EDITE HATSUMI YAMASHIRO KANASHIRO
Projetos de Pesquisa
Unidades Organizacionais
Departamento de Moléstias Infecciosas e Parasitárias, Faculdade de Medicina
LIM/48 - Laboratório de Imunologia, Hospital das Clínicas, Faculdade de Medicina
LIM/48 - Laboratório de Imunologia, Hospital das Clínicas, Faculdade de Medicina
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- LOOSE AND COMPACT AGGLOMERATES OF 50 NM MICROVESICLES DERIVED FROM GOLGI AND ENDOPLASMIC RETICULUM MEMBRANES IN PRE- AND IN -APOPTOTIC MYCOPLASMA INFECTED HELA CELLS: HOST-PARASITE INTERACTIONS UNDER THE TRANSMISSION ELECTRON MICROSCOPE(2015) SESSO, Antonio; YAMASHIRO-KANASHIRO, Edite Hatsumi; ORII, Noemia Mie; TANIWAKI, Noemi Nosomi; KAWAKAMI, Joyce; CARNEIRO, Sylvia Mendes
- Oral administration of buparvaquone nanostructured lipid carrier enables in vivo activity against Leishmania infantum(2022) MONTEIRO, Lis Marie; LOBENBERG, Raimar; BARBOSA, Eduardo Jose; ARAUJO, Gabriel Lima Barros de; SATO, Paula Keiko; KANASHIRO, Edite; ELIODORO, Raissa H. de Araujo; ROCHA, Mussya; FREITAS, Vera Lucia Teixeira de; FOTAKI, Nikoletta; BOU-CHACRA, Nadia AraciLeishmaniasis, a neglected tropical disease, is prevalent in 98 countries with the occurrence of 1.3 million new cases annually. The conventional therapy for visceral leishmaniasis requires hospitalization due to the severe adverse effects of the drugs, which are administered parenterally. Buparvaquone (BPQ) showed in vitro activity against leishmania parasites; nevertheless, it has failed in vivo tests due to its low aqueous solubility. Though, lipid nanoparticles can overcome this holdback. In this study we tested the hypothesis whether BPQ-NLC shows in vivo activity against L. infantum. Two optimized formulations were prepared (V1: 173.9 +/- 1.6 nm, 0.5 mg of BPQ/mL; V2: 232.4 +/- 1.6 nm, 1.3 mg of BPQ/mL), both showed increased solubility up to 73.00-fold, and dissolution up to 83.29%, while for the free drug it was only 2.89%. Cytotoxicity test showed their biocompatibility (CC50 >554.4 mu M). Besides, the V1 dose of 0.3 mg/kg/day for 10 days reduced the parasite burden in 83.4% +/- 18.2% (p <0.05) in the liver. BPQ-NLC showed similar leishmanicidal activity compared to miltefosine. Therefore, BPQ-NLC is a promising addition to the limited therapeutic arsenal suitable for leishmaniasis oral administration treatment.
- In vitro miltefosine and amphotericin B susceptibility of strains and clinical isolates of Leishmania species endemic in Brazil that cause tegumentary leishmaniasis(2023) FERREIRA, Bianca A.; COSER, Elizabeth M.; SABORITO, Cristiele; YAMASHIRO-KANASHIRO, Edite H.; COELHO, Adriano C.; LINDOSE, Jose Angelo L.Tegumentary leishmaniasis encompasses a spectrum of clinical manifestations caused by the parasitic protozoa of the genus Leishmania. In Brazil, there are at least seven Leishmania species that are endemic and responsible for this set of clinical manifestations of the disease. Current treatment is limited to a restricted number of drugs that in general have several drawbacks including parenteral use, toxicity, and severe side effects. Amphotericin B is considered a second-line drug for tegumentary leishmaniasis in Brazil, while miltefosine was recently approved for clinical use in the treatment of this disease. In this study, we investigated the in vitro susceptibility of Leishmania strains representative of the species endemic to Brazil, as well as a panel of thirteen clinical isolates of tegumentary leishmaniasis, to both amphotericin B and miltefosine. A moderate variation in the susceptibility to both drugs was found, where the EC50 values varied from 11.43 to 52.67 mu M for miltefosine and from 12.89 to 62.36 nM for amphotericin B in promastigotes, while for the intracellular amastigotes, values ranged from 1.08 to 9.60 mu M and from 1.69 to 22.71 nM for miltefosine and amphotericin B respectively. Furthermore, the clinical isolates and strains of the subgenus Viannia were evaluated for the presence of Leishmania RNA virus 1 (LRV1), as this is an important factor associated with disease severity and treatment outcome. These findings provide a preclinical dataset of the activity of these drugs against the causative species of tegumentary leishmaniasis in Brazil.
