Miltefosine: A Repurposing Drug against Mucorales Pathogens.

Mucorales are a group of non-septated filamentous fungi widely distributed in nature, frequently associated with human infections, and are intrinsically resistant to many antifungal drugs. For these reasons, there is an urgent need to improve the clinical management of mucormycosis. Miltefosine, which is a phospholipid analogue of alkylphosphocholine, has been considered a promising repurposing drug to be used to treat fungal infections. In the present study, miltefosine displayed antifungal activity against a variety of Mucorales species, and it was also active against biofilms formed by these fungi. Treatment with miltefosine revealed modifications of cell wall components, neutral lipids, mitochondrial membrane potential, cell morphology, and the induction of oxidative stress. Treated Mucorales cells also presented an increased susceptibility to SDS. Purified ergosterol and glucosylceramide added to the culture medium increased miltefosine MIC, suggesting its interaction with fungal lipids. These data contribute to elucidating the effect of a promising drug repurposed to act against some relevant fungal pathogens that significantly impact public health.

Pandemic Response Box® library as a source of antifungal drugs against Scedosporium and Lomentospora species.

Scedosporium and Lomentospora species are opportunistic filamentous fungi that cause localized and disseminated infections in immunocompetent and immunocompromised patients. These species are considered resistant fungi due to their low susceptibility to most current antifungal agents used in healthcare settings. The search for new compounds that could work as promising candidate antifungal drugs is an increasing field of interest. In this context, in the present study we screened the Pandemic Response Box® library (Medicines for Malaria Venture [MMV], Switzerland) to identify compounds with antifungal activity against Scedosporium and Lomentospora species. An initial screening of the drugs from this collection at 5 µM was performed using a clinical Scedosporium aurantiacum isolate according to the EUCAST protocol. Compounds with activity against this fungus were also tested against four other species (S. boydii¸ S. dehoogii, S. apiospermum and L. prolificans) at concentrations ranging from 0.078 to 10 µM. Seven compounds inhibited more than 80% of S. aurantiacum growth, three of them (alexidine, amorolfine and olorofim) were selected due to their differences in mechanism of action, especially when compared to drugs from the azole class. These compounds were more active against biofilm formation than against preformed biofilm in Scedosporium and Lomentospora species, except alexidine, which was able to decrease preformed biofilm about 50%. Analysis of the potential synergism of these compounds with voriconazole and caspofungin was performed by the checkerboard method for S. aurantiacum. The analysis by Bliss methodology revealed synergistic effects among selected drugs with caspofungin. When these drugs were combined with voriconazole, only alexidine and amorolfine showed a synergistic effect, whereas olorofim showed an antagonistic effect. Scanning electron microscopy revealed that alexidine induces morphology alterations in S. aurantiacum biofilm grown on a catheter surface. Reactive oxygen species production, mitochondrial activity and surface components were analyzed by fluorescent probes when S. aurantiacum was treated with selected drugs and revealed that some cell parameters are altered by these compounds. In conclusion, alexidine, amorolfine and olorofim were identified as promising compounds to be studied against scedosporiosis and lomentosporiosis.

Promising Antifungal Molecules against Mucormycosis Agents Identified from Pandemic Response Box®: In Vitro and In Silico Analyses.

Mucormycosis is considered concerning invasive fungal infections due to its high mortality rates, difficult diagnosis and limited treatment approaches. Mucorales species are highly resistant to many antifungal agents and the search for alternatives is an urgent need. In the present study, a library with 400 compounds called the Pandemic Response Box® was used and four compounds were identified: alexidine and three non-commercial molecules. These compounds showed anti-biofilm activity, as well as alterations in fungal morphology and cell wall and plasma membrane structure. They also induced oxidative stress and mitochondrial membrane depolarization. In silico analysis revealed promising pharmacological parameters. These results suggest that these four compounds are potent candidates to be considered in future studies for the development of new approaches to treat mucormycosis.

NeuroAIreh@b: an artificial intelligence-based methodology for personalized and adaptive neurorehabilitation.

