Navegação por Autores IPEN "CABRAL, FERNANDA V."

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  • IPEN-DOC 23242

    CABRAL, FERNANDA V. ; RIBEIRO, MARTHA S. ; SAUTER, ISMAEL P.; VELIZ, MAURO J.C.. Comparison of protocols in methylene blue-mediated photodynamic inactivation on cutaneous leishmaniasis in a murine model using real time bioluminescence. In: WORLD CONGRESS ON LEISHMANIASIS, 6th, May 16-20, 2017, Toledo, Spain. Abstract... 2017. p. 749-749.

    Abstract: 1 Background Leishmaniasis is a chronic disease developed by parasites of the genus Leishmania. Cutaneous leishmaniasis (CL) has several different forms including destructive and ulcerated lesions. The available treatments are limited because of side effects, resistance and toxicity. Photodynamic therapy (PDT) has been explored as an alternative treatment once it is less expensive and no reports about resistance have been described. The aim of this work was to evaluate two protocols of methylene blue (MB)-mediated PDT on CL induced in mice using real time bioluminescence. In addition, we monitored lesion size progression and hyperalgesia. 2 Methods Promastigotes of L. (L) amazonensis transgenic line expressing luciferase were used. Twelve BALB/c mice were infected in the left footpad with 1.106 promastigotes. After 4 weeks, mice were randomly assigned to experimental groups (n=4): Control (non-treated), G1 and G2 submitted to one and two PDT sessions, respectively. The second session was performed 24h after the first. PDT was performed using a red LED (λ= 660±22 nm), MB (100μM) and 150J/cm² fluence. Disease progression was evaluated once a week by measuring lesion size with a caliper and hyperalgesia with von Frey filaments. Parasite burden was evaluated through luciferase detection by bioimaging, every day, in the first 96h and then for the next 4 weeks. Prior to imaging, mice received 75mg/kg luciferin, intraperitoneally. Results were quantified with proper software, expressing the number of photons/s/cm2. Statistically significant differences were considered when p<0.05. 3 Results Statistically significant lesion size reduction was observed after 2 weeks post PDT compared to G1 and control groups. In 4 weeks, G2 demonstrated to be clinically more effective and no ulcer signs were detected. Hyperalgesia revealed a positive outcome after 2 weeks, with 50% nociceptive stimulus decrease. In 3 weeks there was a 75% reduction in G2, suggesting that 2 sessions were more efficient. Parasite burden was lower in the first 24h and 72h in both treated groups. Besides, G2 demonstrated statistically significant parasite burden reduction 96h after PDT compared to control. 4 Conclusions The present study demonstrated that PDT reduced lesion size and hyperalgesia in BALB/c mice, suggesting that therapy probably influenced the inflammatory process. However, further studies are required to deep understanding about PDT mechanisms in cutaneous leishmaniasis treatment.

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  • IPEN-DOC 29637

    YOSHIMURA, TANIA M. ; CABRAL, FERNANDA V. ; SELLERA, FABIO P.; POZZO, LORENA ; RIBEIRO, MARTHA S. . Could light-based technologies improve stem cell therapy for skin wounds?: A systematic review and meta-analysis of preclinical studies. Photochemistry and Photobiology, v. 99, n. 2, p. 519-528, 2023. DOI: 10.1111/php.13702

    Abstract: Several diseases or conditions cause dermatological disorders that hinder the process of skin repair. The search for novel technologies has inspired the combination of stem cell (SC) and light-based therapies to ameliorate skin wound repair. Herein, we systematically revised the impact of photobiomodulation therapy (PBM) combined with SCs in animal models of skin wounds and quantitatively evaluated this effect through a meta-analysis. For inclusion, SCs should be irradiated in vitro or in vivo, before or after being implanted in animals, respectively. The search resulted in nine eligible articles, which were assessed for risk of bias. For the meta-analysis, studies were included only when PBM was applied in vivo, five regarding wound closure, and three to wound strength. Overall, a positive influence of SC + PBM on wound closure (mean difference: 9.69; 95% CI: 5.78–13.61, P < 0.00001) and strength (standardized mean difference: 1.7, 95% CI: 0.68–2.72, P = 0.001) was detected, although studies have shown moderate to high heterogeneity and a lack of information regarding some bias domains. Altogether, PBM seems to be an enabling technology able to be applied postimplantation of SCs for cutaneous regeneration. Our findings may guide future laboratory and clinical studies in hopes of offering wound care patients a better quality of life.

    Palavras-Chave: skin diseases; therapy; stem cells; biological repair

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  • IPEN-DOC 29539

    SALVEGO, CAMILA de A. ; CABRAL, FERNANDA V. ; SEABRA, AMEDEA B.; SANTANA, BIANCA de M.; RIBEIRO, MARTHA S. . Cytotoxic effects of chitosan nanoparticles containing S-nitrosoglutathione in triple-negative breast cancer cells. In: ANNUAL MEETING OF THE BRAZILIAN SOCIETY FOR BIOCHEMISTRY AND MOLECULAR BIOLOGY (SBBq), 51st; CONGRESS OF BRAZILIAN BIOPHYSICAL SOCIETY (SBBf)/LATIN AMERICAN FEDERATION OF BIOPHYSICAL SOCIETIES (Lafebs), 46th, September 5-8, 2022, Águas de Lindóia, SP. Abstract... São Paulo, SP: Sociedade Brasileira de Bioquímica e Biologia Molecular - SBBq, 2022. p. 237-237.

