The Response of Paracoccidioides lutzii to the Interaction with Human Neutrophils.

The fungal pathogen Paracoccidioides lutzii causes systemic mycosis Paracoccidioidomycosis (PCM), which presents a broad distribution in Latin America. Upon infection, the fungus undergoes a morphological transition to yeast cells and provokes an inflammatory granulomatous reaction with a high number of neutrophils in the lungs. In this work, we employed proteomic analysis to investigate the in vitro response of the fungus to the interaction with human neutrophils. Proteomic profiling of P. lutzii yeast cells harvested at 2 and 4 h post interaction with human polymorphonuclear cells allowed the identification of 505 proteins differentially accumulated. The data indicated that P. lutzii yeast cells underwent a shift in metabolism from glycolysis to Beta oxidation, increasing enzymes of the glyoxylate cycle and upregulating enzymes related to the detoxification of oxidative and heat shock stress. To our knowledge, this is the first study employing proteomic analysis in the investigation of the response of a member of the Paracoccidioides genus to the interaction with neutrophils.

Cheminformatics-driven discovery of hit compounds against Paracoccidioides spp.

Aims: The development of safe and effective therapies for treating paracoccidioidomycosis using computational strategies were employed to discover anti-Paracoccidioides compounds. Materials & methods: We 1) collected, curated and integrated the largest library of compounds tested against Paracoccidioides spp.; 2) employed a similarity search to virtually screen the ChemBridge database and select nine compounds for experimental evaluation; 3) performed an experimental evaluation to determine the minimum inhibitory concentration and minimum fungicidal concentration as well as cytotoxicity; and 4) employed computational tools to identify potential targets for the most active compounds. Seven compounds presented activity against Paracoccidioides spp. Conclusion: These compounds are new hits with a predicted mechanisms of action, making them potentially attractive to develop new compounds.

Proteomic identification of metabolic changes in Paracoccidioides brasiliensis induced by a nitroheteroarylchalcone.

Aim: To access the metabolic changes caused by a chalcone derivative (LabMol-75) through a proteomic approach. Methods: Proteomic analysis was performed after 9 h of Paracoccidioides brasiliensis yeast (Pb18) cell incubation with the LabMol-75 at MIC. The proteomic findings were validated through in vitro and in silico assays. Results: Exposure to the compound led to the downregulation of proteins associated with glycolysis and gluconeogenesis, ß-oxidation, the citrate cycle and the electron transport chain. Conclusion: LabMol-75 caused an energetic imbalance in the fungus metabolism and deep oxidative stress. Additionally, the in silico molecular docking approach pointed to this molecule as a putative competitive inhibitor of DHPS.

Exposure of Paracoccidioides brasiliensis to Mebendazole Leads to Inhibition of Fungal Energy Production.

Paracoccidioidomycosis (PCM) is a fungal disease caused by organisms of the genus Paracoccidioides spp. The treatment of the disease is lengthy and includes several adverse effects. Various methodologies focus on the search for new treatments against fungal disease, including the repositioning of drugs. Our group showed the fungicidal effect of mebendazole in P. brasiliensis cells. Thus, understanding the effect of exposing fungal cells to mebendazole is significant for further studies in order to demonstrate it as a potential drug for the treatment of PCM. A proteomic analysis of P. brasiliensis exposed to mebendazole was carried out. Analyses showed that exposure strongly affected the pathways related to energy production, such as glycolysis, fermentation, and the electron transport chain. The quantification of adenosine triphosphate (ATP) and mitochondrial activity demonstrated that the drug alters the electron chain, resulting in an increase in oxidative stress. Enzymes such as superoxide dismutase (SOD) and cytochrome c oxidase (Cyt C) were repressed in cells exposed to mebendazole. The concentration of ethanol produced by the cells under treatment demonstrated that the attempt to produce energy through fermentation is also arrested. Thus, the drug inhibits fungal growth through changes in energy metabolism, making it a promising compound for use in the treatment of PCM.

Molecular Interactions of the Copper Chaperone Atx1 of Paracoccidioides brasiliensis with Fungal Proteins Suggest a Crosstalk between Iron and Copper Homeostasis.

