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1.
Braz. j. biol ; 82: e234500, 2022. tab, graf
Artigo em Inglês | LILACS-Express | LILACS | ID: biblio-1153464

RESUMO

Abstract This work aimed to obtain aspartic proteases of industrial and biotechnological interest from the stomach of the crevalle jack fish (Caranx hippos). In order to do so, a crude extract (CE) of the stomach was obtained and subjected to a partial purification by salting-out, which resulted in the enzyme extract (EE) obtainment. EE proteases were characterized physicochemically and by means of zymogram. In addition, the effect of chemical agents on their activity was also assessed. By means of salting-out it was possible to obtain a purification of 1.6 times with a yield of 49.4%. Two acid proteases present in the EE were observed in zymogram. The optimum temperature and thermal stability for EE acidic proteases were 55 ºC and 45 °C, respectively. The optimum pH and pH stability found for these enzymes were pH 1.5 and 7.0, respectively. Total inhibition of EE acid proteolytic activity was observed in the presence of pepstatin A. dithiothreitol (DTT) and Ca2+ did not promote a significant effect on enzyme activity. In the presence of heavy metals, such as Al3+, Cd2+ and Hg2+, EE acidic proteases showed more than 70% of their enzymatic activity. The results show that it is possible to obtain, from the stomach of C. hippos, aspartic proteases with high proteolytic activity and characteristics that demonstrate potential for industrial and biotechnological applications.


Resumo Este trabalho objetivou obter proteases aspárticas de interesse industrial e biotecnológico a partir do estômago do peixe xaréu (Caranx hippos). Para isso, foi obtido um extrato bruto do estômago, o qual foi submetido a uma purificação parcial por salting-out onde se obteve o extrato enzimático (EE). As proteases do EE foram caracterizadas físico-quimicamente e através de zimograma. Além disso, o efeito de agentes químicos sobre sua atividade também foi avaliado. Através de salting-out foi possível obter uma purificação de 1,6 vezes com rendimento de 49,4%. Foram observadas duas proteases ácidas presentes no EE através de zimograma. A temperatura ótima e a estabilidade térmica para as proteases ácidas do EE foram de 55 ºC e 45 °C, respectivamente. O pH ótimo e a estabilidade ao pH encontrados para estas enzimas foram o pH 1,5 e 7,0, respectivamente. Observou-se a inibição total da atividade proteolítica ácida do EE na presença de pepstatina A. O ditiotreitol (DTT) e o Ca2+ não promoveram efeito significativo na atividade enzimática. Na presença de metais pesados, como Al3+, Cd2+ e Hg2+, o EE manteve mais de 70% de atividade enzimática do EE. Os resultados mostram que é possível obter, a partir do estômago de C. hippos, proteases aspárticas com alta atividade proteolítica e características que demonstram potencial para aplicações industriais e biotecnológicas.

2.
Braz. j. biol ; 81(2): 268-277, Jan.-May 2021. tab, graf
Artigo em Inglês | LILACS-Express | LILACS | ID: biblio-1153357

RESUMO

Abstract This study evaluated the genotoxicity of lyophilized glycolic extract of Theobroma cacao Linné seeds (TCL), using the micronucleus assay in bone marrow of mice. The interaction between TCL and doxorubicin (DXR) was also analyzed. Experimental groups were evaluated 24-48 h after treatment with N-Nitroso-N-ethylurea (NEU: 50 mg/kg), DXR (5 mg/kg), NaCl (145 mM), TCL (0.5-2 g/kg), and TCL (2 g/kg) in combination with DXR (antigenotoxic assays). Analysis of micronucleated polychromatic erythrocytes (MNPCEs) showed no significant differences between all the treatment doses of TCL and NaCl control. Mice experimentally treated with DXR and NEU significantly induced MNPCEs. However, a significant reduction of MNPCEs was also observed when TCL was administered in combination with the chemotherapeutic agent DXR. The analysis of the PCE/NCE ratio revealed no significant differences between the NaCl control, all doses of TCL, and DXR. However, there were significant differences in the PCE/NCE ratio between positive NEU control and all other treatments. The PCE/NCE ratio observed after treatment with TCL and DXR showed significant differences and intermediate values to controls (NaCl and NEU). This study suggests absence of genotoxicity and cytotoxicity of TCL, regardless of dose, sex, and time. TCL reduced genotoxic effects induced by DXR, suggesting potential antigenotoxic effects.


