Daniel Graziani - Academia.edu (original) (raw)

Papers by Daniel Graziani

Research paper thumbnail of Extrato etanólico de açafrão (Curcuma longa L.) reduz apoptose e promove proliferação de células de osteossarcoma canino

Ciência Animal Brasileira, 2022

Extrato etanólico de açafrão (Curcuma longa L.) reduz apoptose e promove proliferação de células ... more Extrato etanólico de açafrão (Curcuma longa L.) reduz apoptose e promove proliferação de células de osteossarcoma canino Turmeric ethanol extract (Curcuma longa L.) reduces apoptosis and promotes canine osteosarcoma cell proliferation

Research paper thumbnail of Turmeric ethanol extract (Curcuma longa L.) reduces apoptosis and promotes canine osteosarcoma cell proliferation

Ciência Animal Brasileira, 2022

Extrato etanólico de açafrão (Curcuma longa L.) reduz apoptose e promove proliferação de células ... more Extrato etanólico de açafrão (Curcuma longa L.) reduz apoptose e promove proliferação de células de osteossarcoma canino Turmeric ethanol extract (Curcuma longa L.) reduces apoptosis and promotes canine osteosarcoma cell proliferation Abstract Curcuma longa L., also known as turmeric, has been widely studied for its various therapeutic properties, including antineoplastic action. The ethanolic extract of the plant contains several phenolic compounds, especially curcumin. Osteosarcoma is a predominant bone tumor in dogs and humans, characterized by high metastatic potential and an unfavorable prognosis. The aim of this study was to investigate the effects of turmeric ethanol extract on canine osteosarcoma cells from established culture. The cells were cultured and treated with different curcumin concentrations (0, 10 μM, 20 μM, 50 μM, 100 μM, and 1000 μM) and exposure times (24h, 48h, and 72h). We first performed tetrazolium reduction technique (MTT) assay and calculated IC 50. An immunocytochemistry assay was performed after extract treatment to verify the expression of mutated p53 and therefore study the proliferative potential of malignant cells; Bcl-2 and Ki-67 were used to assess apoptosis and the degree of malignancy, respectively. The extract enhanced the proliferation of canine osteosarcoma cells, reaching 3,819.74% at 50 μM of curcumin. The extract also significantly altered the expression of mutated p53 and Ki-67 proteins but not that of Bcl-2, suggesting that it did not induce this antiapoptotic pathway. Overall, these results are prerequisite to better understanding how natural compounds such as turmeric ethanolic extract affect cell proliferation and could be used to treat various diseases.

Research paper thumbnail of Efeitos celulares e comportamentais de peptídeos bioativos de baixo peso molecular extraídos de Phaseolus vulgaris

pré-requisito para a defesa de produto final-Nível Doutorado.

Research paper thumbnail of Nasal biocompatible powder of Geraniol oil complexed with cyclodextrins for neurodegenerative diseases: physicochemical characterization and in vivo evidences of nose to brain delivery

Journal of Controlled Release, Jul 1, 2021

Recently, many studies have shown that plant metabolites, such as geraniol (GER), may exert anti-... more Recently, many studies have shown that plant metabolites, such as geraniol (GER), may exert anti-inflammatory effects in neurodegenerative diseases and, in particular, Parkinson's disease (PD) models. Unfortunately, delivering GER to the CNS via nose-to-brain is not feasible due to its irritant effects on the mucosae. Therefore, in the present study β-cyclodextrin (βCD) and its hydrophilic derivative hydroxypropyl-beta-cyclodextrin (HPβCD) were selected as potential carriers for GER nose-to-brain delivery. Inclusion complexes were formulated and the biocompatibility with nasal mucosae and drug bioavailability into cerebrospinal fluid (CSF) were studied in rats. It has been demonstrated by DTA, FT-IR and NMR analyses that both the CDs were able to form 1:1 GER-CD complexes, arising long-term stable powders after the freeze-drying process. GER-HPβCD-5 and GER-βCD-2 complexes exhibited comparable results, except for morphology and solubility, as demonstrated by SEM analysis and phase solubility study, respectively. Even though both complexes were able to directly and safely deliver GER to CNS, GER-βCD-2 displayed higher ability in releasing GER in the CSF. In conclusion, βCD complexes can be considered a very promising tool in delivering GER into the CNS via nose-to-brain route, preventing GER release into the bloodstream and ensuring the integrity of the nasal mucosa.

Research paper thumbnail of Extrato rico em Curcumina do Brasil Central Curcuma longa protege células endoteliais de veias umbilicais humanas sob estresse oxidativo

ABCS Health Sciences, Dec 16, 2022

Borges et al. Curcumin-rich extract from Central Brazil Curcuma longa protects human umbilical ve... more Borges et al. Curcumin-rich extract from Central Brazil Curcuma longa protects human umbilical vein endothelial cells under oxidative stress. ABCS Health Sci.

Research paper thumbnail of Estudo Das Características Epidemiológicas Da Leishmaniose Tegumentar Americana No Estado De Goiás, Brasil, 2007-2009

Revista de Patologia Tropical, Jan 13, 2014

A leishmaniose tegumentar americana é um grave problema de saúde pública em vários países do mund... more A leishmaniose tegumentar americana é um grave problema de saúde pública em vários países do mundo. O presente trabalho visou realizar o levantamento descritivo das características epidemiológicas da doença no estado de Goiás entre os anos de 2007 e 2009. Este trabalho consistiu em uma pesquisa retrospectiva descritiva com base nos dados notificados e cadastrados no Sistema de Informação de Agravos de Notificação (SINAN). O estudo mostrou a distribuição dos casos autóctones por município, evidenciando a expansão da doença no estado. Assim, foram notificados 1.285 casos confirmados de leishmaniose tegumentar americana, dos quais 848 eram autóctones em 130 municípios. Em 2007, foram notificados 233 casos autóctones em 75 municípios; em 2008, 276 casos em 81 municípios e, em 2009, 349 casos em 85 municípios. Estes dados sugerem uma leve tendência de aumento de casos e uma ampla distribuição geográfica. Houve predominância de casos que envolviam indivíduos habitantes da área urbana, do sexo masculino, de raça parda e escolaridade inferior ao ensino médio. O resultado da pesquisa sugere a necessidade de uma investigação mais profunda das causas dessas infecções, o que demonstra a importância deste estudo, uma vez que oferece dados que podem subsidiar programas voltados à informação e profilaxia deste tipo de infecção.