article 8 Citação(ões) na Scopus Leptospirosis Infection in Sheep and Characterization of the Renal Inflammatory Response.(2011) CARVALHO, Sonia Maria de; GONCALVES, Larissa Maria Feitosa; MACEDO, Nicodemos Alves de; GOTO, Hiro; SILVA, Silvana Maria Medeiros de Sousa; MINEIRO, Ana Lys Bezerra Barradas; KANASHIRO, Edite Hatsumi Yamashiro; COSTA, Francisco Assis LimaCarvalho S.M., Gon alves L.M.F., Macedo N.A., Goto H., Silva S.M.M.S., Mineiro A.L.B.B., Kanashiro E.H.Y. & Costa F.A.L. 2011. [Leptospirosis Infection in Sheep and Characterization of the Renal Inflammatory Response.] Infeccao por leptospiras em ovinos e caracterizacao da resposta inflamatoria renal. Pesquisa Veterinaria Brasileira 31(8):637-642. Departamento de Clinica e Cirurgia Veterinaria, Centro de Ciencias Agrarias, Universidade Federal do Piaui, Campus Socopo, Teresina, PI 64046-550, Brazil. E-mail: fassisle@gmail.com Lepitospirosis is a serious worldwide distribution disease which affects man and other animals. The infection is generally asymptomatic in animals. In cases whose symptoms are present, symptoms are similar to other infections. In the present study serum samples from 119 sheep and their kidneys were collected during their slaughter in outdoor markets in the city of Teresina, Piaui, Brazil. The Microscopic Agglutination Test (MAT) obtained 34 positive serological samples for one or more Leptospira spp. serovar with occurrence rate of 28.6% of leptospiral antibodies. There were 23 cases of infection for a single serovar, and 11 cases with coagglutination for two or more serovars. Autumnalis had the highest occurrence (29.4%) among the pathogenic serovars. The histopathological analysis of 36 kidney fragments revealed tubulo-interstitial alterations in 33 (91.7%) positive animals. Tubular lesions were observed in 20 (55.5%) positive animals. The Warthin Starry staning revealed the presence of Leptospira in 8 (22.20%) of the 36 positive samples. The immunoperoxidase staining revealed the presence of Leptospira in 12 (60%) of 20 positive samples. The inflammatory infiltrate in the positive animals was significantly more evident in the cortical-medullar and cortical regions than in the medullar region (p=0.000), however, there was no difference between positive and negative animals. The presence of hyaline casts in the proximal tubules was significantly higher in positive animals compared to the negative ones (p=0.0001). Discrete lesion was observed in glomeruli. In conclusion, the results from this study showed that sheep which are positive for Leptospira present tubulo-intersticial renal lesions with the presence of Leptospira in the tubules, conferring to these animals the condition of asymptomatic carriers.- Susceptibility to paromomycin in clinical isolates and reference strains of Leishmania species responsible for tegumentary leishmaniasis in Brazil(2021) COSER, Elizabeth M.; FERREIRA, Bianca A.; YAMASHIRO-KANASHIRO, Edite H.; LINDOSO, Jose Angelo L.; COELHO, Adriano C.Treatment of tegumentary leishmaniasis in Brazil is limited to pentavalent antimonial, amphotericin B and pentamidine. These drugs, administered parenterally, cause several side effects and have a varied clinical response, depending on the species of Leishmania. Urgent expansion of the therapeutic arsenal against the disease is