    Abstract: INTRODUCTION: Breast cancer is the most common type of cancer affecting women worldwide. Among the treatments, radiation therapy (RT) is frequently chosen as a primary strategy; however, it demands high doses of ionizing radiation to achieve a curative dose. To enhance RT effectiveness, an external agent can be used to sensitize cells before the treatment, allowing a dose reduction. Nitric oxide (NO) is an essential molecule linked to several organic processes, besides being described as a potential radiosensitizer of tumor cells by different mechanisms, including oxidative stress. However, NO have a short half-life in biological conditions, making it difficult to achieve anticancer effects. To overcome this, NO donors can be encapsulated into polymer-based nanoparticles, ensuring a sustained NO releasing. OBJECTIVES: To evaluate the cytotoxicity induced by chitosan nanoparticles containing S-nitrosoglutathione (GSNO-CS NPs) in 4T1 cells (murine triple-negative breast cancer). MATERIALS AND METHODS: Cells were cultivated, seeded in 96-well plates (2 x 10 4 cells/well), and incubated at 37ºC with 5% of CO2 for 24 h. Both CS NPs and CS NPs containing GSNO encapsulated were added to the plates at different concentrations (0-2.4 mg/ml CS NPs, 0-6 mM GSNO) and incubated for 24 h. Cytotoxic effects were evaluated through Resazurin fluorometric assay in both groups. DISCUSSION AND RESULTS: Our results showed a 65% reduction in cell viability for GSNO-CS NPs groups treated at 6 mM, while only 30% of cells were killed when treated by CS NPs group. CONCLUSION: Our data suggest that GSNO-CS NPs were able to promote cytotoxicity effects, thus inducing oxidative stress in triple-negative breast cancer cells. Next steps involve the use of these nanoparticles before RT to evaluate its radiosensitizer effect.

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  • IPEN-DOC 26934

    CABRAL, FERNANDA V. ; SILVA, CAMILA R. ; SAUTER, ISMAEL P.; SABINO, CAETANO P. ; YOSHIMURA, TANIA M. ; CORTEZ, MAURO; RIBEIRO, MARTHA S. . Efeito da terapia fotodinâmica no tratamento de leishmaniose cutânea em um modelo murino. Anais da SBBN, v. 3, p. 67-77, 2016.

    Abstract: Leishmanioses são doenças parasitárias desenvolvidas por protozoários do gênero Leishmania. A forma cutânea abrange lesões destrutivas e ulceradas com diversas limitações no tratamento. Terapias alternativas são fundamentais devido à grande relevância da doença e elevada incidência. Nesse contexto, a terapia fotodinâmica (PDT) tem sido introduzida devido ao baixo custo, baixa toxicidade, praticidade e sem relatos de resistência na literatura. Neste trabalho, dezesseis camundongos BALB/c foram infectados com 1.106 parasitos de Leishmania(L) amazonensisno membro posterior esquerdo e acompanhados por 4 semanas até o surgimento da lesão. Após esse período, os animais foram submetidos à PDT usando um diodo emissor de luz (λ=660 ±22 nm) e azul de metileno (100 μM), com fluências de 50, 100 e 150J/cm² e acompanhados durante 3 semanas. Animais controle não receberam tratamento. O tamanho da lesão e escala de dor foram mensurados utilizando um paquímetro e filamentos von Frey, respectivamente. A quantificação da carga parasitária foi realizada através do método de diluição limitante. Os resultados demonstraram que, nas semanas 2 e 3 após tratamento, ocorreu diminuição da lesão e alívio de dor nos animais que receberam 150J/cm², sugerindo que a PDT promoveu melhora clínica através da modulação do processo inflamatório.

    Palavras-Chave: photosensitivity; methylene blue; parasites; parasitic diseases; biological models; epithelium; light sources; light emitting diodes; therapy; skin diseases

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  • IPEN-DOC 24631

    CABRAL, FERNANDA V. ; PELLEGRINO, MILENA T.; SEABRA, AMEDEA B.; RIBEIRO, MARTHA S. . Efeitos da terapia fotodinâmica antimicrobiana associada a nanopartículas doadoras de óxido nítrico em leishmaniose cutânea. In: ANAIS DA SOCIEDADE BRASILEIRA DE BIOCIENCIAS NUCLEARES, 09-11 de outubro, 2017, São Paulo, SP. Resumo... Rio de Janeiro, RJ: Sociedade Brasileira de Biociências Nucleares, 2017. p. 94-94.

    Abstract: Introdução: A leishmaniose cutânea (LC) é uma doença crônica desenvolvida por parasitas do gênero Leishmania que promove lesões destrutivas e ulceradas. Os tratamentos disponíveis são limitados devido a efeitos colaterais, resistência e toxicidade. A inativação fotodinâmica (PDI) envolve a geração de estresse oxidativo e tem sido explorada como tratamento alternativo para LC. Além disso, estudos indicam que a administração de doadores de NO exógenos representa uma estratégia interessante contra LC. Objetivos: O propósito deste trabalho foi explorar os efeitos da PDI mediada por azul de metileno (AM) em associação com doadores de NO encapsulados em nanopartículas de quitosana (CSNP) em LC induzida em modelo murino utilizando bioluminescência em tempo real. Métodos: Projeto aprovado pela CEUA/IPEN 189/17. Utilizamos promastigotas de linhagem transgênica de Leishmania (L.) amazonensis expressando luciferase. Dezesseis camundongos BALB/c foram infectados na pata esquerda com 1.106 promastigotas. Após 4 semanas, os animais foram distribuídos aleatoriamente em 4 grupos experimentais (n=4): Controle (não tratado), PDI, PDI+CSNP e CSNP. A PDI foi administrada em duas sessões espaçadas de 24 horas e as CSNPs (80 μM) foram aplicadas imediatamente após a segunda sessão de PDI. A PDI foi realizada utilizando-se um LED vermelho (λ=660±22 nm), AM (100μM), irradiância de 100 mW/cm2 e exposição radiante de 150 J/cm². A carga parasitária foi analisada através da detecção da luciferase por bioimageamento nas primeiras 96 horas após o tratamento e nas 4 semanas subsequentes. Estatística: análise de variância com teste Tukey como pós-teste e diferenças entre grupos foram consideradas estatisticamente significativas quando p <0,05. Resultados: O grupo teste apresentarou redução significativa na carga parasitária em relação ao controle durante o experimento. 24 h após os tratamentos, a carga parasitária no grupo PDI+CSNP (1,1.106 ±9,6.105) reduziu em relação ao controle (1,3.107±1,1 .107) e continuou significativamente menor em 48 horas (1,8.106±1,4.106, PDI+CSNP e 2,7.107±1,4.107, controle), mas não foram observadas diferenças estatisticamente significantes quando comparada aos outros grupos teste. Após 72 h, todos os grupos tratados foram semelhantes com redução significativa em relação ao controle (controle 2,6.107±1,4.107, PDI+CSNP 4,6.106±3,4.106 e CSNP 1,9.107±4,8.107). A carga parasitária nos grupos teste permaneceu aproximadamente menor que o controle após 1, 2, 3 e 4 semanas pós-tratamento. Conclusão: Sob as condições utilizadas neste estudo, concluímos que as CSNPs não foram capazes de aumentar a eficiência da PDI mediada por azul de metileno em CL induzida por L. (L) amazonensis em camundongos.

