Features of the microalga Raphidocelis subcapitata: physiology and applications.

The microalga Raphidocelis subcapitata was isolated from the Nitelva River (Norway) and subsequently deposited in the collection of the Norwegian Institute of Water Research as "Selenastrum capricornutum Printz". This freshwater microalga, also known as Pseudokirchneriella subcapitata, acquired much of its notoriety due to its high sensitivity to different chemical species, which makes it recommended by different international organizations for the assessment of ecotoxicity. However, outside this scope, R. subcapitata continues to be little explored. This review aims to shed light on a microalga that, despite its popularity, continues to be an "illustrious" unknown in many ways. Therefore, R. subcapitata taxonomy, phylogeny, shape, size/biovolume, cell ultra-structure, and reproduction are reviewed. The nutritional and cultural conditions, chronological aging, and maintenance and preservation of the alga are summarized and critically discussed. Applications of R. subcapitata, such as its use in aquatic toxicology (ecotoxicity assessment and elucidation of adverse toxic outcome pathways) are presented. Furthermore, the latest advances in the use of this alga in biotechnology, namely in the bioremediation of effluents and the production of value-added biomolecules and biofuels, are highlighted. To end, a perspective regarding the future exploitation of R. subcapitata potentialities, in a modern concept of biorefinery, is outlined. KEY POINTS: • An overview of alga phylogeny and physiology is critically reviewed. • Advances in alga nutrition, cultural conditions, and chronological aging are presented. • Its use in aquatic toxicology and biotechnology is highlighted.

Effect of natural agents on the bond strength to eroded dentin.

OBJECTIVE: This study evaluated the effect of bromelain and propolis extract on the bond strength (BS) of a universal adhesive system to eroded dentin. MATERIALS AND METHODS: Sixty human molars with exposed dentin were halved, with one half protected by composite resin and the other subjected to erosive treatment followed by remineralization. After the erosive treatment, the composite resin was removed, and the teeth were randomly assigned to three groups (n = 20): Adhesive-Control System; Br-10%; Pr-16%. Following the treatments, composite resin blocks were built on the dentin surfaces and sticks of 0.9 mm2 were obtained and stored in distilled water at 37°C for 24 h and 6 months. After these periods, the sticks underwent bond strength testing and the data were analyzed using 2-way ANOVA, Bonferroni test, p < 0.05. Fracture patterns were observed using light microscope and scanning electron microscopy. RESULTS: Irrespective of the substrate and aging duration, propolis demonstrated higher BS (p < 0.05) compared to the other treatments. Eroded dentin exhibited greater removal of the smear layer and dentinal tubules with a larger diameter than sound dentin, especially when treated with bromelain, resulting in the formation of resin tags. CONCLUSIONS: Propolis consistently promoted the highest bond strength, irrespective of aging or substrate. Eroded dentin treated with propolis, or bromelain exhibited a higher prevalence of non-adhesive fractures and resin tag formation. CLINICAL SIGNIFICANCE: Propolis shows promise for enhancing the longevity of adhesive restorations in eroded dentin due to its ability to promote high bond strength.

Incorporation of bioactive glass-ceramic into coconut oil for remineralization of incipient carious lesions.

This study evaluated the efficacy of incorporating different concentrations of bioactive glass-ceramic (Biosilicate) into coconut oil on the remineralizing potential and surface roughness of white spot lesions. Fragments (6 x 6 x 2mm) of bovine teeth were sectioned and initial microhardness (KHN) and surface roughness (Ra) readings were obtained. The samples were submitted to cariogenic challenge to form white spot lesions and were separated into six groups (n=13): 1) Artificial Saliva (AS); 2) Coconut Oil (CO); 3) CO+2% Biosilicate (CO+2%Bio); 4) CO+5% Biosilicate (CO+5%Bio); 5) 2% Biosilicate Suspension (2% Bio) and 6) 5% Biosilicate Suspension (5% Bio). The treatments for 1 cycle/day were: immersion into the treatments for 5 minutes, rinsing in distilled water, and storage in artificial saliva at 37ºC. After 14 days, KHN and Ra readings were taken. The surface roughness alteration ((Ra) was analyzed (Kruskal-Wallis, Dunn's post-test, p<0.05). CO+2%Bio had higher (p = 0.0013) (Ra followed by CO+5%Bio (p = 0.0244) than AS. The relative KHN and remineralization potential were analyzed (ANOVA, Tukey, p<0.05), and 5% Bio treatment presented a higher relative microhardness than all other groups (p>0.05). The remineralizing potential of all the treatments was similar (p > .05). When Biosilicate was added, the pH of the suspensions increased and the alkaline pH remained during the analysis. Biosilicate suspension is more efficient than the incorporation of particles into coconut oil at white spot lesion treatment. In addition to the benefits that coconut oil and Biosilicate present separately, their association can enhance the remineralizing potential of Biosilicate.

