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The escalating prevalence of antibiotic-resistant bacteria poses a grave threat to human health, necessitating the exploration of novel alternatives to conventional antibiotics. This study investigated the impact of extracts derived from the supernatant of four lactic acid bacteria strains on factors contributing to the pathogenicity of three Staphylococcus aureus strains. The study evaluated the influence of lactic acid bacteria supernatant extracts on the growth, biofilm biomass formation, biofilm metabolic activity, and biofilm integrity of the S. aureus strains. Additionally, the impact on virulence factors (hemolysin and coagulase) was examined. Gas chromatography coupled with mass spectrometry was used to identify the bioactive compounds in the extracts, while molecular docking analyses explored potential interactions. Predominantly, the extracts contain eight 2,5-diketopiperazines, which are cyclic forms of peptides. The extracts demonstrated inhibitory effects on biofilm formation, the ability to disrupt mature biofilms, and reduce the biofilm cell metabolic activity of the S. aureus strains. Furthermore, they exhibited the ability to inhibit α-hemolysin production and reduce coagulase activity. An in silico docking analysis reveals promising interactions between 2,5-diketopiperazines and key proteins (SarA and AgrA) in S. aureus, confirming their antivirulence and antibiofilm activities. These findings suggest that 2,5-diketopiperazines could serve as a promising lead compound in the fight against antibiotic-resistant S. aureus.
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BACKGROUND: Drugs used for the treatment of diseases associated with chronic inflammation, such as cancer and rheumatoid arthritis have the potential to cause undesirable side-effects, which might result in patients ending treatment prematurely. However, plants are a viable option for the treatment of inflammatory diseases. In this study, we assessed the in vivo and in vitro anti-inflammatory activity, and the antitumor effects of the chloroform extract of Salvia ballotiflora (ECL). The pro-apoptotic effects of ECL in CT26 cells were also determined. METHODS: The chloroform extract of Salvia ballotiflora (ECL) was standardized using 19-deoxyicetexone (DEOX) as a phytochemical marker. The anti-inflammatory activity of ECL was determined on acute and chronic inflammatory models using the TPA-induced mouse ear edema assay. The antitumor activity of ECL was evaluated by the subcutaneous inoculation of CT26 cells on the back of Balb/c mice. In vitro CT26 cell death induced by ECL was determined by Annexin V/propidium iodide staining assay using flow cytometry. ECL and the diterpenes isolated from the chloroform extract included 19-deoxyicetexone (DEOX), icetexone (ICT), and 7,20-dihydroanastomosine (DAM), which were tested in LPS-stimulated J774A.1 macrophages to quantify pro-inflammatory cytokine levels. The in vitro anti-arthritic activity of ECL was determined using the bovine serum protein (BSP) denaturation assay. RESULTS: ECL exerted anti-inflammatory activities in acute (84% of inhibition, 2 mg/ear) and chronic models (62.71%, at 100 mg/kg). ECL showed antitumor activity at 200 mg/kg and 300 mg/kg, reducing tumor volume by 30 and 40%, respectively. ECL (9.5 µg/mL) induced in vitro apoptosis in CT26 cells by 29.1% (48 h of treatment) and 93.9% (72 h of treatment). ECL (10 µg/ml) decreased levels of NO (53.7%), pro-inflammatory cytokines IL-6 (44.9%), IL-1ß (71.9%), and TNF-α (40.1%), but increased the production of the anti-inflammatory cytokine IL-10 (44%). The diterpenes DEOX, ICT, and DAM decreased levels of NO (38.34, 47.63, 67.15%), IL-6 (57.84, 60.45, 44.26%), and TNF-α (38.90, 31.30, 32.83%), respectively. ECL showed in vitro antiarthritic activity (IC50 = 482.65 µg/mL). CONCLUSIONS: ECL exhibited anti-inflammatory and anti-tumor activities. Furthermore, the diterpenes DEOX, DAM, and ICT showed anti-inflammatory activity by reducing levels of NO, TNF-α, and IL-6.