Resumo Este estudo avaliou a genotoxicidade do extrato glicólico liofilizado de sementes de Theobroma cacao Linné (TCL), usando o ensaio do micronúcleo em medula óssea de camundongos. A interação entre TCL e doxorrubicina (DXR) foi também analisada. Grupos experimentais foram avaliados 24-48 h após tratamento com N-Nitroso-N-etilureia (NEU: 50 mg/kg), DXR (5 mg/kg), NaCl (145 mM), TCL (0,5-2 g/kg), e TCL (2 g/kg) em combinação com DXR (ensaio antigenotóxico). As análises de eritrócitos policromáticos micronucleados (EPCMNs) não mostraram diferenças significantes entre todas as doses de tratamento do TCL e o controle NaCl. Camundongos experimentalmente tratados com DXR e NEU induziram significativamente EPCMNs. Contudo, uma redução significante de EPCMNs foi também observada quando TCL foi administrada em combinação com o agente quimioterapêutico DXR. As análises da relação EPC/ENC (eritrócito policromático/eritrócito normocromático) revelaram ausência de diferenças significantes entre o controle NaCl, todas as doses de TCL e DXR. Contudo, houve diferenças significantes na relação EPC/ENC entre o controle positivo NEU e todos os outros tratamento. A relação ECP/ENC observada após o tratamento com TCL e DXR mostrou diferenças significantes e valores intermediários aos controles (NaCl e NEU). Este estudo sugere ausência de genotoxicidade e citotoxicidade de TCL, independentemente da dose, sexo e tempo. TCL reduziu os efeitos genotóxicos induzidos por DXR, sugerindo potencial efeitos antigenotóxicos.

3.
Braz J Biol ; 82: e234500, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33787732

RESUMO

This work aimed to obtain aspartic proteases of industrial and biotechnological interest from the stomach of the crevalle jack fish (Caranx hippos). In order to do so, a crude extract (CE) of the stomach was obtained and subjected to a partial purification by salting-out, which resulted in the enzyme extract (EE) obtainment. EE proteases were characterized physicochemically and by means of zymogram. In addition, the effect of chemical agents on their activity was also assessed. By means of salting-out it was possible to obtain a purification of 1.6 times with a yield of 49.4%. Two acid proteases present in the EE were observed in zymogram. The optimum temperature and thermal stability for EE acidic proteases were 55 ºC and 45 °C, respectively. The optimum pH and pH stability found for these enzymes were pH 1.5 and 7.0, respectively. Total inhibition of EE acid proteolytic activity was observed in the presence of pepstatin A. dithiothreitol (DTT) and Ca2+ did not promote a significant effect on enzyme activity. In the presence of heavy metals, such as Al3+, Cd2+ and Hg2+, EE acidic proteases showed more than 70% of their enzymatic activity. The results show that it is possible to obtain, from the stomach of C. hippos, aspartic proteases with high proteolytic activity and characteristics that demonstrate potential for industrial and biotechnological applications.


Assuntos
Peptídeo Hidrolases , Animais , Concentração de Íons de Hidrogênio , Temperatura
4.
Br J Nutr ; : 1-12, 2020 Aug 18.
Artigo em Inglês | MEDLINE | ID: mdl-32807252

RESUMO

Protein supplementation may be beneficial for patients with chronic liver disease (CLD). This study compared the effects of whey protein isolate (WP) and casein (CA) supplementation on nutritional status and immune parameters of CLD patients who were randomly assigned to take 20 g of WP or CA twice per d as a supplement for 15 d. Body composition, muscle functionality and plasmatic immunomarkers were assessed before and after supplementation. Patients were also classified according to the model for end-stage liver disease (MELD) into less (MELD < 15) and more (MELD ≥ 15) severe disease groups. Malnutrition, determined by the Subjective Global Assessment at baseline, was observed in 57·4 % and 54·2 % of patients in the WP and CA groups, respectively (P = 0·649). Protein intake was lower at baseline in the WP group than in the CA group (P = 0·035), with no difference after supplementation (P = 0·410). Both the WP and CA MELD < 15 groups increased protein intake after supplementation according to the intragroup analysis. No differences were observed in body composition, muscle functionality, most plasma cytokines (TNF, IL-6, IL-1ß and interferon-γ), immunomodulatory proteins (sTNFR1, sTNFR2, brain-derived neurotrophic factor and glial cell line-derived neurotrophic factor) or immunomodulatory hormones (adiponectin, insulin and leptin) after supplementation in the WP groups at the two assessed moments. WP supplementation increased the levels of interferon-γ-induced protein-10/CXCL10 (P = 0·022), eotaxin-1/CCL11 (P = 0·031) and monocyte chemoattractant protein-1/CCL2 (P = 0·018) and decreased IL-5 (P = 0·027), including among those in the MELD ≥ 15 group, for whom IL-10 was also increased (P = 0·008). Thus, WP consumption by patients with CLD impacted the immunomodulatory responses when compared with CA with no impact on nutritional status.