CS2/KOH System-Promoted Stereoselective Synthesis of (E)-Alkenes from Diarylalkynes and a "Hidden" Zinin-Type Reduction of Nitroarenes into Arylamines.

In this work, we present the CS2/KOH system as a practical and efficient reductive medium for obtaining (E)-alkenes from alkynes through a highly stereoselective semireduction reaction. This cost-effective system enabled successful semireduction reactions of diverse alkynes using water as a hydrogen source, yielding moderate to excellent yields. The versatility of this protocol is further demonstrated through the synthesis of relevant compounds such as pinosylvin and resveratrol precursors, along with the notable anticancer agent DMU-212. Furthermore, during the reaction scope investigation, we serendipitously disclosed that this reductive system was also able to promote a Zinin-type reaction to reduce nitroarenes into arylamines.

Effect of artificial aging on mechanical and physical properties of CAD-CAM PMMA resins for occlusal splints.

PURPOSE: This study aimed to assess and compare the color stability, flexural strength (FS), and surface roughness of occlusal splints fabricated from heat-cured acrylic resin, milled polymethyl methacrylate (PMMA)-based resin, and 3D-printed (PMMA) based-resin. MATERIALS AND METHODS: Samples of each type of resin were obtained, and baseline measurements of color and surface roughness were recorded. The specimens were divided into three groups (n = 10) and subjected to distinct aging protocols: thermomechanical cycling (TMC), simulated brushing (SB), and control (without aging). Final assessments of color and surface roughness and three-point bending test (ODM100; Odeme) were conducted, and data were statistically analyzed (2-way ANOVA, Tukey, P <.05). RESULTS: Across all resin types, the most significant increase in surface roughness (Ra) was observed after TMC (P < .05), with the 3D-printed resin exhibiting the lowest Ra (P < .05). After brushing, milled resin displayed the highest Ra (P < .05) and greater color alteration (ΔE00) compared to 3D-printed resin. The most substantial ΔE00 was recorded after brushing for all resins, except for heat-cured resin subjected to TMC. Regardless of aging, milled resin exhibited the highest FS (P < .05), except when compared to 3D-printed resin subjected to TMC. Heat-cured resin exposed to TMC demonstrated the lowest FS, different (P < .05) from the control. Under control conditions, milled resin exhibited the highest FS, different (P < .05) from the brushed group. 3D-printed resin subjected to TMC displayed the highest FS (P < .05). CONCLUSION: Among the tested resins, 3D-printed resin demonstrated superior longevity, characterized by minimal surface roughness and color alterations. Aging had a negligible impact on its mechanical properties.

Palmelloid-like phenotype in the alga Raphidocelis subcapitata exposed to pollutants: A generalized adaptive strategy to stress or a specific cellular response?

This work focuses on the formation of palmelloid-like phenotype in the freshwater alga Raphidocelis subcapitata (formerly known as Pseudokirchneriella subcapitata and Selenastrum capricornutum), when exposed to adverse conditions generated by the presence of organic [the antibiotic erythromycin (ERY) and the herbicide metolachlor (MET)] or inorganic [the heavy metals, cadmium (Cd) and zinc (Zn)] pollutants, at environmentally relevant concentrations. This alga in absence of stress or when exposed to ERY or Zn, up to 200 µg/L, essentially showed a single-nucleus state, although algal growth was reduced or stopped. R. subcapitata "switched" to a multinucleated state (palmelloid-like morphology) and accumulated energy-reserve compounds (neutral lipids) when stressed by 100-200 µg/L MET or 200 µg/L Cd; at these concentrations of pollutants, growth was arrested, however, the majority of the algal population (≥83 %) was alive. The formation of palmelloid-like phenotype, at sub-lethal concentrations of pollutants, was dependent on the pollutant, its concentration and exposure time. The multinucleated structure is a transitory phenotype since R. subcapitata population was able to revert to a single-nucleus state, with normal cell size, within 24-96 h (depending on the impact of the toxic in the alga), after being transferred to fresh OECD medium, without pollutants. The obtained results indicate that the formation of a palmelloid-like phenotype in R. subcapitata is dependent on the mode of action of toxics and their concentration, not constituting a generalized defense mechanism against stress. The observations here shown contribute to understanding the different strategies used by the unicellular alga R. subcapitata to cope with severe stress imposed by organic and inorganic pollutants.