Paracoccidioides spp. are endemic fungi from Latin America that cause Paracoccidioidomycosis, a systemic disease. These fungi present systems for high-affinity metal uptake, storage, and mobilization, which counteract host nutritional immunity and mitigate the toxic effects of metals. Regarding Cu mobilization, the metallochaperone Atx1 is regulated according to Cu bioavailability in Paracoccidioides spp., contributing to metal homeostasis. However, additional information in the literature on PbAtx1 is scarce. Therefore, in the present work, we aimed to study the PbAtx1 protein-protein interaction networks. Heterologous expressed PbAtx1 was used in a pull-down assay with Paracoccidioides brasiliensis cytoplasmic extract. Nineteen proteins that interacted with PbAtx1 were identified by HPLC-MSE. Among them, a relevant finding was a Cytochrome b5 (PbCyb5), regulated by Fe bioavailability in Aspergillus fumigatus and highly secreted by P. brasiliensis in Fe deprivation. We validated the interaction between PbAtx1-PbCyb5 through molecular modeling and far-Western analyses. It is known that there is a relationship between Fe homeostasis and Cu homeostasis in organisms. In this sense, would PbAtx1-PbCyb5 interaction be a new metal-sensor system? Would it be supported by the presence/absence of metals? We intend to answer those questions in future works to contribute to the understanding of the strategies employed by Paracoccidioides spp. to overcome host defenses.

3-phenacylideneoxindoles as a new class of antifungal compounds against Paracoccidioides spp.

Aims: Considering the need to identify new compounds with antifungal action, the activity of five 3-phenacylideneoxindoles compounds was evaluated. Materials & methods: The compounds were synthesized, and their antifungal activity was elucidated through minimum inhibitory concentration tests and interaction assay with other antifungals. Potential targets of compounds were predicted in silico. Results: 3-phenacylideneoxindoles compounds inhibited fungal growth with minimum inhibitory concentration and minimum fungicidal concentration ranging from 3.05 to 12.26 µM. The compounds demonstrated high selectivity index and presented a synergistic effect with itraconazole. In silico prediction revealed the pentafunctional AROM polypeptide, enolase, superoxide dismutase, catalase and kinases as proteins targets of the compound 4a. Conclusion: The results demonstrate that 3-phenacylideneoxindoles is a potential new class of antifungal compounds for paracoccidioidomycosis treatment.

Patients affected by paracoccidioidomycosis (PCM) require long-term treatment, which commonly influences their adherence. In addition, only three drugs are in clinical use, which indicates the relevance of research in identifying new drugs for treating PCM. Thus, five drugs were tested in the laboratory to verify whether they could prevent the growth of the fungus without being toxic to humans. In addition, whether these compounds in combination with drugs used to treat PCM could be even more potent was evaluated. All compounds tested efficiently inhibited the growth of Paracoccidioides, the fungus that causes PCM. One drug was identified that, combined with itraconazole, decreased the required dose of both the discovered compound and itraconazole needed to inhibit fungal growth. Using computational tools, this work suggests how the new drug could act against the fungus. The results demonstrate a potential new treatment option, but more studies are needed to confirm the safety of these drugs.

Midias sociais para atração de pacientes em clínicas odontológicas privadas durante pandemia de Covid 19 / Social media for attracting patients in private dental clinics during the Covid 19 pandemic

Verificar a rotina dos cirurgiões-dentistas (CD) e a utilização das mídias sociais, como meio de divulgação profissional e de atração de pacientes em clínicas odontológicas privadas, durante a pandemia de Covid-19. Este estudo transversal qualitativo inclui questionários respondidos por CD atuantes em clínicas privadas na região sudeste do Brasil. Um questionário virtual, elaborado através da plataforma Google Forms, abordou informações sobre o perfil dos profissionais e os aspectos dos métodos publicitários e mídias sociais utilizados para o alcance de pacientes durante o período de pandemia de Covid-19. Os dados obtidos foram tabulados e submetidos à análise estatística descritiva (%). No total, 102 CD participaram do estudo, sendo que 96 questionários seguiram os critérios de inclusão para a análise dos dados. A divulgação social como meio de exposição profissional foi uma ação realizada por 75 (78,13%) CD da amostra. Considerando esses profissionais, 74 (98,67%) utilizam redes sociais e domínios virtuais para tal finalidade, sendo que 71 (95,95%) CD usufruem do Instagram. Grande parte dos participantes (71,62%) relatou não possuir assessoria de marketing especializada para fazer publicações de conteúdo profissional nas redes sociais, embora a maioria publique este tipo de conteúdo mais de uma vez por semana (58,11%). Uma grande parcela dos participantes do estudo (67,71%) notou que a pandemia de Covid-19 procovou diminuição na quantidade de pacientes nas clínicas odontológicas. A inclusão de outros equipamentos de proteção individual foi a conduta mais seguida pelos CD (56,25%) para evitar a transmissão da doença neste período. Conclui-se que os CD participantes acreditam que a pandemia de Covid-19 promoveu um impacto negativo na atração de pacientes em clínicas odontológicas privadas na região sudeste brasileira, ainda que a maioria destes profissionais tenham incluído outros equipamentos de proteção individual como medida de segurança e utilizem frequentemente redes sociais para divulgação de conteúdo profissional, tendo o Instagram como a principal mídia social. No entanto, a assessoria de marketing especializada em publicações de conteúdo profissional nas redes sociais ainda é um recurso pouco utilizado no meio odontológico(AU)