Cognitive impairments are a prevalent consequence of acquired brain injury, dementia, and age-related cognitive decline, hampering individuals' daily functioning and independence, with significant societal and economic implications. While neurorehabilitation represents a promising avenue for addressing these deficits, traditional rehabilitation approaches face notable limitations. First, they lack adaptability, offering one-size-fits-all solutions that may not effectively meet each patient's unique needs. Furthermore, the resource-intensive nature of these interventions, often confined to clinical settings, poses barriers to widespread, cost-effective, and sustained implementation, resulting in suboptimal outcomes in terms of intervention adaptability, intensity, and duration. In response to these challenges, this paper introduces NeuroAIreh@b, an innovative cognitive profiling and training methodology that uses an AI-driven framework to optimize neurorehabilitation prescription. NeuroAIreh@b effectively bridges the gap between neuropsychological assessment and computational modeling, thereby affording highly personalized and adaptive neurorehabilitation sessions. This approach also leverages virtual reality-based simulations of daily living activities to enhance ecological validity and efficacy. The feasibility of NeuroAIreh@b has already been demonstrated through a clinical study with stroke patients employing a tablet-based intervention. The NeuroAIreh@b methodology holds the potential for efficacy studies in large randomized controlled trials in the future.

Treatment of canine cutaneous leishmaniasis by Leishmania (Viannia) braziliensis in dogs with furazolidone and ß-cyclodextrin: case report.

Euthanasia of animals is not accepted as a control for cutaneous leishmaniasis caused by Leishmania (Viannia) braziliensis and drugs used in humans for the treatment of leishmaniasis are not allowed for animals in Brazil. Miltefosine was authorized for dogs infected by Leishmania infantum with variable results for L. braziliensis. Thus, nine dogs infected with Leishmania (V.) braziliensis were treated by a combination of furazolidone and ß-cyclodextrin. The nine dogs were mongrels, weighing between 4-17 kg and 3-10 years old. These dogs had ulcerous lesions in different regions such as scrotal tissue, auricular pavilion and nostrils. Serological, molecular and protozoal culture techniques were used for laboratory diagnosis. The treatment used furazolidone + ß-cyclodextrin complex (1: 2) at a concentration of 60 mg/mL given orally at a dose of 15 mg/kg every 12 hours. The re-epithelialization of lesions occurred between 35 and 41 days of treatment. During fourteen months the animals were monitored and there was no reactivation of lesions or growth of the protozoan in a culture medium of the biopsies. This study demonstrated that treatment with FZD and CD is effective in reducing the cutaneous lesions caused by L. braziliensis in dogs.

Identification of Promising Antifungal Drugs against Scedosporium and Lomentospora Species after Screening of Pathogen Box Library.

Fungal infections have been increasing during the last decades. Scedosporium and Lomentospora species are filamentous fungi most associated to those infections, especially in immunocompromised patients. Considering the limited options of treatment and the emergence of resistant isolates, an increasing concern motivates the development of new therapeutic alternatives. In this context, the present study screened the Pathogen Box library to identify compounds with antifungal activity against Scedosporium and Lomentospora. Using antifungal susceptibility tests, biofilm analysis, scanning electron microscopy (SEM), and synergism assay, auranofin and iodoquinol were found to present promising repurposing applications. Both compounds were active against different Scedosporium and Lomentospora, including planktonic cells and biofilm. SEM revealed morphological alterations and synergism analysis showed that both drugs present positive interactions with voriconazole, fluconazole, and caspofungin. These data suggest that auranofin and iodoquinol are promising compounds to be studied as repurposing approaches against scedosporiosis and lomentosporiosis.

Detection of Leishmania (Viannia) braziliensis in bats from Espírito Santo, Brazil (2018-2019).

Studying the occurrence of Leishmania in bats would help toward clarifying the role of these animals in epidemiological cycles. Therefore, the objective of this study was to detect Leishmania spp. in bats from the Brazilian State of Espírito Santo. In total, 105 bats from 11 species which had been collected from 17 municipalities of Espírito Santo were obtained from the IDAF Rabies Diagnostic Laboratory between December 2018 and June 2019. DNA was extracted from the bone marrow, skin, and spleen samples and analysed using conventional PCR, with primers specific for the kinetoplastic DNA minicircle of the parasitic species Leishmania (Viannnia) braziliensis, Leishmania (Leishmania) infantum, and Leishmania (Leishmania) amazonensis. To identify the Leishmania species, the positive PCR products were analysed using PCR-RFLP with the HaeIII enzyme and 8% polyacrylamide gel electrophoresis. Positivity for L. (V.) braziliensis was observed in five (4.76%) bats: namely, one male Eumops glaucinus from Vitória (in a skin sample), one male Artibeus lituratus from Vila Velha and one female Eumops glaucinus from Vitória (both in bone marrow samples), and one male A. lituratus from Vitória and one male Molossus rufus from Guarapari (both in the spleen samples). To our best knowledge, this is the first report of the detection of Leishmania in bats from Espírito Santo as well as of Leishmania DNA in the bone marrow of bats.