Research paper thumbnail of Análise molecular de isolados do gênero leishmania e clínica de pacientes portadores de leishmaniose tegumentar americana atendidos no Hospital Universitário de Brasília

Leishmaniasis is a serious public health problem in Brazil and in several countries worldwide. Th... more Leishmaniasis is a serious public health problem in Brazil and in several countries worldwide. The different clinical manifestations may vary since a localized lesion until multiple lesions distributed throughout the body. Factors inherent to the different species of the parasite contribute to the clinical diversity and pathogenesis of the disease. The aim of this work was to study through molecular methodologies of Leishmania's isolates and relate them with clinical aspects of the patient's sources. The identification was performed by PCR miniexon gene, PCR-RFLP and sequencing of ITS1 from rDNA gene. In this study 38 isolates from 36 patients were analyzed and were identified: 22 L. (V.) braziliensis, 3 L. (V.) panamensis, 1 L. (V.

Research paper thumbnail of Déficit Da Memória Na Doença De Alzheimer Visualizada Por Outra Perspectiva

Research paper thumbnail of Behavioral effects of Bj-PRO-7a, a proline-rich oligopeptide from Bothrops jararaca venom

Brazilian Journal of Medical and Biological Research, 2019

The heptapeptide Bj-PRO-7a, isolated and identified from Bothrops jararaca (Bj) venom, produces a... more The heptapeptide Bj-PRO-7a, isolated and identified from Bothrops jararaca (Bj) venom, produces antihypertensive and other cardiovascular effects that are independent on angiotensin converting enzyme inhibition, possibly relying on cholinergic muscarinic receptors subtype 1 (M 1 R). However, whether Bj-PRO-7a acts upon the central nervous system and modifies behavior is yet to be determined. Therefore, the aims of this study were: i) to assess the effects of acute administration of Bj-PRO-7a upon behavior; ii) to reveal mechanisms involved in the effects of Bj-PRO-7a upon locomotion/exploration, anxiety, and depression-like behaviors. For this purpose, adult male Wistar (WT, wild type) and spontaneous hypertensive rats (SHR) received intraperitoneal injections of vehicle (0.9% NaCl), diazepam (2 mg/kg), imipramine (15 mg/kg), Bj-PRO-7a (71, 213 or 426 nmol/kg), pirenzepine (852 nmol/kg), a-methyl-DL-tyrosine (200 mg/kg), or chlorpromazine (2 mg/kg), and underwent elevated plus maze, open field, and forced swimming tests. The heptapeptide promoted anxiolytic and antidepressant-like effects and increased locomotion/exploration. These effects of Bj-PRO-7a seem to be dependent on M 1 R activation and dopaminergic receptors and rely on catecholaminergic pathways.

Research paper thumbnail of Behavioral effects of a low molecular weight peptide fraction from Phaseolus vulgaris in rats

Brazilian Journal of Medical and Biological Research, 2022

Seminal studies stated that bean proteins are efficient neuronal tracers with affinity for brain ... more Seminal studies stated that bean proteins are efficient neuronal tracers with affinity for brain tissue. A low molecular weight peptide fraction (o3kDa) from Phaseolus vulgaris (PV3) was previously reported to be antioxidant, non-cytotoxic, and capable of reducing reactive oxygen species and increasing nitric oxide in cells. We evaluated the effects of PV3 (5, 50, 100, 500, and 5000 mg/kg) on behavior and the molecular routes potentially involved. Acute and chronic PV3 treatments were performed before testing Wistar rats: i) in the elevated plus-maze (EPM) to assess the anxiolytic-like effect; ii) in the open field (OF) to evaluate locomotion and exploration; and iii) for depression-like behavior in forced swimming (FS). Catecholaminergic involvement was tested using the tyrosine hydroxylases (TH) enzyme inhibitor, a-methyl-DL-tyrosine (AMPT). Brain areas of chronically treated groups were dissected to assess: i) lipid peroxidation (LPO); ii) carbonylated proteins (CP); iii) superoxide dismutase (SOD) and catalase (CAT) enzymatic activities. Neuronal nitric oxide synthases (nNOS) and argininosuccinate synthase (ASS) protein expression was evaluated by western blotting. Acute treatment with PV3 increased the frequency and time spent in the EPM open arms, suggesting anxiolysis. PV3 increased crossing episodes in the OF. These PV3 effects on anxiety and locomotion were absent in the chronically treated group. Acute and chronic PV3 treatments reduced the immobility time in the FS test, suggesting an antidepressant effect. TH inhibition by AMPT reverted acute PV3 effects. PV3 decreased LPO and CP levels and SOD and CAT activities, whereas nNOS and ASS were reduced in few brain areas. In conclusion, PV3 displayed central antioxidant actions that are concomitant to catecholaminergic-dependent anxiolytic and antidepressant effects.

Research paper thumbnail of Phylogenetic Inferences Based on Distinct Molecular Markers Confirm a Novel Babesia Species (Babesia goianiaensis nov. sp.) in Capybaras (Hydrochoerus hydrochaeris) and Associated Ticks

Microorganisms, Aug 6, 2023

Inferences Based on Distinct Molecular Markers Confirm a Novel Babesia Species (Babesia goianiaen... more Inferences Based on Distinct Molecular Markers Confirm a Novel Babesia Species (Babesia goianiaensis nov. sp.) in Capybaras (Hydrochoerus hydrochaeris) and Associated Ticks.