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Anti-Inflamatórios não Esteroides/isolamento & purificação , Antineoplásicos Fitogênicos/isolamento & purificação , Diterpenos/farmacologia , Extratos Vegetais/farmacologia , Salvia/química , Animais , Anti-Inflamatórios não Esteroides/farmacologia , Anti-Inflamatórios não Esteroides/toxicidade , Antineoplásicos Fitogênicos/farmacologia , Antineoplásicos Fitogênicos/toxicidade , Apoptose/efeitos dos fármacos , Artrite/tratamento farmacológico , Linhagem Celular Tumoral , Clorofórmio , Citocinas/imunologia , Diterpenos/isolamento & purificação , Diterpenos/toxicidade , Edema/tratamento farmacológico , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Óxido Nítrico/metabolismo , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/toxicidade , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Trypanosoma cruzi is the agent of Chagas disease, an infection that affects around 8 million people worldwide. The search for new anti-T. cruzi drugs are relevant, mainly because the treatment of this disease is limited to two drugs. The objective of this study was to investigate the trypanocidal and cytotoxic activity and elucidate the chemical profile of extracts from the roots of the Lonchocarpus cultratus. Roots from L. cultratus were submitted to successive extractions with hexane, dichloromethane, and methanol, resulting in LCH, LCD, and LCM extracts, respectively. Characterization of extracts was done using 1H-RMN, 13C-RMN, CC and TLC. Treatment of T. cruzi forms (epimastigotes, trypomastigotes, and amastigotes) with crescent concentrations of LCH, LCD, and LCM was done for 72, 48, and 48 h, respectively. After this, the percentage of inhibition and IC50/LC50 were calculated. Benznidazole was used as a positive control. Murine macrophages were treated with different concentrations of both extracts for 48 h, and after, the cellular viability was determined by the MTT method and CC50 was calculated. The chalcones derricin and lonchocarpine were identified in the hexane extract, and for the first time in the genus Lonchocarpus, the presence of a dihydrolonchocarpine derivative was observed. Other chalcones such as isocordoin and erioschalcone B were detected in the dichloromethane extract. The dichloromethane extract showed higher activity against all tested forms of T. cruzi than the other two extracts, with IC50 values of 10.98, 2.42, and 0.83 µg/mL, respectively; these values are very close to those of benznidazole. Although the dichloromethane extract presented a cytotoxic effect against mammalian cells, it showed selectivity against amastigotes. The methanolic extract showed the lowest anti-T. cruzi activity but was non-toxic to peritoneal murine macrophages. Thus, the genus Lonchocarpus had demonstrated in the past action against epimastigotes forms of T. cruzi but is the first time that the activity against infective forms is showed, which leading to further studies with in vivo tests.
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This scoping review study aimed to map the evidence about solvents' use for gutta-percha dissolution and removal during endodontic retreatments. The study protocol followed the Joanna Briggs Institute guidelines, available online (https://osf.io/5vy8n/). Reporting was based on PRISMA Extension for Scoping Reviews. We selected dentistry studies that considered the effectiveness of solvents in gutta-percha dissolution in endodontic retreatments and compared their performance to the use of instrumentation techniques without solvents. The search and study screening were performed in PubMed and Scopus databases by two independent researchers. A descriptive analysis considered the study design, method/technique used for obturation, method/technique used for instrumentation during retreatment, solvent solutions tested, exposure time, and main findings. A total of 41 studies were included. Despite that, most studies suggested that solvents' use may complicate root canal cleanliness, regardless of the type of instrumentation used, and facilitate the presence of gutta-percha remnants in the root surface. Thus, the use of solvents should be avoided and its use should only be considered if the previous working length was not possible to access without it. Despite that, high heterogeneity was observed, further studies are still encouraged comparing the performance and effects of different solvents in different clinical scenarios.
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Guta-Percha/química , Materiais Restauradores do Canal Radicular/química , Tratamento do Canal Radicular , Solventes/química , Guta-Percha/farmacologia , Humanos , Retratamento , Materiais Restauradores do Canal Radicular/farmacologiaRESUMO
Resumen La palta (Persea americana) es uno de los frutos con bastante abundancia en Bolivia, esto se debe a su capacidad de producirse en climas templados y cálidos, lo que trae consigo múltiples beneficios para la salud, pues hay evidencia cientifica que sugiere que la palta podría tener efectos para inhibir o destruir el desarrollo de múltiples microorganismos. Así mismo se pudo evidenciar que los extractos Clorofórmicos y Etanólicos de la semilla de la palta si tiene un efecto bacteriostático y bactericida contra cepas de M. tuberculosis, por inducir la liberación de radicales libres, sin embargo estos extractos también han demostrado tener eficacia contra otras cepas bacterianas, micóticas y parasitarias y algunos virus.
Abstract The avocado (Persea americana) is one of the most abundant fruits in Bolivia, due to its capacity to be produced in temperate and warm climates, bringing multiple health benefits, as scientific evidence suggests the avocado may have effects in inhibiting or destroying the growth of multiple microorganisms. Likewise, it has been shown that the chlorophormic and ethanolic extracts of avocado seed have a bacteriostatic and bactericidal effect against strains of M. tuberculosis, by inducing the release of free radicals; furthermore, these extracts have also been shown to be effective against other bacterial, fungal and parasitic strains and some viruses.
