in Tennessee Collabera – A HR recruitment, search and training company located in New Jersey, USA. Munoz Engineering P.C. – Provides engineering, land surveying and building and construction services, based in New York, USA. Ahmed Almazrouei Group – Engineering and Design Services, General Trading, Property Management, Facilities Management, Oil & Gas, Educational and Food retail business headquartered in Abu Dhabi, UAE. Omnix Int’l – A provider of business analytics, BPM, mobility solutions, information management, ERP implementation and IT networking services to public and private sectors, based in Dubai, UAE. Westmoreland Mechanical Testing and Research, Inc. – A material testing facility for the additive manufacturing, aerospace, automotive, composites, and medical and power generation industries, based in Pennsylvania, USA. CPFL Energia – A large energy company with businesses in distribution, generation, commercialization of electricity and services serving 9.6 million customers in Brazil. Here are extracts of the stolen data belonging to the above organizations which are now publicly available. Here’re the screens as captured from the Maze Group forum where the hackers announced their successful campaigns of locking down victims’ systems: Maze ransomware operators have a history of first stealing the data before locking their target devices and demanding ransom. They capitalize on the reputational consequences of their target as their strategy is “steal, lock and inform.” Suspected threat actors appear to be Russian-based APT28 (Confidence Level: Moderate) and TA2101/APT29 ((Confidence Level: Moderate). The IP addresses <Details Masked> mentioned in Cognizant attack vector was seen exploiting two old vulnerabilities CVE-2016-7255 and CVE-2018-8453. Both these Win32k Elevations of Privilege Vulnerabilities targeting Windows server was exploited by suspected Russian state sponsored threat actor APT28 in the past. The threat actor group APT28 leverages TTPs like obfuscated files or information, PowerShell, exploitation of remote services for lateral movement, credential stealing through spear-phishing links and data-staged techniques. These similar TTPs were seen in the Cognizant hack due to Maze ransomware. The mentioned IP addresses like <Details Masked> belong to Russian origin which gives us the glimpse that Russian threat actor groups (possibly APT28) are behind the attack. On similar lines, the threat actor group TA2101 had leveraged Maze Ransomware targeting German, Italian and U.S. organizations with malicious emails carrying samples of Maze ransomware in November 2019. TA2101 has been actively using Maze ransomware for attacks. Maze Ransomware usually deploy phishing emails with MS Office attachments and fake/phishing websites laced with Exploit Kits. Recipients should be cautious when handling emails from unknown sources, downloading attachment, or clicking on suspicious links as Maze operators utilizes social engineering tactics aggressively. To strengthen security controls, software and applications should contain the most updated patch. Avoid pirated and counterfeit software as they could be laced with malware. Maze tends to use known vulnerabilities like the Pulse VPN CVE-2019-11510 to break in and this means employees working from home must be mindful when accessing sensitive company information. More actions can be taken to protect your organization from Maze ransomware. Hong Leong Building, 16 Raffles Quay, Floor #09-01 & #10-01, Singapore 048581 Goodworks Co work, Plot no 72 and 73, 3rd Floor, Akshay Tech Park, EPIP Zone, Whitefield, Bangalore, Karnataka. Otemachi One Tower, 6th Floor, 1-2-1 Otemachi, Chiyoda-ku, Tokyo, 100-0004 Tokyo, Japan 501 Fifth Avenue Suite 805, New York, NY 10017 10F, 373 Gangnam-daero, Seocho-gu, Seoul, Korea 06621 Unit 20 270 Blackburn Road, Glen Waverley, VIC, 3150 9F, Second Building, No.96, Sec. 2, Zhongshan N. Rd., Taipei, Taiwan 14th Floor, HM Town building, 412 Nguyen Thi Minh Khai, Ward 5, District 3, Ho Chi Minh City Unit JLT-PH2-RET-5, Cluster R, Jumeirah Lakes Towers, Dubai, UAE Page unavailable.This page either does not exist or is currently unavailable From here you can either hit the "back" button on your browser to return to the previous page, or visit the ABCNews.com Home Page You can also search for something on our site below coordinator of the National Council of Evangelical Youth in Brazil shares his hopes for the LWF’s upcoming Assembly in Krakow which he will attend as youth delegate for his church (LWI) - A passionate supporter of the Evangelical Youth network in his native Brazil Natan Schumann will be a youth delegate from his church to the Lutheran World Federation (LWF) Thirteenth Assembly in Krakow Born in the town of Sapiranga in southern Brazil where his family has lived for several generations he was baptized into the Evangelical Church of the Lutheran Confession in Brazil (IECLB) were initially surprised by his interest and involvement in the life of the church sparked by his participation in confirmation classes as a teenager But gradually his enthusiasm and his commitment to the Evangelical Youth movement had a positive influence on the rest of his family who began to accompany him to worship services and other activities Even after he moved to Porto Alegre to pursue studies in international relations they continued to support his involvement and encouraged him to follow his passion As well as preparing for the Krakow Assembly as coordinator of the National Council of Evangelical Youth (CONAJE) is busy with plans for the next national congress which will take place next July in Domingos Martins He is excited about the way this biennial event enables youth from across the country to forge stronger ties and to strengthen their witness to their Christian faith in an increasingly secular society in Brazil Alongside your role with the church in Brazil you were also a member of the LWF delegation to the COP conference in Glasgow in 2021 - what motivates you most in your work at both national and global level What gives me most hope is to see young people welcomed and recognized for their work alongside people of other generations It encourages me when I see you youth assigned important and representative roles but because the community has faith in their gifts and their potential to serve God by assuming leadership roles I enjoy working together with people from all generations and I learn from the experiences they share I dream of a church where relationships between different generations are based on mutual respect and broader cooperation What are your expectations as you look ahead to the LWF 13th Assembly I am excited to represent the IECLB at this Assembly young people play a vital role in the daily building up of the church We want to convey the idea that young people are not only the future I hope that this Assembly will be a space to reflect and promote greater recognition of youth leadership in LWF decision-making Encouraging youth participation has been a priority for the LWF for several decades since the 1984 Assembly - what do you see as the current challenges we still face challenges in youth participation also within the Lutheran churches in our Latin America and Caribbean region More work is needed to increase youth participation and ensure that their voices are heard at all levels of the LWF it was agreed to present a proposal to formulate an intergenerational justice policy for the LWF This proposal aims to ensure that decision-making processes are inclusive and fair to all generations We hope that this proposal will be considered and that concrete measures will be put in place to move towards greater intergenerational justice in the LWF What impact do you think the Assembly can have in promoting stronger youth leadership The LWF has already succeeded in meeting the youth quota in terms of registered participants globally for the Krakow Assembly It is an important step in ensuring that youth voices are heard at this global event Intergenerational justice and youth leadership will be an important part of the discussions and we look forward to promoting greater youth engagement and participation in all levels of decision-making the LWF reaffirms its commitment to build a united body where all generations have a voice and feel represented in the global Lutheran community What does it mean for you and your church to be a part of the global communion of churches I would like to emphasize the importance of continuing to work together to promote inclusive leadership and stronger intergenerational justice We need to create spaces where generations meet The diversity of experiences and perspectives enriches the Church and helps us face today's challenges I hope that this Assembly will be a starting point for greater youth participation in the LWF and greater attention to intergenerational justice in our global Lutheran community The Lutheran World Federation is a global body that shares the work and love of Christ in the world we profile church leaders and staff as they discuss topical issues and set out ideas for building peace and justice in the world ensuring the churches and communion grow in witness and strength [email protected]+41 (0)22 791 60 00 This website is using a security service to protect itself from online attacks. The action you just performed triggered the security solution. There are several actions that could trigger this block including submitting a certain word or phrase, a SQL command or malformed data. You can email the site owner to let them know you were blocked. Please include what you were doing when this page came up and the Cloudflare Ray ID found at the bottom of this page. Methods: Various techniques have been recently developed to address this problem, including cost-sensitive strategies, methods that oversample the minority class, undersample the majority one, or combinations of both. Here, we investigate the most effective approaches for improving the performance of ML algorithms for known resonant asteroidal databases. Results: Cost-sensitive methods either improved or had not affect the outcome of ML methods and should always be used, when possible. The methods that showed the best performance for the studied databases were SMOTE oversampling plus Tomek undersampling, SMOTE oversampling, and Random oversampling and undersampling. Discussion: Testing these methods first could save significant time and efforts for future studies with imbalanced asteroidal databases. Volume 10 - 2023 | https://doi.org/10.3389/fspas.2023.1196223 Introduction: Machine learning (ML) applications for studying asteroid resonant dynamics are a relatively new field of study Results from several different approaches are currently available for asteroids interacting with the z2 one challenge when using ML to the databases produced by these studies is that there is often a severe imbalance ratio between the number of asteroids in librating orbits and the rest of the asteroidal population This imbalance ratio can be as high as 1:270 which can impact the performance of classical ML algorithms that were not designed for such severe imbalances Methods: Various techniques have been recently developed to address this problem methods that oversample the minority class we investigate the most effective approaches for improving the performance of ML algorithms for known resonant asteroidal databases Results: Cost-sensitive methods either improved or had not affect the outcome of ML methods and should always be used The methods that showed the best performance for the studied databases were SMOTE oversampling plus Tomek undersampling Discussion: Testing these methods first could save significant time and efforts for future studies with imbalanced asteroidal databases have been used to study the population of asteroids interacting with the ν6 secular resonance Because the class distribution is often unbalanced most ML algorithms will underperform and will need to be modified to prevent predicting the majority class in all circumstances measures like classification accuracy lose relevance and new methods for evaluating predictions on unbalanced data and in Section 2 of the Supplementary Materials As is often the case in machine learning, there are many approaches to imbalanced classification, and it is often challenging to know a priori which method is best suited for a given problem. Here, we tested 19 different methods available in the imblearn library developed by Lemaître et al. (2017) on five databases of labeled asteroids interacting with four resonances Our main goal was to investigate the use of imbalanced classification methods in resonant asteroid dynamics and to identify what methods one should try first to improve the performance of standard ML models when applied to imbalanced datasets for problems in asteroid dynamics As far as we know, this is the first application of imbalanced classification in this area. While some of the techniques studied in this work have been applied in other astronomical areas, like solar flares forecasting (Ribeiro and Gradvohl, 2021) there is simply no precedent application in our field Here we also aim to provide a basic framework We will start our analysis by reviewing the currently available in the literature The methods used to study a population of asteroids interacting with a resonance vary but a preliminary classification of resonant asteroids is often performed by analyzing plots of the resonant arguments of asteroids interacting with the resonances The resonant argument of a resonance is a combination of the asteroid and planet angles associated with the given resonance in the case of the ν6 secular resonance FIGURE 1. Time behavior of the resonant angle ϖ − ϖ6 for an asteroid in a circulating orbit (top left panel), in a switching orbit (top right panel) in an antialigned orbit (bottom left panel) and in an aligned orbit (bottom right panel). Adapted from the Figure 1 of Carruba et al. (2022b) Datasets with proper elements and labels for these four resonances are currently available in repositories listed in the cited articles and at a GitHub repository prepared for this work with a 0 label for asteroids in circulating orbits Here we will focus our attention on the use of imbalance learning to predict the labels of asteroids in the librating classes which are the ones that are of most interest for asteroid dynamical studies we converted the available databases into dual ones for which 0 identifies orbits that are not in the librating class of interest and 1 is associated with the relevant librating class such as the one for the aligned configuration of the ν6 secular resonance there is a severe imbalance between the two classes and one could expect to encounter the problems and limitations of conventional machine learning methods We report the number of asteroids in class 0 and the imbalance ratios between class 0 and class 1 for the asteroid databases studied in this work AA ν6 stands for the antialigned configuration of this resonance with oscillations of the resonant argument around the 180° equilibrium point with oscillations around the 0° equilibrium point An (a, sin(i)) projection of the asteroids in the z2 and aligned ν6 databases After setting up the databases that we are going to study in this work with the procedure discussed in the previous section we aim to identify the optimal machine learning (ML) method to fit each database we will use the area under curve score (AUC) of the receiver operating characteristic (ROC) curve The ROC curve is a is a graphical representation of the performance of a binary classification model It plots the True Positive Rate (TPR) on the y-axis and the False Positive Rate (FPR) on the x-axis where the true positives are data correctly identified by the model to belong to the desired class while false negatives are data miss-classified as belonging to the class of interest Other used names for TPR are sensitivity or recall where false positives are outcomes where the model incorrectly predicts the positive class and true negatives are data in the negative class correctly identified by the model The plot displays the percentage of correct predictions for the positive class (y-axis) versus the percentage of errors for the negative class (x-axis) The best possible classifier would have a TPR of 1 and a FPR of 0 By analyzing the true positives and false positives for different threshold levels a curve that runs from the bottom left to the top right and bows toward the top left can be formed A classifier that is unable to distinguish between positive and negative classes will draw a diagonal line from a FPR of 0 to a TPR of 1 (coordinate (0,1)) or forecast all negative classes Models indicated by points below this line have worse than no skill The performance of a model can be quantitatively estimated using the ROC area under curve (ROC AUC) with 1.0 being the value for a perfect classifier For the ROC AUC score we are using the default cut-off point in probability between the positive and negative classes at 0.5 As an example of an ROC curve we display in Figure 3 the case of predictions from a no-skill classifier, which is unable to distinguish between positive and negative classes, and a Logistic Regression model Cox (1958), used to predict classes for a binary problem with equal weights for the two classes. Interested readers can find more details on how to produce this example in chapter 7 of Brownlee (2020) The logistic regression model had an ROC AUC of 0.90 while the no-skill model had a score of 0.50 FIGURE 3. ROC Curve of a Logistic Regression Model and a no-skill classifier. Adapted from figure 7.1 of Brownlee (2020), available from Jason Brownlee, Imbalanced Classification with Python, https://machinelearningmastery.com/imbalanced-classification-with-python/ while in Section 1 of the supplementary materials we report the full set of hyper-parameters for each model We use the ROC AUC score to assess the validity of each model Since the methods are stochastic in nature we applied each model ten times to provide an estimate of the ROC AUC score’s value and of its error defined as the mean and the standard deviation of the ten outcomes Errors were generally very small and of the order of 10–3 The optimal machine learning model for the five datasets studied in this work (second column The third column shows the Imbalance ratio of the database The fourth