Larvicidal activity in vitro of essential oils against Cochliomyia hominivorax.

Cochliomyia hominivorax, from the family Calliphoridae, is responsible for causing most myiases in domesticated animals in Brazil. As alternatives to conventional treatments, essential oils (EOs) could possibly be used as natural control of ectoparasites. The aim of this study was to evaluate the in vitro activity of EOs from Salvia sclarea L., Rosmarinus officinalis L., Lavandula hybrida Reverchon, Citrus bergamia Risso, Citrus paradisi L., Juniperus virginiana L., Copaifera reticulata Ducke, Illicium verum Hook f., Pelargonium roseum, Cymbopogon winterianus Jowitt, Cymbopogon flexuosus Steud, Eugenia caryophyllus Spreng, Cinnamomum cassia Presl, Thymus vulgaris L. and Origanum vulgare L. against third-instar larvae of C. hominivorax. To evaluate the mortality rate, filter paper tests were performed at different concentrations and verified after 24 h and 48 h. Essential oils of T. vulgaris, O. vulgare and I. verum induced 100% mortality in the screening test, and their main active components (thymol, carvacrol and anethol, respectively) were also tested alone. In this new test, lethal concentrations (LC50) after 24 h and 48 h, respectively, were 407.1 and 314.2 µg.cm-² for T. vulgaris, 540.9 and 253.8 µg.cm-² for O. vulgare, 314.2 µg.cm-² (after 48 h) for I. verum, 255.6 and 102.3 µg.cm-² for thymol, 970.5 and 931.1 µg.cm-2 for carvacrol and 559.4 µg.cm-2 (after 48 h) for anethol. These results showed the potential of these EOs and their main components for development of ectoparasiticides for veterinary use, especially T. vulgaris EO and thymol.

Complementary Supramolecular Functionalization Enhances Antifouling Surfaces: A Ureidopyrimidinone-Functionalized Phosphorylcholine Polymer.

Fibrosis of implants remains a significant challenge in the use of biomedical devices and tissue engineering materials. Antifouling coatings, including synthetic zwitterionic coatings, have been developed to prevent fouling and cell adhesion to several implantable biomaterials. While many of these coatings need covalent attachment, a conceptually simpler approach is to use a spontaneous self-assembly event to anchor the coating to a surface. This could simplify material processing through highly specific molecular recognition. Herein, we investigate the ability to utilize directional supramolecular interactions to anchor an antifouling coating to a polymer surface containing a complementary supramolecular unit. A library of controlled copolymerization of ureidopyrimidinone methacrylate (UPyMA) and 2-methacryloyloxyethyl phosphorylcholine (MPC) was prepared and their UPy composition was assessed. The MPC-UPy copolymers were characterized by 1H NMR, Fourier transform infrared (FTIR), and gel permeation chromatography (GPC) and found to exhibit similar mol % of UPy as compared to feed ratios and low dispersities. The copolymers were then coated on an UPy elastomer and the surfaces were assessed for hydrophilicity, protein absorption, and cell adhesion. By challenging the coatings, we found that the antifouling properties of the MPC-UPy copolymers with more UPy mol % lasted longer than the MPC homopolymer or low UPy mol % copolymers. As a result, the bioantifouling nature could be tuned to exhibit spatio-temporal control, namely, the longevity of a coating increased with UPy composition. In addition, these coatings showed nontoxicity and biocompatibility, indicating their potential use in biomaterials as antifouling coatings. Surface modification employing supramolecular interactions provided an approach that merges the simplicity and scalability of nonspecific coating methodology with the specific anchoring capacity found when using conventional covalent grafting with longevity that could be engineered by the supramolecular composition itself.

Isotopic Tracing of Nucleotide Sugar Metabolism in Human Pluripotent Stem Cells.