To verify the routine of Dental Surgeons (DS) and the use of social media as a means of professional dissemination and patient attraction in private dental clinics during the Covid-19 pandemic. This cross-sectional qualitative study included questionnaires answered by Dental Surgeons working in private clinics in the southeastern region of Brazil. A virtual questionnaire, which was developed through the Google Forms platform, addressed information about the profile of professionals and aspects of advertising methods and social media used to reach patients during the Covid-19 pandemic period. The data were tabulated and submitted to descriptive statistical analysis (%). A total of 102 Dental Surgeons participated in the study, with 96 questionnaires meeting the inclusion criteria for data analysis. Social disclosure as a means of professional exposure was an action performed by 75 (78.13%) Dental Surgeons of the survey. Considering these professionals, 74 (98.67%) use social networks and virtual domains for this purpose and 71 (95.95%) Dental Surgeons use Instagram. A large portion of the participants (71.62%) reported not having a specialized marketing consultancy to make professional contente publications on social media, although most publish this type of content more than once a week (58.11%). A large portion of study participants (67.71%) noted that the Covid-19 pandemic caused a decrease in the amount of patients in the dental clinics. The inclusion of other personal protective equipment was the conduct most followed by the Dental Surgeons (56.25%) to avoid the transmission of the disease in this period. It can be concluded that the participating Dental Surgeons believe that the Covid-19 pandemic promoted a negative impact on attracting patients in private dental clinics in the southeastern region of Brazil, in spite of most of these professionals have included other personal protective equipment as safety measure and frequently use social networks for dissemination of professional content, with Instagram as main social media. However, a marketing consultancy specialized in publishing professional content on social networks is still a resource which is little used in the dental environment(AU)

Proteomic alterations in Paracoccidioides brasiliensis caused by exposure to curcumin.

Paracoccidioides spp. are the etiological agent of paracoccidioidomycosis, a disease that causes skin lesions and affect the lungs and other organs. The current management of the disease is long and has several side effects that often lead the patient to give up the treatment, sequelae and even death. The search for new forms of treatment that minimize these drawbacks is very important. Thus, natural compounds are targets of great interest. Curcumin is one of the main components of the tubers of Curcuma longa, presenting medicinal effects well described in the literature, including the antifungal effect on Paracocidioides brasiliensis. Nevertheless, the mechanisms related to the antifungal effect of such compound are still unknown, so the objective of the present research is to understand what changes occur in the metabolism of P. brasiliensis after exposure to curcumin and to identify the main targets of the compound. Proteomic analysis as based on nanoUPLC-MS analysis and the functional classification of the identified proteins. The main metabolic processes that were being regulated were biologically validated through assays such as fluorescence microscopy, EPR and phagocytosis. Proteomic analysis revealed that curcumin regulates several metabolic processes of the fungus, including important pathways for energy production, such as the glycolytic pathway, beta oxidation and the glyoxylate cycle. Protein synthesis was down-regulated in fungi exposed to curcumin. The electron transport chain and the tricarboxylic acid cycle were also down-regulated, indicating that both the mitochondrial membrane and the mitochondrial activity were compromised. Plasma membrane and cell wall structure were altered following exposure to the compound. The fungus' ability to survive the phagocytosis process by alveolar macrophages was reduced. Thus, curcumin interferes with several metabolic pathways in the fungus that causes paracoccidioidomycosis. BIOLOGICAL SIGNIFICANCE: The challenges presented by the current treatment of paracoccidioidomycosis often contributing to patients' withdrawal from treatment, leading to sequelae or even death. Thus, the search for new treatment options against this disease is growing. The discovery that curcumin is active against Paracoccidioides was previously reported by our study group. Here, we clarify how the compound acts on the fungus causing its growth inhibition and decreased viability. Understanding the mechanisms of action of curcumin on P. brasiliensis elucidates how we can seek new alternatives and which metabolic pathways and molecular targets we should focus on in this incessant search to bring the patient a treatment with fewer adverse effects.