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Native agave fructans were modified by an acylation reaction with lauric acid. Native and modified fructans were characterized using NMR, FTIR and various physicochemical and functional properties at different pHs were evaluated. NMR and FTIR spectra demonstrated the incorporation of lauric acid in the molecular structure of fructans. Modified agave fructans exhibited a color, moisture and water activity similar to native fructans, but properties such as solubility, swelling capacity, emulsifying activity and foam capacity were significantly modified by the acylation reaction mainly when the samples were analyzed at different pHs. The thermogram of the acylated fructans evidenced significant changes in thermal properties when compared with native fructans and acylated fructans were able to form micellar aggregates. In general, modified fructans showed improved functional properties in comparison with native fructans representing an important opportunity to improve the functionality of the foods in which it is incorporated.
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Agave/química , Frutanos/química , Tensoativos/química , Acilação , Domínio Catalítico , Emulsões , Esterificação , Temperatura Alta , Concentração de Íons de Hidrogênio , Ácidos Láuricos/química , Estrutura Molecular , Espectroscopia de Prótons por Ressonância Magnética , Solubilidade , Espectroscopia de Infravermelho com Transformada de Fourier , Tensão Superficial , Água/químicaRESUMO
RESUMEN Objetivos: Evaluar la actividad citotóxica de la fracción clorofórmica del extracto metanólico de Piper aduncum (PAMoCl) y su efecto en el ciclo celular en dos líneas celulares de cáncer gástrico: AGS y KATO III. Materiales y métodos: El efecto citotóxico de PAMoCl se evaluó en las líneas celulares: AGS y KATO III. Se probaron concentraciones de PAMoCl: 1,25; 2,5; 5; 10; 20; 40; 80 y 160 µg/mL. Para evaluar la viabilidad celular se usó el reactivo resazurina. En el ensayo de ciclo celular las células fueron tratadas con 19,62 µg/mL y 39,23 µg/mL de PAMoCl para AGS, así como 87,49 µg/mL y 160 µg/mL para KATO III. Después de 24 horas ambas líneas celulares fueron analizadas por citometría de flujo. Resultados: PAMoCl mostró actividad citotóxica con una inhibición del crecimiento celular en un 50% (IC50) de 39,23 µg/mL y 87,49 µg/mL a las 24 horas y un IC50 de 49,47 µg/mL y 64,68 µg/mL a las 48 horas frente a las líneas celulares AGS y KATO III, respectivamente. Además, se observó que PAMoCl tiene efecto a nivel del ciclo celular: provoca una acumulación de células en la fase G2/M. Conclusiones: PAMoCl contiene metabolitos secundarios con actividad citotóxica que tienen efecto en la fase G2/M del ciclo celular, en dos líneas celulares de cáncer gástrico tanto primario como metastásico. Los resultados de este estudio permitirán profundizar en la búsqueda de principios activos presentes en PAMoCl que tengan mayor eficacia en la eliminación de células de cáncer gástrico, pero con menor toxicidad en células sanas.
ABSTRACT Objectives: To evaluate the cytotoxic activity of the chloroform fraction of the Piper aduncum methanolic extract (PAMoCl) and its effect on the cell cycle in two gastric cancer cell lines: AGS and KATO III. Materials and methods: The cytotoxic effect of PAMoCl was evaluated in cell lines AGS and KATO III. The following PAMoCl concentrations were tested, 1.25, 2.5, 5, 10, 20, 40, 80 and 160 μg/mL. Resazurine was used to evaluate cell viability. In the cell cycle assay, the cells were treated with 19.62 μg/mL and 39.23 μg/mL of PAMoCl for AGS as well as 87.49 μg/mL and 160 μg/mL for KATO III. After 24 hours both cell lines were analyzed by flow cytometry. Results: PAMoCl showed cytotoxic activity, inhibiting cell growth by 50%. It presented a (IC50) of 39.23 μg/mL and 87.49 μg/mL at 24 hours and a (IC50) of 49.47 μg/mL and 64.68 μg/mL at 48 hours against AGS and KATO III cell lines, respectively. In addition, it was observed that PAMoCl has an effect on the cell cycle, it causes an accumulation of cells in the G2/M phase. Conclusions: PAMoCl contains secondary metabolites with cytotoxic activity that have an effect on the G2/M phase of the cell cycle, in two gastric cancer cell lines, both primary and metastatic. The results of this study will allow us to deepen the search for more effective active ingredients found in PAMoCl for eliminating gastric cancer cells, but with less toxicity for healthy cells.