Metabolism not only produces energy necessary for the cell but is also a key regulator of several cellular functions, including pluripotency and self-renewal. Nucleotide sugars (NSs) are activated sugars that link glucose metabolism with cellular functions via protein N-glycosylation and O-GlcNAcylation. Thus, understanding how different metabolic pathways converge in the synthesis of NSs is critical to explore new opportunities for metabolic interference and modulation of stem cell functions. Tracer-based metabolomics is suited for this challenge, however chemically-defined, customizable media for stem cell culture in which nutrients can be replaced with isotopically labeled analogs are scarcely available. Here, we established a customizable flux-conditioned E8 (FC-E8) medium that enables stem cell culture with stable isotopes for metabolic tracing, and a dedicated liquid chromatography mass-spectrometry (LC-MS/MS) method targeting metabolic pathways converging in NS biosynthesis. By 13C6-glucose feeding, we successfully traced the time-course of carbon incorporation into NSs directly via glucose, and indirectly via other pathways, such as glycolysis and pentose phosphate pathways, in induced pluripotent stem cells (hiPSCs) and embryonic stem cells. Then, we applied these tools to investigate the NS biosynthesis in hiPSC lines from a patient affected by deficiency of phosphoglucomutase 1 (PGM1), an enzyme regulating the synthesis of the two most abundant NSs, UDP-glucose and UDP-galactose.

Insecticidal activity of essential oil of Cannabis sativa against the immature and adult stages of Ctenocephalides felis felis.

Essential oil (EO) of Cannabis sativa (C. sativa) was evaluated against the egg, larval, pupal, and adult stages of the flea Ctenocephalides felis felis. The chemical composition of EO was determined by gas chromatography with flame ionization and mass spectrometry. EO mainly comprised γ-elemene (16.2%) and caryophyllene oxide (14.2%) as major compounds. To evaluate the mortality of flea stages in vitro, filter paper tests were performed at different concentrations. EO of C. sativa showed insecticidal activity (100% mortality at the highest concentrations) for flea control at egg, larval, pupal, and adult stages, with lethal concentrations (LC50) of 32.45; 91.61; 466.41 and 927.92 µg/cm2, respectively. EO of C. sativa indicated the potential for the development of ectoparasiticide for veterinary use, especially for fleas in egg and larval stages.

Surgically assisted maxillary expansion with or without pterygoid disjunction alters maxillomandibular positioning.

PURPOSE: This study aimed to analyze alterations in mandibular positioning after surgically assisted maxillary expansion (SARME) with and without pterygoid disjunction (PD). METHODS: Cone-beam computed tomography scans of 24 healthy individuals (18-45 years old) with transverse deficiency, superior to 5 mm, underwent SARME with or without PD. The aspects prospectively assessed were (1) alignment and position of the head (ITK-Snap and 3D Slicer software); (2) McNamara's and Steiner-Tweed-Wits' cephalometric analysis (Dolphin Imaging®); and (3) colorimetric evaluation based on 3D correspondence analysis (3D Slicer software). RESULTS: A decrease in 1-NA and 1-SN angles as well as an increased occlusal plane in both groups was observed. Superior-inferior and anteroposterior spatial displacements of the chin were statistically significant in the PD group. Altered colorimetric patterns were also observed in the PD group. CONCLUSIONS: This study found more evident tooth inclination in the group without PD; mandibular alterations were more evident in the PD group. Further studies with 3D analysis are strongly recommended for more comprehensive results.

Incorporation of bioactive glass-ceramic into coconut oil for remineralization of incipient carious lesions

Abstract This study evaluated the efficacy of incorporating different concentrations of bioactive glass-ceramic (Biosilicate) into coconut oil on the remineralizing potential and surface roughness of white spot lesions. Fragments (6 x 6 x 2mm) of bovine teeth were sectioned and initial microhardness (KHN) and surface roughness (Ra) readings were obtained. The samples were submitted to cariogenic challenge to form white spot lesions and were separated into six groups (n=13): 1) Artificial Saliva (AS); 2) Coconut Oil (CO); 3) CO+2% Biosilicate (CO+2%Bio); 4) CO+5% Biosilicate (CO+5%Bio); 5) 2% Biosilicate Suspension (2% Bio) and 6) 5% Biosilicate Suspension (5% Bio). The treatments for 1 cycle/day were: immersion into the treatments for 5 minutes, rinsing in distilled water, and storage in artificial saliva at 37ºC. After 14 days, KHN and Ra readings were taken. The surface roughness alteration ((Ra) was analyzed (Kruskal-Wallis, Dunn's post-test, p<0.05). CO+2%Bio had higher (p = 0.0013) (Ra followed by CO+5%Bio (p = 0.0244) than AS. The relative KHN and remineralization potential were analyzed (ANOVA, Tukey, p<0.05), and 5% Bio treatment presented a higher relative microhardness than all other groups (p>0.05). The remineralizing potential of all the treatments was similar (p > .05). When Biosilicate was added, the pH of the suspensions increased and the alkaline pH remained during the analysis. Biosilicate suspension is more efficient than the incorporation of particles into coconut oil at white spot lesion treatment. In addition to the benefits that coconut oil and Biosilicate present separately, their association can enhance the remineralizing potential of Biosilicate.