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  • IPEN-DOC 27775

    SOUZA, TIAGO H.S.; ANDRADE, CAMILA G.; CABRAL, FERNANDA V. ; SARMENTO-NETO, JOSE F.; REBOUÇAS, JULIO S.; SANTOS, BEATE S.; RIBEIRO, MARTHA S. ; FIGUEIREDO, REGINA C.B.Q.; FONTES, ADRIANA. Efficient photodynamic inactivation of Leishmania parasites mediated by lipophilic water-soluble Zn(II) porphyrin ZnTnHex-2-PyP4+. BBA - General Subjects, v. 1865, n. 7, p. 1-10, 2021. DOI: 10.1016/j.bbagen.2021.129897

    Abstract: Background Photodynamic inactivation (PDI) is emerging as a promising alternative for cutaneous leishmaniasis (CL). The chemotherapy currently used presents adverse effects and cases of drug resistance have been reported. ZnTnHex-2-PyP4+ is a porphyrin with a high potential as a photosensitizer (PS) for PDI, due to its photophysical properties, structural stability, and cationic/amphiphilic character that can enhance interaction with cells. This study aimed to investigate the photodynamic effects mediated by ZnTnHex-2-PyP4+ on Leishmania parasites. Methods ZnTnHex-2-PyP4+ stability was evaluated using accelerated solvolysis conditions. The photodynamic action on promastigotes was assessed by (i) viability assays, (ii) mitochondrial membrane potential evaluation, and (iii) morphological analysis. The PS-promastigote interaction was studied. PDI on amastigotes and the cytotoxicity on macrophages were also analyzed. Results ZnTnHex-2-PyP4+, under submicromolar concentration, led to immediate inactivation of more than 95% of promastigotes. PDI promoted intense mitochondrial depolarization, loss of the fusiform shape, and plasma membrane wrinkling in promastigotes. Fluorescence microscopy revealed a punctate PS labeling in the parasite cytoplasm. PDI also led to reductions of ca. 64% in the number of amastigotes/macrophage and 70% in the infection index after a single treatment session. No noteworthy toxicity was observed on mammalian cells. Conclusions ZnTnHex-2-PyP4+ is stable against demetallation and more efficient as PS than the ethyl analogue ZnTE-2-PyP4+, indicating readiness for evaluation in in vivo studies as an alternative approach to CL. General significance This report highlighted promising photodynamic effects mediated by ZnTnHex-2-PyP4+ on Leishmania parasites, opening up perspectives for applications in CL pre-clinical assays and PDI of other microorganisms.

    Palavras-Chave: inactivation; photosensitivity; photochemistry; antimicrobial agents; therapy; parasites; parasitic diseases; porphyrins

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  • IPEN-DOC 23093

    CABRAL, FERNANDA V. . Estudo da terapia fotodinâmica no tratamento de Leishmaniose cutânea em modelo murino / Evaluation of photodynamic therapy on cutaneous Leishmaniasis in a murine model . 2017. Dissertação (Mestrado em Tecnologia Nuclear) - Instituto de Pesquisas Energéticas e Nucleares - IPEN-CNEN/SP, São Paulo. 100 p. Orientador: Martha Simões Ribeiro. DOI: 10.11606/D.85.2017.tde-05052017-145507

    Abstract: Leishmaniose é uma zoonose desenvolvida por protozoários do gênero Leishmania. A doença se manifesta sob a forma visceral e cutânea cujo tratamento apresenta diversas limitações como alto custo, elevada toxicidade dos fármacos e altos índices de recidiva. A leishmaniose cutânea abrange lesões destrutivas e ulceradas que podem evoluir para condições mais graves culminando em óbito dos hospedeiros acometidos. Tratamentos alternativos tem sido implementados com a finalidade de proporcionar acessibilidade financeira e menores efeitos colaterais aos pacientes. A terapia fotodinâmica se insere nesse contexto devido à praticidade, custo reduzido, mínima toxicidade e sem relatos de resistência descritos na literatura. O objetivo desse trabalho foi avaliar os efeitos da terapia fotodinâmica em leishmaniose cutânea induzida em camundongos BALB/c infectados com leishmaniose cutânea. A otimização da PDT também foi avaliada pela administração de doadores de óxido nítrico (S-nitroso-MSA) encapsulados em nanopartículas de quitosana (CSNPs), já que esse composto é altamente reativo e potencialmente tóxico para o parasito. Camundongos BALB/c foram infectados no membro posterior esquerdo com 1.106 promastigotas de Leishmania (L) amazonensis que expressam o gene da luciferase e acompanhados por 4 semanas até o surgimento da lesão. Após esse período, os animais foram distribuídos em 6 grupos (n=4): Controle (não tratado), PDT1 (submetidos à uma sessão de PDT), PDT2 (duas sessões ), PDT1NPNO e PDT2NPNO (uma sessão e duas sessões de PDT em associação com S-nitroso-MSACPNPS, respectivamente), e NPNO (somente S-nitroso-MSA-CP NPS). A segunda sessão foi realizada 24 h após a primeira. A PDT foi efetuada usando um diodo emissor de luz (LED, &lambda;=660 &plusmn; 22 nm) e azul de metileno (100 &mu;M), com densidade de energia de 150 J/cm2. O progresso da doença foi avaliado por meio do tamanho da lesão e escala de dor utilizando um paquímetro e filamentos von Frey, respectivamente. A carga parasitária foi quantificada por intermédio do bioimageamento nas primeiras 96 h após o tratamento e nas 4 semanas seguintes. Os resultados obtidos demonstraram redução na carga parasitária durante o período experimental, com exceção do grupo PDT1NPNO. Houve redução parasitária significante em 72 h e 96 h para os grupos PDT2, PDT2NPNO e NPNO. A maior redução da lesão foi observada para o grupo PDT2 bem como menor sensibilidade ao estímulo doloroso. Nossos resultados indicam efeitos benéficos da PDT em duas sessões, sugerindo que pode ter ocorrido modulação do processo inflamatório. Entretanto, o uso das nanopartículas nas condições utilizadas nesse experimento não foi capaz de otimizar a eficiência da PDT nos animais infectados com Leishmania (L) amazonensis.

