Magnetite-levan nanoparticles for lectin purification: A single-step strategy for protein isolation from the seeds extract of the plant Cratylia mollis.

The ability to reversibly bind carbohydrates is an incredible property from lectins. Such characteristic has led these molecules to be employed in several applications involving medical research and biotechnology. Generally, these proteins follow several steps towards purification. Here, the synthesis, physical characterization, and use of levan-coated magnetite nanoparticles (MNPs-levan) for lectin isolation is described. Canavalia ensiformis and Cratylia mollis were used as sources of Concanavalin A and Cramoll, respectively, that were purified by using MNPs-levan. Mass spectrometry, SDS-PAGE, and hemagglutinating activity were employed to assess the efficiency of the process. Moreover, by using mass spectrometry approaches, a novel lectin, similar to Canavalin, was also identified for C. mollis, corroborating the advantages of using nanoparticles over microparticles. MNPs-levan could also be recycled, making this a low-cost, scalable process that can be efficiently employed over crude samples.

Evaluation of marginal sealing quality of restorations with low shrinkage composite resins.

BACKGROUND: This study compared the quality of marginal sealing in the gingival wall of class II preparations of two low-shirinkage resins of the bulk fill type with a conventional resin isolated or associated with a glass ionomer cement (GIC). MATERIAL AND METHODS: 40 human molars were divided into 4 groups and 80 occlusal-mesial and occlusal-distal restorations were performed with the following materials: SureFil SDR flow, Filtek Bulk Fill Posterior, Z250 resins and Riva Light Cure GIC. 40 restorations were evaluated in Scanning Electron Microscopy (SEM) with Elemental Microanalysis Spectrometry (EDS) initially and the remainder after a period of 6 months of aging in a 37 ± 5°C oven. An average of the silver penetration at each restoration was obtained in the two evaluations and the results were statistically analyzed in a descriptive and inferential way, through the paired t-Student and one-way ANOVA F-test. RESULTS: There were no significant statistical differences between the materials with respect to silver nanoinfiltration, except for the Bulk Fill Posterior/3M ESPE resin compared to the GIC and conventional resin in the final evaluation. CONCLUSIONS: The low shrinkage resins showed a similar behavior in relation to the marginal sealing quality observed in the GIC or composite resin with the incremental technique, also presenting the advantage of simplicity in the technique of confection of the restorations and reduction of the time of work. Key words:Resin composites, Bulk fill, dental restorations, marginal quality, adhesion.

Magnetite-levan nanoparticles for lectin purification: a single-step strategy for protein isolation from the seeds extract of the plant Cratylia mollis

The ability to reversibly bind carbohydrates is an incredible property from lectins. Such characteristic has led these molecules to be employed in several applications involving medical research and biotechnology. Generally, these proteins follow several steps towards purification. Here, the synthesis, physical characterization, and use of levan-coated magnetite nanoparticles (MNPs-levan) for lectin isolation is described. Canavalia ensiformis and Cratylia mollis were used as sources of Concanavalin A and Cramoll, respectively, that were purified by using MNPs-levan. Mass spectrometry, SDS-PAGE, and hemagglutinating activity were employed to assess the efficiency of the process. Moreover, by using mass spectrometry approaches, a novel lectin, similar to Canavalin, was also identified for C. mollis, corroborating the advantages of using nanoparticles over microparticles. MNPs-levan could also be recycled, making this a low-cost, scalable process that can be efficiently employed over crude samples.

Ultrastructural Assessment of 2-(acridin-9-ylmethylene)-N-phenylhydrazinecarbothioamide activity on human breast adenocarcinoma cells.

The aim of the present study was to investigate ultrastructural changes induced by (Z)-2-(acridin-9-ylmethylene)-N-phenylhydrazinecarbothioamide (APHCA) treatment on human breast adenocarcinoma cancer cells MCF-7, besides the evaluation of phosphatidylserine externalization and DNA fragmentation in treated cells. Cell viability analysis demonstrated concentration and time-manner cytotoxicity. Treated MCF-7 cells did not expose phosphatidylserine residues to the external plasma membrane surface and DNA fragmentation was not visualized by electrophoresis. Light microscopy showed compromised cell density and presence of vacuolization after APHCA treatment with 60µM. Scanning and transmission electron microscopies revealed hallmarks of autophagy, namely the presence of membrane bebbling and autophagosomes, besides shrunken cells and cell debris in treated MCF-7 cells. However, more specific tests such as the quantification of mammalian autophagy proteins are necessary to determine the kind of death that is trigged by APHCA.

ULTRASTRUCTURAL CHANGES IN Schistosoma mansoni MALE WORMS AFTER in vitro INCUBATION WITH THE ESSENTIAL OIL OF Mentha x villosa Huds.