column displays the values of the ROC AUC score for the best-performing methods we displayed in the fifth column values of the ROC AUC score after the methods were optimized by assigning higher weights to the imbalanced classes Models that use the ExtraTrees classifier can be further optimized by assigning proper weights to each class This method is called a cost-sensitive approach We briefly mentioned the confusion matrix when we introduced the concept of ROC AUC score we can distinguish between the actual negatives and positives The confusion matrix has the structure shown below: since there are much fewer members of class 1 than class 0 the cost of predicting a False Negative is much higher than that of predicting a False Positive Failing to detect a real member of class 1 may produce a sample that is even more imbalanced than the original class predicting a False Positive has a smaller impact This imbalance can be corrected by assigning different weights to False Negatives and False Positives in the case of a 1:100 ratio of examples in the minority class we can define a cost matrix where the cost of a False Positive event is 1 while that of a False Negative is 100 as shown below: Tree-based algorithms can automatically assign different weights to different classes based on the heuristic approach by selecting the option class_weight = “balanced.” We further optimized the tree-based algorithms for the z2, M1:2, and aligned ν6 databases, and our results are displayed in the fifth and sixth columns of Table 2 We define Δ(score) as the difference between the ROC AUC scores of the models with and without class weights Its error is computed using standard error propagation formulas We can see that Δ(score) depends on the imbalance ratio It is negligible for the balanced z2 dataset and it is maximum for the more imbalanced aligned ν6 database Methods for correcting class imbalance depend on how they handle the minority and majority classes Methods that increase the population of the minority class are classified as oversampling Approaches that decrease the majority class are called undersampling both oversampling and undersampling can be used at the same time SMOTE works by picking instances in the feature space that are close together and drawing a new sample at a position along that line initially a random case from the minority class is picked and then k of its nearest neighbors are determined (usually k = 5) A randomly determined neighbor is selected and a new example is constructed in the feature space at a randomly chosen position between the two instances one of which is a method that selects pairs of samples that have the minimal Euclidean distance in feature space to each other In a binary classification problem with classes 0 and 1 this means that a pair would have an example from each class and be closest neighbors throughout the dataset Examples of applications of both SMOTE and Tomek to the ν6 antialigned database are shown in Figure 4 The first method oversamples the minority class so that it has the same number of objects as the majority one while Tomek undersamples the majority class to reduce the imbalance Left panel: application of SMOTE to the ν6 antialigned database: Right panel: application of Tomek links to the same data and in Section 2 of the Supplementary materials Imbalanced classification algorithms can provide an increase in performance of up to ≃ 1% Results of imbalanced models for the ν6 aligned dual database (1: 268 imbalance) Dependence of the Δ(score) versus the Imbalance ratio for the three most successful models: a combination of Random oversampling and undersampling and SMOTE oversampling plus Tomek undersampling we applied imbalanced classification methods to study five available datasets for asteroids interacting with four different resonances we transformed the datasets from multi-labels to dual classes with a label 1 assigned to asteroids in librating configurations to allow the application of standard methods of imbalanced classification we used genetic algorithms to identify which machine learning methods and what combination of their hyper-parameters a cost-sensitive approach was also used for further optimization using the optimal models identified with the genetic algorithms Our findings show that cost-sensitive optimization methods should always be considered when feasable since they either enhance the model’s performance or have no effect Imbalanced classification methods are recommended for datasets with severe imbalance Although each database is unique and should be studied independently to determine the most effective imbalance method the experience gained in this study suggests that testing SMOTE oversampling plus Tomek undersampling could be a good strategy for dealing with imbalanced datasets for asteroids’ resonant problems Testing these methods first could save considerable time and efforts for future studies in this area Any additional data not found in these sources can be obtained from the first author upon reasonable request All authors contributed to the study conception and design The first draft of the manuscript was written by VC and all authors commented on later versions of the manuscript All authors read and approved the final manuscript All authors contributed to the article and approved the submitted version We thank the Brazilian National Research Council (CNPq grants 304168/2021-17 and 153683/2018-0) for supporting VC and BM research ML is thankful for the support from the São Paulo Science Foundation (FAPESP GC acknowledge support by the Coordination of Superior Level Staff Improvement (CAPES) – Finance Code 001 We are grateful to two reviewers for comments and suggestions that greatly improved the quality of this work. This is a publication from the MASB (Machine learning Applied to Small Bodies, https://valeriocarruba.github.io/Site-MASB/) research group. Questions regarding this paper can also be sent to the group email address: bWxhc2IyMDIxQGdtYWlsLmNvbQ== The evolution of intelligence: The nervous system as a model of its environment Google Scholar Imbalanced classification with Python: Choose better metrics Google Scholar Optimization of artificial neural networks models applied to the identification of images of asteroids’ resonant arguments CrossRef Full Text | Google Scholar Artificial neural network classification of asteroids in the M1:2 mean-motion resonance with Mars CrossRef Full Text | Google Scholar Identifying the population of stable ν6 resonant asteroids using large data bases CrossRef Full Text | Google Scholar Identification of asteroid groups in the z1 and z2 nonlinear secular resonances through genetic algorithms CrossRef Full Text | Google Scholar Smote: Synthetic minority over-sampling technique CrossRef Full Text | Google Scholar “Model selection of SVMS using GA approach,” in 2004 IEEE International Joint Conference on Neural Networks (IEEE Cat Google Scholar The regression analysis of binary sequences CrossRef Full Text | Google Scholar “A fuzzy citation-knn algorithm for multiple instance learning,” in 2015 IEEE International Conference on Fuzzy Systems (FUZZ-IEEE) CrossRef Full Text | Google Scholar The condensed nearest neighbor rule (corresp) CrossRef Full Text | Google Scholar Outline for a logical theory of adaptive systems CrossRef Full Text | Google Scholar Knežević Proper element catalogs and asteroid families CrossRef Full Text | Google Scholar Lemaître Imbalanced-learn: A python toolbox to tackle the curse of imbalanced datasets in machine learning Google Scholar Lourenço Genetic optimization of asteroid families’ membership CrossRef Full Text | Google Scholar Google Scholar Machine learning techniques applied to solar flares forecasting CrossRef Full Text | Google Scholar Two modifications of cnn in IEEE transactions on systems Google Scholar Martins BS and Alves AA (2023) Imbalanced classification applied to asteroid resonant dynamics Received: 29 March 2023; Accepted: 26 April 2023;Published: 11 May 2023 Copyright © 2023 Carruba, Aljbaae, Caritá, Lourenço, Martins and Alves. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use *Correspondence: V. Carruba, dmFsZXJpby5jYXJydWJhQHVuZXNwLmJy Volume 13 - 2022 | https://doi.org/10.3389/fmicb.2022.946286 Fungal secondary metabolites constitute a rich source of yet undiscovered bioactive compounds Their production is often silent under standard laboratory conditions but the production of some compounds can be triggered simply by altering the cultivation conditions The usage of an organic salt – ionic liquid – as growth medium supplement can greatly impact the biosynthesis of secondary metabolites leading to higher diversity of compounds accumulating extracellularly specifically cholinium-based ionic liquids can support the discovery of bioactive secondary metabolites across three model species: Neurospora crassa Enriched organic extracts obtained from medium supernatant revealed high diversity in metabolites The supplementation led apparently to increased levels of either 1-aminocyclopropane-1-carboxylate or α-aminoisobutyric acid The extracts where bioactive against two major foodborne bacterial strains: Staphylococcus aureus and Escherichia coli crassa cultures showed greater bactericidal potential compared to control extracts derived from non-supplemented cultures An untargeted mass spectrometry analysis using the Global Natural Product Social Molecular Networking tool enabled to capture the chemical diversity driven by the ionic liquid stimuli fumigatus; whereas an unexpected richness of cyclic (depsi)peptides with N Further studies are required to understand if the identified peptides are the major players of the bioactivity of N and to decode their biosynthesis pathways as well This study examines if ionic liquids supplements can support discovery of bioactive secondary metabolites in fungi Three model fungi – Neurospora crassa and two choline-based ionic liquids – choline chloride (ChoCl) and choline decanoate (ChoDec) we focused on compounds accumulating extracellularly The antibacterial activity of the ensuing crude extracts was evaluated against two major foodborne bacterial strains Staphylococcus aureus and Escherichia coli To characterize the chemical landscape of the extracts their amino acid composition and an untargeted mass spectrometry analysis using the online platform Global Natural Product Social Molecular Networking – GNPS – were applied An unexpected richness of peptide-based structures could be putatively associated with N Compounds used in preparation of minimal media were purchased from Sigma-Aldrich except for NaCl and MgSO4⋅7H2O (Panreac) phosphoric acid (Fisher Scientific) and NaNO3 (ACROS organics) The standard chemicals [1-aminocyclopropane-1-carboxylate (ACC) and α-aminoisobutyric acid (Aib)] and chromatographic solvents were of highest analytical grade and purchased from Sigma Aldrich and Fisher Scientific Water was obtained from a Milli-Q system (Millipore) Choline Chloride (>98%; ChoCl) was purchased from Sigma Aldrich and Choline Decanoate (>95%; ChoDec) from Iolitec crassa (FGSC 2489) were obtained from the Fungal Genetics Stock Center All strains were cultivated on DG18 (Oxoid) agar plates Asexual spores (conidia) were harvested using a NaCl (0.85% w/v) and Tween-20 (0.1% w/v) sterile solution and collected after passing through three layers of miracloth The harvested spores were washed twice with a sterile NaCl solution (0.85% w/v) and finally resuspended in the NaCl solution (0.85% w/v) or in a cryoprotective saline solution containing 10% (v/v) glycerol to be stored at −20°C or −80°C nidulans were cultivated in liquid minimal medium containing glucose (10.0 g⋅L–1) 5% (v/v) nitrate salts solution [NaNO3 (120.0 g⋅L–1) MgSO4⋅7H2O (10.4 g⋅L–1) and KH2PO4 (30.4 g⋅L–1)] and 0.1% (v/v) trace elements solution [ZnSO4⋅7H2O (22.0 g⋅L–1) MnCl2⋅4H2O (5.0 g⋅L–1) FeSO4⋅7H2O (5.0 g⋅L–1) CoCl2⋅6H2O (1.7 g⋅L–1) CuSO4⋅5H2O (1.6 g⋅L–1) Na2MoO4⋅2H2O (1.5 g⋅L–1) and Na4EDTA (50.0 g⋅L–1)] The pH was adjusted to 6.5 with NaOH and the medium sterilized in an autoclave (15 min; 110°C) Neurospora crassa was cultivated in liquid minimal medium containing K2PO4 (1 g⋅L–1) and glucose (10 g⋅L–1) dissolved in distilled water The pH was adjusted to 7 with 10% phosphoric acid and the medium sterilized in an autoclave (10 min; 110°C) Filter sterilized salts solution [1% (v/v) HCl 37% (10 μL) and FeSO4.7H2O (100 mg)] was added after autoclaving The minimal inhibitory concentrations (MICs) were determined as described previously (Petkovic et al., 2010) Final concentrations of ionic liquids in growth media ranged from 100 μM up to maximum solubility Each liquid medium (1 mL) was inoculated with 106 spores and divided into four wells (0.2 mL each) of a 96 well microtiter plate Fungal growth (or lack thereof) was determined at the end of incubation gauging by eye the formation of mycelium (turbidity) The lowest concentration that inhibited the formation of mycelium was defined as the MIC Values should not be interpreted as absolute ones but as an indication of the inhibitory and the fungicidal upper concentration limits Fungal cultures (100 mL) were initiated from 106 spores per mL in the respective minimal medium Liquid cultures were incubated in the dark at 30°C (N fumigatus) with orbital agitation of 200 rpm the ionic liquid supplement was added at 50% (i.e. Negative conditions (without ionic liquid supplement) were prepared in parallel Cultures were grown for 10 more days under agitation (100 rpm) the media supernatants were separated from mycelia using vacuum assisted filtration with miracloth (Merck Millipore Calbiochem) Neurospora crassa filtrates required the use of protease inhibitors (cOmplete Protease Inhibitor Cocktail Waters) as preliminary tests showed degradation of untreated extracts (data not shown) The mycelia and filtrates were frozen immediately in liquid nitrogen and lyophilized Lyophilized filtrates were homogenized in 10 mL Milli-Q water, extracted three times with ethyl acetate (1:1) and the combined ethyl acetate fractions dried under soft nitrogen flow. Peptide enrichment was achieved using the Sep-Pak plus C18 cartridge (Waters) as previously described (Krause et al., 2006) The samples were re-dissolved in 10 mL of MeOH/H2O (1/2 v/v) and loaded with a syringe into a conditioned cartridge The cartridge was washed with 10 mL of Milli-Q water and 10 mL MeOH/H2O (1/2 The retained compounds were eluted with 10 mL of MeOH to a pre-weighed glass tube and dried under soft nitrogen flow; crude extracts Conditioning of the cartridge was done successively with 10 mL of MeOH were chromatographically separated using a Waters Acquity chromatographer with Photodiode Array detector A Symmetry C18 column (250 × 4.6 mm) packed with end-capped particles (5 μm pore size 100 Å) (Waters Corporation) Data were acquired using Empower 2 software Samples were injected using a 10 μL loop operated in full loop mode at a flow rate of 0.9 mL⋅min–1 consisted of a solution of 0.1% trifluoracetic acid in water (v/v) (TFA set to a linear gradient of 99.5 to 0% of solvent A during 30 min and additional 10 min to re-equilibrate the column The chromatographic profiles of the samples were obtained at the wavelength of 205 nm Sample fractionation was performed with a Fraction collector III (Waters) connected to the Acquity chromatographer (Waters) using the same conditions described above The collected fractions were dried under nitrogen flow and kept at 4°C (short term) or −20°C (long term) until further analysis and the standard mixture of amino acid hydrolyzates were derivatized following the manufacturer’s instructions The obtained derivatives were separated on an AccQTag Ultra column (100 mm × 2.1 mm 1.7 μm) by reversed phase ultra-performance liquid chromatography (UPLC) and the mobile phase flow rate was maintained at 0.7 mL⋅min–1 Eluent A was 5% AccQTag Ultra concentrate solvent A and eluent B was 100% AccQTag Ultra solvent B The separation gradient was 0–0.54 min (99.9% A) 8.05–8.64 min (10.0% A) and 8.73–10.50 min (99.9% A) Two microliters (2 μL) of sample were injected for analysis using a 10 μL loop The FLR detector was set at 266 and 473 nm of excitation and emission wavelengths Calibration curves of each standard were used to quantify amino acids the values are represented as the relative % of total amount of amino acids The total area of peaks was used to determine the overall % of identification The extracts were assessed for their antimicrobial activity against gram-positive bacteria Staphylococcus aureus NCTC8325 and gram-negative bacteria Escherichia coli TOP 10, following the standard methodology implemented by the Clinical and Laboratory Standards Institute (Clinical and Laboratory Standards Institute [CLSI], 2018) bacteria were grown until approximately 1 to 2 × 108 CFU⋅mL–1 in Mueller Hinton Broth (MHB two-fold serial dilutions were performed to obtain final extracts concentrations between 1,000 and 62.5 μg⋅mL–1 Plates were incubated at 37°C for 24 h in a Bioscreen C analyzer (Oy Growth Curves Ab Ltd) taking hourly absorbance measurements (600 nm) All tests were done in triplicate; abiotic (medium alone) and biotic controls (each bacterium without extract) were included for each replicate 100 μL of each sample were mixed with 10 μL of 5 mg⋅mL–1 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) (Sigma Aldrich) in PBS (96-well microtiter plates) and incubated (dark 100 μL 10% SDS in 0.01 M HCl were added to each well and plates incubated for 2 h in the dark at room temperature Absorbance was measured at 560 and 700 nm using Tecan Infinite 200 Microplate (Männedorf values at 560 nm were subtracted from the values at 700 nm A second aliquot of 50 μL was used to label the cells with propidium iodide (20 μM PI Biotium) and SYTO9 (3 μM; Alfagene) and further incubated for 15 min at room temperature with agitation Fluorescence intensity was measured with a FLUOstar OPTIMA Microplate Reader (BMG⋅Labtech) using a 488/20 nm excitation filter (for both SYTO9 and PI) and a 528/20 nm (SYTO9 emission wavelength) and 645/40 nm (PI emission wavelength) emission filter The signal from the staining solution (SYTO9/PI) was subtracted from all data to minimize cross-signal background Microscopy assessment of the live/dead staining was done on a Leica DM 6000B upright microscope equipped with an Andor iXon 885 EMCCD camera and controlled with the MetaMorph V5.8 software using the 100 × 1.4 NA oil immersion objective plus a 1.6× optovar the fluorescence filter sets FTIC + TX2 and Contrast Phase optics Images were analyzed by FIJI software (Fiji Is Just ImageJ) IC50 (half maximal inhibitory concentration of a compound) values were calculated from dose response curves constructed by plotting cell viability (MTT data) versus extract concentration (μg⋅mL–1) using the Logit regression model (dose effect analysis tool of XLSTAT) samples were separated at a flow rate of 300 nL⋅min–1 using a linear gradient of 0.05% (v/v) TFA in water (Solvent A) and 0.05% (v/v) TFA in ACN/MeOH (1:1 Gradient program for solvent B was 65% for 5 min then 100% until 75 min and last 65% from 76 to 81 min MS data was acquired in positive mode over a mass range 300–1,250 m/z (for N The 30 most intense ions were selected to perform fragmentation with high sensitivity mode using the automatically adjusted system of rolling collision energy MS/MS scans were acquired over a mass range 100–1500 m/z with an accumulation time set at 50 ms; raw data files Input data (MS/MS spectra in .mgf format) were uploaded and search parameters were set as follows: peptide tolerance of 0.02 Da and fragment mass tolerance of 0.01 Da with M + H ionization with a charge up to 2 Decoy was activated; generates p-values associated with the identifications Hits were validated (p-value < 0.05) upon further inspection of the number of scored peaks vs Data were analyzed using standard statistical software (Origin v8.5 Software Results are expressed as mean value ± standard deviation The statistical significance of values between conditions was evaluated by One-Way ANOVA test Differences were considered significant when the p-value ≤0.05 Minimal inhibitory concentrations of the cholinium-based ionic liquids (choline chloride ChoDec) used as media supplements for each fungal strain Ionic liquid supplements triggered a metabolic shift in the fungal cultures Chromatographic analyses of the metabolic footprint of A Crude extracts are from cultures grown for 10 days in either choline chloride (ChoCl) or choline decanoate (ChoDec) supplemented media and from cultures without ionic liquid supplementation (i.e. Truncated parts of the chromatogram from N crassa cultures (C) correspond to the elution of protease inhibitors The y-axis scale represents the base peak intensity fumigatus control extracts show low levels of Aib with a slight increase when the culture is supplemented with ChoDec (at 80% of MIC) an increasing trend in either NPAAs upon ChoDec supplementation was observed but the overall amounts of Aib and ACC are substantially lower compared to the other two fungi Total amino acid hydrolysis discloses the presence of two non-proteinogenic residues in N Scatter plot depicting individual values of percentage (%) of total amount of 1-aminocyclopropane-1-carboxylate ACC (●) and α-aminoisobutyric acid Aib (■) obtained from total hydrolysis of the crude extracts derived from cultures grown in media with or without (i.e. The y-axis scales are not normalized to allow easier visualization of the amount of either non-proteinogenic amino acid in each condition Ionic liquid-exposure altered the pattern of the overall amino acid content, suggestive of an altered peptidome profile (Supplementary Table 1) no meaningful alterations were found (pair-wise ANOVA) in the detected amounts of each amino acid with or without media supplementation possibly consequence of high variability between the biological replicates crassa 45–65% of the peaks could not be assigned to any of the amino acid standards the values were lower: 4–7% (negative and ChoCl supplemented extracts) and 27% (ChoDec supplemented extracts) Despite these inherent technical fragilities this analysis provides an estimation of the amino acid profiles of each sample and excitingly point to the existence of peptides containing ACC and/or Aib in either crude extract from ionic liquid supplemented cultures fumigatus extracts were selected for subsequent analyses focusing antibacterial efficacy and compositional signature (LC-MS/MS) fumigatus crude extracts depict antibacterial activity against S Cell viability measured by MTT assay of extracts derived from cultures grown in media with or without (i.e. Quantification of bacterial viability through the normalized green/red ratio (i.e. Statistically significant differences are depicted; *p ≤ 0.05 ****p ≤ 0.0001 fumigatus crude extracts led to significant lysis of S aureus grown in the absence of extract (A) and in the presence of crude extracts derived from cultures grown in media with or without (i.e. Images of bacteria at concentrations near the measured IC50 for each crude extract are shown Cells were stained with SYTO9 (green) and PI (red) denoting live and dead cells crassa crude extracts from media supplemented or not (negative control) with choline chloride (ChoCl) or choline decanoate (ChoDec) the two most intense peaks in the total ion chromatogram are depicted crassa extracts derived from ionic liquid supplemented cultures suggests the accumulation among other metabolites of macrolides and peptides The results show the production of antimicrobial compounds in N crassa cultures under ionic liquid supplementation likely associated to production of metabolites otherwise cryptic The hypothesis that these antimicrobial compounds support N crassa competitiveness in specific niches deserves further consideration the supplementation did not increase the antibacterial activity of A the chemical landscape of either extract was further analyzed using an untargeted MS metabolomics approach therefore all compounds identification discussed below remain putative these observations further support the hypothesis that growth medium supplementation with ChoCl triggered production of peptaibiotics in N the fractions were analyzed by NMR but their chemical complexity and low abundance of each constituent of the mixture hindered stringent spectral assignments (data not shown) irrespectively of their detection in the hydrolyzates of the crude extracts/fractions This is likely due to the lack of similar structures in the GNPS and NRPro databases it reveals that the chemical space of either extract remains to be fully disclosed specifically the GNPS tool exposed the most promising candidates – cyclic (depsi)peptides of N setting foundations for their isolation and identification in the near future The data attained highlight the capacity of N crassa to generate a rich portfolio of cyclic peptide-based metabolites only four putative NRPS genes have been assigned has been linked to the produced metabolite to date Preliminary tests suggest that three of these genes suffered upregulation in the supplemented medium compared to control (data not shown) ionic-liquid supplementation shows matchless potential to link each NRPS to its peptide-product(s) The datasets presented in this study can be found in online repositories The names of the repository/repositories and accession number(s) can be found below: EBI – MTBLS5072 CSP supervised the interpretation of data and prepared the final version of the manuscript All authors have made substantial contributions to the acquisition and contributed to the drafting of the manuscript This work was supported by the FCT – Fundação para a Ciência e a Tecnologia through MOSTMICRO-ITQB R&D Unit (UIDB/04612/2020) and LS4FUTURE Associated Laboratory (LA/P/0087/2020) as well as the projects PTDC/CTA-AMB/6587/2020 and AAC 01/SAICT/2016 (CERMAX This work was also partially supported by the Portuguese Platform of BioImaging (PPBI) (PPBI-POCI-01-0145-FEDER-022122) co-funded by national funds from Orçamento de Estado (OE) and by European funds from Fundo Europeu de Desenvolvimento Regional (FEDER) and IM are grateful to FCT for the fellowship PD/BD/113989/2015 PD/BD/135481/2018 and for the working contract financed by national funds under Norma Transitória D.L We acknowledge the use of microscope at the Bacterial Imaging Cluster (ITQB-NOVA) We are thankful to Pedro Lamosa and Maria C Leitão (ITQB NOVA) for support with the NMR and chromatographic analyses we acknowledge Paula Alves (ITQB alumni) for collection of preliminary data that inspired this study The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fmicb.2022.946286/full#supplementary-material Transcriptomic and metabolomic profiling of ionic liquid stimuli unveils enhanced secondary metabolism in Aspergillus nidulans Sensitive and rapid method for amino acid quantitation in malaria biological samples using AccQ Tag ultra performance liquid chromatography-electrospray ionization-MS/MS with multiple reaction monitoring Reproducible molecular networking of untargeted mass spectrometry data using GNPS “Trends in antimicrobial resistance in healthcare-associated infections: a global concern,” in Encyclopedia of Infection and Immunity VelB/VeA/LaeA complex coordinates light signal with fungal development and secondary metabolism secondary metabolism and light-dependent carotenoid biosynthesis by the velvet complex of Neurospora crassa Google Scholar Current strategies to induce secondary metabolites from microbial biosynthetic cryptic gene clusters CrossRef Full Text | Google Scholar Genomics-driven discovery