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Neoplasias Gástricas , Ciclo Celular , Linhagem Celular , Clorofórmio , Piper , Metástase NeoplásicaRESUMO
We report the catalytic activity for the complexes-cis-[RuCl2 (dppb)(bipy)] (A), and [η6 -(p-cymene)Ru (dppb)Cl]PF6 (B), wherein dppb = 1,4-bis(diphenylphosphine)butane, and bipy = 2,2'-bipyridine-for the synthesis of CDCl3 from CHCl3 using D2 O as deuterium source. H/D exchange reactions were performed using a chloroform/D2 O, 1:2 molar ratio, vigorously stirred, at room temperature. One mole of KOH was dissolved in D2 O fraction and catalytic complexes from 0.002 to 0.05 mmol were dissolved in chloroform. The H/D exchange reactions were monitored using 13 C nuclear magnetic resonance sequences without proton decoupling. The reaction using 0.01 mmol of compound A reached approximately 55% of H/D conversion in 1 h. In the same time, the reactions with 0.002 mmol of compound A and without catalyst show approximately 28% and 3% H/D exchange, respectively. Without the catalysts, the H/D exchange was only 12.0% in 5 h. For compound B, 55% H/D conversion was observed in 1 h, only when 0.05 mmol was used, which is much higher catalyst concentration. After the isolation of the chloroform fraction and two more addition of D2 O, it was possible to obtain 95.0% H/D exchange in approximately 3 h, using 0.01 mmol of the compound A. Therefore, compound A is an efficient catalyst for a rapid and straightforward synthesis of CDCl3 from CHCl3 at room temperature and using D2 O as deuterium source.
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Developing simple and cost-effective methods for soil microbial biomass carbon (MBC) measurement eases routine laboratory analysis and enables large numbers of soil samples to be measured in a relatively short period of time. Thus, the objective of this study was to develop a microwave-assisted biocidal-extraction (MWE) method which does not employ CHCl3 as biocide and K2SO4 as C-extractor, to estimate MBC. First, the microorganisms of soil samples are killed using microwave (MW) irradiation at energy level of 800 J g-1 soil as biocide followed by microwave irradiation extraction (MWE) at 562 W (120 J g-1 soil for 1 min), using deionized water as solvent. Microbial biomass of carbon from two contrasting soils microwaved with 80, 100, and 140 J g-1 soil did not differ from those obtained by using the chloroform fumigation-extraction (CFE) method with 0.5 mol L-1 K2SO4 as extractant. To evaluate the robustness of the MWE method, twenty-six soil samples, from cultivated and non-cultivated areas, with clay contents from 70-690 g kg-1, organic carbon from 5.52 to 50.82 g C kg-1 and pH values from 3.9 to 6.8 were analyzed for MBC using MWE and CFE methods. There was a linear regression (MW = - 17.87 + 0.92*K2SO4; R2 = 0.705; p < 0.001) between MWE and CFE. The biocidal microwave-assisted extraction method using 120 J g-1 soil for 1 min is a cleaner method for evaluating MBC, because it does not require chloroform, potassium sulfate salt and takes a shorter time to extract a set of soil samples.
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Biomassa , Micro-Ondas , Microbiologia do Solo , Solo/química , Agricultura/métodos , Carbono/análiseRESUMO
This paper focuses on developing, fabricating, and characterizing composite polycaprolactone (PCL) membranes reinforced with titanium dioxide nanoparticles (NPs) elaborated by using two solvents; acetic acid and a mixture of chloroform and N,N-dimethylformamide (DMF). The resulting physical, chemical, and mechanical properties of the composite materials are studied by using experimental characterization techniques such as scanning electron microscopy (SEM), differential scanning calorimetry (DSC), X-Ray diffraction (XRD), Fourier-transform infrared (FTIR) analysis, contact angle (CA), uniaxial and biaxial tensile tests, and surface roughness measurements. Experimental results show that the composite material synthesized by sol-gel and chloroform-DMF has a better performance than the one obtained by using acetic acid as a solvent.
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In recent years, the rapid advances of culture-independent methods and new molecular tools have revolutionized our understanding of microbial biodiversity and ecological functions. DNA extraction from microbial communities is a critical step in this process and several methods have been proposed and used, but the influence of the extraction method on the outcome and ultimately on ecological inferences from the results is not yet precisely determined. Here, we compared two of the most commonly used extraction methods in aquatic microbial ecology, and investigated whether the two methods yielded comparable results for community ecology analyses. We extracted DNA from 15 different shallow lakes with phenol:chloroform, a classical and widely used extraction method, and with the PowerSoil DNA isolation Kit, often suggested as the standard DNA extraction method, with some adaptations for aquatic environments. We found that although only 5% of all OTUs showed significant differences in pairwise comparisons (using the 15 lakes as replicates), these OTUs accounted for >35% (on average) of the relative abundance. Diversity and richness did not differ significantly between the two extraction methods, but the beta-dispersion of the communities indicated that the organic extraction yielded more homogeneous communities, while the kit extraction generated variability. Consequently, we conclude that despite the small number of OTUs with significant differences, their impact on the community composition obtained was not negligible, and therefore the results from these two extraction methods were not comparable.