Research paper thumbnail of Phylogenetic Inferences Based on Distinct Molecular Markers Confirm a Novel Babesia Species (Babesia goianiaensis nov. sp.) in Capybaras (Hydrochoerus hydrochaeris) and Associated Ticks

Microorganisms

Piroplasmids (order Piroplasmida) are a diverse group of tick-borne protozoa that may cause disea... more Piroplasmids (order Piroplasmida) are a diverse group of tick-borne protozoa that may cause disease in animals and occasionally in humans. Novel Piroplasmida clades and species have been found in wild animals from Brazil based on the phylogenetic assessment of near-complete 18S rRNA, mitochondrial and heat-shock protein genes. For instance, a putative novel Babesia species has been detected in capybaras and Amblyomma ticks in three Brazilian states. The present work aimed to describe, using phylogenetic assessments based on distinct molecular markers, this novel Babesia species in capybaras and associated ticks (Amblyomma sculptum and Amblyomma dubitatum) sampled in Goiânia city, Goiás state, midwestern Brazil. While the phylogenetic analysis based on both near-complete 18S rRNA and hsp-70 genes positioned the sequences obtained from capybara blood samples into a new clade sister to the Babesia sensu stricto clade, the phylogenetic inference based on the COX-3 amino acid positioned ...

Research paper thumbnail of Oxidonitrergic and antioxidant effects of a low molecular weight peptide fraction from hardened bean (Phaseolus vulgaris) on endothelium

Brazilian Journal of Medical and Biological Research, 2021

About 3000 tons of beans are not used in human food due to hardening. Several studies on bean-der... more About 3000 tons of beans are not used in human food due to hardening. Several studies on bean-derived bioactive peptides have shown potential to treat some diseases, including those relying on oxidative dysfunctions. We assessed the effects of peptides extracted from hardened bean Phaseolus vulgaris (PV) on reactive oxygen species (ROS) and nitric oxide (NO) production, cytotoxic and cytoprotective effects in endothelial cells, and oxidonitrergic-dependent vasodilating effects. Extract was composed by peptide fraction o3 kDa (PV3) from hardened common bean residue. PV3 sequences were obtained and analyzed with bioinformatics. Human umbilical vein endothelial cells were treated with 10, 20, 30, and 250 mg/mL PV3. Oxidative stress was provoked by 3% H 2 O 2. Cytotoxicity and cytoprotective effects were evaluated by MTT assay, whereas, ROS and NO were quantified using DHE and DAF-FM fluorescent probes by confocal microscopy. NO-and endotheliumdependent vasodilating effects of PV3 were assessed in isolated aortic rings. We found 35 peptides with an average mass of 1.14 kDa. There were no cell deaths with 10 and 20 mg/mL PV3. PV3 at 30 mg/mL increased cell viability, while cytotoxicity was observed only with 250 mg/mL PV3. PV3 at 10 mg/mL was able to protect cells from oxidative stress. PV3 also increased NO release without causing cell death. It also reduced relative ROS production induced by H 2 O 2. PV3 vasodilating effects relied on endothelium-dependent NO release. PV3 obtained from low-commercial-value bean displays little cytotoxicity and exerts antioxidant effects, whereas it increases endothelial NO release.

Research paper thumbnail of Turmeric ethanol extract (Curcuma longa L.) reduces apoptosis and promotes canine osteosarcoma cell proliferation

Ciência Animal Brasileira

Curcuma longa L., also known as turmeric, has been widely studied for its various therapeutic pro... more Curcuma longa L., also known as turmeric, has been widely studied for its various therapeutic properties, including antineoplastic action. The ethanolic extract of the plant contains several phenolic compounds, especially curcumin. Osteosarcoma is a predominant bone tumor in dogs and humans, characterized by high metastatic potential and an unfavorable prognosis. The aim of this study was to investigate the effects of turmeric ethanol extract on canine osteosarcoma cells from established culture. The cells were cultured and treated with different curcumin concentrations (0, 10 μM, 20 μM, 50 μM, 100 μM, and 1000 μM) and exposure times (24h, 48h, and 72h). We first performed tetrazolium reduction technique (MTT) assay and calculated IC50. An immunocytochemistry assay was performed after extract treatment to verify the expression of mutated p53 and therefore study the proliferative potential of malignant cells; Bcl-2 and Ki-67 were used to assess apoptosis and the degree of malignancy,...

Research paper thumbnail of Ventromedial medullary pathway mediating cardiac responses evoked from periaqueductal gray

Autonomic Neuroscience, 2020

Periaqueductal gray (PAG) is a midbrain region that projects to areas controlling behavioral and ... more Periaqueductal gray (PAG) is a midbrain region that projects to areas controlling behavioral and autonomic outputs and is involved in the behavioral and physiological components of defense reactions. Since Raphe Pallidus (RPa) is a medial medullary region comprising sympathetic premotor neurons governing heart function, it is worth considering the PAG-RPa path. We assessed: i) whether PAG projects to RPa; ii) the amplitude of cardiac responses evoked from PAG; iii) whether cardiovascular responses evoked from PAG rely on RPa. Experiments conducted in Wistar rats (± 300 g) were approved by Ethics Committee CEUA-UFG (092/18). Firstly, (n = 3), monosynaptic retrograde tracer Retrobeads was injected into RPa; PAG slices were analyzed. Other two groups (n = 6 each) were anesthetized with urethane (1.4 g/kg) and chloralose (120 mg/ kg) and underwent craniotomy, tracheostomy, catheterization of femoral artery and vein and of cardiac left ventricle. In one group, we injected the GABA A receptor antagonist, bicuculline methiodide (BMI-40 pmol/ 100 nL) into lateral/dorsolateral PAG. Another group was injected (100 nL) with the GABA A receptor agonist muscimol (20 mM) into RPa, 20 min before BMI into PAG. The results were: i) retrogradely labelled neurons were found in PAG; ii) PAG activation by BMI caused positive chronotropism and inotropism, which were accompanied by afterload increases; iii) RPa inhibition with Muscimol reduced heart rate, arterial and ventricular pressures; iv) the subsequent PAG activation still increased arterial pressure, cardiac chronotropy and inotropy, but these responses were significantly attenuated. In conclusion, PAG activation increases cardiac chronotropy and inotropy, and these responses seem to rely on a direct pathway reaching ventromedial medullary RPa neurons.

