Effects of Qualea grandiflora Extract on the Expression of MMP-14 and HIF-1α in Cultured Fibroblasts and Preosteoblasts

Abstract Background: Qualea grandiflora (QG) (Vochysiaceae), also known as "pau-ferro", "pau-terra" or "pau-de-tucano", is a very common deciduous tree in the Brazilian Cerrado used in traditional medicine to treat inflammations, ulcers, diarrhea, and infections. There are reports in the scientific literature that demonstrate the medicinal effects of the bark and leaf of the QG. However, studies involving this plant are rather imited. Aim of the study: To perform the phytochemical analysis of the QG hydroalcoholic extract (HAE) of leaves, and to investigate it effects on fibroblast and preosteoblasts. Methods: Phytochemical analysis was done by HPLC-DAD. Murine NIH/3T3 fibroblasts and MC3T3-E1 preosteoblasts cell lines (ATCC) were used for the experiments. Cell viability was assessed by the MTT colorimetric assay and the expression of MMP-14 and HIF-1α by immunofluorescence. Results and conclusion: The following compounds were identified by HPLC-DAD, such as quinic acid, ethyl galate, ellagic acid derivatives as O-methylellagic acid O-galloyl, O-methylellagic acid O-deoxyhexoside, galloyl derivatives, flavonol glycoside as kaempferol-O-deoxyhexoside, quercetin-O-deoxyhexoside, myricetin-O-deoxyhexoside and the pentacyclic triterpene arjunglucoside. Cell viability results demonstrated no cytotoxic effects in the studied concentrations. We found in QG HAE some compounds with therapeutic properties that can increase the expression of MMP-14 and HIF-1α, in fibroblasts and preosteoblasts. These data suggest that QG HAE has an action on these two molecules widely involved in physiological conditions, such as collagen remodeling, bone development and growth and pathological processes as HIF signaling in cancer metastasis.

Concentração inibitória mínima e concentração bactericida mínima de extratos hidroalcoólicos das folhas de Myracrodruon urundeuva All. e Qualea grandiflora Mart. sobre Streptococcus mutans e Lactobacillus casei / Minimum inhibitory concentration and minimum bactericidal concentration of hydroalcoholic extracts of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves on Streptococcus mutans and Lactobacillus casei

A cárie dentária é uma doença bucal de alta prevalência e impactante em países em desenvolvimento. É causada pela presença de biofilme dentário rico em bactérias acidogênicas e acidúricas, como Streptococcus mutans e Lactobacillus casei. Neste sentido, a fitoterapia tem sido aplicada na odontologia devido ao seu conhecido efeito antimicrobiano, tendo potencial para prevenir doenças como a cárie dentária. Portanto, o presente estudo tem como objetivo testar o potencial antimicrobiano de extratos bruto e etanólico das folhas de Myracrodruon urundeuva (M. urundeuva.) e Qualea grandiflora (Q. grandiflora) sobre S.mutans e L. casei. Para tal, determinaram-se a Concentração Inibitória Mínima (CIM) e a Concentração Bactericida Mínima (CBM). A CIM foi definida como a menor concentração do agente antimicrobiano capaz de inibir 100% o crescimento microbiano (absorbância) em relação aos controles negativos. Para a CBM, alíquotas foram removidas dos poços que não apresentaram nenhuma absorbância (viabilidade, concentrações CIM) e semeadas em placas de ágar BHI, incubadas por 24 h a 37°C em estufa de CO2 5%. A CBM foi determinada considerando a menor concentração dos extratos capaz de impedir o crescimento bacteriano visível. Cepas de S. mutans (ATCC 21175) e L. casei (ATTC 334) foram ativadas em BHI e caldo Rogosa, respectivamente. A CIM foi determinada pela técnica de diluição em microplacas de 96 poços (100 l de extrato + 80 l BHI/Rogosa + 20 l da bactéria diluída em BHI/Rogosa equivalente a 5x105 UFC/mL), as quais foram incubadas por 24 h (S. mutans) e 48 h (L. casei) a 37°C em estufa de CO2 5%. Os extratos de M. urundeuva e Q. grandiflora inicialmente foram diluídos em BHI/Rogosa variando as concentrações entre 2 mg/ml a 0,00012207 mg/ml e os mesmos extratos diluídos em álcool foram avaliados nas concentrações entre 20 mg/ml a 0,00244 mg/ml para S. mutans e L. casei. Não foi possível determinar a CIM e a CBM para os extratos diluídos no BHI/Rogosa. Foram utilizados como controle positivo a clorexidina e como controles negativos BHI/Rogosa com e sem álcool a 5%. As CIMs (CBMs) da M. urundeuva e Q. grandiflora, diluídas em álcool, e clorexidina contra S. mutans foram 2,5 mg/ml (2,5 mg/ml), 5,0 mg/ml (--) e 0,00468 mg/ml (0,00937 mg/ml), respectivamente. Em relação ao L. casei, as CIMs (CBMs) da M. urundeuva e Q. grandiflora, diluídas em álcool, e da clorexidina foram 0,156 mg/ml (0,312 mg/ml), 0,156 0,625 mg/ml (0,312 0,625mg/ml) e 0,00468 mg/ml (0,3 mg/ml), respectivamente. Como conclusão, nosso estudo mostrou que L. casei (ATTC 334) é mais susceptível aos extratos que S. mutans (ATCC 21175) e o extrato M. urundeuva apresenta melhor efeito antimicrobiano que a Q. grandiflora em S. mutans (ATCC 21175), porém os dois extratos apresentam efeito similar sobre L. casei (ATTC 334) e ambos foram inferiores à CHX.(AU)