of PKS-NRPS hybrid metabolites from Aspergillus nidulans Fungal natural products galaxy: biochemistry and molecular genetics toward blockbuster drugs discovery Biologically Active Secondary Metabolites from the Fungi Big effects from small changes: possible ways to explore nature’s chemical diversity doi: 10.1002/1439-7633(20020703)3:7<619::AID-CBIC619>3.0.CO;2-9 Designing Small Molecules for Biodegradability PubMed Abstract | CrossRef Full Text | Google Scholar Brückner peptaibiotics Containing the rare 1-aminocyclopropanecarboxylic acid Complexity generation in fungal polyketide biosynthesis: a spirocycle-forming P450 in the concise pathway to the antifungal drug griseofulvin Identification and characterization of the echinocandin b biosynthetic gene cluster from Emericella rugulosa NRRL 11440 Relationship between secondary metabolism and fungal development In vitro inhibition of 1,3-β-Glucan synthase by glycolipids from convolvulaceous species Clinical and Laboratory Standards Institute [CLSI] (2018) Performance Standards for Antimicrobial Susceptibility Testing PA: Clinical and Laboratory Standards Institute Google Scholar Peptaibiomics: towards a myriad of bioactive peptides containing C(alpha)-dialkylamino acids The occurrence of peptaibols and structurally related peptaibiotics in fungi and their mass spectrometric identification via diagnostic fragment ions A new pregnane steroid from cultures of Aspergillus versicolor Impact of non-proteinogenic amino acids in the discovery and development of peptide therapeutics The Rare Amino Acid Building Block 3-(3-furyl)-Alanine in the Formation of Non-ribosomal Peptides CrossRef Full Text | Google Scholar a key regulator of bacteria-triggered production of fungal secondary metabolites Secondary metabolism: regulation and role in fungal biology Structure determination of neoefrapeptins A to N: peptides with insecticidal activity produced by the fungus Geotrichum candidum a Novel Cyclic Peptide from the Toxic Cyanobacterium Nostoc sp Chemical Modifications Designed to Improve Peptide Stability: Incorporation of Non-Natural Amino Acids Gil-De-la-fuente Aspergillus Metabolome Database for Mass Spectrometry Metabolomics Marine natural product peptides with therapeutic potential: chemistry Temperature during conidiation affects stress tolerance and trypacidin accumulation in the conidia of the airborne pathogen Aspergillus fumigatus Identification and predictions regarding the biosynthesis pathway of polyene macrolides produced by Streptomyces roseoflavus Men-myco-93-63 Ionic liquids chemical stress triggers sphingoid base accumulation in aspergillus nidulans Plasma membrane permeabilisation by ionic liquids: a matter of charge Combination of isotope labeling and molecular networking of tandem mass spectrometry data to reveal 69 unknown metabolites produced by Penicillium nordicum The biosynthetic logic of polyketide diversity PubMed Abstract | CrossRef Full Text | Google Scholar Google Scholar PubMed Abstract | CrossRef Full Text | Google Scholar Fungal Secondary Metabolism - From Biochemistry to Genomics ProteoWizard: open source software for rapid proteomics tools development SMURF: genomic mapping of fungal secondary metabolite clusters Efomycins K and L from a termite-associated Streptomyces sp M56 and their putative biosynthetic origin rapid and selective analysis of peptaibiotics/peptaibols by SPE/LC-ES-MS Fungal cell wall: emerging antifungals and drug resistance The fumagillin biosynthetic gene cluster in Aspergillus fumigatus encodes a cryptic terpene cyclase involved in the formation of β-trans-bergamotene Recent advances in silent gene cluster activation in Streptomyces Regulation and Role of Fungal Secondary Metabolites Epigenetic genome mining of an endophytic fungus leads to the pleiotropic biosynthesis of natural products New 19-Residue Peptaibols from Trichoderma Clade Viride Proteomic alterations induced by ionic liquids in Aspergillus nidulans and Neurospora crassa Activation of Microbial Silent Gene Clusters: Genomics Driven Drug Discovery Approaches CrossRef Full Text | Google Scholar Refining and expanding nonribosomal peptide synthetase function and mechanism Current challenges of research on filamentous fungi in relation to human welfare and a sustainable bio-economy: a white paper Dereplication of microbial metabolites through database search of mass spectra Ways of assembling complex natural products on modular nonribosomal peptide synthetases doi: 10.1002/1439-7633(20020603)3:6<490::AID-CBIC490<3.0.CO;2-N The discovery of fungal polyene macrolides via a postgenomic approach reveals a polyketide macrocyclization by trans-acting thioesterase in fungi New Potent Serine Protease Inhibitors from the Marine Sponge of the Genus Theonella Microbial communication leading to the activation of silent fungal secondary metabolite gene clusters Natural Products as Sources of New Drugs over the Nearly Four Decades from 01/1981 to 09/2019 Diversity of linear non-ribosomal peptide in biocontrol fungi PubMed Abstract | CrossRef Full Text | Google Scholar Production of prostaglandins and leukotrienes by pathogenic fungi Determination of 1-aminocycopropane-1-carboxylic acid (ACC) to assess the effects of ACC deaminase-containing bacteria on roots of canola seedlings Novel biocompatible cholinium-based ionic liquids—toxicity and biodegradability Exploring fungal activity in the presence of ionic liquids Isolation of aerucyclamides C and D and structure revision of microcyclamide 7806A: Heterocyclic ribosomal peptides from Microcystis aeruginosa PCC 7806 and their antiparasite evaluation Secondary metabolites in fungus-plant interactions The effects of improving low dietary protein utilization on the proteome of lamb tissues Automatic Annotation and Dereplication of Tandem Mass Spectra of Peptidic Natural Products “Secondary Metabolism and Antimicrobial Metabolites of Aspergillus,” in New and Future Developments in Microbial Biotechnology and Bioengineering: Aspergillus System Properties and Applications a new polyene macrolide antifungal antibiotic Cytoscape: a software environment for integrated models of biomolecular interaction networks Cyclodepsipeptides: A Rich Source of Biologically Active Compounds for Drug Research In vitro antifungal and fungicidal activities and erythrocyte toxicities of cyclic lipodepsinonapeptides produced by Pseudomonas syringae pv Therapeutic design of peptide modulators of protein-protein interactions in membranes cytotoxic macrolides isolated from the sea hare Dolabella auricularia Proposed minimum reporting standards for chemical analysis Chemical Analysis Working Group (CAWG) Metabolomics Standards Initiative (MSI) and Other Cytotoxic Metabolites from the Filamentous Fungus Trichothecium sp Chemical classification of cyclic depsipeptides How histone deacetylase inhibitors alter the secondary metabolites of Botryosphaeria mamane an endophytic fungus isolated from Bixa orellana Oxylipins as developmental and host–fungal communication signals Sharing and community curation of mass spectrometry data with Global Natural Products Social Molecular Networking Structural Diversity and Biological Activities of Cyclic Depsipeptides from Fungi Mass spectral molecular networking of living microbial colonies World Health Organization [WHO] (2021) Global Antimicrobial Resistance and Use Surveillance System (GLASS) Report: 2021 Google Scholar Identification of elaiophylin derivatives from the marine-derived actinomycete Streptomyces sp Cholesterol import by Aspergillus fumigatus and its influence on antifungal potency of sterol biosynthesis inhibitors A new antiviral pregnane from a gorgonian-derived Cladosporium sp Goldman GH and Silva Pereira C (2022) Untargeted Metabolomics Sheds Light on the Secondary Metabolism of Fungi Triggered by Choline-Based Ionic Liquids Copyright © 2022 Sequeira, Rothkegel, Domingos, Martins, Leclercq, Renaut, Goldman and Silva Pereira. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY) *Correspondence: Cristina Silva Pereira, c3BlcmVpcmFAaXRxYi51bmwucHQ= †These authors have contributed equally to this work and share first authorship Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher 94% of researchers rate our articles as excellent or goodLearn more about the work of our research integrity team to safeguard the quality of each article we publish Get quality reporting directly into your inbox As soon as the officers from Brazil’s environmental investigation agency spotted well-known bow maker Marco Antonio Raposo Nascimento preparing to board a flight to Europe out of São Paulo’s international airport in April X-rays showed Raposo’s luggage was stuffed with 114 horsehair bows and another 120 wooden rods that could be used to make more Tests showed all of them were made of brazilwood an endangered hardwood that is considered a symbol of Brazil in an image he posted to his Instagram page and fined 105,300 Brazilian reais ($20,172) according to an arrest report obtained by reporters Two men who were with him fled before they could be apprehended one of them abandoning another suitcase full of wood in his hurry to get away Raposo is among a select group of Brazilian artisans who make bows for stringed musical instruments like violins and cellos out of brazilwood bows crafted from the slow-growing hardwood have made strings sing for centuries in concert halls and opera theaters around the world Sticks of brazilwood cost less than $8 in Brazil but once crafted into bows they can sell for more than 300 times as much in the U.S. Raposo’s bows are sold by music stores in the U.S But as supplies of brazilwood have dwindled many of Brazil’s bow makers — known as archetiers — have turned to illegally harvested wood is now one of the most threatened biomes in the world At present only the trade in raw brazilwood is regulated under the Convention on International Trade in Endangered Species (CITES) though rules governing the export of newly made bows are being drawn up who has been fined at least eight times during his career and stopped at the São Paulo airport carrying suspect wood at least twice Raposo conceded he did not have the proper documentation for the wood he was carrying when he was stopped at the airport in May but he claimed the “error was not intentional” and that the wood was of legal origin He said all sanctions against his company had been withdrawn or are still pending and that he had never been found to be dealing in wood from protected areas he has paid two of the fines and is appealing the rest “I have never been part of any type of criminal scheme,” Raposo wrote in an email Reporters tracked down the identities of Raposo and the 32 other bowmakers targeted in Operation Dó-Ré-Mi — who have never been publicly named before — by cross-referencing court cases and official data obtained through freedom of information requests They include several other prominent Brazilian archetiers who IBAMA believes have been trafficking brazilwood overseas “It is evident they have formed a large gang specialized in extracting brazilwood from protected areas and trading it abroad,” IBAMA officials wrote in multiple reports related to Operation Dó-Ré-Mi calling for the urgent involvement of the Public Prosecutor's Office and the federal police to “deepen investigations into the destructive actions of these criminals.” officers from the Federal Police and IBAMA raided 37 properties connected to what they described as a “criminal group” of bowmakers as part of the operation The police declined to confirm if these included any of the people and companies identified by reporters because their investigation is ongoing As law enforcement has tried to crack down on trafficking of brazilwood authorities have also sought to tighten regulations on how it’s legally traded signatories to the CITES treaty voted to extend restrictions to include brazilwood bows and instruments meaning Brazil’s archetiers will need to obtain permits to export newly made bows for sale overseas Plans to regulate the bow trade have faced fierce opposition from the classical music industry the music director of the London Symphony Orchestra and pianist Martha Argerich arguing the restrictions would spell the end of the bow making profession and destroy 300 years of culture But ecologists say the sale of brazilwood bows must be banned if the species is to be saved there will be someone who facilitates this sale,” said André Freitas a professor at Unicamp's Institute of Biology “The price you pay is the extinction of a species Over the course of a two-decade career, Raposo has become a public face of the Brazilian bow industry. According to a report quoting him in Brazilian media he started selling bows made by a friend in Europe in 2004 while working as a flight attendant Raposo set up a civil society organization dedicated to preserving brazilwood Raposo’s lawyers said his organization works with local government and research institutes to encourage reforestation Raposo said he no longer sits on the organization’s board but noted that it has planted more than 400,000 brazilwood seedlings in the state of Espirito Santo and about 130,000 more in Bahia since it was founded Raposo has also become a vocal advocate for commercial brazilwood cultivation In 2009 he appeared as an interviewee in “A Árvore da Música,” or “The Music Tree,” a documentary film about saving brazilwood “For us to continue this beautiful work [manufacturing bows] we have to have a new conscience,” he said Musicians and other Brazilian bow makers were shocked to hear about the investigations into Raposo when contacted by reporters "He is quite famous," said one archetier from São Paulo who asked not to be named as he did not want to be associated with any controversial dealings saying they met over a discussion about brazilwood conservation Raposo admitted to police that he had attempted to smuggle the brazilwood out of the country but said he had resorted to transporting it illegally only after the permits he applied for were delayed Raposo was released after a custody hearing, but investigations into him continue. In November, a regional tribunal agreed to allow the Federal Public Prosecutor's Office to tap his phones as part of their investigation This wasn’t Raposo’s first brush with the law IBAMA officers had stopped him the previous year on the way to Switzerland with a suitcase filled with over 200 bows though they could not immediately confirm they were made of brazilwood public attorney Guilherme Rocha Gopfert noted that Raposo made a "large number of international trips,” which "seem to be aimed precisely at the sale of illegally exported products.” Raposo denied he was part of any criminal scheme All sanctions against his company “have either been withdrawn or are still pending,” he said in an email attaching several IBAMA certificates clearing his company to trade where they can retail for up to $2,500 each though there is no indication that any of the sellers are aware these could be made from illicit wood OCCRP was shown five bows made by his company some of which sell for more than 1,400 British pounds The shop’s owner said he had known Raposo for more than 25 years describing his bows as “excellent.” He said he was always “keenly aware of deforestation” being an issue so had discussed it with Raposo and was glad to hear of his planting initiatives which is why it's quite distressing to hear your information about Marco now,” said the owner who OCCRP is not naming as there is no indication he knew of the allegations against Raposo Raposo’s company was also fined after IBAMA investigators discovered irregularities in its paperwork during an inspection of its office said the company had failed to log all of the wood it held and had entered “partially misleading” information in its records and shouted at the inspectors that they were “mangy dogs,” one of them wrote in a report on the visit "I have never received such rude treatment with such offensive words … during the hundreds of inspections that I have participated in,” it said adding that Raposo "sees himself as more qualified than anyone in or outside the government Raposo said he respects “each and every employee of IBAMA and/or any other government body Raposo’s company was fined another 1.8 million Brazilian reais ($356,000) for storing wood without a valid license in June 2022 two months after he was arrested trying to sneak the brazilwood through the airport (He still has an opportunity to appeal this fine.) "It is more profitable to sell our wood than to sell cocaine It is an absurd profit and a much lower risk,” said Eugenio Ricas the superintendent of the Federal Police of Espírito Santo the epicenter of Brazil’s bow manufacturing industry “Each bow costs between 20 and 40 reais ($8) [to make] and then it [is sold] in Europe or the U.S you are arrested and there is no social acceptance You are seen as a drug dealer and will spend five years behind bars those who extract wood are socially accepted People don't see them as dangerous criminals.” IBAMA believes that most of the brazilwood confiscated during Operation Dó-Ré-Mi originated in a protected area in the south of Bahia state A local man and his daughter admitted to investigators that they had been acting as “mules” or middlemen to smuggle it out of the region The wood was then sent to bowmakers in Espírito Santo "IBAMA's perception is that a large part of the market for brazilwood bows is based on wood extracted in a criminal way from conservation units or forests in private areas,” said a document from an inspection by the agency of one of the 33 companies identified by reporters Documents from IBAMA show the 33 companies and people have faced charges ranging from buying wood without the proper paperwork to trying to conceal its origins but the agency has never publicly named them before Almost 150,000 bows and sticks of brazilwood have been confiscated and 77 fines worth nearly 54.7 million Brazilian reais ($10.5 million) have been handed out since Operation Dó-Ré-Mi began IBAMA frequently refers to the bowmakers as a “gang” in its reports though it does not explain how they are connected Six of them are members of the Association of Craftsmen and String Instrument Companies United for the Preservation of the Brazilian Atlantic Forest including its president and five directors But reporters could find no proof that the 33 bowmakers have all been working together to smuggle