5.
Braz J Biol ; 2020 Jul 20.
Artigo em Inglês | MEDLINE | ID: mdl-32696851

RESUMO

This study evaluated the genotoxicity of lyophilized glycolic extract of Theobroma cacao Linné seeds (TCL), using the micronucleus assay in bone marrow of mice. The interaction between TCL and doxorubicin (DXR) was also analyzed. Experimental groups were evaluated 24-48 h after treatment with N-Nitroso-N-ethylurea (NEU: 50 mg/kg), DXR (5 mg/kg), NaCl (145 mM), TCL (0.5-2 g/kg), and TCL (2 g/kg) in combination with DXR (antigenotoxic assays). Analysis of micronucleated polychromatic erythrocytes (MNPCEs) showed no significant differences between all the treatment doses of TCL and NaCl control. Mice experimentally treated with DXR and NEU significantly induced MNPCEs. However, a significant reduction of MNPCEs was also observed when TCL was administered in combination with the chemotherapeutic agent DXR. The analysis of the PCE/NCE ratio revealed no significant differences between the NaCl control, all doses of TCL, and DXR. However, there were significant differences in the PCE/NCE ratio between positive NEU control and all other treatments. The PCE/NCE ratio observed after treatment with TCL and DXR showed significant differences and intermediate values to controls (NaCl and NEU). This study suggests absence of genotoxicity and cytotoxicity of TCL, regardless of dose, sex, and time. TCL reduced genotoxic effects induced by DXR, suggesting potential antigenotoxic effects.

6.
Blood Rev ; 43: 100651, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32014342

RESUMO

L-asparaginase is an enzyme used as a biopharmaceutical to treat acute lymphoblastic leukemia. Several adverse effects have been related to L-asparaginase use, so the scientific community has searched for novel proteoforms of L-ASNase. However, some critical characteristics must be considered for a novel L-ASNase source to be effective as an antitumour drug. Accordingly, this article provides a critical analysis of the parameters and methods applied to estimate L-ASNase concentration, measure the L-ASNase and GLNase activities and kinetics, evaluate the enzyme preparations purity and define the antitumour activity against leukemic cells in vitro. Among the main features, the proposed new enzyme needs to present high affinity for L-asparagine; low percentage of glutaminase activity in relation to L-ASNase; high enzyme stability and half-life and mainly antileukemic activity when a low protein amount is applied. These parameters are discussed in an attempt to guide the consideration of an enzyme as a promising biopharmaceutical against ALL.

7.
Eur Arch Paediatr Dent ; 21(2): 193-202, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31471856

RESUMO

AIM: The object of this study is to evaluate the oral health-related quality of life (OHRQoL) of caregivers of individuals with Cerebral Palsy (CP). METHODS: Ninety-eight caregivers of individuals with CP (SG) and 196 caregivers of individuals without special needs (CG) completed the reduced version of the Oral Health Impact Profile (OHIP-14). In addition, a questionnaire with demographic and socioeconomic questions was used. Groups were matched according to gender, schooling and family income. Fisher's exact tests, Chi square and Mann-Whitney, and bi/multivariate logistic regression were used (p < 0.05). RESULTS: The impact on OHRQoL was similar for SG and CG (OHIP-14 total score). For SG, the high negative impact was in the following OHIP domains: "Functional limitation", "Physical disability", "Psychological incapacity" and "Disadvantages". For SG, a higher number of caregivers was single, unemployed, received government financial aid. There was a statistically significant association between the number of children a caregiver has and a negative impact on OHRQoL. CONCLUSIONS: Being a caregiver of children with or without CP had a negative impact of similar magnitude on ORHQoL. The negative impact on OHRQoL of caregivers of children with CP was associated with having a high number of children; the higher the number of children, the greater the negative impact on their OHRQoL.


Assuntos
Paralisia Cerebral , Qualidade de Vida , Cuidadores , Estudos de Casos e Controles , Criança , Humanos , Saúde Bucal , Inquéritos e Questionários
8.
Appl Radiat Isot ; 153: 108823, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31400649

RESUMO

This work aimed at assessing the neutron radiation field with activation foils around of the PETtrace biomedical cyclotron. Activation foils of gold, gold covered with cadmium, indium and nickel was used to evaluate the thermal, epithermal and fast neutron contributions, respectively. The irradiations of the activation foils were performed using the intermittent irradiation method to conciliate the measurements with the routine 18F production. The evaluated points showed the contributions of the thermal, epithermal and fast neutrons due to interaction with the vault room, cyclotron accelerator and main cyclotron components.


Assuntos
Nêutrons , Tomografia por Emissão de Pósitrons , Síncrotrons
9.
Comput Methods Programs Biomed ; 176: 195-209, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31200906

RESUMO

BACKGROUND AND OBJECTIVE: The orthopedic Maxillary Expansion (ME) procedure is used for treating the transverse maxillary deficiency. This pathology consists in a smaller transverse dimension in the maxilla and leads to malocclusion. The treatment takes advantage of the existence of the midpalatal suture (MPS), which corresponds to the junction at the palatine bones of its horizontal portions. The technique employs a device, conventionally a palatal expander attached to the posterior teeth, to separate the two maxillary bones in the MPS. The objective of this study was to analyze, using the Finite Element Method, the biomechanical behavior of the MPS when an expansion is applied. METHODS: A Computer Tomography image of the maxilla was reconstructed, the suture geometry was modeled with different interdigitation levels and types of hyrax devices. A total of 12 geometric models (three levels for interdigitation and four types of hyrax devices) were prepared and analyzed taking into account the chewing forces and the expansion displacement. For each case, maximum principal stresses on the maxilla (bone), and equivalent stresses on the expander device (stainless steel) were observed. In the MPS, maximum principal stresses and directional displacement were evaluated. RESULTS: The results showed that the interdigitation does not have an important influence on the deformation behavior of the maxilla but it affects the stress distribution. In addition, the type of expander device and anchorage have a direct relationship with the treatment effectiveness; larger deformation in the expansion direction was obtained with skeletal when compared to dental anchorage. CONCLUSIONS: A study that allows a better understanding of the oral biomechanics during the application of ME was presented. To our knowledge, it is the first study based on computational simulations that takes into account bone structures, like maxilla and part of the skull, to analyze the interdigitation influence on the MPS behavior when exposed to a ME.