Dental caries is an oral disease of high prevalence and impact in developing countries. It is caused by the presence of a dental biofilm rich in acidogenic and aciduric bacteria, such as Streptococcus mutans and Lactobacillus casei. Accordingly, phytotherapy has been applied in dentistry due to its known antimicrobial effect, having potential to prevent diseases such as dental caries. Therefore, the present study aims to test the antimicrobial potential of crude and ethanolic extracts of Myracrodruon urundeuva (M. urundeuva) and Qualea grandiflora (Q. grandiflora) leaves on S. mutans and L. casei. For this, the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) were determined. MIC was defined as the lowest concentration of the antimicrobial agent capable of inhibiting 100% the microbial growth in comparison to the negative controls. For MBC, aliquots were removed from the wells that did not show any absorbance (viability, concentrations than MIC) and seeded on BHI agar plates, incubated for 24 h at 37°C and 5% CO2. The MBC was determined considering the lowest concentration of extracts capable of preventing visible bacterial growth. Strains of S. mutans (ATCC 21175) and L. casei (ATTC 334) were activated in BHI and Rogosa broth. MIC was determined by the dilution technique in 96-wells microplates (100 l of extract + 80 l BHI/ Rogosa + 20 l of bacterium diluted in BHI/Rogosa equivalent to 5x105 CFU/ml), which were incubated for 24 h (S. mutans) and 48 h (L. casei) at 37°C and 5% CO2. The extracts were firstly diluted in BHI/Rogosa varying the concentrations between 2 mg/mL and 0.00012207 mg/ml; the same extracts were also diluted in alcohol at concentrations ranging from 20 mg/ml to 0.00244 mg/ml and both tested against S. mutans and L. casei. It was not possible to determine the MIC and MBC for the extracts diluted in BHI/Rogosa. Chlorhexidine was used as positive control, while BHI/Rogosa with or without 5% alcohol were used as negative controls. The MICs (MBCs) of M. urundeuva and Q. grandiflora diluted in alcohol, and clorexidine against S. mutans were 2.5 mg/ml (2.5 mg/ml), 5.0 mg/ml (--) and 0.00468 mg/ml (0.00937 mg/ml), respectively. In respect to L. casei, the MICs (MBCs) of M. urundeuva and Q. grandiflora, diluted in alcohol, and chlorhexidine were 0.156 mg/ml (0.312 mg/ml), 0.156 0.625 mg/ml (0.312 0.625 mg/ml) and 0.00468 mg/ml (0.3 mg/ml), respectively. In conclusion, our study showed that L. casei (ATTC 334) is more susceptible than S. mutans (ATCC 21175) to the extracts and the extract of M. urundeuva has a better antimicrobial effect than Q. grandiflora against S. mutans (ATCC 21175), but both extracts have similar effect on L. casei (ATTC 334) and they were inferior to CHX.(AU)