Resumo Este estudo avaliou a eficácia da incorporação de diferentes concentrações de vitrocerâmica bioativa (biosilicato) ao óleo de coco no potencial remineralizante e na rugosidade superficial de lesões de manchas brancas. Fragmentos (6 x 6 x 2mm) de dentes bovinos foram seccionados e as leituras iniciais de microdureza (KHN) e rugosidade superficial (Ra) foram obtidas. As amostras foram submetidas ao desafio cariogênico para formação de lesões de manchas brancas e foram separadas em seis grupos (n=13): 1) Saliva Artificial (AS); 2) Óleo de Coco (CO); 3) CO+2% Biosilicato (CO+2%Bio); 4) CO+5% Biosilicato (CO+5%Bio); 5) Suspensão de Biosilicato 2% (2% Bio) e 6) Suspensão de Biosilicato 5% (5% Bio). Os tratamentos de 1 ciclo/dia foram: imersão nos tratamentos por 5 minutos, enxágue em água destilada e armazenamento em saliva artificial a 37ºC. Após 14 dias, foram feitas as leituras de KHN e Ra. A alteração da rugosidade superficial ((Ra) foi analisada (Kruskal-Wallis, pós-teste de Dunn, p<0,05). CO+2%Bio apresentou maior (p = 0,0013) (Ra seguido de CO+5%Bio (p = 0,0244) do que AS. O KHN relativo e o potencial de remineralização foram analisados (ANOVA, Tukey, p<0,05), e o tratamento 5% Bio apresentou uma microdureza relativa maior do que todos os outros grupos (p>0,05). A suspensão de biosilicato é mais eficiente que a incorporação de partículas ao óleo de coco no tratamento de lesões de mancha branca. Além dos benefícios que o óleo de coco e o Biosilicato apresentam separadamente, sua associação pode amplificar o potencial remineralizante do Biosilicato.

Life and death of Pseudokirchneriella subcapitata: physiological changes during chronological aging.

The green alga Pseudokirchneriella subcapitata is widely used in ecotoxicity assays and has great biotechnological potential as feedstock. This work aims to characterize the physiology of this alga associated with the aging resulting from the incubation of cells for 21 days, in the OECD medium, with continuous agitation and light exposure, in a batch mode. After inoculation, cells grow exponentially during 3 days, and the culture presents a typical green color. In this phase, "young" algal cells present, predominantly, a lunate morphology with the chloroplast occupying a large part of the cell, maximum photosynthetic activity and pigments concentration, and produce starch as a reserve material. Between the 5th and the 12th days of incubation, cells are in the stationary phase. The culture becomes less green, and the cells stop dividing (≥ 99% have one nucleus) and start to age. "Old" algal cells present chloroplast shrinkage, an abrupt decline of chlorophylls content, and photosynthetic capacity (Fv/Fm and ɸPSII), accompanied by a degradation of starch and an increase of neutral lipids content. The onset of the death phase occurs after the 12th day and is characterized by the loss of cell membrane integrity of some algae (cell death). The culture stays, progressively, yellow, and the majority of the population (~93%) is composed of live cells, chronologically "old," with a significant drop in photosynthetic activity (decay > 75% of Fv/Fm and ɸPSII) and starch content. The information here achieved can be helpful when exploring the potential of this alga in toxicity studies or in biotechnological applications. KEY POINTS: • Physiological changes of P. subcapitata with chronological aging are shown • "Young" algae exhibit a semilunar shape, high photosynthetic activity, and accumulated starch • "Old"-live algae show reduced photosynthetic capacity and accumulated lipids.

Effect of thermal processing on the antigenicity of allergenic milk, egg and soy proteins.