INTRODUCTION: The essential oil Mentha x villosa (MVEO) has a wide range of actions, including antibacterial, antifungal, antiprotozoal and schistosomicidal actions. The present study aimed to investigate the ultrastructural changes of MVEO on the tegument of adult Schistosoma mansoni. MATERIALS AND METHODS: Different concentrations of MVEO were tested on S. mansoni adult worms in vitro. Ultrastructural changes on the tegument of these adult worms were evaluated using scanning electron microscopy (SEM) and transmission electron microscopy (TEM). RESULTS: The MVEO caused the death of all worms at 500 µg mL(-1) after 24 h. After 24h of 500 µg mL(-1) MVEO treatment, bubble lesions were observed over the entire body of worms and they presented loss of tubercles in some regions of the ventral portion. In the evaluation by TEM, S. mansoni adult worms treated with MVEO, 500 µg mL(-1), presented changes in the tegument and vacuoles in the syncytial matrix region. Glycogen granules close to the muscle fibers were visible. CONCLUSION: The ability of MVEO to cause extensive ultrastructural damage to S. mansoni adult worms correlates with its schistosomicidal effects and confirms earlier findings with S. mansoni.

Different cell death responses induced by eupomatenoid-5 in MCF-7 and 786-0 tumor cell lines.

Natural products remain an important source of new drugs, including anticancer drugs. Recently, our group reported the anticancer activity of eupomatenoid-5 (eup-5), a neolignan isolated from Piper regnellii (Miq.) C. DC. var. regnellii leaves. In vitro studies demonstrated that MCF-7 (breast) and 786-0 (kidney) were among the cancer cell lines most sensitive to eup-5 treatment. The current results demonstrate that mitochondrial membrane depolarization and generation of reactive oxygen species are implicated in eup-5-mediated cytotoxic effects on these cancer cells lines. In MCF-7 cells, eup-5 led to phosphatidylserine externalization and caspase activation, whereas the same did not occur in 786-0 cells. Scanning electron microscopy revealed a reduction of microvilli density, as well as cell morphology alterations. Moreover, treated MCF-7 cells exhibited well-characterized apoptosis alterations, while treated 786-0 cells exhibited characteristics of programmed necroptosis process. These findings support the possibility that different mechanisms may be targeted by eup-5 in cell death response.

Ultrastructural study on the morphological changes to male worms of Schistosoma mansoni after in vitro exposure to allicin.

INTRODUCTION: Garlic has a wide range of actions, including antibacterial, antiviral, antifungal, antiprotozoal and anthelmintic actions. This antiparasitic activity has been attributed to allicin, which is the main constituent of garlic. The present study aimed to investigate the in vitro activity of allicin on the tegument of adult Schistosoma mansoni worms using scanning electron microscopy. METHODS: Swiss Webster mice were infected with S. mansoni cercariae (100 per mouse) and sacrificed 50 days later to acquire the adult worms. These worms were collected by perfusion and placed in RPMI medium 1,640 at 37°C before transferring to RPMI media containing 0 (control), 5, 10, 15 and 20mg/mL of allicin, where they were incubated for 2h. The worms were fixed in 2.5% glutaraldehyde solution, washed twice, post-fixed in osmium tetroxide, washed twice and then dehydrated with ascending grades of ethanol. The samples were air-dried, mounted on stubs, gold coated in an ion sputtering unit and viewed using a scanning electron microscope. RESULTS: A concentration of 5mg/mL caused wrinkling in the tegument; a concentration of 10mg/mL resulted in changes to tubercles and loss or modification of spines. With 15 and 20mg/mL increasing damage to the tegument could be seen, such as vesicle formation and the presence of ulcers. CONCLUSIONS: These findings demonstrate the effect of allicin on adult S. mansoni worms and indicate that most of the changes occur at concentrations greater than that normally indicated for treatment.

ULTRASTRUCTURAL CHANGES IN Schistosoma mansoni MALE WORMS AFTER in vitro INCUBATION WITH THE ESSENTIAL OIL OF Mentha x villosa Huds

Introduction: The essential oil Mentha x villosa (MVEO) has a wide range of actions, including antibacterial, antifungal, antiprotozoal and schistosomicidal actions. The present study aimed to investigate the ultrastructural changes of MVEO on the tegument of adult Schistosoma mansoni. Materials and Methods: Different concentrations of MVEO were tested on S. mansoni adult worms in vitro. Ultrastructural changes on the tegument of these adult worms were evaluated using scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Results: The MVEO caused the death of all worms at 500 μg mL-1 after 24 h. After 24h of 500 μg mL-1 MVEO treatment, bubble lesions were observed over the entire body of worms and they presented loss of tubercles in some regions of the ventral portion. In the evaluation by TEM, S. mansoni adult worms treated with MVEO, 500 μg mL-1, presented changes in the tegument and vacuoles in the syncytial matrix region. Glycogen granules close to the muscle fibers were visible. Conclusion: The ability of MVEO to cause extensive ultrastructural damage to S. mansoni adult worms correlates with its schistosomicidal effects and confirms earlier findings with S. mansoni.