therefore necessary. Pammomycin is an aminoglycoside antibiotic that has already been approved for the treatment of visceral leishmaniasis in Southeast Asia. Here, we provide an in vitro evaluation of the activity of paromomycin in fifteen clinical isolates from patients with tegumentary leishmaniasis at a reference center for the treatment of the disease. Furthermore, the in vitro susceptibility to this drug in reference strains of Leishmania species that are endemic in Brazil has also been evaluated. Among the clinical isolates, nine were typed as Leishmania (Viannia) braziliensis, five as L. (Leishmania) amazonensis and one as L. (V.) guyanensis. Although never exposed to paromomycin, we found variable susceptibility among these isolates and reference strains in promastigotes and intracellular amastigotes, with the drug being more active in the amastigote form of the parasite. This study provides a preclinical dataset that is useful for the evaluation of pammomycin in the treatment of tegumentary leishmaniasis caused by species that are endemic in Brazil.
- Specific label-free and real-time detection of oxidized low density lipoprotein (oxLDL) using an immunosensor with three monoclonal antibodies(2014) CABRAL-MIRANDA, Gustavo; YAMASHIRO-KANASHIRO, E. H. G.; GIDLUND, Magnus; SALES, M. Goreti F.Increased levels of plasma oxLDL, which is the oxidized fraction of Low Density Lipoprotein (LDL), are associated with atherosclerosis, an inflammatory disease, and the subsequent development of severe cardiovascular diseases that are today a major cause of death in modern countries. It is therefore important to find a reliable and fast assay to determine oxLDL in serum. A new immunosensor employing three monoclonal antibodies (mAbs) against oxLDL is proposed in this work as a quick and effective way to monitor oxLDL. The oxLDL was first employed to produce anti-oxLDL monoclonal antibodies by hybridoma cells that were previously obtained. The immunosensor was set-up by self-assembling cysteamine (Cyst) on a gold (Au) layer (4 mm diameter) of a disposable screen-printed electrode. Three mAbs were allowed to react with N-hydroxysuccinimide (NHS) and ethyl(dimethylaminopropyl) carbodiimide (EDAC), and subsequently incubated in the Au/Cys. Albumin from bovine serum (BSA) was immobilized further to ensure that other molecules apart from oxLDL could not bind to the electrode surface. All steps were followed by various characterization techniques such as electrochemical impedance spectroscopy (EIS) and square wave voltammetry (SWV). The analytical operation of the immunosensor was obtained by incubating the sensing layer of the device in oxLDL for 15 minutes, prior to EIS and SWV. This was done by using standard oxLDL solutions prepared in foetal calf serum, in order to simulate patient's plasma with circulating oxLDL. A sensitive response was observed from 0.5 to 18.0 mu g mL(-1). The device was successfully applied to determine the oxLDL fraction in real serum, without prior dilution or necessary chemical treatment. The use of multiple monoclonal antibodies on a biosensing platform seemed to be a successful approach to produce a specific response towards a complex multi-analyte target, correlating well with the level of oxLDL within atherosclerosis disease, in a simple, fast and cheap way.
