Effect of prednisolone in a kindling model of epileptic seizures in rats on cytokine and intestinal microbiota diversity.

Epilepsy is a neurological disease characterized by spontaneous and recurrent seizures. Epileptic seizures can be initiated and facilitated by inflammatory mechanisms. As the dysregulation of the immune system would be involved in epileptogenesis, it is suggested that anti-inflammatory medications could impact epileptic seizures. These medications could potentially have a side effect by altering the structure and composition of the intestinal microbiota. These changes can disrupt microbial homeostasis, leading to dysbiosis and potentially exacerbating intestinal inflammation. We hypothesize that prednisolone may affect the development of epileptic seizures, potentially influencing the diversity of the intestinal microbiota and the regulation of pro-inflammatory cytokines in intestinal tissue. This study aimed to evaluate the effects of prednisolone treatment on epileptic seizures and investigate the effect of this drug on the bacterial diversity of the intestinal microbiota and markers of inflammatory processes in intestinal tissue. We used Male Wistar rat littermates (n = 31, 90-day-old) divided into four groups: positive control treated with 2 mg/kg of diazepam (n = 6), negative control treated with 0.9 g% sodium chloride (n = 6), and the remaining two groups were subjected to treatment with prednisolone, with one receiving 1 mg/kg (n = 9) and the other 5 mg/kg (n = 10). All administrations were performed intraperitoneally (i.p.) over 14 days. To induce the chronic model of epileptic seizures, we administered pentylenetetrazole (PTZ) 25 mg/kg i.p. on alternate days. Seizure latency (n = 6 - 10) and TNF-α and IL-1ß concentrations from intestinal samples were measured by ELISA (n = 6 per group), and intestinal microbiota was evaluated with intergenic ribosomal RNA (rRNA) spacer (RISA) analysis (n = 6 per group). The prednisolone treatment demonstrated an increase in the latency time of epileptic seizures and TNF-α and IL-1ß concentrations compared to controls. There was no statistically significant difference in intestinal microbiota diversity between the different treatments. However, there was a strong positive correlation between microbial diversity and TNF-α and IL-1ß concentrations. The administration of prednisolone yields comparable results to diazepam on increasing latency between seizures, exhibiting promise for its use in clinical studies. Although there were no changes in intestinal microbial diversity, the increase in the TNF-α and IL-1ß cytokines in intestinal tissue may be linked to immune system signaling pathways involving the intestinal microbiota. Additional research is necessary to unravel the intricacies of these pathways and to understand their implications for clinical practice.

Reduction of seizures and inflammatory markers by betamethasone in a kindling seizure model.

Epilepsy is a chronic disease characterized by an ongoing predisposition to seizures. Although inflammation has emerged as a crucial factor in the etiology of epilepsy, no approaches to anti-inflammatory treatment have been clinically proven to date. Betamethasone (a corticosteroid drug used in the clinic for its anti-inflammatory and immunosuppressive effects) has never been evaluated in attenuating the intensity of seizures in a kindling animal model of seizures. Using a kindling model in male wistar rats, this study evaluated the effect of betamethasone on the severity of seizures and levels of pro-inflammatory interleukins. Seizures were induced by pentylenetetrazole (30 mg/kg) on alternate days for 15 days. The animals were divided into four groups: a control group treated with saline, another control group treated with diazepam (2 mg/kg), and two groups treated with betamethasone (0.125 and 0.250 mg/kg, respectively). Open field test was conducted. Betamethasone treatments were effective in reducing the intensity of epileptic seizures. There were lower levels of Tumor Necrosis Factor-α and interleukin-1ß in the cortex, compared to the saline group, on the other hand, levels in the hippocampus remained similar to the control groups. There was no change in the levels of interleukin-6 in the evaluated structures. Serum inflammatory mediators remained similar. Lower quantities of inflammatory mediators in the central nervous system may have been the key to the reduced severity of seizures on the Racine scale.

Antioxidant and in vitro cytogenotoxic properties of Amburana cearensis (Allemão) A.C.Sm. leaf extract.