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Bactérias/isolamento & purificação , Fracionamento Químico/métodos , DNA Bacteriano/isolamento & purificação , Lagos/microbiologia , Bactérias/química , Bactérias/genética , Biodiversidade , DNA Bacteriano/genética , Hidrobiologia , MicrobiotaRESUMO
The aim of this study was to evaluate the effects of different levels of hydrated wheat fiber replacing meat and fat in beef burgers on technological characteristics, sensory acceptance and hunger satisfaction. The different levels of hydrated wheat fiber (1â¯g fiber: 6â¯g water) were 0, 1.25, 2.5, 3.75 and 5.0â¯g of fiber/80â¯g burger portion. Results showed that the greater the addition of hydrated wheat fiber, the lower the protein (Pâ¯<â¯.0001) and lipid (Pâ¯=â¯.0006) content and consequently the greater the reduction in caloric value. Burgers with up to 3.75â¯g fiber/80â¯g portion showed the same (Pâ¯>â¯.05) sensory acceptance as the Control burgers (those without added fiber). Sandwiches comprised of burgers with 2.5 and 5.0â¯g fiber/80â¯g portion caused the same (Pâ¯>â¯.05) hunger satisfaction (satiety feeling) as those comprised of Control burgers for up to 3â¯h after consumption. Burgers containing 3.75â¯g fiber/80â¯g burger may represent an interesting alternative for people who want to reduce caloric intake and/or increase the proportion of insoluble fiber in their diet.
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Fibras na Dieta/análise , Produtos da Carne/análise , Saciação , Animais , Brasil , Bovinos , Comportamento do Consumidor , Substitutos da Gordura , Humanos , Lipídeos , Valor Nutritivo , Suínos , TriticumRESUMO
The golden mussel, Limnoperna fortunei, is a mollusk native to Southeast Asia and a highly invasive species in South American countries such as Brazil, Uruguay, and Argentina. In order to better understand the biological behavior of the species and develop alternative control methods, genetic studies involving the optimization of DNA isolation procedures are of utmost importance.The objective of the present study was to develop a simple, reproducible, free of contaminants, and cheap protocol to extract DNA from L. fortunei using the adductor muscle of the mussel as the source. Four DNA extraction protocols were compared: extraction with SDS and proteinase K (P1); extraction with SDS, proteinase K and phenol (P2); TRIzol extraction (P3); and NaCl, SDS and RNase extraction (P4). DNA concentration (ng μL-1) and purity (at 260/280 nm) were measured using a spectrophotometer. DNA purity and amplification were verified by electrophoresis and PCR, respectively. P1 resulted in samples with low DNA concentrations or without any DNA, as revealed by the quantification and purity analysis; P2 had low efficiency, given the absence of DNA in most of the samples subjected to electrophoresis. On the other hand, P3 showed contamination with proteins, as indicated by an absorbance of <1.8 and by the low-quality electrophoresis results. Finally, P4 resulted in well-defined bands, absorbance between 1.8 and 2.0, and successful amplification by PCR. In conclusion, the extraction protocol P4 is a practical, fast, free of contaminants, and efficient method for the isolation of L. fortunei DNA.(AU)
O mexilhão dourado, Limnoperna fortunei é um molusco originário do sudeste da Ásia, altamente invasor em países Sul-Americanos como Brasil, Uruguai e Argentina. Para compreender melhor o comportamento biológico da espécie e criar alternativas de controle é indispensável a realização de estudos genéticos, onde a otimização dos procedimentos de isolamento do DNA é fundamental.O objetivo desse estudo foi obter um protocolo simples, reproduzível, não contaminante e barato para a extração do DNA de L. fortunei. Foram comparados quatro protocolos experimentais de extração de DNA, utilizando como material biológico o músculo abdutor: extração por SDS e proteinase K (P1), extração por SDS, proteinase K e fenol (P2), extração por Trizol (P3) e extração por NaCl (P4). A quantificação (ng μL-1) e a pureza (260/280 nm) do DNA foram obtidas por espectrofotometria. A integridade e a amplificação do DNA foram verificadas através de eletroforese e PCR, respectivamente. P1 demonstrou baixas concentrações e ausência de DNA nas amostras, identificado pela quantificação e teste de integridade. P2 apresentou baixa eficácia, visualizada pela ausência de DNA na maioria das amostras na eletroforese. Por outro lado, P3 exibiu sinais de contaminação por proteínas, identificado pela razão de absorbância <1.8 e pela baixa qualidade da eletroforese. Finalmente, P4 mostrou um padrão na formação das bandas, absorbância entre 1,8 2,0 e sucesso na amplificação pela PCR. Conclui-se que o protocolo de extração P4 mostrou-se como um método prático, rápido, não contaminante e eficiente para obtenção do DNA de L. fortunei.(AU)