Assuntos
Maxila/patologia , Técnica de Expansão Palatina/instrumentação , Palato/fisiologia , Fenômenos Biomecânicos , Simulação por Computador , Análise de Elementos Finitos , Humanos , Imageamento Tridimensional/métodos , Modelos Anatômicos , Estresse Mecânico , Dente
10.
Bone ; 125: 112-121, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31100533

RESUMO

Short-chain fatty acids (SCFAs) exert a variety of immune and metabolic functions by binding to G-protein-coupled receptors, mainly free fatty acid receptor 2 (FFAR2). However, the effects of SCFAs and FFARs on bone remodeling, especially in alveolar bone, have been less explored. In this study, we investigated the influence of the SCFA/FFAR2 axis on alveolar bone. Bone samples from wild-type (WT) and FFAR2-deficient mice (FFAR2-/-) were analyzed using micro-CT, histology and qPCR. WT and FFAR2-/- animals received a high-fiber diet (HFD) reported to increase circulating levels of SCFAs. Additionally, we analyzed the effects of SCFAs and a synthetic FFAR2 agonist, phenylacetamide-1 (CTMB), on bone cell differentiation. The participation of histone deacetylase inhibitors (iHDACs) in the effects of SCFAs was further assessed in vitro. CTMB treatment was also evaluated in vivo during orthodontic tooth movement (OTM). FFAR2-/- mice exhibited deterioration of maxillary bone parameters. Consistent with this, FFAR2-/- mice exhibited a significant increase of OTM and changes in bone cell numbers and in the expression of remodeling markers. The HFD partially reversed bone loss in the maxillae of FFAR2-/- mice. In WT mice, the HFD induced changes in the bone markers apparently favoring a bone formation scenario. In vitro, bone marrow cells from FFAR2-/- mice exhibited increased differentiation into osteoclasts, while no changes in osteoblasts were observed. In line with this, differentiation of osteoclasts was diminished by SCFAs and CTMB. Moreover, CTMB treatment significantly reduced OTM. Pretreatment of osteoclasts with iHDACs did not modify the effects of SCFAs on these cells. In conclusion, SCFAs function as regulators of bone resorption. The effects of SCFAs on osteoclasts are dependent on FFAR2 activation and are independent of the inhibition of HDACs. FFAR2 agonists may be useful to control bone osteolysis.


Assuntos
Ácidos Graxos Voláteis/farmacologia , Osteoclastos/efeitos dos fármacos , Osteoclastos/metabolismo , Receptores Acoplados a Proteínas-G/metabolismo , Animais , Células da Medula Óssea/citologia , Células da Medula Óssea/efeitos dos fármacos , Células da Medula Óssea/metabolismo , Reabsorção Óssea/tratamento farmacológico , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Osteoblastos/efeitos dos fármacos , Osteoblastos/metabolismo , Receptores Acoplados a Proteínas-G/genética , Microtomografia por Raio-X
11.
J Appl Microbiol ; 126(5): 1426-1437, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30762925

RESUMO

AIMS: The aim of this study was to find new eukaryotic sources of the l-asparaginase (l-ASNase), since the prokaryotic sources of the enzyme are well-reported as causing allergic hypersensitivity reactions in a significant number of patients. This report describes screening for l-ASNase production by filamentous fungi isolated from the Brazilian Caatinga, and the optimization of fermentation parameters to increase fungal growth and improve yield in the production of l-ASNase. METHODS AND RESULTS: Thirty-two filamentous fungi were investigated in this study. When Aspergillus terreus strain S-18 was cultured in a proline-enriched medium, intracellular l-ASNase was expressed in concurrence with reduced l-glutaminase (l-GLUase) and protease activities. Fermentation conditions were then optimized in a 5-l bioreactor system to produce a maximum volumetric yield of 108 U total of l-ASNase activity. CONCLUSIONS: The work reported here represents the first attempt to produce l-ASNase by filamentous fungi isolated from Brazil and offers a promising alternative eukaryotic source for l-ASNase production. SIGNIFICANCE AND IMPACT OF THE STUDY: In order to minimize the side effects caused by bacterial l-ASNase, the search of eukaryotic micro-organism for l-ASNase was carried out in fungi. This study demonstrates the diversity of filamentous fungi isolated from the Brazilian Caatinga Biome and the importance of knowledge of the microbial metabolism to obtain high concentrations of biotechnological products.