- Targeting Leishmania amazonensis amastigotes through macrophage internalisation of a hydroxymethylnitrofurazone nanostructured polymeric system(2017) MONTEIRO, Lis Marie; LOBENBERG, Raimar; FERREIRA, Elizabeth Igne; COTRIM, Paulo Cesar; KANASHIRO, Edite; ROCHA, Mussya; CHUNG, Man Chin; BOU-CHACRA, NadiaDextran-coated poly (n-butyl cyanoacrylate) nanoparticles (PBCA-NPs) were prepared and were evaluated for enhanced delivery of a promising anti-Leishmania drug candidate, hydroxymethylnitrofurazone (NFOH), to phagocytic cells. Currently available chemotherapy for leishmaniasis, such as pentavalent antimonials, presents low safety and efficacy. Furthermore, widespread drug resistance in leishmaniasis is rapidly emerging. To overcome these drawbacks, the use of nanosized delivery systems can reduce systemic drug toxicity and increase the drug concentration in infected macrophages, therefore improving treatment of leishmaniasis. PBCA-NPs containing NFOH (PBCA-NFOH-NPs) were prepared by an anionic emulsion polymerisation method. The z-average and polydispersity index (PDI) were determined by photon correlation spectroscopy, the zeta potential by microelectrophoresis and the entrapment efficiency by HPLC. Cytotoxicity was determined using macrophages from BALB/c mice. Efficacy tests were performed using Leishmania amazonensis promastigotes and amastigotes. The z-average of PBCA-NFOH-NPs was 151.5 +/- 61.97 nm, with a PDI of 0.104 +/- 0.01, a zeta potential of -10.1 +/- 6.49 mV and an entrapment efficiency of 64.47 +/- 0.43%. Efficacy in amastigotes revealed IC50 values of 0.33 mu M and 31.2 mu M for the nanostructured and free NFOH, respectively (95-fold increase). The cytotoxicity study indicated low toxicity of the PBCA-NFOH-NPs to macrophages. The selectivity index was 370.6, which is 49-fold higher than free NFOH (7.6). Such findings indicated that improved efficacy could be due to NP internalisation following site-specific drug delivery and reactivation of immune protective reactions by the NP components. Thus, PBCA-NFOH-NPs have the potential to significantly improve the treatment of leishmaniasis, with reduced systemic side effects.
- Lipid nanoparticles for amphotericin delivery in the treatment of American tegumentary leishmaniasis(2020) SOUZA, Regina Maia de; MARANHAO, Raul Cavalcante; TAVARES, Elaine Rufo; FILIPPIN-MONTEIRO, Fabiola Branco; NICODEMO, Antonio Carlos; MORIKAWA, Aleksandra Tiemi; KANASHIRO, Edite Hatsumi Yamashiro; AMATO, Valdir SabbagaLeishmaniasis occurs in the five continents and represents a serious public health challenge, but is still a neglected disease, and the current pharmacological weaponry is far from satisfactory. Triglyceride-rich nanoparticles mimicking chylomicrons (TGNP) behave metabolically like native chylomicrons when injected into the bloodstream. Previously we have shown that TGNP as vehicle to amphothericin B (AB) for treatment of fungi infection showed reduced renal toxicity and lower animal death rates compared to conventional AB. The aim of the current study was to test the tolerability and effectiveness of the TGNP-AB preparation in a murine model of Leishmania amazonensis infection. The in vitro assays determined the cytotoxicity of TGNP-AB, AB, and TGNP in macrophages and promastigote forms and the leishmanicidal activity in infected macrophages. The in vivo toxicity tests were performed in healthy mice with increasing doses of TGPN-AB and AB. Then, animals were treated with 2.5 mg/kg/day of AB, 17.5 mg/kg/day of TGNP-AB, or TGNP three times a week for 4 weeks. TGNP-AB formulation was less cytotoxic for macrophages than AB. TGNP-AB was more effective than AB against the promastigotes forms of the parasite and more effective in reducing the number of infected macrophages and the number of amastigotes forms per cell. TGNP-AB-treated animals showed lower hepatotoxicity. In addition, TGNP-AB group showed a marked reduction in lesion size on the paws and parasitic load. The TGNP-AB preparation attained excellent leishmanicidal activity with remarkable lower drug toxicity at very high doses that, due to the toxicity-buffering properties of the nanocarrier, become fully tolerable.