Research paper thumbnail of A peptide fraction from hardened common beans (Phaseolus vulgaris) induces endothelium-dependent antihypertensive and renal effects in rats

Current Research in Food Science

Beans reached the research spotlight as a source of bioactive compounds capable of modulating dif... more Beans reached the research spotlight as a source of bioactive compounds capable of modulating different functions. Recently, we reported antioxidant and oxidonitrergic effect of a low molecular weight peptide fraction (<3 kDa) from hardened bean (Phaseolus vulgaris) in vitro and ex vivo, which necessitate further in vivo assessments. This work aimed to evaluate the hypotensive effect and the involved physiological mechanisms of the hardened common bean peptide (Phaseolus vulgaris) in normotensive (Wistar) and hypertensive (SHR) animals. Bean flour was combined with a solution containing acetonitrile, water and formic acid (25: 24: 1). Protein extract (PV3) was fractioned (3 kDa membrane). We assessed PV3 effects on renal function and hemodynamics of wistar (WT-normotensive) and spontaneously hypertensive rats (SHR) and measured systemic arterial pressure and flow in aortic and renal beds. The potential endothelial and oxidonitrergic involvements were tested in isolated renal artery rings. As results, we found that PV3: I) decreased food consumption in SHR, increased water intake and urinary volume in WT, increased glomerular filtration rate in WT and SHR, caused natriuresis in SHR; II) caused NO-and endothelium-dependent vasorelaxation in renal artery rings; III) reduced arterial pressure and resistance in aortic and renal vascular beds; IV) caused antihypertensive effects in a dose-dependent manner. Current findings support PV3 as a source of bioactive peptides and raise the potential of composing nutraceutical formulations to treat renal and cardiovascular diseases.

Research paper thumbnail of Extrato etanólico de açafrão (Curcuma longa L.) reduz apoptose e promove proliferação de células de osteossarcoma canino

Ciência Animal Brasileira

Resumo A Curcuma longa L., planta conhecida popularmente como açafrão, tem sido amplamente estuda... more Resumo A Curcuma longa L., planta conhecida popularmente como açafrão, tem sido amplamente estudada por suas diversas propriedades terapêuticas, incluindo a ação antineoplásica. O extrato etanólico da planta contém diversos compostos fenólicos, com destaque para a curcumina. O osteossarcoma é um tumor ósseo predominante em cães e humanos, caracterizado por apresentar alto potencial metastático e prognóstico desfavorável. Procurou-se investigar os efeitos de diferentes concentrações de curcumina do extrato etanólico de açafrão sobre células de osteossarcoma canino de cultura estabelecida. As células foram cultivadas e submetidas ao tratamento com extrato com diferentes concentrações de curcumina (0, 10 μM, 20 μM, 50 μM, 100 μM e 1000 μM) e tempos de exposição (24h, 48h e 72h) pelo EEA. Inicialmente, foram realizados: técnica de redução do tetrazólio (MTT) e cálculo da IC50. Posteriormente, após o tratamento com o extrato, realizou-se o ensaio de imunocitoquímica para verificar a expr...

Research paper thumbnail of Extrato rico em Curcumina do Brasil Central Curcuma longa protege células endoteliais de veias umbilicais humanas sob estresse oxidativo

ABCS Health Sciences

Introdução: Em doenças vasculares, a interrupção do fluxo sanguíneo locale subsequente reperfusão... more Introdução: Em doenças vasculares, a interrupção do fluxo sanguíneo locale subsequente reperfusão de oxigênio pode causar efeitos deletérios e danos irreparáveis às células. Curcuma (Curcuma longa L.) neutraliza radicais livres além de apresentar efeitos preventivos e terapêuticos. Objetivo: Caracterizar os compostos bioativos e a capacidade antioxidante do extrato etanólico de cúrcuma (EEC); avaliar seu efeito nas células endoteliais da veia umbilical humana, e analisar a expressão de vias de sinalização celular. Métodos: As células foram expostas a diferentes concentrações de EEC por 24, 48 e 72 horas. Utilizamos o teste de Folin-Ciocalteau, análise por HPLC-Fluorescência e método DPPH para determinar os compostos fenólicos, conteúdo de curcumina e ação antioxidante, respectivamente; o método de redução de tetrazólio para viabilidade celular, a citotoxicidade e a concentração que inibe 50% da viabilidade celular; e a técnica de imunocitoquímica para analisar a expressão de caspase...

Research paper thumbnail of Déficit Da Memória Na Doença De Alzheimer Visualizada Por Outra Perspectiva

Research paper thumbnail of Nasal biocompatible powder of Geraniol oil complexed with cyclodextrins for neurodegenerative diseases: physicochemical characterization and in vivo evidences of nose to brain delivery

Journal of Controlled Release, 2021

Recently, many studies have shown that plant metabolites, such as geraniol (GER), may exert anti-... more Recently, many studies have shown that plant metabolites, such as geraniol (GER), may exert anti-inflammatory effects in neurodegenerative diseases and, in particular, Parkinson's disease (PD) models. Unfortunately, delivering GER to the CNS via nose-to-brain is not feasible due to its irritant effects on the mucosae. Therefore, in the present study β-cyclodextrin (βCD) and its hydrophilic derivative hydroxypropyl-beta-cyclodextrin (HPβCD) were selected as potential carriers for GER nose-to-brain delivery. Inclusion complexes were formulated and the biocompatibility with nasal mucosae and drug bioavailability into cerebrospinal fluid (CSF) were studied in rats. It has been demonstrated by DTA, FT-IR and NMR analyses that both the CDs were able to form 1:1 GER-CD complexes, arising long-term stable powders after the freeze-drying process. GER-HPβCD-5 and GER-βCD-2 complexes exhibited comparable results, except for morphology and solubility, as demonstrated by SEM analysis and phase solubility study, respectively. Even though both complexes were able to directly and safely deliver GER to CNS, GER-βCD-2 displayed higher ability in releasing GER in the CSF. In conclusion, βCD complexes can be considered a very promising tool in delivering GER into the CNS via nose-to-brain route, preventing GER release into the bloodstream and ensuring the integrity of the nasal mucosa.