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Animais , Perna (Organismo)/genética , DNA/isolamento & purificação , DNA/síntese química , Bioacumulação/legislação & jurisprudência , Reação em Cadeia da Polimerase/veterinária , Dodecilsulfato de Sódio , Ribonucleases/administração & dosagem , Endopeptidase K/administração & dosagemRESUMO
The golden mussel, Limnoperna fortunei, is a mollusk native to Southeast Asia and a highly invasive species in South American countries such as Brazil, Uruguay, and Argentina. In order to better understand the biological behavior of the species and develop alternative control methods, genetic studies involving the optimization of DNA isolation procedures are of utmost importance.The objective of the present study was to develop a simple, reproducible, free of contaminants, and cheap protocol to extract DNA from L. fortunei using the adductor muscle of the mussel as the source. Four DNA extraction protocols were compared: extraction with SDS and proteinase K (P1); extraction with SDS, proteinase K and phenol (P2); TRIzol extraction (P3); and NaCl, SDS and RNase extraction (P4). DNA concentration (ng μL-1) and purity (at 260/280 nm) were measured using a spectrophotometer. DNA purity and amplification were verified by electrophoresis and PCR, respectively. P1 resulted in samples with low DNA concentrations or without any DNA, as revealed by the quantification and purity analysis; P2 had low efficiency, given the absence of DNA in most of the samples subjected to electrophoresis. On the other hand, P3 showed contamination with proteins, as indicated by an absorbance of <1.8 and by the low-quality electrophoresis results. Finally, P4 resulted in well-defined bands, absorbance between 1.8 and 2.0, and successful amplification by PCR. In conclusion, the extraction protocol P4 is a practical, fast, free of contaminants, and efficient method for the isolation of L. fortunei DNA.
O mexilhão dourado, Limnoperna fortunei é um molusco originário do sudeste da Ásia, altamente invasor em países Sul-Americanos como Brasil, Uruguai e Argentina. Para compreender melhor o comportamento biológico da espécie e criar alternativas de controle é indispensável a realização de estudos genéticos, onde a otimização dos procedimentos de isolamento do DNA é fundamental.O objetivo desse estudo foi obter um protocolo simples, reproduzível, não contaminante e barato para a extração do DNA de L. fortunei. Foram comparados quatro protocolos experimentais de extração de DNA, utilizando como material biológico o músculo abdutor: extração por SDS e proteinase K (P1), extração por SDS, proteinase K e fenol (P2), extração por Trizol (P3) e extração por NaCl (P4). A quantificação (ng μL-1) e a pureza (260/280 nm) do DNA foram obtidas por espectrofotometria. A integridade e a amplificação do DNA foram verificadas através de eletroforese e PCR, respectivamente. P1 demonstrou baixas concentrações e ausência de DNA nas amostras, identificado pela quantificação e teste de integridade. P2 apresentou baixa eficácia, visualizada pela ausência de DNA na maioria das amostras na eletroforese. Por outro lado, P3 exibiu sinais de contaminação por proteínas, identificado pela razão de absorbância <1.8 e pela baixa qualidade da eletroforese. Finalmente, P4 mostrou um padrão na formação das bandas, absorbância entre 1,8 2,0 e sucesso na amplificação pela PCR. Conclui-se que o protocolo de extração P4 mostrou-se como um método prático, rápido, não contaminante e eficiente para obtenção do DNA de L. fortunei.
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Animais , DNA , Bioacumulação/legislação & jurisprudência , Perna (Organismo)/genética , Dodecilsulfato de Sódio , Endopeptidase K/administração & dosagem , Reação em Cadeia da Polimerase/veterinária , Ribonucleases/administração & dosagemRESUMO
SUMMARY Some chalcone compounds are synthesized and their characterization was done by spectroscopic techniques such as IR, NMR and mass spectrometry. Some physicochemical properties such as acoustical properties, refractive index, conductance and partition coefficient have been studied for these synthesized compounds in N, N-dimethyl formamide and chloroform at 303.15 K. The studied properties are useful in QSAR studies and applications of these compounds in various other fields. It is observed that these parameters are affected by solvent and substitutions present in compounds.
RESUMEN Se sintetizaron algunos compuestos del tipo chalcona y su caracterizaron mediante técnicas espectroscópicas tales como IR, RMN y espectrometría de masa. Algunas propiedades fisicoquímicas tales como propiedades acústicas, índice de refracción, conductancia y coeficiente de reparto se estudiaron, para los compuestos sintetizados, en N,N-dimetil formamida y cloroformo a 303,15 K. Las propiedades estudiadas son útiles en estudios QSAR y en aplicaciones de estos compuestos en otros campos. Se observa que estos parámetros se ven afectados por el disolvente y las sustituciones presentes en los compuestos.