Amburana cearensis leaves have been used in folk medicine to treat respiratory diseases and inflammations. This study aimed to evaluate the biological potential of A. cearensis leaves by antioxidant and in vitro cytogenotoxic analyses of ethanolic crude extract (EE) and its fractions in healthy human cells. The EE was obtained by percolation, followed by fractionation using dichloromethane, cyclohexane, ethyl acetate (EtOAc), and methanol (MeOH) as organic solvents. Extract and all fractions were evaluated for their antioxidant potential by DPPH and reducing power tests. In vitro cytotoxic activity was determined in human peripheral blood mononuclear cells by MTT assay for the extract, EtOAc and MeOH fractions. In turn, the genotoxic activity was determined in human lymphocytes by the Cytokinesis Block Micronucleus assay only for the EtOAc fraction. Only EtOAc fraction was analyzed via gas chromatography coupled to mass spectrometry due to its higher biological activity. Considering the antioxidant potential, the EtOAc fraction was most effective in DPPH (EC50 43.37 µg/mL) and reducing power (EC50 89.80 µg/mL) assays. GC-MS analysis of the EtOAc fraction led to the identification of guaiacol, 2,3-dihydro-benzofuran, 2-methoxy-4-vinylphenol, isovanillic acid methyl ester, 4-hydroxybenzaldehyde, and 4-(ethoxymethyl)-phenol. The EE (400-1000 µg/mL), EtOAc (≤150 µg/mL) and MeOH (50 and 150-600 µg/mL) fractions were not cytotoxic by MTT test. Additionally, the EtOAc fraction (100-400 µg/mL) did not induce significant genotoxic damage. Concentrations of the EtOAc fraction with antioxidant activity showed no cytotoxicity, nor genotoxicity potential, indicating them as a nontoxic natural antioxidant source.

Contribuições da psicanálise no manejo de crianças com transtornos do espectro autista / Contributions of psychoanalysis in the management of children with autistic spectrum disorders / Aportaciones del psicoanálisis en el tratamiento de niños con trastornos del espectro autista

Em todo o mundo, as pessoas com Transtorno do Espectro Autista são discriminadas e têm seus direitos violados. Estima-se que uma em cada 160 crianças possuem o distúrbio e os estudos indicam o aumento globalizado da prevalência do Espectro. A psicanálise é conhecida por ser uma clínica do sujeito, e em crianças com TEA, se coloca no lugar de respeito ao jeito de ser de cada indivíduo, independentemente de sua patologia. As intervenções psicossociais com base nas teorias psicanalíticas são uma forma de reduzir as angústias vividas pelo sujeito, às dificuldades de comunicação e convívio social, melhorando a qualidade de vida. Desse modo, a presente pesquisa buscou compreender como a técnica psicanalítica pode contribuir no manejo de crianças com TEA. Para isto foi realizado um estudo de caráter descritivo e qualitativo com 12 psicanalistas que atendem crianças com o espectro. As entrevistas e dados sociodemográficos foram analisados com auxílio do IRAMUTEQ e do SPSS 21.0, respectivamente. Após a análise, as palavras puderam ser agrupadas em dois grandes eixos. O primeiro caracterizando o processo de evolução, onde a importância da família é ponto chave, e o segundo, relacionado ao lugar do psicanalista na direção da análise, caracterizando as especificidades do tratamento, do manejo e linguagem desta demanda. Pode- se perceber que as entrevistas explanaram as vivências da prática clínica, além de um tratamento possível e que mostra resultados no atendimento psicanalítico de crianças diagnosticadas ou em processo de diagnóstico do TEA.