Research paper thumbnail of Extrato etanólico de açafrão (Curcuma longa L.) reduz apoptose e promove proliferação de células de osteossarcoma canino

Ciência Animal Brasileira, 2022

Extrato etanólico de açafrão (Curcuma longa L.) reduz apoptose e promove proliferação de células ... more Extrato etanólico de açafrão (Curcuma longa L.) reduz apoptose e promove proliferação de células de osteossarcoma canino Turmeric ethanol extract (Curcuma longa L.) reduces apoptosis and promotes canine osteosarcoma cell proliferation

Research paper thumbnail of Turmeric ethanol extract (Curcuma longa L.) reduces apoptosis and promotes canine osteosarcoma cell proliferation

Ciência Animal Brasileira, 2022

Extrato etanólico de açafrão (Curcuma longa L.) reduz apoptose e promove proliferação de células ... more Extrato etanólico de açafrão (Curcuma longa L.) reduz apoptose e promove proliferação de células de osteossarcoma canino Turmeric ethanol extract (Curcuma longa L.) reduces apoptosis and promotes canine osteosarcoma cell proliferation Abstract Curcuma longa L., also known as turmeric, has been widely studied for its various therapeutic properties, including antineoplastic action. The ethanolic extract of the plant contains several phenolic compounds, especially curcumin. Osteosarcoma is a predominant bone tumor in dogs and humans, characterized by high metastatic potential and an unfavorable prognosis. The aim of this study was to investigate the effects of turmeric ethanol extract on canine osteosarcoma cells from established culture. The cells were cultured and treated with different curcumin concentrations (0, 10 μM, 20 μM, 50 μM, 100 μM, and 1000 μM) and exposure times (24h, 48h, and 72h). We first performed tetrazolium reduction technique (MTT) assay and calculated IC 50. An immunocytochemistry assay was performed after extract treatment to verify the expression of mutated p53 and therefore study the proliferative potential of malignant cells; Bcl-2 and Ki-67 were used to assess apoptosis and the degree of malignancy, respectively. The extract enhanced the proliferation of canine osteosarcoma cells, reaching 3,819.74% at 50 μM of curcumin. The extract also significantly altered the expression of mutated p53 and Ki-67 proteins but not that of Bcl-2, suggesting that it did not induce this antiapoptotic pathway. Overall, these results are prerequisite to better understanding how natural compounds such as turmeric ethanolic extract affect cell proliferation and could be used to treat various diseases.

Research paper thumbnail of Efeitos celulares e comportamentais de peptídeos bioativos de baixo peso molecular extraídos de Phaseolus vulgaris

pré-requisito para a defesa de produto final-Nível Doutorado.

Research paper thumbnail of Nasal biocompatible powder of Geraniol oil complexed with cyclodextrins for neurodegenerative diseases: physicochemical characterization and in vivo evidences of nose to brain delivery

Journal of Controlled Release, Jul 1, 2021

Recently, many studies have shown that plant metabolites, such as geraniol (GER), may exert anti-... more Recently, many studies have shown that plant metabolites, such as geraniol (GER), may exert anti-inflammatory effects in neurodegenerative diseases and, in particular, Parkinson's disease (PD) models. Unfortunately, delivering GER to the CNS via nose-to-brain is not feasible due to its irritant effects on the mucosae. Therefore, in the present study β-cyclodextrin (βCD) and its hydrophilic derivative hydroxypropyl-beta-cyclodextrin (HPβCD) were selected as potential carriers for GER nose-to-brain delivery. Inclusion complexes were formulated and the biocompatibility with nasal mucosae and drug bioavailability into cerebrospinal fluid (CSF) were studied in rats. It has been demonstrated by DTA, FT-IR and NMR analyses that both the CDs were able to form 1:1 GER-CD complexes, arising long-term stable powders after the freeze-drying process. GER-HPβCD-5 and GER-βCD-2 complexes exhibited comparable results, except for morphology and solubility, as demonstrated by SEM analysis and phase solubility study, respectively. Even though both complexes were able to directly and safely deliver GER to CNS, GER-βCD-2 displayed higher ability in releasing GER in the CSF. In conclusion, βCD complexes can be considered a very promising tool in delivering GER into the CNS via nose-to-brain route, preventing GER release into the bloodstream and ensuring the integrity of the nasal mucosa.

Research paper thumbnail of Extrato rico em Curcumina do Brasil Central Curcuma longa protege células endoteliais de veias umbilicais humanas sob estresse oxidativo

ABCS Health Sciences, Dec 16, 2022

Borges et al. Curcumin-rich extract from Central Brazil Curcuma longa protects human umbilical ve... more Borges et al. Curcumin-rich extract from Central Brazil Curcuma longa protects human umbilical vein endothelial cells under oxidative stress. ABCS Health Sci.

Research paper thumbnail of Estudo Das Características Epidemiológicas Da Leishmaniose Tegumentar Americana No Estado De Goiás, Brasil, 2007-2009

Revista de Patologia Tropical, Jan 13, 2014

A leishmaniose tegumentar americana é um grave problema de saúde pública em vários países do mund... more A leishmaniose tegumentar americana é um grave problema de saúde pública em vários países do mundo. O presente trabalho visou realizar o levantamento descritivo das características epidemiológicas da doença no estado de Goiás entre os anos de 2007 e 2009. Este trabalho consistiu em uma pesquisa retrospectiva descritiva com base nos dados notificados e cadastrados no Sistema de Informação de Agravos de Notificação (SINAN). O estudo mostrou a distribuição dos casos autóctones por município, evidenciando a expansão da doença no estado. Assim, foram notificados 1.285 casos confirmados de leishmaniose tegumentar americana, dos quais 848 eram autóctones em 130 municípios. Em 2007, foram notificados 233 casos autóctones em 75 municípios; em 2008, 276 casos em 81 municípios e, em 2009, 349 casos em 85 municípios. Estes dados sugerem uma leve tendência de aumento de casos e uma ampla distribuição geográfica. Houve predominância de casos que envolviam indivíduos habitantes da área urbana, do sexo masculino, de raça parda e escolaridade inferior ao ensino médio. O resultado da pesquisa sugere a necessidade de uma investigação mais profunda das causas dessas infecções, o que demonstra a importância deste estudo, uma vez que oferece dados que podem subsidiar programas voltados à informação e profilaxia deste tipo de infecção.