Estimación del contenido de biomasa, fijación de carbono y servicios ambientales, en un área de bosque primario en el resguardo indígena piapoco chigüiro-chátare de barrancominas, departamento del guainía (colombia) / Estimated biomass content, carbon sequestration and environmental services,in a forest area in the shelter primary indian piapoco chigüiro-chátare, barrancominas, guainia department (colombia)

En Barrancominas (Guainía, Colombia) las diferentes formas culturales de trabajo y producción de la comunidad Piapoco, incluyen la tumba y quema de bosques a orillas del río, para ampliación de frontera agrícola, establecimiento de cultivos ilícitos y potreros; lo cual contribuye en el aumento de las concentraciones de carbono en la atmósfera que, a la vez, repercute en el cambio climático global. En consecuencia, el presente estudio quiso estimar el contenido de biomasa, fijación de carbono y los servicios ambientales, en un área de bosque primario en el Resguardo Indígena Chigüiro-Chátare, empleando una metodología no destructiva. Se estimó tanto la biomasa aérea, como el contenido de carbono (C) del estrato arbóreo del bosque por hectárea y la tasa de producción de hojarasca de las especies maderables: Mure, Pendare y Arenillo blanco. La biomasa aérea fue estimada a partir de los parámetros medidos en campo como altura y DAP y densidades de la madera de cada especie. Además, se analizó la dominancia de las especies para de esta forma conocer la representatividad del aporte de biomasa aérea y contenido de carbono en el bosque.

In Barrancominas (Guainía) different cultural forms of work and production of the Piapoco community, includes the slash and burn of forest along the river, to extend the farming, establishment of illicit crops and pastures contribute to increase carbon concentrations in the atmosphere which in turn affect global climate change. Thus this study wanted to value the biomass content, carbon fixation and ecosystem services in an area of primary forest in the indigenous community of Chigüiro-Chátare using a nondestructive method or indirect. In this study the aerial biomass was estimated as well as the carbon (C) content of the arboreal stratum of forest per hectare and production rate of dead leaves of three species forestry Mure, Pendare and Arenillo trees. The aerial biomass was estimated from the measured parameters in field such as: height, DAP and wood densities of each species. In addition, the species dominance was analyzed in order to know the representativeness of the biomass contribution and carbon content in the forest.

Pollen morphology of Vochysiaceae tree species in the State of Santa Catarina, Southern Brazil / Morfología del polen de las especies de árboles de Vochysiaceae en el estado de Santa Catarina, sur de Brasil

Tropical Vochysiaceae includes mainly trees, and also shrubs and subshrubs. Three genera and seven species are present in the Brazilian state of Santa Catarina. The pollen morphology of six species of trees, belonging to three genera of the Vochysiaceae A. St-Hil. family, was studied. Herbaria samples were obtained, processed and treated by standard methods. The pollen grain morphology of Callisthene, Qualea and Vochysia is distinct. Medium sized pollen grains occur in Vochysia species, and small ones in Callisthene and Qualea. Specific characteristics were considered at species level [C. castellanosii H. F. Martins, C. kuhlmannii H. F.Martins, Qualea cordata Spreng var. cordata, Q. cryptantha (Spreng) Warm. var. cryptantha, Vochysia magnifica Warm. and V. tucanorum Mart.]. The presence of a fastigium (vestibulum) and a thin space devoid of nexine fixing the boundary of the apertural area is characteristic of Qualea and Vochysia species only. Rev. Biol. Trop. 62 (3): 1209-1215. Epub 2014 September 01.

Las Vochysiaceae tropicales incluyen principalmente árboles, arbustos y subarbustos. Tres géneros y siete especies están presentes en el estado brasileño de Santa Catarina. La morfología del polen de las seis especies de árboles, pertenecientes a tres géneros de la Vochysiaceae A. St.- Hil . familia, fue considerado en el presente trabajo. La morfología de los granos de polen de Callisthene Mart., Qualea Aubl. y Vochysia (Aubl.) Juss. es distinta. Los granos de polen de tamaño medio se producen en las especies de Vochysia y pequeños en Callisthene y Qualea. Características particulares fueron considerados a nivel de especie [C. castellanosii H.F. Martins, C. kuhlmannii H.F. Martins, Qualea cordata Spreng var. cordata, Q. cryptantha (Spreng) Warm. var. cryptantha, Vochysia magnifica Warm. and V. tucanorum Mart.]. La presencia de un fastigium (vestibulum) y de un espacio delgado que carece de nexina limita la zona apertural y es característica solo de las especies de Qualea y Vochysia.