Iron Deprivation Modulates the Exoproteome in Paracoccidioides brasiliensis.

Fungi of the Paracoccidioides genus are the etiological agents of the systemic mycosis paracoccidioidomycosis and, when in the host, they find a challenging environment that is scarce in nutrients and micronutrients, such as Fe, which is indispensable for the survival of the pathogen. Previous studies have shown that fungi of this genus, in response to Fe deprivation, are able to synthesize and capture siderophores (Fe3+ chelators), use Fe-containing host proteins as a source of the metal, and use a non-canonical reductive pathway for Fe3+ assimilation. Despite all of these findings, there are still gaps that need to be filled in the pathogen response to metal deprivation. To contribute to the knowledge related to this subject, we obtained the exoproteome of Paracoccidioides brasiliensis (Pb18) undergoing Fe deprivation and by nanoUPLC-MSE. One hundred forty-one proteins were identified, and out of these, 64 proteins were predicted to be secreted. We also identified the regulation of several virulence factors. Among the results, we highlight Cyb5 as a secreted molecule of Paracoccidioides in the exoproteome obtained during Fe deprivation. Cyb5 is described as necessary for the Fe deprivation response of Saccharomyces cerevisiae and Aspergillus fumigatus. Experimental data and molecular modeling indicated that Cyb5 can bind to Fe ions in vitro, suggesting that it can be relevant in the arsenal of molecules related to iron homeostasis in P. brasiliensis.

Bioluminescence imaging in Paracoccidioides spp.: a tool to monitor the infectious processes.

The genus Paracoccidioides comprises the species complex causing paracoccidioidomycoses (PCM). These fungi are a serious public health problem due to the long-term drug therapy, follow-up treatment, and frequent sequelae generated by the infection, such as pulmonary fibrosis. In this sense, the objective of this work was to generate bioluminescent reporter strains of Paracoccidioides spp. harboring a thermostable, red-shifted luciferase gene under the control of different constitutive promoters. The strains were generated by the integration of a species-specific codon-optimized luciferase gene upon actin or enolase promoter's control. The insertion of the constructs in Paracoccidioides brasiliensis and Paracoccidioides lutzii yeast cells were performed through Agrobacterium tumefaciens-mediated transformation. The results demonstrated the presence of several transformants harboring the luciferase gene. These transformants were further confirmed by the expression of luciferase and by the presence of the hygromycin resistance gene. Moreover, the luciferase activity could be detected in in vitro bioluminescence assays and in vivo models of infection. In general, this work presents the methodology for the construction of bioluminescent strains of Paracoccidioides spp., highlighting potential promoters and proposing an in vivo model, in which those strains could be used for the systemic study of PCM.

Agenesias dentárias atípicas: relato de caso clínico / Atypical dental agenesis: clinical case report

A agenesia dentaria é uma anomalia de desenvolvimento caracterizada pela determinação congênita de menor numero de dentes, podendo estar associada a síndromes genéticas ou ocorrer isoladamente. Embora seja considerada uma das anomalias mais frequentes quando envolve terceiros molares, segundo pré-molares e incisivos laterais, sua ocorrência em dentes estáveis, tais como o canino e o primeiro molar permanente é rara. Desta forma, este relato apresentará o caso de uma paciente do sexo feminino, 8 anos idade, que iniciou tratamento de rotina na Clínica de Odontopediatria da UNIFENAS. Na anamnese não foi relatada pela responsável a ocorrência de alterações sistêmicas, nem queixa ou histórico odontológico relevante. No exame clínico odontológico, notou-se o não irrompimento do primeiro molar permanente superior direito (16). Diante dos exames de imagem, foi confirmada a agenesia do dente 16 e também do canino permanente superior direito (13). A agenesia, sobretudo de dentes estáveis e relevantes no arco dentário, pode comprometer o desenvolvimento adequado da oclusão, mastigação, fonação e estética. Desta maneira, é importante o diagnóstico precoce desta ocorrência com o objetivo de favorecer a elaboração de um adequado plano de tratamento e, minimizar as sequelas destas agenesias atípicas(AU)