Research paper thumbnail of Análise molecular de isolados do gênero leishmania e clínica de pacientes portadores de leishmaniose tegumentar americana atendidos no Hospital Universitário de Brasília

Leishmaniasis is a serious public health problem in Brazil and in several countries worldwide. Th... more Leishmaniasis is a serious public health problem in Brazil and in several countries worldwide. The different clinical manifestations may vary since a localized lesion until multiple lesions distributed throughout the body. Factors inherent to the different species of the parasite contribute to the clinical diversity and pathogenesis of the disease. The aim of this work was to study through molecular methodologies of Leishmania's isolates and relate them with clinical aspects of the patient's sources. The identification was performed by PCR miniexon gene, PCR-RFLP and sequencing of ITS1 from rDNA gene. In this study 38 isolates from 36 patients were analyzed and were identified: 22 L. (V.) braziliensis, 3 L. (V.) panamensis, 1 L. (V.

Research paper thumbnail of Déficit Da Memória Na Doença De Alzheimer Visualizada Por Outra Perspectiva

Research paper thumbnail of Behavioral effects of Bj-PRO-7a, a proline-rich oligopeptide from Bothrops jararaca venom

Brazilian Journal of Medical and Biological Research, 2019

The heptapeptide Bj-PRO-7a, isolated and identified from Bothrops jararaca (Bj) venom, produces a... more The heptapeptide Bj-PRO-7a, isolated and identified from Bothrops jararaca (Bj) venom, produces antihypertensive and other cardiovascular effects that are independent on angiotensin converting enzyme inhibition, possibly relying on cholinergic muscarinic receptors subtype 1 (M 1 R). However, whether Bj-PRO-7a acts upon the central nervous system and modifies behavior is yet to be determined. Therefore, the aims of this study were: i) to assess the effects of acute administration of Bj-PRO-7a upon behavior; ii) to reveal mechanisms involved in the effects of Bj-PRO-7a upon locomotion/exploration, anxiety, and depression-like behaviors. For this purpose, adult male Wistar (WT, wild type) and spontaneous hypertensive rats (SHR) received intraperitoneal injections of vehicle (0.9% NaCl), diazepam (2 mg/kg), imipramine (15 mg/kg), Bj-PRO-7a (71, 213 or 426 nmol/kg), pirenzepine (852 nmol/kg), a-methyl-DL-tyrosine (200 mg/kg), or chlorpromazine (2 mg/kg), and underwent elevated plus maze, open field, and forced swimming tests. The heptapeptide promoted anxiolytic and antidepressant-like effects and increased locomotion/exploration. These effects of Bj-PRO-7a seem to be dependent on M 1 R activation and dopaminergic receptors and rely on catecholaminergic pathways.

Research paper thumbnail of Behavioral effects of a low molecular weight peptide fraction from Phaseolus vulgaris in rats

Brazilian Journal of Medical and Biological Research, 2022

Seminal studies stated that bean proteins are efficient neuronal tracers with affinity for brain ... more Seminal studies stated that bean proteins are efficient neuronal tracers with affinity for brain tissue. A low molecular weight peptide fraction (o3kDa) from Phaseolus vulgaris (PV3) was previously reported to be antioxidant, non-cytotoxic, and capable of reducing reactive oxygen species and increasing nitric oxide in cells. We evaluated the effects of PV3 (5, 50, 100, 500, and 5000 mg/kg) on behavior and the molecular routes potentially involved. Acute and chronic PV3 treatments were performed before testing Wistar rats: i) in the elevated plus-maze (EPM) to assess the anxiolytic-like effect; ii) in the open field (OF) to evaluate locomotion and exploration; and iii) for depression-like behavior in forced swimming (FS). Catecholaminergic involvement was tested using the tyrosine hydroxylases (TH) enzyme inhibitor, a-methyl-DL-tyrosine (AMPT). Brain areas of chronically treated groups were dissected to assess: i) lipid peroxidation (LPO); ii) carbonylated proteins (CP); iii) superoxide dismutase (SOD) and catalase (CAT) enzymatic activities. Neuronal nitric oxide synthases (nNOS) and argininosuccinate synthase (ASS) protein expression was evaluated by western blotting. Acute treatment with PV3 increased the frequency and time spent in the EPM open arms, suggesting anxiolysis. PV3 increased crossing episodes in the OF. These PV3 effects on anxiety and locomotion were absent in the chronically treated group. Acute and chronic PV3 treatments reduced the immobility time in the FS test, suggesting an antidepressant effect. TH inhibition by AMPT reverted acute PV3 effects. PV3 decreased LPO and CP levels and SOD and CAT activities, whereas nNOS and ASS were reduced in few brain areas. In conclusion, PV3 displayed central antioxidant actions that are concomitant to catecholaminergic-dependent anxiolytic and antidepressant effects.

Research paper thumbnail of Phylogenetic Inferences Based on Distinct Molecular Markers Confirm a Novel Babesia Species (Babesia goianiaensis nov. sp.) in Capybaras (Hydrochoerus hydrochaeris) and Associated Ticks

Microorganisms, Aug 6, 2023

Inferences Based on Distinct Molecular Markers Confirm a Novel Babesia Species (Babesia goianiaen... more Inferences Based on Distinct Molecular Markers Confirm a Novel Babesia Species (Babesia goianiaensis nov. sp.) in Capybaras (Hydrochoerus hydrochaeris) and Associated Ticks.