    Palavras-Chave: photosensitivity; skin diseases; parasitic diseases; loaders; clinical trials; mice; in vivo; in vitro; radiotherapy; nitrous acid; sulfuric acid; nanomaterials; light emitting diodes

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  • IPEN-DOC 25289

    CABRAL, FERNANDA V. ; PELEGRINO, MILENA T.; DIMMER, JESICA A.; SEABRA, AMEDEA B.; RIBEIRO, MARTHA S. . Evaluation of methylene blue-mediated photodynamic inactivation in association with encapsulated nitric oxide donor in chitosan nanoparticles on Leishmania (L.) amazonensis. An in vitro study. In: DAI, TIANHONG (Ed.) LIGHT-BASED DIAGNOSIS AND TREATMENT OF INFECTIOUS DISEASES, January 27 - February 01, 2018, San Francisco, CA, USA. Abstract... Bellingham, WA, USA: Society of Photo-optical Instrumentation Engineers, 2018. (SPIE Proceedings Series, 10479).

    Abstract: Cutaneous leishmaniasis (CL) is a chronic disease developed by parasites of the genus Leishmania that promotes destructive lesions. The available treatments are limited because of resistance and toxicity. Reactive oxygen species and nitric oxide (NO) are potentially toxic to these parasites. Photodynamic inactivation (PDI) has been explored as an alternative treatment once no reports about resistance have been described. Additionally, several studies indicate that the administration of exogenous NO donors represents an interesting strategy against CL. The aim of this work was to explore the effects of methylene blue (MB)-mediated PDI in association with encapsulated NO donors (S-nitroso-MSA) in chitosan nanoparticles (CSNPs) on Leishmania (L.) amazonensis. S-nitroso-MSA-CSNPs were tested with L. (L) amazonensis transgenic line expressing luciferase (La-LUC) at 25 μM, 50 μM, 75 μM, and 100 μM. PDI was perfomed using a red LED (λ= 66022 nm) at fluences of 12.5, 25, 37.5 and 50 J/cm² and MB at 100 μM. The association of both therapies was performed using 25 μM of S-nitroso-MSA-CSNP immediately after PDI at 25 J/cm² fluence. Results demonstrated a 50% decrease in La-LUC metabolic activity with 25 μM S-nitroso-MSA-CSNP and a 70% reduction with 25 J/cm² fluence when the tests were performed separately. However, the association with S-nitroso-MSA-CSNP showed 97% reduction of the parasite burden. The present study demonstrates that encapsulated S-nitroso-MSA-CSNPs were able to improve the effects of PDI on Leishmania (L.) amazonensis, which suggests that both therapies combined could be a potential alternative treatment for CL.

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  • IPEN-DOC 23565

    SILVA, CAMILA R. ; CABRAL, FERNANDA V. ; CAMARGO, CLAUDINEI F.M. de ; NUNEZ, SILVIA C. ; YOSHIMURA, TANIA M. ; LUNA, ARTHUR C. de L.; MARIA, DURVANEI A.; RIBEIRO, MARTHA S. . Exploring the effects of low-level laser therapy on fibroblasts and tumor cells following gamma radiation exposure. Journal of Biophotonics, v. 9, n. 11-12, p. 1157-1166, 2016. DOI: 10.1002/jbio.201600107

    Abstract: Ionizing radiation (IR) induces DNA damage and low-level laser therapy (LLLT) has been investigated to prevent or repair detrimental outcomes resulting from IR exposure. Few in vitro studies, however, explore the biological mechanisms underlying those LLLT benefits. Thus, in this work, fibroblasts and tumor cells are submitted to IR with doses of 2.5 Gy and 10 Gy. After twenty-four-h, the cells are exposed to LLLT with fluences of 30 J cm–2, 90 J cm–2, and 150 J cm–2. Cellular viability, cell cycle phases, cell proliferation index and senescence are evaluated on days 1 and 4 after LLLT irradiation. For fibroblasts, LLLT promotes – in a fluence-dependent manner – increments in cell viability and proliferation, while a reduction in the senescence was observed. Regarding tumor cells, no influences of LLLT on cell viability are noticed. Whereas LLLT enhances cell populations in S and G2/M cell cycle phases for both cellular lines, a decrease in proliferation and increase in senescence was verified only for tumor cells. Putting together, the results suggest that fibroblasts and tumor cells present different responses to LLLT following exposure to gamma-radiation, and these promising results should stimulate further investigations. Senescence of tumor cells and fibroblasts on the 4th day after ionizing radiation (IR) and low-level laser therapy (LLLT) exposures. The number of senescent cells increased significantly for tumor cells (a) while for fibroblasts no increment was observed (b). The blue collor indicates senescence activity.