Dental agenesis is a developmental anomaly characterized by the congenital determination of fewer teeth, may be associated with genetic syndromes or occur in an isolated form. Although it is considered one of the most frequent anomalies when it involves third molars, second lateral premolars and incisors, the occurrence in stable teeth, such as the permanent canine and the permanent first molar is rare. This report will present the case of an 8-year-old female patient who started routine treatment at the Pediatric Dentistry Clinic of UNIFENAS. In the anamnesis, the responsible person did not report the occurrence of systemic alterations, or a relevant dental complaint or history. On dental clinical examination, it was noticed the non-rupture of the permament maxillar right first molar (16). In view of the imaging tests, the agenesis of tooth 16 and also of the permanent maxillary right canine (13) was confirmed. Agenesis, especially of stable and relevant teeth in the dental arch, may compromise the proper development of occlusion, chewing, phonation and aesthetics. Thus, it is of paramount importance to early diagnosis of this occurrence in order to favor the development of an appropriate treatment plan in order to minimize the sequelae of these atypical agenesis(AU)

A structure-based approach for the discovery of inhibitors against methylcitrate synthase of Paracoccidioides lutzii.

Paracoccidioidomycosis (PCM) is a systemic mycosis, endemic in Latin America, caused by fungi of the genus Paracoccidioides. The treatment of PCM is complex, requiring a long treatment period, which often results in serious side effects. The aim of this study was to screen for inhibitors of a specific target of the fungus that is absent in humans. Methylcitrate synthase (MCS) is a unique enzyme of microorganisms and is responsible for the synthesis of methylcitrate at the beginning of the propionate degradation pathway. This pathway is essential for several microorganisms, since the accumulation of propionyl-CoA can impair virulence and prevent the development of the pathogen. We performed the modeling and molecular dynamics of the structure of Paracoccidioides lutzii MCS (PlMCS) and performed a virtual screening on 89,415 compounds against the active site of the enzyme. The compounds were selected according to the affinity and efficiency criteria of in vitro tests. Six compounds were able to inhibit the enzymatic activity of recombinant PlMCS but only the compound ZINC08964784 showed fungistatic and fungicidal activity against Paracoccidioides spp. cells. The analysis of the interaction profile of this compound with PlMCS showed its effectiveness in terms of specificity and stability when compared to the substrate (propionyl-CoA) of the enzyme. In addition, this compound did not show cytotoxicity in mammalian cells, with an excellent selectivity index. Our results suggest that the compound ZINC08964784 may become a promising alternative antifungal against Paracoccidioides spp. Communicated by Ramaswamy H. Sarma.

Atraso de escolaridade e dificuldade de socialização de pacientes com fissura labiopalatina não sindrômica / Scholar delay and socialization dificulty of patients with non syndromic cleft lip and palate

Dentre as principais consequências da fissura labial e/ou palatina não sindrômica (FL/ PNS) estão dificuldades com fonação e autoestima, a primeira uma questão funcional e a segunda um problema social derivado não raro de contextos de bullying que, dentre outros, podem levar o indivíduo à evasão escolar. O objetivo deste estudo foi avaliar o atraso de escolaridade e a dificuldade de socialização de pacientes com FL/PNS quando comparados a uma população não afetada da mesma faixa etária de 7 a 20 anos, atendidos na Universidade José do Rosário Vellano ­UNIFENAS, campus de Alfenas. Os sujeitos foram agrupados em duas categorias de indivíduos, o grupo caso composto por indivíduos com FL/PNS em tratamento no Centro Pró-Sorriso da UNIFENAS; e o grupo controle composto por indivíduos sem FL/PNS em tratamento nas clínicas de Odontopediatria e Integrada da UNIFENAS. Os resultados demonstraram que a proporção de pacientes com FL/PNS atrasados na escola foi de quase 5 vezes maior que o número de pacientes sem fissuras (p<0,01). Constatou-se que a presença da FL/PNS pode ser o ponto de partida para outros contribuintes, com interferências psicológicas e/ou sociais, interferindo negativamente no processo de socialização (bullying) do paciente (p=0,0018). Portanto devem ser tratadas com abordagem multidisciplinar, incluindo diversos profissionais, dentre eles pedagogos, psicólogos e odontólogos(AU)