Research paper thumbnail of Phylogenetic Inferences Based on Distinct Molecular Markers Confirm a Novel Babesia Species (Babesia goianiaensis nov. sp.) in Capybaras (Hydrochoerus hydrochaeris) and Associated Ticks

Microorganisms

Piroplasmids (order Piroplasmida) are a diverse group of tick-borne protozoa that may cause disea... more Piroplasmids (order Piroplasmida) are a diverse group of tick-borne protozoa that may cause disease in animals and occasionally in humans. Novel Piroplasmida clades and species have been found in wild animals from Brazil based on the phylogenetic assessment of near-complete 18S rRNA, mitochondrial and heat-shock protein genes. For instance, a putative novel Babesia species has been detected in capybaras and Amblyomma ticks in three Brazilian states. The present work aimed to describe, using phylogenetic assessments based on distinct molecular markers, this novel Babesia species in capybaras and associated ticks (Amblyomma sculptum and Amblyomma dubitatum) sampled in Goiânia city, Goiás state, midwestern Brazil. While the phylogenetic analysis based on both near-complete 18S rRNA and hsp-70 genes positioned the sequences obtained from capybara blood samples into a new clade sister to the Babesia sensu stricto clade, the phylogenetic inference based on the COX-3 amino acid positioned ...

Research paper thumbnail of Oxidonitrergic and antioxidant effects of a low molecular weight peptide fraction from hardened bean (Phaseolus vulgaris) on endothelium

Brazilian Journal of Medical and Biological Research, 2021

About 3000 tons of beans are not used in human food due to hardening. Several studies on bean-der... more About 3000 tons of beans are not used in human food due to hardening. Several studies on bean-derived bioactive peptides have shown potential to treat some diseases, including those relying on oxidative dysfunctions. We assessed the effects of peptides extracted from hardened bean Phaseolus vulgaris (PV) on reactive oxygen species (ROS) and nitric oxide (NO) production, cytotoxic and cytoprotective effects in endothelial cells, and oxidonitrergic-dependent vasodilating effects. Extract was composed by peptide fraction o3 kDa (PV3) from hardened common bean residue. PV3 sequences were obtained and analyzed with bioinformatics. Human umbilical vein endothelial cells were treated with 10, 20, 30, and 250 mg/mL PV3. Oxidative stress was provoked by 3% H 2 O 2. Cytotoxicity and cytoprotective effects were evaluated by MTT assay, whereas, ROS and NO were quantified using DHE and DAF-FM fluorescent probes by confocal microscopy. NO-and endotheliumdependent vasodilating effects of PV3 were assessed in isolated aortic rings. We found 35 peptides with an average mass of 1.14 kDa. There were no cell deaths with 10 and 20 mg/mL PV3. PV3 at 30 mg/mL increased cell viability, while cytotoxicity was observed only with 250 mg/mL PV3. PV3 at 10 mg/mL was able to protect cells from oxidative stress. PV3 also increased NO release without causing cell death. It also reduced relative ROS production induced by H 2 O 2. PV3 vasodilating effects relied on endothelium-dependent NO release. PV3 obtained from low-commercial-value bean displays little cytotoxicity and exerts antioxidant effects, whereas it increases endothelial NO release.

Research paper thumbnail of Turmeric ethanol extract (Curcuma longa L.) reduces apoptosis and promotes canine osteosarcoma cell proliferation

Ciência Animal Brasileira

Curcuma longa L., also known as turmeric, has been widely studied for its various therapeutic pro... more Curcuma longa L., also known as turmeric, has been widely studied for its various therapeutic properties, including antineoplastic action. The ethanolic extract of the plant contains several phenolic compounds, especially curcumin. Osteosarcoma is a predominant bone tumor in dogs and humans, characterized by high metastatic potential and an unfavorable prognosis. The aim of this study was to investigate the effects of turmeric ethanol extract on canine osteosarcoma cells from established culture. The cells were cultured and treated with different curcumin concentrations (0, 10 μM, 20 μM, 50 μM, 100 μM, and 1000 μM) and exposure times (24h, 48h, and 72h). We first performed tetrazolium reduction technique (MTT) assay and calculated IC50. An immunocytochemistry assay was performed after extract treatment to verify the expression of mutated p53 and therefore study the proliferative potential of malignant cells; Bcl-2 and Ki-67 were used to assess apoptosis and the degree of malignancy,...

Research paper thumbnail of Ventromedial medullary pathway mediating cardiac responses evoked from periaqueductal gray

Autonomic Neuroscience, 2020

Periaqueductal gray (PAG) is a midbrain region that projects to areas controlling behavioral and ... more Periaqueductal gray (PAG) is a midbrain region that projects to areas controlling behavioral and autonomic outputs and is involved in the behavioral and physiological components of defense reactions. Since Raphe Pallidus (RPa) is a medial medullary region comprising sympathetic premotor neurons governing heart function, it is worth considering the PAG-RPa path. We assessed: i) whether PAG projects to RPa; ii) the amplitude of cardiac responses evoked from PAG; iii) whether cardiovascular responses evoked from PAG rely on RPa. Experiments conducted in Wistar rats (± 300 g) were approved by Ethics Committee CEUA-UFG (092/18). Firstly, (n = 3), monosynaptic retrograde tracer Retrobeads was injected into RPa; PAG slices were analyzed. Other two groups (n = 6 each) were anesthetized with urethane (1.4 g/kg) and chloralose (120 mg/ kg) and underwent craniotomy, tracheostomy, catheterization of femoral artery and vein and of cardiac left ventricle. In one group, we injected the GABA A receptor antagonist, bicuculline methiodide (BMI-40 pmol/ 100 nL) into lateral/dorsolateral PAG. Another group was injected (100 nL) with the GABA A receptor agonist muscimol (20 mM) into RPa, 20 min before BMI into PAG. The results were: i) retrogradely labelled neurons were found in PAG; ii) PAG activation by BMI caused positive chronotropism and inotropism, which were accompanied by afterload increases; iii) RPa inhibition with Muscimol reduced heart rate, arterial and ventricular pressures; iv) the subsequent PAG activation still increased arterial pressure, cardiac chronotropy and inotropy, but these responses were significantly attenuated. In conclusion, PAG activation increases cardiac chronotropy and inotropy, and these responses seem to rely on a direct pathway reaching ventromedial medullary RPa neurons.