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A nano-composite from biologically obtained chitin nanofillers homogenously dispersed in a poly(ε-caprolactone) matrix was successfully achieved by an ultrasonication-assisted non-toxic and non-aqueous methodology. For this purpose, biological chitin was obtained from lactic acid fermentation of shrimp wastes and converted into chitin whiskers by acidic hydrolysis in a novel process at low temperature (4°C) that enhanced the distribution and yield. Additionally, the polyester matrix was enzymatically produced in a non-toxic compressed fluid (1,1,1,2-tetrafluoroethane at 25bar and 65°C) medium. The homogeneous distribution of the nanofiller in the matrix was corroborated by confocal and atomic force microscopies. Films of the nanocomposite were physicochemically characterized to assess its adequate properties. Additionally, the qualitative viability of human fibroblasts and osteoblasts cells was studied on the produced nanocomposite films showing good biocompatibility.
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Quitina/química , Nanocompostos/química , Nanopartículas/química , Poliésteres/química , Adulto , Animais , Candida/enzimologia , Criança , Quitina/isolamento & purificação , Fibroblastos , Química Verde , Humanos , Hidrocarbonetos Fluorados/química , Hidrólise , Lactobacillus plantarum/química , Lipase/química , Osteoblastos , Tamanho da Partícula , Penaeidae/químicaRESUMO
Ultrasonic velocity, density and viscosity of some synthesized chalcones were measured in N,N-dimethyl formamide and chloroform at different temperatures (298.15 to 318.15 K). From these experimental data, various acoustical parameters such as specific impedance (Z), adiabatic compressibility (k s), Rao's molar sound function (Rm), intermolecular free path length (Lf), solvation number (Sn), internal pressure (Π) have been calculated in order to understand the molecular interactions in the studied solutions. The results are interpreted in terms of molecular interactions occurring in the solutions.
La velocidad ultrasónica, la densidad y la viscosidad de soluciones de algunas chalconas sintéticas se midieron en N, N-dimetilformamida y cloroformo a diferentes temperaturas (desde 298,15 hasta 318,15 K). A partir de estos datos experimentales, se calcularon diversos parámetros acústicos tales como la impedancia específica (Z), la compresibilidad adiabática (k s), la función de sonido molar de Rao (Rm), la longitud de trayecto libre intermolecular (Lf), el número de solvatación (Sn) y la presión interna (Π), para comprender las interacciones moleculares en las soluciones estudiadas. Los resultados se interpretan en términos de las posibles interacciones moleculares que ocurren en las soluciones.
RESUMO
Chloroform is an organic solvent used as an intermediate in the synthesis of various fluorocarbons. Despite its widespread use in industry and agriculture, exposure to chloroform can cause illnesses such as cancer, especially in the liver and kidneys. The aim of the study was to analyze the effects of chloroform on redox imbalance and pulmonary inflammatory response in adult C57BL/6 mice. Forty animals were divided into 4 groups (N = 10): female (FCG) and male (MCG) controls, and females (FEG) and males (MEG) exposed to chloroform (7.0 ppm) 3 times/d for 20 minutes for 5 days. Total and differential cell counts, oxidative damage analysis, and protein carbonyl and antioxidant enzyme catalase (CAT) activity measurements were performed. Morphometric analyses included alveolar area (Aa) and volume density of alveolar septa (Vv) measurements. Compared to FCG and MCG, inflammatory cell influx, oxidative damage to lipids and proteins, and CAT activity were higher in FEG and MEG, respectively. Oxidative damage and enzyme CAT activity were higher in FEG than in FCG. The Aa was higher in FEG and MEG than in FCG and MCG, respectively. The Vv was lower in FEG and MEG than in FCG and MCG, respectively. This study highlights the risks of occupational chloroform exposure at low concentrations and the intensity of oxidative damage related to gender. The results validate a model of acute exposure that provides cellular and biochemical data through short-term exposure to chloroform.