Among the main consequences of Non Syndromic Cleft Lip and Palate (NSCLP) are the difficulties with phonation and self estime, the first being a functional issue na the later being social that is derived from, not rarely, bullying contexts, that among other things, may lead na individual to school evasion. The objective of this study was to avaluate the levels of scholarity of patients with NSCLP when compared to a non affected population of the same age in individuals from 7 to 20 years old, attended the Pediatric and Integrated Pediatric Clinic of UNIFENAS, Alfenas campus. The subjects were grouped into two categories of individuals, the case group was composed of individuals with FL/PNS with treatment at the ProSmile center at UNIFENAS. The control group was composed of individuals without FL/PNS in treatment at the clinics of pediatric and integrated denistry at UNIFENAS. The results demonstrated the number of patients with FL/PNS that presented scholar delay were almost 5 times the number of patients that didn't present FL/PNS (p<0,01). The presence of NFL/PNS may be the starting point for other contributors with psychological and/or social interferences, interfering negatively with the socialization process (bullying) of the patient (p=0,0018). They should be treated with a multidisciplinary manner, including multiple professionals, among them pedagogues, psychologist and dentist(AU)

In vitro and in silico analysis reveals antifungal activity and potential targets of curcumin on Paracoccidioides spp.

The search for new compounds with activity against Paracoccidioides, etiologic agents of Paracoccidioidomycosis (PCM), is extremely necessary due to the current scenario of the available therapeutic arsenal. Treatment is restricted to three classes of antifungals with side effects. Curcumin is a polyphenol with antifungal effects that is extracted from Curcuma longa. The present work aimed to evaluate the activity of curcumin in different species of Paracoccidioides and to evaluate the potential molecular targets of curcumin using computational strategies. In addition, interactions with classic antifungals used in the treatment of PCM were evaluated. Curcumin inhibits the growth of Paracoccidioides spp. exerting a fungicidal effect. The combination of curcumin with amphotericin B, co-trimoxazole, and itraconazole showed a synergistic or additive interaction. Molecular targets as superoxide dismutase, catalase, and isocitrate lyase were proposed based on in silico approaches. Curcumin affects the fungal plasma membrane and increases the production of reactive oxygen species. Therefore, curcumin is a good alternative for the treatment of PCM.

De Elton Mayo a Ivar Oddone: redescobrir a instrução ao sósia / From Elton Mayo to Ivar Oddone: rediscovering the instruction to the double

O presente texto foi publicado originalmente como prefácio de uma obra de Yves Clot em homenagem a Ivar Oddone. Decidimos publicá-lo no formato de artigo, com a autorização do autor, por considerarmos que traz importantes e atuais contribuições para o campo do trabalho em intercruzamento com diversas teorias: Clínica da Atividade, Ergologia, Ergonomia, Saúde do Trabalhador, dentre outras. Neste texto, Yves Clot retoma aspectos significativos da obra de Ivar Oddone e tece reflexões sobre aspectos originais dessa, nem sempre considerados pelos estudiosos da área. Assim, faz uma análise comparativa com as propostas apresentadas por Elton Mayo, as quais se sagraram como importante ferramenta para o desenvolvimento do capitalismo industrial sob a suposta égide do enaltecimento do aspecto humano nas situações de trabalho. Ao contrário das concepções e práticas de Mayo, que foram efetivamente voltadas para o aumento da produtividade das empresas e, portanto, para a exploração dos trabalhadores, Oddone buscou reconhecer e valorizar o saber dos operários, com o intuito de que esse se configurasse como um contrapoder ao ampliar o seu poder de agir na situação de trabalho, aspectos que são amplamente discutidos por Clot no texto que ora apresentamos.¹