The detection of allergenic proteins and the influence of processing on the structure and antigenicity of these proteins are relevant topics. Using commercial enzyme-linked immunosorbent assay kits, this study aimed to evaluate the degradation profiles of milk, egg and soy proteins during the processing of semisweet biscuits. The formulations were baked under different conditions according to a complete factorial experiment that included a three-level temperature factor and a six-level time factor. ß-lactoglobulin and egg white proteins were severely degraded, the degradation of casein was intermediate, and soy proteins were the most stable. Complete allergen protein degradation was found under only the extreme baking conditions, which resulted in products that were not sensorily acceptable. Residual levels of the proteins were detected after baking, indicating that this thermal processing reduced but did not eliminate the antigenicity of these proteins; thus, baking cannot be considered a strategy to protect allergic consumers.

Perspective on the biotechnological production of bacterial siderophores and their use.

Iron (Fe) is an essential element in several fundamental cellular processes. Although present in high amounts in the Earth's crust, Fe can be a scarce element due to its low bioavailability. To mitigate Fe limitation, microorganism (bacteria and fungi) and grass plant biosynthesis and secret secondary metabolites, called siderophores, with capacity to chelate Fe(III) with high affinity and selectivity. This review focuses on the current state of knowledge concerning the production of siderophores by bacteria. The main siderophore types and corresponding siderophore-producing bacteria are summarized. A concise outline of siderophore biosynthesis, secretion and regulation is given. Important aspects to be taken into account in the selection of a siderophore-producing bacterium, such as biological safety, complexing properties of the siderophores and amount of siderophores produced are summarized and discussed. An overview containing recent scientific advances on culture medium formulation and cultural conditions that influence the production of siderophores by bacteria is critically presented. The recovery, purification and processing of siderophores are outlined. Potential applications of siderophores in different sectors including agriculture, environment, biosensors and the medical field are sketched. Finally, future trends regarding the production and use of siderophores are discussed. KEY POINTS : • An overview of siderophore production by bacteria is critically presented • Scientific advances on factors that influence siderophores production are discussed • Potential applications of siderophores, in different fields, are outlined.

Pluripotent Stem Cell Differentiation Toward Functional Basal Stratified Epithelial Cells.

In this chapter, an efficient feeder-free protocol of differentiating human pluripotent stem cells (hPSCs) toward the epidermal lineage to generate induced epidermal keratinocytes (iKCs) is described. The iKCs are able to terminally differentiate supra-basally. This hPSC-to-iKC differentiation can serve as a useful model to study epidermal development and disease as well as for therapeutic applications.

Imidazolium salts as an alternative for anti-Leishmania drugs: Oxidative and immunomodulatory activities.

In this study we explored the previously established leishmanicidal activity of a complementary set of 24 imidazolium salts (IS), 1-hexadecylimidazole (C16Im) and 1-hexadecylpyridinium chloride (C16PyrCl) against Leishmania (Leishmania) amazonensis and Leishmania (Leishmania) infantum chagasi. Promastigotes of L. amazonensis and L. infantum chagasi were incubated with 0.1 to 100 µM of the compounds and eight of them demonstrated leishmanicidal activity after 48 h - C10MImMeS (IC50 L. amazonensis = 11.6), C16MImPF6(IC50 L. amazonensis = 6.9), C16MImBr (IC50 L. amazonensis = 6), C16M2ImCl (IC50 L. amazonensis = 4.1), C16M4ImCl (IC50 L. amazonensis = 1.8), (C10)2MImCl (IC50 L. amazonensis = 1.9), C16Im (IC50 L. amazonensis = 14.6), and C16PyrCl (IC50 L. amazonensis = 4).The effect of IS on reactive oxygen species production, mitochondrial membrane potential, membrane integrity and morphological alterations of promastigotes was determined, as well as on L. amazonensis-infected macrophages. Their cytotoxicity against macrophages and human erythrocytes was also evaluated. The IS C10MImMeS, C16MImPF6, C16MImBr, C16M2ImCl, C16M4ImCl and (C10)2MImCl, and the compounds C16Im and C16PyrCl killed and inhibited the growth of promastigote forms of L. amazonensis and L. infantum chagasi in a concentration-dependent manner, contributing to a better understanding of the structure-activity relationship of IS against Leishmania. These IS induced ROS production, mitochondrial dysfunction, membrane disruption and morphological alterations in infective forms of L. amazonensis and killed intracellular amastigote forms in very low concentrations (IC50 amastigotes ≤ 0.3), being potential drug candidates against L. amazonensis.