Assuntos
Asparaginase , Aspergillus , Reatores Biológicos/microbiologia , Asparaginase/análise , Asparaginase/metabolismo , Aspergillus/química , Aspergillus/enzimologia , Aspergillus/metabolismo , Brasil , Microbiologia Ambiental , Fermentação , Florestas , Microbiota
12.
J Dent Res ; 98(4): 476-484, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30681895

RESUMO

Apical periodontitis is an inflammatory disorder that results from the host immune response to microbial infection through the dental pulp, leading to alveolar bone destruction. The nod-like receptor 12 (NLRP12) is an atypical intracellular sensor of the NLR family that is involved in the negative regulation of several inflammatory conditions and also osteoclastogenesis. However, the role of NLRP12 in the regulation of immune response and bone loss induced by bacterial infection remains unclear. Here we investigated the development of apical periodontitis in wild-type (WT) and NLRP12 knockout (NLRP12-/-) mice by using micro-computed tomography together with histological, immunohistochemical, and molecular analyses. We found that NLRP12-/- mice are highly susceptible to apical periodontitis induced by bacterial infection, which is associated with an elevated infiltration of neutrophils and macrophages, periapical lesion extension, and alveolar bone destruction. Furthermore, NLRP12-/- mice showed a high expression of inflammatory cytokines ( Il1b, Il6, and Tnfa) and the osteoclastogenic markers ( Rankl and Acp5) in the periapical tissues. Consistent with this observation, NLRP12-/- mice showed an increased number of tartrate-resistant acid phosphatase-positive cells lining the apical periodontitis site, which was associated with augmented expression of the osteoclast effector genes, Ctsk and Mmp9. Mechanistically, NLRP12-deficient preosteoclasts showed elevated IκB-α degradation and p65 phosphorylation when stimulated with receptor activator of nuclear factor (NF)-κB ligand (RANKL). Similarly, increased IκB-α degradation was observed in the periapical tissue of NLRP12-/- mice. Furthermore, our in vitro study showed that preosteoclasts from NLRP12-/- mice exhibited higher RANKL-induced osteoclastogenesis, which was synergistically amplified by interleukin-1ß and tumor necrosis factor α (mimicking an inflammatory periapical milieu). In conclusion, our data show that NLRP12 exhibits a protective role in the periapical bone destruction by attenuating inflammation and osteoclastogenesis through negative regulation of the NF-κB pathway.


Assuntos
Periodontite Periapical , Ligante RANK , Animais , Peptídeos e Proteínas de Sinalização Intracelular , Camundongos , Camundongos Endogâmicos NOD , Osteoclastos , Microtomografia por Raio-X
13.
J Dent Res ; 98(2): 148-156, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30359170

RESUMO

Periodontal diseases are initiated by bacteria that accumulate in a biofilm on the tooth surface and affect the adjacent periodontal tissue. Systemic diseases such as diabetes, rheumatoid arthritis (RA), and systemic lupus erythematosus (SLE) increase susceptibility to destructive periodontal diseases. In human studies and in animal models, these diseases have been shown to enhance inflammation in the periodontium and increase the risk or severity of periodontitis. All 3 systemic diseases are linked to a decrease in bacterial taxa associated with health and an increase in taxa associated with disease. Although there is controversy regarding the specific oral bacterial changes associated with each disease, it has been reported that diabetes increases the levels of Capnocytophaga, Porphyromonas, and Pseudomonas, while Prevotella and Selenomonas are increased in RA and Selenomonas, Leptotrichia, and Prevotella in SLE. In an animal model, diabetes increased the pathogenicity of the oral microbiome, as shown by increased inflammation, osteoclastogenesis, and periodontal bone loss when transferred to normal germ-free hosts. Moreover, in diabetic animals, the increased pathogenicity could be substantially reversed by inhibition of IL-17, indicating that host inflammation altered the microbial pathogenicity. Increased IL-17 has also been shown in SLE, RA, and leukocyte adhesion deficiency and may contribute to oral microbial changes in these diseases. Successful RA treatment with anti-inflammatory drugs partially reverses the oral microbial dysbiosis. Together, these data demonstrate that systemic diseases characterized by enhanced inflammation disturb the oral microbiota and point to IL-17 as key mediator in this process.