This text was initially published as a preface written by Yves Clot in honor of Ivar Oddone. We decided to publish it as an article, authorized by the author, as we believe that it brings important and current contributions to the field of work, in interseccion with several theories: Clinic of Activity, Ergology, Ergonomics, Worker's Health, among others. In this text, Yves Clot takes up highlights of Ivar Oddone's work and produces reflections about its original aspects, not always considered by experts in the field. Thus, he makes a comparative analysis with those aspects and the proposals presented by Elton Mayo, which have become an important tool for the development of industrial capitalism under the supposed appreciation of the human sides in work situations. Contrary to Mayo's conceptions and practices, which were aimed at increasing the productivity of companies and, therefore, at the exploitation of workers, Oddone sought to recognize and value the knowledge of workers, intending that this was used as a kind of counterpower, by expanding its power to act in the work situation, points that are widely discussed by Clot in the text that we present now.

In silico identification of glycosylphosphatidylinositol-anchored proteins in Paracoccidioides spp.

Aim: To predict glycosylphosphatidylinositol (GPI)-anchored proteins in the genome of Paracoccidioides brasiliensis and Paracoccidioides lutzii. Materials & methods: Five different bioinformatics tools were used for predicting GPI-anchored proteins; we considered as GPI-anchored proteins those detected by at least two in silico analysis methods. We also performed the proteomic analysis of P. brasiliensis cell wall by mass spectrometry. Results: Hundred GPI-anchored proteins were predicted in P. brasiliensis and P. lutzii genomes. A series of 57 proteins were classified in functional categories and 43 conserved proteins were reported with unknown functions. Four proteins identified by in silico analyses were also identified in the cell wall proteome. Conclusion: The data obtained in this study are important resources for future research of GPI-anchored proteins in Paracoccidioides spp. to identify targets for new diagnostic tools, drugs and immunological tests.

Interacting with Hemoglobin: Paracoccidioides spp. Recruits hsp30 on Its Cell Surface for Enhanced Ability to Use This Iron Source.

Paracoccidioides spp. are thermally dimorphic fungi that cause paracoccidioidomycosis and can affect both immunocompetent and immunocompromised individuals. The infection can lead to moderate or severe illness and death. Paracoccidioides spp. undergo micronutrients deprivation within the host, including iron. To overcome such cellular stress, this genus of fungi responds in multiple ways, such as the utilization of hemoglobin. A glycosylphosphatidylinositol (GPI)-anchored fungal receptor, Rbt5, has the primary role of acquiring the essential nutrient iron from hemoglobin. Conversely, it is not clear if additional proteins participate in the process of using hemoglobin by the fungus. Therefore, in order to investigate changes in the proteomic level of P. lutzii cell wall, we deprived the fungus of iron and then treated those cells with hemoglobin. Deprived iron cells were used as control. Next, we performed cell wall fractionation and the obtained proteins were submitted to nanoUPLC-MSE. Protein expression levels of the cell wall F1 fraction of cells exposed to hemoglobin were compared with the protein expression of the cell wall F1 fraction of iron-deprived cells. Our results showed that P. lutzii exposure to hemoglobin increased the level of adhesins expression by the fungus, according to the proteomic data. We confirmed that the exposure of the fungus to hemoglobin increased its ability to adhere to macrophages by flow cytometry. In addition, we found that HSP30 of P. lutzii is a novel hemoglobin-binding protein and a possible heme oxygenase. In order to investigate the importance of HSP30 in the Paracoccidioides genus, we developed a Paracoccidioides brasiliensis knockdown strain of HSP30 via Agrobacterium tumefaciens-mediated transformation and demonstrated that silencing this gene decreases the ability of P. brasiliensis to use hemoglobin as a nutrient source. Additional studies are needed to establish HSP30 as a virulence factor, which can support the development of new therapeutic and/or diagnostic approaches.

Prospecting for new catechol-O-methyltransferase (COMT) inhibitors as a potential treatment for Parkinson's disease: a study by molecular dynamics and structure-based virtual screening.