Research paper thumbnail of A peptide fraction from hardened common beans (Phaseolus vulgaris) induces endothelium-dependent antihypertensive and renal effects in rats

Current Research in Food Science

Beans reached the research spotlight as a source of bioactive compounds capable of modulating dif... more Beans reached the research spotlight as a source of bioactive compounds capable of modulating different functions. Recently, we reported antioxidant and oxidonitrergic effect of a low molecular weight peptide fraction (<3 kDa) from hardened bean (Phaseolus vulgaris) in vitro and ex vivo, which necessitate further in vivo assessments. This work aimed to evaluate the hypotensive effect and the involved physiological mechanisms of the hardened common bean peptide (Phaseolus vulgaris) in normotensive (Wistar) and hypertensive (SHR) animals. Bean flour was combined with a solution containing acetonitrile, water and formic acid (25: 24: 1). Protein extract (PV3) was fractioned (3 kDa membrane). We assessed PV3 effects on renal function and hemodynamics of wistar (WT-normotensive) and spontaneously hypertensive rats (SHR) and measured systemic arterial pressure and flow in aortic and renal beds. The potential endothelial and oxidonitrergic involvements were tested in isolated renal artery rings. As results, we found that PV3: I) decreased food consumption in SHR, increased water intake and urinary volume in WT, increased glomerular filtration rate in WT and SHR, caused natriuresis in SHR; II) caused NO-and endothelium-dependent vasorelaxation in renal artery rings; III) reduced arterial pressure and resistance in aortic and renal vascular beds; IV) caused antihypertensive effects in a dose-dependent manner. Current findings support PV3 as a source of bioactive peptides and raise the potential of composing nutraceutical formulations to treat renal and cardiovascular diseases.

Research paper thumbnail of Extrato etanólico de açafrão (Curcuma longa L.) reduz apoptose e promove proliferação de células de osteossarcoma canino

Ciência Animal Brasileira

Resumo A Curcuma longa L., planta conhecida popularmente como açafrão, tem sido amplamente estuda... more Resumo A Curcuma longa L., planta conhecida popularmente como açafrão, tem sido amplamente estudada por suas diversas propriedades terapêuticas, incluindo a ação antineoplásica. O extrato etanólico da planta contém diversos compostos fenólicos, com destaque para a curcumina. O osteossarcoma é um tumor ósseo predominante em cães e humanos, caracterizado por apresentar alto potencial metastático e prognóstico desfavorável. Procurou-se investigar os efeitos de diferentes concentrações de curcumina do extrato etanólico de açafrão sobre células de osteossarcoma canino de cultura estabelecida. As células foram cultivadas e submetidas ao tratamento com extrato com diferentes concentrações de curcumina (0, 10 μM, 20 μM, 50 μM, 100 μM e 1000 μM) e tempos de exposição (24h, 48h e 72h) pelo EEA. Inicialmente, foram realizados: técnica de redução do tetrazólio (MTT) e cálculo da IC50. Posteriormente, após o tratamento com o extrato, realizou-se o ensaio de imunocitoquímica para verificar a expr...

Research paper thumbnail of Extrato rico em Curcumina do Brasil Central Curcuma longa protege células endoteliais de veias umbilicais humanas sob estresse oxidativo

ABCS Health Sciences

Introdução: Em doenças vasculares, a interrupção do fluxo sanguíneo locale subsequente reperfusão... more Introdução: Em doenças vasculares, a interrupção do fluxo sanguíneo locale subsequente reperfusão de oxigênio pode causar efeitos deletérios e danos irreparáveis às células. Curcuma (Curcuma longa L.) neutraliza radicais livres além de apresentar efeitos preventivos e terapêuticos. Objetivo: Caracterizar os compostos bioativos e a capacidade antioxidante do extrato etanólico de cúrcuma (EEC); avaliar seu efeito nas células endoteliais da veia umbilical humana, e analisar a expressão de vias de sinalização celular. Métodos: As células foram expostas a diferentes concentrações de EEC por 24, 48 e 72 horas. Utilizamos o teste de Folin-Ciocalteau, análise por HPLC-Fluorescência e método DPPH para determinar os compostos fenólicos, conteúdo de curcumina e ação antioxidante, respectivamente; o método de redução de tetrazólio para viabilidade celular, a citotoxicidade e a concentração que inibe 50% da viabilidade celular; e a técnica de imunocitoquímica para analisar a expressão de caspase...

Research paper thumbnail of Déficit Da Memória Na Doença De Alzheimer Visualizada Por Outra Perspectiva

Research paper thumbnail of Nasal biocompatible powder of Geraniol oil complexed with cyclodextrins for neurodegenerative diseases: physicochemical characterization and in vivo evidences of nose to brain delivery

Journal of Controlled Release, 2021

Recently, many studies have shown that plant metabolites, such as geraniol (GER), may exert anti-... more Recently, many studies have shown that plant metabolites, such as geraniol (GER), may exert anti-inflammatory effects in neurodegenerative diseases and, in particular, Parkinson's disease (PD) models. Unfortunately, delivering GER to the CNS via nose-to-brain is not feasible due to its irritant effects on the mucosae. Therefore, in the present study β-cyclodextrin (βCD) and its hydrophilic derivative hydroxypropyl-beta-cyclodextrin (HPβCD) were selected as potential carriers for GER nose-to-brain delivery. Inclusion complexes were formulated and the biocompatibility with nasal mucosae and drug bioavailability into cerebrospinal fluid (CSF) were studied in rats. It has been demonstrated by DTA, FT-IR and NMR analyses that both the CDs were able to form 1:1 GER-CD complexes, arising long-term stable powders after the freeze-drying process. GER-HPβCD-5 and GER-βCD-2 complexes exhibited comparable results, except for morphology and solubility, as demonstrated by SEM analysis and phase solubility study, respectively. Even though both complexes were able to directly and safely deliver GER to CNS, GER-βCD-2 displayed higher ability in releasing GER in the CSF. In conclusion, βCD complexes can be considered a very promising tool in delivering GER into the CNS via nose-to-brain route, preventing GER release into the bloodstream and ensuring the integrity of the nasal mucosa.