Esteroide e Triterpenos de espécies de Qualea - Bioatividade sobre Mycobacterium tuberculosis / Sterol and Triterpenes from Qualea species: Bioactivity on Mycobacterium tuberculosis

O extrato clorofórmico das cascas de Qualea parviflora (Vochysiaceae) foi fracionado em cromatografia em coluna usando sílica gel para fornecer triterpenos (lupeol, lupenona, betulina, ácido epibetulínico, e friedelina) e um esteroide (β-sitosterol). Os compostos β- sitosterol, lupenona e lupeol foram identificados também em Q. grandiflora e Q. multiflora, enquanto friedelina foi detectada apenas em Q. multiflorautilizando a cromatografia gasosa acoplada à espectrometria de massas. A atividade anti Mycobacterium tuberculosis do extrato clorofórmico e dos compostos isolados foi determinada por MABA e o valor da CIM variou de 250,0 a 31,2 μg/mL. Esta investigação constitui o primeiro relato do estudo químico e antitubercular de compostos apolares das espécies de Qualea.

The chloroform extract of bark of the tropical tree Qualea parviflora (Vochysiaceae) was fractionated by column chromatography on silica gel, yielding triterpenes (lupeol, lupenone, betulin, epibetulinic acid and friedelin) and a steroid (β-sitosterol). β-sitosterol, lupenone and lupeol were also identified in Q. grandiflora and Q. multiflora, while friedelin was detected only in Q. Multiflora, by means of gas chromatography-mass spectrometry. The anti-Mycobacterium tuberculosis activity of the chloroform extract and isolated compounds was assayed by MABA and MIC values ranged from 250.0 to 31.2 μg/mL. This study is the first to investigate the chemistry and antitubercular activity of apolar compounds from Qualeaspecies.

Atividade antibacteriana de plantas úteis e constituintes químicos da raiz de Copernicia prunifera / Antibacterial activity of useful plants and chemical constituents of the roots of Copernicia prunifera

Extratos etanólicos de Qualea grandiflora e Copernicia prunifera e extrato hexânico de Dipteryx lacunifera foram avaliados quanto a atividade antibacteriana, utilizando ensaios de difusão a partir de orifício e concentração inibitória mínima (CIM), frente a cepas Gram positivas e Gram-negativas, incluindo espécies multidroga resistentes. O extrato de Q. grandiflora apresentou atividade moderada para as cepas de Staphylococcus epidermidis (CIM = 500 µg/mL) e atividade fraca sobre as demais bactérias Gram-positivas testadas e inativo sobre bactérias Gram-negativas. Os resultados obtidos com S. epidermidis apesar de moderados são importantes, uma vez que este microorganismo é o principal causador de bacteremias e sepse associada com dispositivos médicos implantados. gama-Tocoferol e a mistura de sitosterol e estigmasterol foram isolados do extrato etanólico de raiz de C. prunifera e as estruturas destes compostos foram identificadas com base na análise dos dados espectrais de RMN e comparação com a literatura.

Ethanol extracts of Qualea grandiflora and Copernicia prunifera and hexane extract of Dipteryx lacunifera were evaluated by diffusion in agar and minimum inhibitory concentration (MIC) against Gram-positive and Gram-negative bacteria including multiresistant drug strains. The extract of Q. grandiflora presented moderate activity for Staphylococcus epidermidis (MIC = 500 µg/mL) and weak activity against other Gram-positive strains and inactive for Gram-negative species. The results obtained for S. epidermidis despite being moderate are important because this pathogen is often recovered from bacteremia and sepsis from infections of implanted devices. gamma-Tocopherol and the mixture of sitosterol and stigmasterol were isolated from the ethanol extract of the roots of C. prunifera. The structures of these compounds were identified by NMR spectroscopy and comparison with literature data.