    Palavras-Chave: gamma radiation; tumor cells; ionizing radiations; laser radiation; mammary glands; neoplasms; tumor cells; therapy; radiotherapy; levels

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  • IPEN-DOC 29091

    CABRAL, FERNANDA V. ; SELLERA, FABIO P.; RIBEIRO, MARTHA S. . Feline sporotrichosis successfully treated with methylene blue-mediated antimicrobial photodynamic therapy and low doses of itraconazole. Photodiagnosis and Photodynamic Therapy, v. 40, p. 1-3, 2022. DOI: 10.1016/j.pdpdt.2022.103154

    Abstract: Sporotrichosis is a mycotic infection of humans and animals caused by different fungal species of the genus Sporothrix. Feline sporotrichosis presents a broad spectrum of clinical manifestations and its treatment with classic antifungal drugs is often long and frustrating. Methylene blue-mediated antimicrobial photodynamic therapy (MB-APDT) comes to light as an interesting approach against fungal infections, including sporotrichosis. In this case report, a 1-year-old male cat was diagnosed with sporotrichosis, being confirmed by fungal culture. The cat was treated by MB-APDT combined with oral administration of itraconazole. Following 2 weeks after the end of treatment, the animal was clinically cured, and an additional fungal culture was negative for Sporothrix spp., confirming the total remission of sporotrichosis. No side effects and recurrences were observed after a 3-moth follow-up. MB-APDT is a promising strategy against feline sporotrichosis, however large-scale studies are welcome to confirm its potential.

    Palavras-Chave: veterinary medicine; fungal diseases; photodynamic therapy; antimicrobial agents; fungi; cats; methylene blue

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  • IPEN-DOC 29319

    WETTER, NIKLAUS ; FREITAS, ANDERSON Z. ; BERECZKI, ALLAN ; PASCOAL, DIEGO R.C. ; DIPOLD, JESSICA ; PARRA, DUCLERC ; ROSSI, WAGNER ; CABRAL, FERNANDA V. ; RIBEIRO, MARTHA S. . From micro to nano Raman: new developments in the detection of micro and nanoplastics of marine origin from the Amazon basin to Santos. In: ENCONTRO BRASILEIRO DE ESPECTROSCOPIA RAMAN, 7., 5-8 de dezembro, 2022, São Pedro, SP. Resumo... 2022.

    Abstract: The environmental accumulation of nanoplastics formed by material of anthropic origin has raised doubts about their safety, especially to the human body. While microplastics are accidentally consumed, nanoplastics (NPs) are even more concerning as they are much more likely to be absorbed by human body cells. It is known that plastics smaller than 200 nm can penetrate cell membranes and cross the blood-brain barrier. Studies have shown that polystyrene (PS) NPs from the environment carry a high load of toxins capable of compromising human brain cells. Very little is still known about what effects, cytotoxic or not, these plastics have on different organs. Understanding the property-function relationship of nanoparticles in various fields of application involves determining their physicochemical properties, which is still a formidable challenge to date. Our project focuses on the development of a methodology for the detection of micro- and NPs using micro-Raman, TERS (Tip Enhanced Raman Spectroscopy), collinear Raman and AFM, nuclear techniques, as well as a methodology for in vitro evaluation of the toxic effects of these materials through biochemical assays of cytotoxicity and genotoxicity. The project contemplates the determination of the adsorption capacity of metallic ions by NPs and the absorption of micro- and nanoplastics in cell cultures with radioactive tracers, the determination of microplastics in tissues from necropsies of marine animals and gamma spectroscopy of the cellular incorporation of NPs labeled with radioactive isotopes. In figure 1 we show examples of (a) the detection of microplastics from the Santos basin and comparison to Raman reference database (KnowItAll®), (b) detection of NPs in mouse fibroblast cells and (c) detection of very small particles (50 nm) which can be achieved by co-localized techniques of AFM + Raman using special, narrow (50 m), femtosecond laser written gratings on quartz.

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  • IPEN-DOC 29365

    SILVA, ABDENEGO R. da ; CABRAL, FERNANDA V. ; FONTES, ADRIANA; RIBEIRO, MARTHA S. . Light-based antifungal strategy for the control of Candida auris. In: LATIN AMERICA OPTICS AND PHOTONICS CONFERENCE, August 7-11, 2022, Recife, PE. Proceedings... Washington, DC, USA: Optica Publishing Group, 2022. DOI: 10.1364/LAOP.2022.W4A.58

    Abstract: Candida auris is a pathogen that has been attracting worldwide focus due to its high resistance to conventional drugs. This work evaluated the photodynamic inactivation mediated by two phenothiazine dyes against the CBS 10913 strain.

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  • IPEN-DOC 29544

    CABRAL, FERNANDA V. ; SMITH, TERRY K.; RIBEIRO, MARTHA S. . Lipidomics analysis of Leishmania amazonensis following photoxidative stress. In: ANNUAL MEETING OF THE BRAZILIAN SOCIETY FOR BIOCHEMISTRY AND MOLECULAR BIOLOGY (SBBq), 51st; CONGRESS OF BRAZILIAN BIOPHYSICAL SOCIETY (SBBf)/LATIN AMERICAN FEDERATION OF BIOPHYSICAL SOCIETIES (Lafebs), 46th, September 5-8, 2022, Águas de Lindóia, SP. Abstract... São Paulo, SP: Sociedade Brasileira de Bioquímica e Biologia Molecular - SBBq, 2022. p. 272-272.

    Abstract: INTRODUCTION: Antimicrobial photodynamic therapy (APDT) is a well-known light-based technology that has been widely studied as an alternative approach to fight cutaneous leishmaniasis (CL). APDT induces lipid peroxidation in cellular membranes due to the generation of oxidative stress OBJECTIVES: In this study, we evaluated the role of 1,9- dimethylmethylene blue (DMMB)-mediated APDT on a wild-type (WT) and a miltefosine-resistant (MF) strain of Leishmania amazonesis and analyzed several cellular processes to get insights into the underlying mechanisms of APDT. MATERIALS AND METHODS: For this, APDT was carried out using red light (λ= 67012 nm) and promastigotes were exposed to different concentrations of DMMB at 8 J/cm2. Then, we measured mitochondrial potential and intracellular levels of reactive oxygen species (ROS) and analyzed quantitative lipidomics of the main phospholipid classes using electrospray-mass spectrometry. DISCUSSION AND RESULTS: As a result, we observed overproduction of ROS, mitochondrial membrane depolarization, and a rapid lipid remodeling immediately after APDT. Of note, MF showed a higher content in levels of phosphatidylcholine (PC) as compared to the WT line before treatment, which suggests it could be also involved in the MF resistance mechanism. In addition, results showed that after APDT, PC levels were substantially decreased, while new phospholipid species of phosphatidylethanolamine (PE) were increased. CONCLUSION: In conclusion, our data suggest DMMB-mediated APDT promoted a significant lipid peroxidation in the parasite's membrane of both strains, which failed to manage redox imbalance, thus resulting in cellular malfunction and death.