Miltefosine Against Scedosporium and Lomentospora Species: Antifungal Activity and Its Effects on Fungal Cells.

Scedosporium and Lomentospora species are filamentous fungi responsible for a wide range of infections in humans and are frequently associated with cystic fibrosis and immunocompromising conditions. Because they are usually resistant to many antifungal drugs available in clinical settings, studies of alternative targets in fungal cells and therapeutic approaches are necessary. In the present work, we evaluated the in vitro antifungal activity of miltefosine against Scedosporium and Lomentospora species and how this phospholipid analogue affects the fungal cell. Miltefosine inhibited different Scedosporium and Lomentospora species at 2-4 µg/ml and reduced biofilm formation. The loss of membrane integrity in Scedosporium aurantiacum caused by miltefosine was demonstrated by leakage of intracellular components and lipid raft disorganisation. The exogenous addition of glucosylceramide decreased the inhibitory activity of miltefosine. Reactive oxygen species production and mitochondrial activity were also affected by miltefosine, as well as the susceptibility to fluconazole, caspofungin and myoricin. The data obtained in the present study contribute to clarify the dynamics of the interaction between miltefosine and Scedosporium and Lomentospora cells, highlighting its potential use as new antifungal drug in the future.

Seroprevalence and associated factors of infectious bovine rhinotracheitis and bovine viral diarrhea in dairy cows in the Caparaó region, Espírito Santo, Brazil / Soroprevalência e fatores associados à rinotraqueíte infecciosa bovina e diarreia viral bovina em vacas leiteiras na região do Caparaó, Espírito Santo, Brasil

ABSTRACT: This study evaluated the seroprevalence and associated factors of Infectious Bovine Rhinotracheitis (IBR) and Bovine Viral Diarrhea (BVD), and to analyze the possible relationship between IBR, BVD, and the occurrence of mastitis. For this purpose, 854 crossbred dairy cows were evaluated from 69 properties allocated in the 12 municipalities that make up the Caparaó region, Espírito Santo (ES), Brazil. The seroprevalence of IBR and BVD was determined using the indirect ELISA test. Associations between variables were estimated using the prevalence ratio (PR) with a 95% confidence interval. The chi-square test was used to verify the significance of the associations (P < 0.05). The average prevalence of IBR and BVD was 48.59% and 26.46%, respectively. Animals seroreactive for IBR were more likely to develop subclinical mastitis (P < 0.01; PR: 1.27), and cows that were seroreactive for BVD were more likely to develop clinical mastitis (P < 0.01; PR: 2.24). Mechanical milking was considered a factor associated with IBR (P < 0.01; PR: 1.36) and BVD (P < 0.01; PR: 1.25). Reproductive management by natural breeding was considered a factor associated with IBR (P < 0.01; PR: 1.22). Animals seroreactive for BVD were more likely to develop reproductive problems (P < 0.01; PR: 1.34). It was concluded that the agents that cause IBR and BVD are widely disseminated in dairy cattle herds in the municipalities of the Caparaó region, ES, Brazil. The presence of IBR and BVD increased the chances of cows developing subclinical mastitis and clinical mastitis, respectively, and the cows that were mechanically milked were more likely to be seroreactive for IBR and BVD.

RESUMO: O objetivo do presente estudo foi avaliar a soroprevalência e os fatores de risco associados à Rinotraqueíte Infecciosa Bovina (IBR) e Diarreia Viral Bovina (BVD), e analisar a possível relação entre IBR, BVD e à ocorrência de mastite. Para tanto, foram avaliadas 854 vacas leiteiras mestiças de 69 propriedades localizadas nos 12 municípios que compõem a região do Caparaó, Espírito Santo (ES), Brasil. A soroprevalência de IBR e BVD foram determinadas pelo teste ELISA indireto. As associações entre variáveis ​​foram estimadas pela razão de prevalência (PR) com intervalo de confiança de 95%. O teste do qui-quadrado foi utilizado para verificar a significância das associações (P < 0,05). A prevalência média de IBR e BVD foi de 48% e 26%, respectivamente. Os animais sororreagentes para IBR foram mais propensos a desenvolver mastite subclínica (P < 0,01; PR: 1,27), e as vacas sororreagentes para BVD foram mais propensas a desenvolver mastite clínica (P < 0,01; PR: 2,24). À ordenha mecânica foi considerada um fator associado a IBR (P < 0,01; PR: 1,36) e BVD (P < 0,01; PR: 1,25). O manejo reprodutivo por monta natural foi considerado um fator associado IBR (P < 0,01; PR: 1,22). Os animais sororreagentes para BVD foram mais propensos a desenvolverem problemas reprodutivos (P < 0,01; PR: 1,34). Concluiu-se que os agentes causadores de IBR e BVD estão amplamente disseminados em rebanhos leiteiros nos municípios da região de Caparaó, ES, Brasil. A presença de IBR e BVD aumentaram as chances das vacas desenvolverem mastite subclínica e mastite clínica, respectivamente, e as vacas que foram ordenhadas mecanicamente apresentaram maior probabilidade de serem sororreagente para IBR e BVD.