wood The association said several of its members had “been the target of abusive and excessive tax actions” since 2018 describing IBAMA’s operation targeting the bow making industry as “disproportionate.” “The sector continues to be treated as an environmental villain as if we entered the forests and left with trucks loaded with dozens of brazilwood logs,” it said in a statement sent to reporters Among those targeted were master Brazilian archetiers Elias Guasti and José Bottoni — better known internationally as Guasti & Bottoni — who in 2012 were brought in to restore a bow once used by Brazil’s most famous composer IBAMA officers believe Guasti and Bottoni have been smuggling brazilwood that was illegally harvested in protected areas for years they have had more than 1,600 bows and other pieces of wood seized during four raids on their homes Guasti was caught by IBAMA officers trying to courier a package out of Brazil that contained wood restricted for sale abroad where they found the wood he had in storage was full of "cracks knots and signs of rot" and was kept in an area riddled with vermin The poor state of the supplies led officers to suspect the bow makers were using other stocks of illegal wood to make their “excellent quality” bows A fine notice issued for Guasti called for investigators to “find out if these craftsmen receive or received material from suspicious origins with the intermediaries of this gang that trafficks Pau-brasil from the south of Bahia to Aracruz and region.” Federal Police launched an investigation into the case they said the probe was still open but declined to provide further details saying he had received the wood he tried to smuggle via courier as a “donation” from IBAMA after it had been seized from another bowmaker and didn’t realize this meant he was only allowed to use it for non-commercial purposes Guasti said he had simply forgotten to update the paperwork for some of his wood when new rules came in but said he generally had the correct permits and operated within the law saying they had sent 10 armed officers to raid his house which had frightened his elderly mother and children “I think that in my case it was an exaggeration on the part of IBAMA,” he said “I believe I have not committed an infraction.” Most of the bows made in Brazil are sold overseas Reporters found the bows produced by the 33 bowmakers retail for as much as $4,000 in Europe and the U.S. a price range that two professional string players from the U.K said meant they were targeted at professional musicians For example, an IBAMA report on bow maker JB Atelier — which has been targeted at least five times and fined more than 3 million Brazilian reais ($575,000) during Operation Dó-Ré-Mi — found all of its bows were exported to Miami from 2019-2021. From there they went to a local reseller owned by the same family as JB Atelier, Trade Sousa Bows said the company had bought the wood from another company with the proper documentation He called IBAMA’s actions “disproportionate,” saying he operated within the law and the accusations had harmed his business “Being the target of a Federal Police operation is something that generates great discomfort and tarnishes the name and image of the company is already suffering serious consequences,” he wrote in an email There is no concrete data on how much brazilwood is left in Brazil The Atlantic Forest on Brazil’s eastern coast the only place in the world where the wood grows today has only 12.4 percent of its original cover and is considered one of the most threatened biomes in the world "We are already reaching a situation where there is no more Pau-Brazil on the northeast coast,” said Freitas Musicians argue their industry is not to blame for the deforestation saying bow makers have worked to ensure the survival of brazilwood the president of Brazil’s National Association of the Music Industry said that violin bow sellers have planted more than 550,000 seedlings A brazilwood tree in the Rio de Janeiro Botanical Garden In a piece published in French newspaper Le Monde more than 200 musicians railed against CITES’ plans to bring in trade restrictions on brazilwood instruments and bows saying they use only “modest” amounts of wood compared to other industries “Let's not hinder the dissemination of culture in general and music in particular under false pretenses Let's not make the musical world a scapegoat for deforestation,” said the commentary backed by many of the industry’s top names Violinist Emilie Belaud, from the Paris Opera, described the changes as “a direct attack on our main working tool, the bow, an object of great international cultural significance for more than 300 years,” in an online petition that has garnered close to 20,000 signatures. After a heated debate over expanding the CITES rules, the original plans to require musicians to get permits to transport their bows were scrapped. Under the new rules, which will come into force in Brazil in late February 2023, bow makers will only need to get CITES export permits for freshly crafted bows when they are exported for the first time. Anafima, a music industry association that opposes restrictions on brazilwood, recently hosted an online discussion about the new law and the future of bow-making. In a video from the event posted online, one bowmaker acknowledged the industry’s complicity in the illegal wood trade, and urged his fellows to stop buying brazilwood from illicit sources. “We who are in this field of making bows know that there are two businesses: one business is selling bows and the other is selling wood. Let’s be honest, selling natural pau-brasil, selling sticks,” he said. “In the eyes of IBAMA, we are all dealers, gangsters, whether by act or design. In one way or another, we all buy illegal wood in a sense. … We know who cuts the wood, who sells it, and [we must] be tough with this group, because then … IBAMA will stop treating us as gangsters." But for some, the new rules still don’t go far enough. Professor Henrique Dias, from the Department of Forest and Wood Sciences at the Federal University of Espírito Santo, said brazilwood should stop being used to make bows altogether. Otherwise, he said, the entire ecosystem it is part of could break down. Protecting "brazilwood is not just [about] the brazilwood. There are microorganisms that sequester nitrogen, trees that store carbon, flowers that promote pollination, insects and birds that disperse seeds. The very existence of the species sustains other species,” he said. "From the moment you extinguish the link, you are also extinguishing everything that relates to it. If we extinguish a species that gave the country its name, what do we want in terms of conservation?" 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UNESCO10 October 2023Last update:11 October 2023Part of UNESCO's History and Peace Education initiative in the country the event brought together 19  teachers from different municipalities and history educators Various challenges for history teaching were identified  such as the lack of teaching materials insufficient training opportunities in history education and a shortage of ICT equipment for educational purposes particularly for accessing audio-visual materials As Timor-Leste celebrates its 21st year of independence the challenges of  history education have come to the forefront Many history teachers lack specialized training which limits both knowledge and pedagogical skills Our curriculum fails to adequately introduce Timorese history and we face a shortage of teaching materials and guidance We need a  curriculum dedicated to history studies History is now integrated into 'social science studies' for the first two cycles of basic education and 'history and geography studies' for the third cycle of basic education and secondary education  the history curriculum doesn’t cover Timor-Leste's entire history.  Most available references and efforts should be made to facilitate more translations The workshop enhanced teachers’ capacity to provide high-quality history education through diverse pedagogical approaches from the Chega National Centre It emphasized the importance of  archival sources Teachers experienced and discussed historical films underscoring the benefits of learning history through audio-visual materials resonating with emotions.  Participants  concurred that history isn’t a mere recollection of past events but an interpretation and re-evaluation of the past through the lens of the present is crucial to ensure that we do not repeat  conflicts and other atrocities that occurred in Timor-Leste and around the world 200 teachers from 13 municipalities participated in  the workshop series since July 2022.  and the CNC will organize workshops at the municipal level The upcoming events will focus on teachers' proposed activities to reshape history education in Timor-Leste  for peace.  The dates displayed for an article provide information on when various publication milestones were reached at the journal that has published the article activities on preceding journals at which the article was previously under consideration are not shown (for instance submission All content on this site: Copyright © 2025 Elsevier B.V.