All over the world, people with Autistic Spectrum Disorder are discriminated against and have their rights violated. It is estimated that one in every 160 children has the disorder and studies indicate the global increase in the prevalence of the Spectrum. Psychoanalysis is known to be a clinic of the subject, and in children with ASD, it takes the place of respect for the way of being of each individual, regardless of their pathology. Psychosocial interventions based on psychoanalytic theories are a way to reduce the anxieties experienced by the subject, the difficulties in communication and social interaction, improving the quality of life. Thus, this research sought to understand how the psychoanalytic technique can contribute to the management of children with ASD. For this, a descriptive and qualitative study was carried out with 12 psychoanalysts who assist children with the spectrum. The interviews and sociodemographic data were analyzed using IRAMUTEQ and SPSS 21.0, respectively. After analysis, the words could be grouped into two major axes. The first characterizing the process of evolution, where the importance of the family is a key point, and the second, related to the place of the psychoanalyst in the direction of the analysis, characterizing the specificities of the treatment, handling and language of this demand. It can be seen that the interviews explained the experiences of clinical practice, in addition to a possible treatment that shows results in the psychoanalytic care of children diagnosed or in the process of being diagnosed with ASD.

En todo el mundo, las personas con Trastorno del Espectro Autista son discriminadas y se violan sus derechos. Se estima que uno de cada 160 niños tiene el trastorno y los estudios indican el aumento global de la prevalencia del Espectro. El psicoanálisis se sabe que es una clínica del sujeto, y en los niños con TEA, toma el lugar del respeto por la forma de ser de cada individuo, independientemente de su patología. Las intervenciones psicosociales basadas en las teorías psicoanalíticas son una forma de reducir las ansiedades que experimenta el sujeto, las dificultades en la comunicación y la interacción social, mejorando la calidad de vida. Así, esta investigación buscó comprender cómo la técnica psicoanalítica puede contribuir al manejo de niños con TEA. Para ello, se realizó un estudio descriptivo y cualitativo con 12 psicoanalistas que asisten a niños con el espectro. Las entrevistas y los datos sociodemográficos se analizaron con IRAMUTEQ y SPSS 21.0, respectivamente. Después del análisis, las palabras podrían agruparse en dos grandes ejes. La primera caracterizando el proceso de evolución, donde la importancia de la familia es un punto clave, y la segunda, relacionada con el lugar del psicoanalista en la dirección del análisis, caracterizando las especificidades del tratamiento, manejo y lenguaje de esta demanda. . Se puede apreciar que las entrevistas explicaron las experiencias de la práctica clínica, además de un posible tratamiento que muestra resultados en la atención psicoanalítica de niños diagnosticados o en proceso de ser diagnosticados con TEA. PALABRAS CLAVE: Psicoanálisis; Niños; Trastorno del Espectro Autista.

Aerobic exercise, alone or combined with an anti-inflammatory drug, reduces the severity of epileptic seizures and levels of central pro-inflammatory cytokines in an animal model of epileptic seizures.

Epilepsy is a chronic neurological disorder and there is increasing evidence about the role of inflammation in epileptogenesis. These findings have spurred the search for new immunomodulatory approaches that can improve prognosis. Using an animal model of chemically-induced epileptic seizures, we tested exercise alone as non-pharmacological therapy, and exercise combined with an anti-inflammatory drug. Five groups were used: sedentary, diazepam, aerobic exercise alone, aerobic exercise combined with an anti-inflammatory drug, and naive control. Our goal was to compare the severity of the epileptic seizures between groups as well as seizure latency in a pentylenetetrazole-induced paradigm. Cytokine levels (IL-1ß, TNF-α, and IL-10) were measured. Both exercise groups showed a reduction in seizure severity and lower levels of pro-inflammatory cytokines in the cortex, while the levels of cytokines in the hippocampus remained unaffected.

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.

Gut microbiota modulation by prednisolone in a rat kindling model of pentylenetetrazol (PTZ)-induced seizure.