Assuntos
Doenças Cardiovasculares/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Disbiose/microbiologia , Microbiota , Boca/microbiologia , Periodontite/microbiologia , Animais , Bactérias , Doenças Cardiovasculares/complicações , Doenças Cardiovasculares/microbiologia , Diabetes Mellitus Tipo 2/microbiologia , Humanos , Periodontite/complicações , Periodonto
14.
Prep Biochem Biotechnol ; 49(1): 95-104, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30488788

RESUMO

L-Asparaginase amidohydrolase (EC 3.5.1.1) has received significant attention owing to its clinical use in acute lymphoblastic leukemia treatment and non-clinical applications in the food industry to reduce acrylamide (toxic compound) formation during the frying of starchy foods. In this study, a sequential optimization strategy was used to determine the best culture conditions for L-asparaginase production from filamentous fungus Aspergillus terreus CCT 7693 by submerged fermentation. The cultural conditions were studied using a 3-level, central composite design of response surface methodology, and biomass and enzyme production were optimized separately. The highest amount of biomass (22.0 g·L-1) was obtained with modified Czapek-Dox medium containing glucose (14 g·L-1), L-proline (10 g·L-1), and ammonium nitrate (2 g·L-1) fermented at 37.2 °C and pH 8.56; for maximum enzyme production (13.50 U·g-1), the best condition was modified Czapek-Dox medium containing glucose (2 g·L-1), L-proline (10 g·L-1), and inoculum concentration of 4.8 × 108 espore·mL-1 adjusted to pH 9.49 at 34.6 °C. The L-asparaginase production profile was studied in a 7 L bench-scale bioreactor and a final specific activity of 13.81 U·g-1 was achieved, which represents an increase of 200% in relation to the initial non-optimized conditions.


Assuntos
Antineoplásicos/metabolismo , Asparaginase/biossíntese , Aspergillus/metabolismo , Técnicas de Cultura de Células , Fermentação , Biomassa , Reatores Biológicos , Meios de Cultura , Glucose/metabolismo , Concentração de Íons de Hidrogênio , Cinética , Nitratos/metabolismo , Prolina/metabolismo , Temperatura
15.
Environ Pollut ; 242(Pt A): 277-287, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-29990935

RESUMO

The hydrocarbon phenanthrene is an organic compound commonly found in the environment. In aquatic ecosystems, it is highly toxic to organisms, although little is known about its effects on sediment-dwelling organisms. The purpose of this study was to evaluate phenanthrene effects on biochemical, histological, and ontogenetic levels in larvae of the sediment-dwelling invertebrate Chironomus sancticaroli at acute and chronic exposure. Lethal concentrations were estimated and toxicity (acute-96 h and chronic- 8 d) tests were performed at phenanthrene concentrations from 0.12 to 1.2 mg L-1. At acute and chronic exposure, we evaluated acetylcholinesterase (AChE), alpha esterase (EST-α), and beta esterase (EST-ß) activities as well as histological alterations. In the assays with chronic exposure, effects on larval development were estimated using antennae length (instar estimative) and body length (growth estimative). The EST-α showed a significantly increased activity after 48 h at acute exposure to high concentrations of phenanthrene, while EST-ß activity was increased after 48 and 72 h at acute exposure at higher concentrations and at 0.12 mg L-1 at chronic exposure. At acute exposure, the midgut showed alterations such as brush border disruption, gastric caeca regression, and lumen area reduction; the fat body showed nuclear alteration in the trophocytes, while the Malpighian tubules showed brush border reduction and the salivary glands were subject to cytoplasm vacuolation. At chronic exposure, the same alterations were observed, in addition to vacuolar coalescence in the trophocytes of the fat body. Regarding larval development, a reduction of body length was observed with increasing phenanthrene concentrations. Similarly, molting was delayed; in the control group, all larvae were in the fourth instar, while at higher phenanthrene concentrations, larvae were predominantly in the third instar. Phenanthrene had toxic effects on this chironomid, indicating risks for natural populations.


Assuntos
Chironomidae/fisiologia , Larva/efeitos dos fármacos , Fenantrenos/toxicidade , Poluentes Químicos da Água/toxicidade , Acetilcolinesterase , Animais , Bioensaio , Chironomidae/efeitos dos fármacos , Esterases , Larva/fisiologia
16.
Braz. j. biol ; 78(1): 1-12, Feb. 2018. tab, graf
Artigo em Inglês | LILACS | ID: biblio-888851

RESUMO

Abstract Handroanthus impetiginosus has long been used in traditional medicine and various studies have determined the presence of bioactive chemical compounds and potential phytotherapeutics. In this study, the genotoxicity of the lyophilized tincture of H. impetiginosus bark (THI) was evaluated in mouse bone marrow using micronucleus assays. The interaction between THI and genotoxic effects induced by the chemotherapeutic agent, doxorubicin (DXR), was also analyzed. Experimental groups were evaluated 24 to 48 h after treatment with N-nitroso-N-ethylurea (NEU; 50 mg/kg), DXR (5 mg/kg), sodium chloride (NaCl; 150 mM), and THI (0.5-2 g/kg). Antigenotoxic assays were carried out using THI (0.5 g/kg) in combination with NEU or DXR. Analysis of the micronucleated polychromatic erythrocytes (MNPCEs) indicated no significant differences between treatment doses of THI (0.5-2 g/kg) and NaCl. Polychromatic erythrocyte (PCE) to normochromatic erythrocyte (NCE) ratios did not indicate any statistical differences between DXR and THI or NaCl, but there were differences between THI and NaCl. A significant reduction in MNPCEs and PCE/NCE ratios was observed when THI was administered in combination with DXR. This study suggested the absence of THI genotoxicity that was dose-, time-, and gender-independent and the presence of moderate systemic toxicity that was dose-independent, but time- and gender-dependent. The combination of THI and DXR also suggested antigenotoxic effects, indicating that THI reduced genotoxic effects induced by chemotherapeutic agents.