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  • IPEN-DOC 28146

    CABRAL, FERNANDA V. ; SELLERA, FABIO P.; RIBEIRO, MARTHA S. . Methylene blue-mediated antimicrobial photodynamic therapy for canine dermatophytosis caused by Microsporum canis: a successful case report with 6 months follow-up. Photodiagnosis and Photodynamic Therapy, v. 36, p. 1-3, 2021. DOI: 10.1016/j.pdpdt.2021.102602

    Abstract: Dermatophytosis is a superficial skin infection that widely effects companion animals. Miscrosporum canis is one of the most prevalent species isolated from dogs and cats, and because of the serious zoonotic potential, short-term treatment regimens are preferred to prevent the spread of disease either by direct contact or through contamination of the environment. Antimicrobial photodynamic therapy (APDT) has emerged as a promising strategy able to kill effectively a wide range of pathogens in a short period with minimal morbidity . In this case report, a 7-year-old male dog was diagnosed with dermatophytosis caused by M. canis. Methylene blue-mediated antimicrobial photodynamic therapy (MB-APDT) was applied over the lesions in two sessions with an interval of 7 days. The dog successfully healed, achieving a complete clinical cure after 21 days, without reports of recurrence after a follow-up period of 6 months. Therefore, MB-APDT could be a potential ally of small animal clinicians to treat superficial fungal diseases and should be further explored in Veterinary Medicine.

    Palavras-Chave: antimicrobial agents; methylene blue; veterinary medicine; therapy; skin diseases; tinea

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  • IPEN-DOC 26560

    CABRAL, FERNANDA V. ; RIBEIRO, MARTHA S. . Methylene blue-mediated photodynamic therapy on cutaneous leishmaniasis: an in vitro and in vivo study. In: INTERNATIONAL PHOTODYNAMIC ASSOCIATION WORLD CONGRESS, 17th, June 28 - July 4, 2019, Cambridge, Massachusetts, USA. Abstract... 2019.

    Abstract: Cutaneous leishmaniasis (CL) is a neglected disease, which promotes destructive lesions. Difficulties in treatment are related to resistance and toxicity. Methylene blue-mediated photodynamic therapy (MB-PDT) has emerged as a promising treatment considering low cost and no reports about resistance. We evaluated MB-PDT on Leishmania amazonensis in vitro and in vivo using a red LED at different fluences. Our results demonstrated that the best fluence in vitro was not effective in vivo, and a higher dose was necessary to provide better responses in mice. This study reinforces the idea that a well-planned protocol is necessary for a successful MB-PDT on CL.

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  • IPEN-DOC 25800

    DIMMER, JESICA; CABRAL, FERNANDA V. ; SABINO, CAETANO P. ; SILVA, CAMILA R. ; NUNEZ-MONTOYA, SUSANA C.; CABRERA, JOSE L.; RIBEIRO, MARTHA S. . Natural anthraquinones as novel photosentizers for antiparasitic photodynamic inactivation. Phytomedicine, v. 61, n. 152894, p. 1-7, 2019. DOI: 10.1016/j.phymed.2019.152894

    Abstract: Background: Cutaneous leishmaniasis (CL) is a vector-borne disease caused by obligate protist parasites from the genus Leishmania. The potential toxicity as well as the increased resistance of standard treatments has encouraged the development of new therapeutical strategies. Photodynamic inactivation (PDI) combines the use of a photosensitizer and light to generate reactive oxygen species and kill cells, including microorganisms. Vegetal kingdom constitutes an important source of bioactive compounds that deserve to be investigated in the search of naturally occurring drugs with leishmanicidal activity. Purpose: The purpose of this study was to test the antiparasitic activity of PDI (ApPDI) of five natural anthraquinones (AQs) obtained from Heterophyllaea lycioides (Rusby) Sandwith (Rubiacae). To support our results, effect of AQ mediated-PDI on parasite´s morphology and AQ uptake were studied. Cytotoxicity on fibroblasts was also evaluated. Study design/Methods: Two monomers, soranjidiol (Sor) and 5-chlorosoranjidiol (5-ClSor) plus three bi-anthraquinones (bi-AQs), bisoranjidiol (Bisor), 7-chlorobisoranjidiol (7-ClBisor) and Lycionine (Lyc) were selected for this study. Recombinant L. amazonensis promastigote strain expressing luciferase was subjected to AQs and LED treatment. Following irradiation with variable light parameters, cell viability was quantified by bioluminescence. Alteration on parasite's morphology was analyzed by scanning electron microscopy (SEM). In addition, we verified the AQ uptake in Leishmania cells by fluorescence and their toxicity on fibroblasts by using MTT assay. Results: Bisor, Sor and 5-ClSor exhibited photodynamic effect on L. amazonensis. SEM showed that promastigotes treated with Bisor-mediated PDI exhibited a significant alteration in shape and size. Sor and 5-ClSor presented higher uptake levels than bi-AQs (Bisor, Lyc and 7-ClBisor). Finally, Sor and Bisor presented the lowest toxic activity against fibroblasts. Conclusion: Taking together, our results indicate that Sor presents the highest specificity towards Leishmania cells with no toxicity on fibroblasts.