Low-cost synthetic tourniquet training model. / Modelo sintético de baixo custo para treinamento do uso de torniquete.

OBJECTIVE: to present a low-cost model for bleeding control training with the use of a tourniquet and the results of the training evaluation. METHOD: low-cost and easy-to-purchase materials, such as upholstery foam, ethylene-vinyl acetate (EVA) sheet, plastic propulsion pump, saline container, water, school paint, serum equipment, rubber tubing for tourniquet, and a leg and an arm of a ghost mannequin, were used. In the model assembly, we created an active bleeding simulation system, which could only be controlled with the correct application of the tourniquet. The model was submitted to professional and academic evaluation. RESULTS: the model was similar to human anatomy, proved to be practical in the bleeding control training with the use of a tourniquet, and had low cost. CONCLUSION: the model for bleeding control training had excellent acceptability, was considered viable for educational purposes of tourniquet use, and had low cost.

OBJETIVO: apresentar um modelo de baixo custo para treinamento de controle de sangramento com o uso de torniquete e os resultados da avaliação do treinamento. MÉTODO: foram utilizados materiais de fácil aquisição e de baixo custo, como espuma de estofado, placa de etileno-acetato de vinila (EVA), bomba plástica de propulsão, recipiente de soro fisiológico, água, tinta escolar, equipos de soro, tubo de látex para garrote e manequins comerciais de perna e de braço. Na montagem, foi criado um sistema de simulação de sangramento ativo que só podia ser controlado com aplicação correta do torniquete. O modelo foi submetido à avaliação de profissionais e acadêmicos. RESULTADOS: o modelo teve semelhança com a anatomia humana, mostrou-se prático no treinamento da contenção de sangramento com uso de torniquete e teve baixo custo financeiro. CONCLUSÃO: o modelo para treinamento de controle de sangramento teve excelente aceitabilidade, foi considerado viável para fins educacionais do uso de torniquete e teve baixo custo.

Modelo sintético de baixo custo para treinamento do uso de torniquete / Low-cost synthetic tourniquet training model

RESUMO Objetivo: apresentar um modelo de baixo custo para treinamento de controle de sangramento com o uso de torniquete e os resultados da avaliação do treinamento. Método: foram utilizados materiais de fácil aquisição e de baixo custo, como espuma de estofado, placa de etileno-acetato de vinila (EVA), bomba plástica de propulsão, recipiente de soro fisiológico, água, tinta escolar, equipos de soro, tubo de látex para garrote e manequins comerciais de perna e de braço. Na montagem, foi criado um sistema de simulação de sangramento ativo que só podia ser controlado com aplicação correta do torniquete. O modelo foi submetido à avaliação de profissionais e acadêmicos. Resultados: o modelo teve semelhança com a anatomia humana, mostrou-se prático no treinamento da contenção de sangramento com uso de torniquete e teve baixo custo financeiro. Conclusão: o modelo para treinamento de controle de sangramento teve excelente aceitabilidade, foi considerado viável para fins educacionais do uso de torniquete e teve baixo custo.

ABSTRACT Objective: to present a low-cost model for bleeding control training with the use of a tourniquet and the results of the training evaluation. Method: low-cost and easy-to-purchase materials, such as upholstery foam, ethylene-vinyl acetate (EVA) sheet, plastic propulsion pump, saline container, water, school paint, serum equipment, rubber tubing for tourniquet, and a leg and an arm of a ghost mannequin, were used. In the model assembly, we created an active bleeding simulation system, which could only be controlled with the correct application of the tourniquet. The model was submitted to professional and academic evaluation. Results: the model was similar to human anatomy, proved to be practical in the bleeding control training with the use of a tourniquet, and had low cost. Conclusion: the model for bleeding control training had excellent acceptability, was considered viable for educational purposes of tourniquet use, and had low cost.