Resumo Handroanthus impetiginosus tem sido usada durante um longo período pela medicina tradicional e vários estudos têm demonstrados a presença de compostos químicos e potencial fitoterapêutico. Esta pesquisa avaliou a genotoxicidade da tintura da casca liofilizada de H. impetiginosus (THI) usando o ensaio do micronúcleo em medula óssea de camundongos. A interação entre THI e os efeitos genotóxicos induzidos pelo quimioterápico doxorrubicina (DXR) também foram analisados. Grupos experimentais foram analisados a 24-48 h após o tratamento com N-Nitroso-N-etiluréia (NEU; 50 mg/kg), DXR (5 mg/kg), NaCl (150 mM) e THI (0,5-2 g/kg). O ensaio antigenotóxico foi conduzido utilizando THI (0,5 g/kg) em combinação com NEU ou DXR. A análise de eritrócitos policromáticos micronucleados (EPCMNs) não mostrou diferenças significativas entre as doses de tratamento (0,5-2 g/kg) e NaCl. As proporções de eritrócitos policromáticos (EPC)/eritrócitos normocromáticos (ENC) não revelaram diferenças estatísticas entre DXR e THI ou NaCl, porém houve diferenças entre THI e NaCl. Uma redução significativa em EPCMNs e na razão EPC/ENC foi observada quando THI foi administrado em combinação com DXR. Essa pesquisa sugere ausência de genotoxicidade de THI, dose-, tempo- e sexo-independente, e moderada toxicidade sistêmica dose-independente, mas tempo- e sexo-dependente. A associação do THI e DXR também sugere efeitos antigenotóxicos. Por conseguinte, THI pode reduzir os efeitos genotóxicos induzidos pelo quimioterapêutico.


Assuntos
Animais , Camundongos , Dano ao DNA/efeitos dos fármacos , Células da Medula Óssea/citologia , Células da Medula Óssea/efeitos dos fármacos , Extratos Vegetais/farmacologia , Doxorrubicina/toxicidade , Substâncias Protetoras/farmacologia , Testes para Micronúcleos , Células Cultivadas , Tabebuia/química
17.
Int J Oral Maxillofac Surg ; 47(7): 900-907, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29329829

RESUMO

The aim of this study was to evaluate the effectiveness and safety of 5% ethanolamine oleate (EO) foam in the treatment of low-flow venous malformations in the head and neck region. Seventeen consecutive patients (six male, 11 female) and 34 low-flow venous malformations were enrolled. The vascular anomalies ranged between 20mm and 80mm in size. The typical clinical indication was a swelling (88.2%) with a purple colour (85.3%); the most frequent location was the tongue (23.5%). Ethanolamine oleate foam was produced via the Tessari method and applied at 10mg per 1cm to the vascular anomalies. This process resulted in the highest clinical healing score in 64.7% of cases, and half of the patients reported a high level of satisfaction (score >9). In the majority of cases (88.2%), the patients reported that the pain immediately postoperative was mild or moderate. There were direct relationships between vascular anomaly size and the volume of EO applied, the number of sessions, and healing (P<0.05). No recurrence was observed during 6 months of follow-up. This case series showed the effectiveness and safety of 5% EO foam for the treatment of venous malformations in the head and neck region.


Assuntos
Malformações Arteriovenosas/terapia , Cabeça/irrigação sanguínea , Pescoço/irrigação sanguínea , Ácidos Oleicos/uso terapêutico , Soluções Esclerosantes/uso terapêutico , Escleroterapia/métodos , Adolescente , Adulto , Idoso , Criança , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Resultado do Tratamento
18.
Braz J Biol ; 78(1): 1-12, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-28699970

RESUMO

Handroanthus impetiginosus has long been used in traditional medicine and various studies have determined the presence of bioactive chemical compounds and potential phytotherapeutics. In this study, the genotoxicity of the lyophilized tincture of H. impetiginosus bark (THI) was evaluated in mouse bone marrow using micronucleus assays. The interaction between THI and genotoxic effects induced by the chemotherapeutic agent, doxorubicin (DXR), was also analyzed. Experimental groups were evaluated 24 to 48 h after treatment with N-nitroso-N-ethylurea (NEU; 50 mg/kg), DXR (5 mg/kg), sodium chloride (NaCl; 150 mM), and THI (0.5-2 g/kg). Antigenotoxic assays were carried out using THI (0.5 g/kg) in combination with NEU or DXR. Analysis of the micronucleated polychromatic erythrocytes (MNPCEs) indicated no significant differences between treatment doses of THI (0.5-2 g/kg) and NaCl. Polychromatic erythrocyte (PCE) to normochromatic erythrocyte (NCE) ratios did not indicate any statistical differences between DXR and THI or NaCl, but there were differences between THI and NaCl. A significant reduction in MNPCEs and PCE/NCE ratios was observed when THI was administered in combination with DXR. This study suggested the absence of THI genotoxicity that was dose-, time-, and gender-independent and the presence of moderate systemic toxicity that was dose-independent, but time- and gender-dependent. The combination of THI and DXR also suggested antigenotoxic effects, indicating that THI reduced genotoxic effects induced by chemotherapeutic agents.