    Palavras-Chave: parasitic diseases; anthraquinones; antimitotic drugs; therapy; photosensitivity; inactivation; monomers

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  • IPEN-DOC 29884

    SILVA, ABDENEGO R. ; CABRAL, FERNANDA V. ; SILVA, CAMILA R. ; SILVA, DANIELA F.T. ; FREITAS, ANDERSON Z. ; FONTES, ADRIANA; RIBEIRO, MARTHA S. . New insights in phenothiazinium-mediated photodynamic inactivation of candida auris. Journal of Fungi, v. 9, n. 7, p. 1-16, 2023. DOI: 10.3390/jof9070717

    Abstract: n recent years, Candida auris has emerged as a hazardous hospital-acquired pathogen. Its resistance to antifungal treatments makes it challenging, requiring new approaches to manage it effectively. Herein, we aimed to assess the impact of photodynamic inactivation mediated by methylene blue (MB-PDI) or 1,9-dimethyl MB (DMMB-PDI) combined with a red LED against C. auris. To evaluate the photoinactivation of yeasts, we quantified colony-forming units and monitored ROS production. To gain some insights into the differences between MB and DMMB, we assessed lipid peroxidation (LPO) and mitochondrial membrane potential (ΔΨm). After, we verified the effectiveness of DMMB against biofilms by measuring metabolic activity and biomass, and the structures were analyzed through scanning electron microscopy and optical coherence tomography. We also evaluated the cytotoxicity in mammalian cells. DMMB-PDI successfully eradicated C. auris yeasts at 3 μM regardless of the light dose. In contrast, MB (100 μM) killed cells only when exposed to the highest dose of light. DMMB-PDI promoted higher ROS, LPO and ΔΨm levels than those of MB. Furthermore, DMMB-PDI was able to inhibit biofilm formation and destroy mature biofilms, with no observed toxicity in fibroblasts. We conclude that DMMB-PDI holds great potential to combat the global threat posed by C. auris. © 2023 by the authors.

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  • IPEN-DOC 29930

    CABRAL, FERNANDA V. ; CERONE, MICHELA; PERSHEYEV, SAYDULLA; LIAN, CHENG; SAMUEL, IFOR D.W.; RIBEIRO, MARTHA S. ; SMITH, TERRY K.. New insights in photodynamic inactivation of Leishmania amazonensis:: A focus on lipidomics and resistance. PLoS ONE, v. 18, n. 9, p. 1-23, 2023. DOI: 10.1371/journal.pone.0289492

    Abstract: The emergence of drug resistance in cutaneous leishmaniasis (CL) has become a major problem over the past decades. The spread of resistant phenotypes has been attributed to the wide misuse of current antileishmanial chemotherapy, which is a serious threat to global health. Photodynamic therapy (PDT) has been shown to be effective against a wide spectrum of drug-resistant pathogens. Due to its multi-target approach and immediate effects, it may be an attractive strategy for treatment of drug-resistant Leishmania species. In this study, we sought to evaluate the activity of PDT in vitro using the photosensitizer 1,9-dimethyl methylene blue (DMMB), against promastigotes of two Leishmania amazonensis strains: the wild-type (WT) and a lab induced miltefosine-resistant (MFR) strain. The underlying mechanisms of DMMB-PDT action upon the parasites was focused on the changes in the lipid metabolism of both strains, which was conducted by a quantitative lipidomics analysis. We also assessed the production of ROS, mitochondrial labeling and lipid droplets accumulation after DMMB-PDT. Our results show that DMMB-PDT produced high levels of ROS, promoting mitochondrial membrane depolarization due to the loss of membrane potential. In addition, both untreated strains revealed some differences in the lipid content, in which MFR parasites showed increased levels of phosphatidylcholine, hence suggesting this could also be related to their mechanism of resistance to miltefosine. Moreover, the oxidative stress and consequent lipid peroxidation led to significant phospholipid alterations, thereby resulting in cellular dysfunction and parasite death. Thus, our results demonstrated that DMMB-mediated PDT is effective to kill L. amazonensis MFR strain and should be further studied as a potential strategy to overcome antileishmanial drug resistance.

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  • IPEN-DOC 26478

    CABRAL, FERNANDA V. ; PELEGRINO, MILENA T.; SAUTER, ISMAEL P.; SEABRA, AMEDEA B.; CORTEZ, MAURO; RIBEIRO, MARTHA S. . Nitric oxide-loaded chitosan nanoparticles as an innovative antileishmanial platform. Nitric Oxide, v. 93, p. 25-33, 2019. DOI: 10.1016/j.niox.2019.09.007

    Abstract: Leishmaniasis is a neglected tropical disease that demands for new therapeutic strategies due to adverse side effects and resistance development promoted by current drugs. Nitric oxide (NO)-donors show potential to kill Leishmania spp. but their use is limited because of their instability. In this work, we synthesize, characterize, and encapsulate S-nitroso-mercaptosuccinic acid into chitosan nanoparticles (NONPs) and investigate their activity on promastigotes and intracellular amastigotes of Leishmania (Leishmania) amazonensis. Cytotoxicity on macrophages was also evaluated. We verified that NONPs reduced both forms of the parasite in a single treatment. We also noticed reduction of parasitophorous vacuoles as an evidence of inhibition of parasite growth and resolution of infection. No substantial cytotoxicity was detected on macrophages. NONPs were able to provide a sustained parasite killing for both L. (L.) amazonensis infective stages with no toxicity on macrophages, representing a promising nanoplatform for cutaneous leishmaniasis.

    Palavras-Chave: chitin; nanoparticles; nitric oxide; protozoa; parasitic diseases; bioluminescence; fluorescence; macrophages; antimitotic drugs; antimicrobial agents

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  • IPEN-DOC 28137

    CABRAL, FERNANDA V. ; PELEGRINO, MILENA T.; SEABRA, AMEDEA B.; RIBEIRO, MARTHA S. . Nitric-oxide releasing chitosan nanoparticles towards effective treatment of cutaneous leishmaniasis. Nitric Oxide, v. 113-114, p. 31-38, 2021. DOI: 10.1016/j.niox.2021.04.008

    Abstract: Cutaneous leishmaniasis (CL) is a major public health problem caused by Leishmania parasites that produce destructive and disfiguring skin conditions. There is an urgent need for alternative topical therapies due to the limitations of current systemic treatments. Recently, we have synthesized nitric oxide-releasing chitosan nanoparticles (NONPs) and shown their potential in vitro against Leishmania amazonensis. Herein we evaluated the application of NONPs for the treatment of CL on infected BALB/c mice. Mice were treated with topical administration of increasing concentrations of NONPs and disease progression was investigated regarding parasite load, lesion thickness, and pain score. As a result, we observed a dose-dependent NONPs effect. Parasite burden and lesion thickness were substantially lower on animals receiving NONPs at a 2 mM concentration compared to untreated control. Moreover, the clinical presentation of the lesions did not show any visible signs of ulcer, suggesting clinical healing in these animals. This successful outcome was sustained for at least 21 days after therapy even in one single dose. Thus, we demonstrate that NONPs are suitable for topical administration, and represent an attractive approach to treat CL.