Ultrastructural study on the morphological changes to male worms of Schistosoma mansoni after in vitro exposure to allicin / Estudo ultraestrutural das alterações morfológicas de vermes machos de Schistosoma mansoni após exposição in vitro à alicina

INTRODUCTION: Garlic has a wide range of actions, including antibacterial, antiviral, antifungal, antiprotozoal and anthelmintic actions. This antiparasitic activity has been attributed to allicin, which is the main constituent of garlic. The present study aimed to investigate the in vitro activity of allicin on the tegument of adult Schistosoma mansoni worms using scanning electron microscopy. METHODS: Swiss Webster mice were infected with S. mansoni cercariae (100 per mouse) and sacrificed 50 days later to acquire the adult worms. These worms were collected by perfusion and placed in RPMI medium 1,640 at 37°C before transferring to RPMI media containing 0 (control), 5, 10, 15 and 20mg/mL of allicin, where they were incubated for 2h. The worms were fixed in 2.5 percent glutaraldehyde solution, washed twice, post-fixed in osmium tetroxide, washed twice and then dehydrated with ascending grades of ethanol. The samples were air-dried, mounted on stubs, gold coated in an ion sputtering unit and viewed using a scanning electron microscope. RESULTS: A concentration of 5mg/mL caused wrinkling in the tegument; a concentration of 10mg/mL resulted in changes to tubercles and loss or modification of spines. With 15 and 20mg/mL increasing damage to the tegument could be seen, such as vesicle formation and the presence of ulcers. CONCLUSIONS: These findings demonstrate the effect of allicin on adult S. mansoni worms and indicate that most of the changes occur at concentrations greater than that normally indicated for treatment.

INTRODUÇÃO: O alho apresenta uma ampla gama de ações, incluindo antibacteriana, antiviral, antifúngico, antiprotozoário e anti-helmíntico. Esta atividade antiparasitária tem sido atribuída à alicina, que é o principal constituinte do alho. O presente estudo teve como objetivo investigar a ação in vitro da alicina no tegumento de vermes adultos de Schistosoma mansoni utilizando a microscopia eletrônica de varredura. MÉTODOS: Camundongos Swiss Webster foram infectados com cercárias de S. mansoni (100 por camundongo) e sacrificados 50 dias depois para aquisição de vermes adultos. Estes vermes foram coletados por perfusão e colocados em meio RPMI 1.640 a 37°C antes de transferir para o meio RPMI contendo 0 (controle), 5, 10, 15 e 20mg/mL de alicina, onde eles foram incubados por 2h. Os vermes foram fixados em uma solução de glutaraldeído a 2,5 por cento, lavados duas vezes, pós-fixados em tetróxido de ósmio, lavados duas vezes e então desidratados em séries crescentes de etanol. As amostras foram secadas, montadas em stubs, metalizadas em ouro e visualizadas utilizando o microscópio eletrônico de varredura. RESULTADOS: A concentração de 5mg/mL causou o enrugamento do tegumento; a concentração de 10mg/mL resultou em alterações nos tubérculos e perda ou modificações nos espinhos. Com 15 e 20mg/mL crescentes danos no tegumento pode ser visto, tais como formação de vesículas e presença de úlceras. CONCLUSÕES: Esses resultados demonstram os efeitos da alicina nos vermes adultos de S. mansoni e indicam que a maioria das alterações ocorrem numa concentração maior do que a normalmente indicada para o tratamento.

Ultrastructural study on the morphological changes to male worms of Schistosoma mansoni after in vitro exposure to allicin / Estudo ultraestrutural das alterações morfológicas de vermes machos de Schistosoma mansoni após exposição in vitro à alicina

INTRODUÇÃO: O alho apresenta uma ampla gama de ações, incluindo antibacteriana, antiviral, antifúngico, antiprotozoário e anti-helmíntico. Esta atividade antiparasitária tem sido atribuída à alicina, que é o principal constituinte do alho. O presente estudo teve como objetivo investigar a ação in vitro da alicina no tegumento de vermes adultos de Schistosoma mansoni utilizando a microscopia eletrônica de varredura. MÉTODOS: Camundongos Swiss Webster foram infectados com cercárias de S. mansoni (100 por camundongo) e sacrificados 50 dias depois para aquisição de vermes adultos. Estes vermes foram coletados por perfusão e colocados em meio RPMI 1.640 a 37°C antes de transferir para o meio RPMI contendo 0 (controle), 5, 10, 15 e 20mg/mL de alicina, onde eles foram incubados por 2h. Os vermes foram fixados em uma solução de glutaraldeído a 2,5%, lavados duas vezes, pós-fixados em tetróxido de ósmio, lavados duas vezes e então desidratados em séries crescentes de etanol. As amostras foram secadas, montadas em stubs, metalizadas em ouro e visualizadas utilizando o microscópio eletrônico de varredura. RESULTADOS: A concentração de 5mg/mL causou o enrugamento do tegumento; a concentração de 10mg/mL resultou em alterações nos tubérculos e perda ou modificações nos espinhos. Com 15 e 20mg/mL crescentes danos no tegumento pode ser visto, tais como formação de vesículas e presença de úlceras. CONCLUSÕES: Esses resultados demonstram os efeitos da alicina nos vermes adultos de S. mansoni e indicam que a maioria das alterações ocorrem numa concentração maior do que a normalmente indicada para o tratamento.