Parkinson's disease (PD) is a neurodegenerative, chronic, and progressive disease, common in the elderly. The catechol-O-methyltransferase (COMT) is a monomeric enzyme involved in dopamine (DA) degradation, the neurotransmitter in deficit in patients with PD. The reference treatment of PD consists of levodopa (L-dopa) administration, which is the precursor of DA. The inhibition of COMT is an adjuvant treatment in PD since it keeps DA levels constant. The goal of this study was to identify drug candidates capable of inhibiting COMT for the treatment of PD and identify important fragments of these molecules. Initially, we analyzed the flexibility of COMT and defined its main conformations in solution regarding the absence (system I) and presence of the S-adenosyl-L-methionine (SAM) cofactor (system II) through molecular dynamics (MD) simulations. Two regions in these structures were selected for molecular docking, firstly the entire cavity where the cofactor and substrates are bound and secondly the specific biding region of the enzyme substrates. Based on the conformations of the MD, the virtual screening (VS) was performed against FDA Approved and Zinc Natural Products databases aiming at the selection of the best compounds. Subsequently, the absorption, distribution, metabolization, excretion, and toxicity (ADMET) properties, as well as drug-score and drug-likeness indexes of the most promising compounds were analyzed. After a detailed analysis of the compounds selected by structure-based VS, it was possible to highlight the fragments most frequently involved in their stability: 2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole, 9H-Benz(c)indole(3,2,1-ij)(1,5)naphthyridin-9-one and (10R,13S)-10,13-dimethyl-1,2,6,7,8,9,11,12,14,15,16,17dodecahydrocyclopenta[a]phenanthren-3-one. The identification of these potential fragments is essential for the prospection of more specific inhibitors against COMT using the technique of Fragment-based lead discovery (FBLD). Besides, this study allowed us to identify the potential COMT inhibitors through a complete understanding of molecular-level interactions based on the flexibility of this protein.Communicated by Ramaswamy H. Sarma.

The "Little Iron Waltz": The Ternary Response of Paracoccidioides spp. to Iron Deprivation.

Paracoccidioides is a genus of thermodimorphic fungi that causes paracoccidioidomycosis. When in the host, the fungus undergoes several challenges, including iron deprivation imposed by nutritional immunity. In response to the iron deprivation triggered by the host, the fungus responds in a ternary manner using mechanisms of high affinity and specificity for the uptake of Fe, namely non-classical reductive iron uptake pathway, uptake of host iron proteins, and biosynthesis and uptake of siderophores. This triple response resembles the rhythmic structure of a waltz, which features three beats per compass. Using this connotation, we have constructed this review summarizing relevant findings in this area of study and pointing out new discoveries and perspectives that may contribute to the expansion of this "little iron waltz".

IR-780-Albumin-Based Nanocarriers Promote Tumor Regression Not Only from Phototherapy but Also by a Nonirradiation Mechanism.

IR-780 iodide is a fluorescent dye with optical properties in the near-infrared region that has applications in tumor detection and photothermal/photodynamic therapy. This multifunctional effect led to the development of theranostic nanoparticles with both IR-780 and chemotherapeutic drugs such as docetaxel, doxorubicin, and lonidamine. In this work, we developed two albumin-based nanoparticles containing near-infrared IR-780 iodide multifunctional dyes, one of them possessing a magnetic core. Molecular docking with AutoDock Vina studies showed that IR-780 binds to bovine serum albumin (BSA) with greater stability at a higher temperature, allowing the protein binding pocket to better fit this dye. The theoretical analysis corroborates the experimental protocols, where an enhancement of IR-780 was found coupled to BSA at 60 °C, even 30 days after preparation, in comparison to 30 °C. In vitro assays monitoring the viability of Ehrlich ascites carcinoma cells revealed the importance of the inorganic magnetic core on the nanocarrier photothermal-cytotoxic effect. Fluorescence molecular tomography measurements of Ehrlich tumor-bearing Swiss mice revealed the biodistribution of the nanocarriers, with marked accumulation in the tumor tissue (≈3% ID). The histopathological analysis demonstrated strong increase in tumoral necrosis areas after 24 and 72 h after treatment, indicating tumor regression. Tumor regression analysis of nonirradiated animals indicate a IR-780 dose-dependent antitumoral effect with survival rates higher than 70% (animals monitored up to 600 days). Furthermore, an in vivo photothermal therapy procedure was performed and tumor regression was also verified. These results show a novel insight for the biomedical application of IR-780-albumin-based nanocarriers, namely cancer therapy, not only by photoinduced therapy but also by a nonirradiation mechanism. Safety studies (acute oral toxicity, cardiovascular evaluation, and histopathological analysis) suggest potential for clinical translation.