Assuntos
Células da Medula Óssea , Dano ao DNA/efeitos dos fármacos , Doxorrubicina/toxicidade , Extratos Vegetais/farmacologia , Substâncias Protetoras/farmacologia , Tabebuia/química , Animais , Células da Medula Óssea/citologia , Células da Medula Óssea/efeitos dos fármacos , Células Cultivadas , Camundongos , Testes para Micronúcleos
19.
Lupus ; 27(2): 283-289, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-28679308

RESUMO

Oral symptoms in systemic lupus erythematosus (SLE) patients are often unexplored and affect the health-related quality of life. The aims of this study were: (a) to evaluate the oral health condition of SLE patients compared to control subjects without rheumatic diseases; (b) to determine the consequences of oral health condition in the quality of life of these two groups. Individuals with SLE ( n = 75) and without SLE ( n = 78) (control group), paired for gender and age, underwent complete oral examination. Sociodemographic and clinical information was obtained, and interviews were conducted using the Brazilian version of the oral health impact profile. The activity and damage of SLE disease were assessed, respectively, by the systemic lupus erythematosus disease activity index 2000 and the Systemic Lupus International Collaborating Clinics/American College of Rheumatology damage index for systemic lupus erythematosus. When we analysed the oral health condition and hygiene habits of the participants, SLE patients exhibited an increased number of missing teeth despite their higher frequency of tooth brushing. No significant differences were verified in other habits and clinical parameters evaluated such as smoking, flossing, salivary flux, periodontitis, decayed and filled teeth. Patients with SLE presented with worse oral health-related quality of life than controls ( P = 0.011). The significant difference was on individuals' physical disability ( P = 0.002). The determinant of the negative impact on the oral health-related quality of life was prosthesis wearing ( P < 0.05). Overall, the oral health impact profile score was higher in individuals with moderate SLE damage compared to SLE individuals with no damage ( P = 0.043). Patients with SLE had a negative impact of oral condition on their quality of life. The evaluation of the oral health-related quality of life might be useful to monitor the effects of SLE on oral condition.


Assuntos
Lúpus Eritematoso Sistêmico/complicações , Saúde Bucal/tendências , Higiene Bucal/tendências , Qualidade de Vida/psicologia , Adulto , Brasil/epidemiologia , Estudos Transversais , Progressão da Doença , Feminino , Humanos , Lúpus Eritematoso Sistêmico/diagnóstico , Lúpus Eritematoso Sistêmico/epidemiologia , Masculino , Pessoa de Meia-Idade , Saúde Bucal/estatística & dados numéricos , Higiene Bucal/normas , Índice de Gravidade de Doença , Fumar/efeitos adversos , Fumar/epidemiologia
20.
Arch Oral Biol ; 86: 101-107, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29216525

RESUMO

The impact of high-refined carbohydrate (HC) diet on fat accumulation, adipokines secretion and systemic inflammation is well described. However, it remains unclear whether these processes affect bone remodeling. OBJECTIVE: To investigate the effects of HC diet in the alveolar bone and femur parameters. METHODS: BalbC mice were fed with conventional chow or HC diet for 12 weeks. After experimental time maxillae, femur, blood and white adipose tissue samples were collected. RESULTS: The animals feed with HC diet exhibited considerable increase of adiposity index and adipose tissue levels of TNF-α, IL-6, IL-10, IL-1ß, TGF-ß and leptin. Microtomography analysis of maxillary bone revealed horizontal alveolar bone loss and disruption of trabecular bone in mice feed with HC diet. These deleterious effects were correlated with a disturbance in bone cells and an augmented expression of Rankl/Opg ratio. Consistently, similar effects were observed in femurs, which also exhibited a reduction in bone maximum load and stiffness. CONCLUSION: Our data indicates that HC diet consumption disrupts bone remodeling process, favoring bone loss. Underlying mechanisms relies on fat tissue accumulation and also in systemic and local inflammation.


Assuntos
Tecido Adiposo/metabolismo , Processo Alveolar/metabolismo , Dieta da Carga de Carboidratos , Fêmur/metabolismo , Maxila/metabolismo , Processo Alveolar/diagnóstico por imagem , Animais , Biomarcadores/sangue , Remodelação Óssea , Citocinas/sangue , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Fêmur/diagnóstico por imagem , Inflamação/etiologia , Inflamação/metabolismo , Masculino , Maxila/diagnóstico por imagem , Camundongos , Camundongos Endogâmicos BALB C , Reação em Cadeia da Polimerase , Estresse Mecânico , Microtomografia por Raio-X
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