The gut microbiota is a complex community composed by several microorganisms that interact in the maintenance of homeostasis and contribute to physiological processes, including brain function. The relationship of the taxonomic composition of the gut microbiota with neurological diseases such as autism, Parkinson's, Alzheimer's, anxiety, and depression is widely recognized. The immune system is an important intermediary between the gut microbiota and the central nervous system, being one of the communication routes of the gut-brain axis. Although the complexity of the relationship between inflammation and epilepsy has not yet been elucidated, inflammatory processes are similar in many ways to the consequences of dysbiosis and contribute to disease progression. This study aimed to analyze the taxonomic composition of the gut microbiota of rats treated with prednisolone in a kindling model of epilepsy. Male Wistar rats (90 days, n = 24) divided into four experimental groups: sodium chloride solution 0.9 g%, diazepam 2 mg/kg, prednisolone 1 mg/kg, and prednisolone 5 mg/kg administered intraperitoneally (i.p.) for 14 days. The kindling model was induced by pentylenetetrazole (PTZ) 25 mg/kg i.p. on alternate days. The taxonomic profile was established by applying metagenomic DNA sequencing. There was no change in alpha diversity, and the composition of the gut microbiota between prednisolone and diazepam was similar. The significant increase in Verrucomicrobia, Saccharibacteria, and Actinobacteria may be related to the protective activity against seizures and inflammatory processes that cause some cases of epilepsy. Further studies are needed to investigate the functional influence that these species have on epilepsy and the inflammatory processes that trigger it.

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.

Effects of prednisolone on behavioral and inflammatory profile in animal model of PTZ-induced seizure.

Epilepsy is a chronic neurological condition that affects 1%-2% of the world population. Although research about the disease is advancing and a wide variety of drugs is available, about 30 % of patients have refractory epilepsy which cannot be controlled with the most common drugs. This highlights the need for a better understanding of the disorder and new types of treatment for it. Against this backdrop, a growing body of evidence has reported that inflammation may play a role both in the origin and in the progression of seizures. It has shown a tendency to be both the root and the result of epilepsy. This investigation aimed to assess the impact of prednisolone, a steroidal anti-inflammatory drug, in an animal model of pentylenetetrazole (PTZ)-induced seizures, at 1 mg/kg and 5 mg/kg doses. We also examined the degree of seizure severity and the modulation of pro-inflammatory cytokines in the treated animals. Four treatment groups were used (saline, diazepam, prednisolone 1 mg/kg, and prednisolone 5 mg/kg) and, in addition to their own daily treatments, subconvulsant doses of pentylenetetrazole (25 mg/kg) were administered every other day during a test protocol that lasted 14 days. After treatment, the cytokines interleukin 1 beta (IL-1ß), interleukin 6 (IL-6), and tumor necrosis factor alpha (TNF-α) were measured in the animals' sera, hippocampi, and prefrontal cortices. Animals treated with prednisolone presented less severe seizures than the animals in the saline group, and there was a decrease in pro-inflammatory cytokine levels in central structures, but not peripheral ones. In short, an animal model of chemically-induced epileptic seizures was used, in which the animals were treated with doses of prednisolone, and these animals presented less severe seizures than the negative control group (saline), in addition to showing decreased levels of pro-inflammatory cytokines IL-6, IL-1ß and TNF-α, in the hippocampi and prefrontal cortices, but not the sera.

Development of hydrophobic polyurethane/castor oil biocomposites with agroindustrial residues for sorption of oils and organic solvents.

Since pollution of aquifer sources by organic compounds is a threat to the environment and humans, technology's constant advancement in developing sustainable adsorbents using renewable resources with hydrophobic characteristics has become increasingly attractive. In this study, we developed and characterized castor oil-based polyurethane biocomposites reinforced with malt and/or acerola residues for adsorption/separation of various organic oils/solvents. The residues evidenced biocomposite flexibility, increase in adsorption capacity for all organic oils/solvents, with a maximum of 3257.8 ± 55.6% for chloroform in 60 min and hydrophobicity (144 ± 0.2°) in acidic, saline and alkaline environments, and when exposed to UV radiation. These biocomposites can be quickly and repeatedly recovered by simple hand-tightening and solvent release systems (10 times) and vacuum suction filtration (20.5 ± 0.9 s/10 mL). Through the pseudo-first order (PPO) and pseudo-second order (PSO) kinetic models, the engine oil adsorption capacity increased with time from 746.7 ± 77.4% (60 min) to 928.6 ± 16.7% (90 min). These bio-composites are prototypes to be used with malt and acerola bagasse residues, becoming a reference for future works, produced in a sustainable way, with high potential for application in treatment of effluents with organic pollutants.