    Palavras-Chave: parasitic diseases; therapy; bioluminescence; nitric oxide; skin; oligosaccharides

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A pesquisa no RD utiliza os recursos de busca da maioria das bases de dados. No entanto algumas dicas podem auxiliar para obter um resultado mais pertinente.

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Buscar os artigos apresentados em um evento internacional de 2015, sobre loss of coolant, do autor Maprelian.

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✔ Para indexação dos documentos é utilizado o Thesaurus do INIS, especializado na área nuclear e utilizado em todos os países membros da International Atomic Energy Agency – IAEA , por esse motivo, utilize os termos de busca de assunto em inglês; isto não exclui a busca livre por palavras, apenas o resultado pode não ser tão relevante ou pertinente.

✔ 95% do RD apresenta o texto completo do documento com livre acesso, para aqueles que apresentam o significa que e o documento está sujeito as leis de direitos autorais, solicita-se nesses casos contatar a Biblioteca do IPEN, bibl@ipen.br .

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O gerenciamento do Repositório está a cargo da Biblioteca do IPEN. Constam neste RI, até o presente momento 20.950 itens que tanto podem ser artigos de periódicos ou de eventos nacionais e internacionais, dissertações e teses, livros, capítulo de livros e relatórios técnicos. Para participar do RI-IPEN é necessário que pelo menos um dos autores tenha vínculo acadêmico ou funcional com o Instituto. Nesta primeira etapa de funcionamento do RI, a coleta das publicações é realizada periodicamente pela equipe da Biblioteca do IPEN, extraindo os dados das bases internacionais tais como a Web of Science, Scopus, INIS, SciElo além de verificar o Currículo Lattes. O RI-IPEN apresenta também um aspecto inovador no seu funcionamento. Por meio de metadados específicos ele está vinculado ao sistema de gerenciamento das atividades do Plano Diretor anual do IPEN (SIGEPI). Com o objetivo de fornecer dados numéricos para a elaboração dos indicadores da Produção Cientifica Institucional, disponibiliza uma tabela estatística registrando em tempo real a inserção de novos itens. Foi criado um metadado que contém um número único para cada integrante da comunidade científica do IPEN. Esse metadado se transformou em um filtro que ao ser acionado apresenta todos os trabalhos de um determinado autor independente das variáveis na forma de citação do seu nome.

A elaboração do projeto do RI do IPEN foi iniciado em novembro de 2013, colocado em operação interna em julho de 2014 e disponibilizado na Internet em junho de 2015. Utiliza o software livre Dspace, desenvolvido pelo Massachusetts Institute of Technology (MIT). Para descrição dos metadados adota o padrão Dublin Core. É compatível com o Protocolo de Arquivos Abertos (OAI) permitindo interoperabilidade com repositórios de âmbito nacional e internacional.

1. Portaria IPEN-CNEN/SP nº 387, que estabeleceu os princípios que nortearam a criação do RDI, clique aqui.


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O Repositório Digital do IPEN é um equipamento institucional de acesso aberto, criado com o objetivo de reunir, preservar, disponibilizar e conferir maior visibilidade à Produção Científica publicada pelo Instituto, desde sua criação em 1956.

Operando, inicialmente como uma base de dados referencial o Repositório foi disponibilizado na atual plataforma, em junho de 2015. No Repositório está disponível o acesso ao conteúdo digital de artigos de periódicos, eventos, nacionais e internacionais, livros, capítulos, dissertações, teses e relatórios técnicos.

A elaboração do projeto do RI do IPEN foi iniciado em novembro de 2013, colocado em operação interna em julho de 2014 e disponibilizado na Internet em junho de 2015. Utiliza o software livre Dspace, desenvolvido pelo Massachusetts Institute of Technology (MIT). Para descrição dos metadados adota o padrão Dublin Core. É compatível com o Protocolo de Arquivos Abertos (OAI) permitindo interoperabilidade com repositórios de âmbito nacional e internacional.

O gerenciamento do Repositório está a cargo da Biblioteca do IPEN. Constam neste RI, até o presente momento 20.950 itens que tanto podem ser artigos de periódicos ou de eventos nacionais e internacionais, dissertações e teses, livros, capítulo de livros e relatórios técnicos. Para participar do RI-IPEN é necessário que pelo menos um dos autores tenha vínculo acadêmico ou funcional com o Instituto. Nesta primeira etapa de funcionamento do RI, a coleta das publicações é realizada periodicamente pela equipe da Biblioteca do IPEN, extraindo os dados das bases internacionais tais como a Web of Science, Scopus, INIS, SciElo além de verificar o Currículo Lattes. O RI-IPEN apresenta também um aspecto inovador no seu funcionamento. Por meio de metadados específicos ele está vinculado ao sistema de gerenciamento das atividades do Plano Diretor anual do IPEN (SIGEPI). Com o objetivo de fornecer dados numéricos para a elaboração dos indicadores da Produção Cientifica Institucional, disponibiliza uma tabela estatística registrando em tempo real a inserção de novos itens. Foi criado um metadado que contém um número único para cada integrante da comunidade científica do IPEN. Esse metadado se transformou em um filtro que ao ser acionado apresenta todos os trabalhos de um determinado autor independente das variáveis na forma de citação do seu nome.