Assuntos
Carcinógenos/toxicidade , Clorofórmio/toxicidade , Estresse Oxidativo/efeitos dos fármacos , Pneumonia/induzido quimicamente , Alvéolos Pulmonares/efeitos dos fármacos , Mucosa Respiratória/efeitos dos fármacos , Solventes/toxicidade , Animais , Câmaras de Exposição Atmosférica , Biomarcadores/metabolismo , Catalase/metabolismo , Feminino , Imunidade Inata/efeitos dos fármacos , Imunidade nas Mucosas/efeitos dos fármacos , Exposição por Inalação , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Camundongos Endogâmicos C57BL , Oxirredução , Pneumonia/imunologia , Pneumonia/metabolismo , Pneumonia/patologia , Carbonilação Proteica/efeitos dos fármacos , Alvéolos Pulmonares/imunologia , Alvéolos Pulmonares/metabolismo , Alvéolos Pulmonares/patologia , Mucosa Respiratória/imunologia , Mucosa Respiratória/metabolismo , Mucosa Respiratória/patologia , Caracteres Sexuais , Testes de Toxicidade AgudaRESUMO
This study used gas chromatography-mass spectrometry (GC-MS) to detect the products formed during the contact of sodium hypochlorite (NaOCl) with bovine pulp and dentin. For analysis of the products formed in the volatile phase, 11 mg of bovine pulp tissue were placed in contact with 0.5%, 2.5% and 5.25% NaOCl until complete tissue dissolution occurred. The solid phase microextraction (SPME) fiber was exposed inside the container through the cover membrane and immediately injected into the GC-MS system. 30 mg of the of dentin were kept in contact with NaOCl, and then the SPME fiber was exposed inside the container through the cover membrane for adsorption of the products and injected into the GC-MS system. The same protocol was used for the aqueous phase. For analysis of the volatile compounds, the final solution was extracted using pure ethyl ether. The suspended particulate phase of the mixture was aspirated, and ether was separated from the aqueous phase of the solution. The ether containing the products that resulted from the chemical interaction of dentin and pulp with the NaOCl was filtered and then injected into the GC-MS system for analysis of the aqueous phase. The aqueous and volatile phases of both dentin and pulp showed the formation of chloroform, hexachloroethane, dichloromethylbenzene and benzaldehyde. In conclusion, organochlorine compounds are generated during the contact of dentin and pulp with NaOCl at concentrations of 0.5%, 2.5% and 5.25%.
Este estudo utilizou a cromatografia gasosa acoplada a espectrometria de massa (CG-MS) para detectar os produtos que se formaram durante o contato de hipoclorito de sódio (NaOCl) com polpa dental bovina e dentina. Para a análise dos produtos formados na fase volátil, 11 mg de polpa bovina foram colocados em contato com 0,5 % , 2,5 % e 5,25 % de NaOCl, até à dissolução completa dos tecidos. A fibra de microextração em fase sólida (SPME) era exposta dentro do recipiente através da membrana da tampa, por 15 minutos, para a adsorção dos produtos formados e imediatamente injetada no CG-MS para análise. Para a análise da dentina, 30 mg do de amostras foram mantidas em contacto com o NaOCl, por 15 min, e então a fibra de SPME era exposta no interior do recipiente através da membrana de cobertura para a adsorção dos produtos e injectado no sistema de GC-MS. O mesmo protocolo foi utilizado para a fase aquosa. Para a análise dos compostos voláteis, a solução final foi extraída com éter etílico puro. A fase de partículas em suspensão da mistura foi aspirada, e o éter foi separado da fase aquosa da solução. O éter contendo os produtos que resultaram da interacção química da dentina e polpa com hipoclorito de sódio foi filtrado e, em seguida, injectado no sistema GC-MS para análise da fase aquosa. As fases aquosas e voláteis de dentina e polpa mostraram a formação de clorofórmio, hexacloroetano, dichloromethylbenzene e benzaldeído. Compostos organoclorados são gerados durante o contacto da dentina e polpa com hipoclorito de sódio em concentrações de 0,5 % , 2,5 % e 5,25 %.
Assuntos
Animais , Bovinos , Polpa Dentária/química , Dentina/química , Hidrocarbonetos Clorados/análise , Hipoclorito de Sódio/química , Cromatografia Gasosa-Espectrometria de Massas , Microextração em Fase SólidaRESUMO
This study compared, by scanning electron microscopy (SEM), the efficacy of three solvents on the removal of filling materials from dentinal tubules during endodontic retreatment. Forty human maxillary canines with straight canals were prepared according to a crown-down technique and enlarged to a#30 apical file size, before obturation with gutta-percha and a zinc-oxide-eugenol based sealer. The samples were stored for 3 months before being randomly assigned to four groups: chloroform (n=10), orange oil (n=10), eucalyptol (n=10) and control (n=10). Solvents were applied to a reservoir created on the coronal root third using Gates Glidden drills. The total time for retreatment using the solvents was 5 minutes per tooth. Following retreatment the roots were split longitudinally for SEM evaluation. SEM images were digitized, analyzed using Image ProPlus 4.5 software, and the number of dentinal tubules free of filling material from the middle and apical thirds was recorded. No significant difference was found among the solvent groups regarding the number of dentinal tubules free of root filling remnants in the middle and apical root thirds (p>0.05). However, the control group had fewer dentinal tubules free of filling material (p<0.05). Under the tested conditions, it may be concluded that there was no significant difference among the solvents used to obtain dentinal tubules free of filling material remnants.