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.

Detecting face presentation attacks in mobile devices with a patch-based CNN and a sensor-aware loss function.

With the widespread use of biometric authentication comes the exploitation of presentation attacks, possibly undermining the effectiveness of these technologies in real-world setups. One example takes place when an impostor, aiming at unlocking someone else's smartphone, deceives the built-in face recognition system by presenting a printed image of the user. In this work, we study the problem of automatically detecting presentation attacks against face authentication methods, considering the use-case of fast device unlocking and hardware constraints of mobile devices. To enrich the understanding of how a purely software-based method can be used to tackle the problem, we present a solely data-driven approach trained with multi-resolution patches and a multi-objective loss function crafted specifically to the problem. We provide a careful analysis that considers several user-disjoint and cross-factor protocols, highlighting some of the problems with current datasets and approaches. Such analysis, besides demonstrating the competitive results yielded by the proposed method, provides a better conceptual understanding of the problem. To further enhance efficacy and discriminability, we propose a method that leverages the available gallery of user data in the device and adapts the method decision-making process to the user's and the device's own characteristics. Finally, we introduce a new presentation-attack dataset tailored to the mobile-device setup, with real-world variations in lighting, including outdoors and low-light sessions, in contrast to existing public datasets.

Antimicrobial Potential of Essential Oils from Cerrado Plants against Multidrug-Resistant Foodborne Microorganisms.

Foodborne pathogens are a real public health concern in an escalating antimicrobial resistance scenario. Natural products represent a promising source of bioactive molecules, and essential oils have attracted much attention due to their myriad of biological properties, including antibacterial activities. In this context, essential oils obtained from the leaves of Chromolaena squalida, Campomanesia sessiliflora, Myrsine guianensis, Matayba guianensis, Siparuna guianensis, Ocotea minarum and Endlicheria paniculata-species from the Cerrado biome of Midwest Brazil-were extracted and evaluated for their antibacterial activity against a panel of four standard and three clinical multidrug-resistant bacterial strains. All tested oils showed moderate to good activity against at least four bacterial strains, including Salmonella Typhi and oxacillin-resistant Staphylococcus. The essential oils from C. squalida, C. sessiliflora, My. guianensis and Ma. guianensis showed strong inhibition of clinical Staphylococcus strains, which cause bovine mastitis and are related to milk-borne diseases. Their chemical profiles were investigated by gas chromatography coupled to mass spectrometry (GC/MS), which revealed a predominance of mono- and sesquiterpene hydrocarbons, some of which with well-known antimicrobial properties. The essential oil from Cerrado plants proved active against resistant Gram-positive and Gram-negative bacteria, revealing their potentialities for the development of new alternative agents to prevent the spreading of resistant bacterial contamination.

Increased levels of hexacosanoic acid in the brain of Winstar rats: a behavioral study

Introduction: X-linked adrenoleukodystrophy (X-ALD) is a peroxisomal metabolic disorder associated with mutations in the ATP-binding cassette sub-family D member1 (ABCD1) gene. Practically all male patients with X-ALD develop adrenocortical insufficiency during childhood and progressive myelopathy and peripheral neuropathy in adulthood. However, some male patients develop a fatal cerebral demyelinating disease named cerebral adrenoleukodystrophy. Although the exact mechanisms underlying brain damage in X-ALD are still poorly elucidated, it is known that hexacosanoic acid (C26:0) accumulation represents a hallmark in the pathogenesis of this disease. In this study, we examined whether an overload of C26:0 injected in Wistar rats was capable of causing behavioral changes in these animals. Methods: Egg lecithin in ethanol was dried under a nitrogen stream and mixed with C26:0 methyl ester. Male Wistar rats at 2-3 weeks of age were obtained from Universidade Federal do Rio Grande do Sul (UFRGS), divided into 8 groups, and submitted to an open field test. We then analyzed line crossings (locomotion and exploration), rearing (orienting and investigatory responses), grooming (anxiety manifestation), and latency to move for each animal. Results: Animals subjected to C26:0 administration presented fewer crossings and rearing episodes and a higher latency to move 45 minutes after C26:0 injection. The present work yields experimental evidence that C26:0, the main accumulated metabolite in X-ALD, can cause behavioral alterations in rats such as the impairment of locomotion and exploratory capabilities, as well as a reduction in orienting and investigatory responses. Conclusion: Although our results are preliminary, they are extremely important for future studies that investigate C26:0 accumulation and locomotor impairment in patients with X-ALD. (AU)