- The hamster (Mesocricetus auratus) as an experimental model of toxocariasis: histopathological, immunohistochemical, and immunoelectron microscopic findings(2015) SILVA, Ana Maria Goncalves da; CHIEFFI, Pedro Paulo; SILVA, Wellington Luiz Ferreira da; KANASHIRO, Edite Hatsumi Yamashiro; RUBINSKY-ELEFANT, Guita; CUNHA-NETO, Edecio; MAIRENA, Eliane Conti; BRITO, Thales DeToxocariasis is a globally distributed parasitic infection caused by the larval stage of Toxocara spp. The typical natural hosts of the parasite are dogs and cats, but humans can be infected by the larval stage of the parasite after ingesting embryonated eggs in soil or from contaminated hands or fomites. The migrating larvae are not adapted to complete their life cycle within accidental or paratenic hosts like humans and laboratory animals, respectively, but they are capable of invading viscera or other tissues where they may survive and induce disease. In order to characterize hamsters (Mesocricetus auratus) as a model for Toxocara canis infection, histopathological and immunohistochemistry procedures were used to detect pathological lesions and the distribution of toxocaral antigens in the liver, lungs, and kidneys of experimentally infected animals. We also attempted to characterize the immunological parameters of the inflammatory response and correlate them with the histopathological findings. In the kidney, a correlation between glomerular changes and antigen deposits was evaluated using immunoelectron microscopy. The hamster is an adequate model of experimental toxocariasis for short-term investigations and has a good immunological and pathological response to the infection. Lung and liver manifestations of toxocariasis in hamsters approximated those in humans and other experimental animal models. A mixed Th2 immunological response to T. canis infection was predominant. The hamster model displayed a progressive rise of anti-toxocaral antibodies with the formation of immune complexes. Circulating antigens, immunoglobulin, and complement deposits were detected in the kidney without the development of a definite immune complex nephropathy.
- 14 Years of History on Sustainability: Waste Management FMUSP/IMT/SVOC(2018) MARQUES, Fabio Luiz Navarro; SARAIVA, Arlete Araujo; CHAVES, Adilson Caetano Cesario; SILVA, Ana Maria Goncalves da; KANASHIRO, Edite Hatsumi Yamashiro; POMPEU, Eduardo; SIQUEIRA, Gabriela Hase; BERNARDO, Joana Sueli Maziero; APOLINARIO, Lilian; SANCHEZ, Maria Carmen Arroyo; GUIMARAES, Maria Ines Calil Cury; MAUAD, Thais; SOUZA, Felipe Neme; AULER JR., Jose Otavio CostaHigh regulation in environmental protection and sustainability was established in Brazil around 2003. From that time, the board of directors of the Faculty of Medicine, Tropical Medicine Institute, Death Verification Service, and Medical Investigation Laboratories, of the University of Sao Paulo created the Waste Management Committee to organize and implement actions to comply with Brazilian legislation. This paper outlines the main actions taken by the Committee and Board of directors of the institutions, presenting it in ages. During the first five years were developed works to know the passive/regular discharge of infectious agents, chemicals, radioactive compounds, recyclable and common garbage. From this data, were constructed stations for provisory disposal, transportation flow and general rules were established, and training was given to staff and students. During the following five years, or in the second phase of actions plans, efforts were focused on improving the management of the process and increasing the gathering of recyclable materials. Now, in the third phase, actions have been developed aiming to eliminate devices that use mercury and mercury vapor lamps from FMUSP building; furthermore, composting of food and garden material is in the evaluation process. These actions gave to FMUSP two Prize Friend of the Environment (2010 and 2016), offered by Health Secretary of Sao Paulo State, and these experience will be shared in this paper. However, it is necessary to have in mind that this work was done under Brazilian regulations, but certainly, it can be adjusted to all countries.