Comparative anatomy and histochemistry of the leaf blade of two species of Artocarpus.

In Brazil, there are two species of Artocarpus that were introduced: Artocarpus altilis (Parkinson) Fosberg, known as fruta-pão, and Artocarpus heterophyllus Lam., known as jaca. Both are used as food and medicine. The objective of this work was to conduct a comparative anatomical and histochemical study between A. altilis and A. heterophyllus. Techniques of optical, polarized and scanning electron coupled to energy dispersive spectroscopy. The anatomical characterization showed the characters of general occurrence in the family Moraceae and of those that allow the differentiation of A. altilis and A. heterophyllus. The histochemistry revealed the sites of synthesis and/or storage of the metabolites. The chemical microanalyses brought new information about the chemical composition of crystals. The study provides pharmacobotanical data for the quality control of the species.

Anatomy, histochemistry and oxalic acid content of the leaflets of Averrhoa bilimbi and Averrhoa carambola

Abstract Averrhoa bilimbi L. and A. carambola L., Oxalidaceae, are the only two species of the genus Averrhoa L. Their leaves are widely used in folk medicine as an adjuvant in the treatment of diabetes. Some species may contain, for example, calcium oxalate crystals, which may lead to risk in its use when there is predisposition of individuals with reduced renal activity. Therefore, there are still few studies on the content of oxalic acid present in them, highlighting the importance of this investigation. The objective of this work was to conduct a comparative anatomical and histochemical study between the species and determining its content of oxalic acid. Semipermanent histological slides were prepared, following common plant anatomy procedures, for analysis of the leaflets in optical microscopy, polarization and scanning electron microscopy coupled with energy-dispersive X-ray spectrometry. To determine the total, soluble and insoluble oxalate content was used titration with potassium permanganate. The anatomical characterization allowed identifying the characters useful in the differentiation of the species. The histochemistry revealed the location of the metabolites. Chemical microanalyses demonstrated that the crystals are of calcium oxalate. A. carambola presented the highest levels of total oxalate and soluble oxalate. The study assists in the identification and quality control of A. bilimbi and A. carambola and brings new data on its oxalic acid content, which are important, in view of the medicinal use of the species.

Anatomical characterization of ultra-structures, biominerals and histolocalization of metabolites in leaves of Genipa americana

Abstract Inserted in one of the largest families among the Angiosperms, Genipa americana L., Rubiaceae, can be found in all Brazilian territory, presenting great medicinal importance, where several uses have been attributed. In view of this, this work has the purpose of analyzing the ultrastructural, biomineral, phytochemical and histochemical characteristics of the leaves of this native species from Brazil. For this, light microscopy, polarization and scanning electron microscopy techniques were used with X-ray scattering energy, associated to chromatographic and histochemical tests. The anatomical ultrastructural characteristics of the leaves detailed information about the type and arrangement of the cuticle, trichomes, surface and arrangement of the tissues that determine the botanical identity of this species. The phytochemical tests allowed determining their chromatographic pattern and histochemistry to determine the exact storage site of these substances in the leaf. It was observed that the characterization of the crystalline macro-pattern present in the analyzed species, as well as its exact elemental composition, can be considered an important differential diagnosis factor. The results characterize the leaves of this species in different aspects, being a native species and pharmacologically promising, with different popular uses and proven pharmacological activities, and more in depth studies is needed.

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.

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.