Evaluation of micronuclei frequency in Tradescantia pallida pollen mother cells treated with ethanolic extracts isolated from Cryptocarya mandioccana, Cryptocarya moschata and Pterogyne nitens

Although herbal extracts contain several classes of compounds with pharmacological activity, they also present toxic substances with mutagenic effects. The aim of the present study was to verify the mutagenicity of Cryptocarya moschata, Cryptocarya mandioccana and Pterogyne nitens using micronucleus assay in pollen mother cells (tetrads) in Tradescantia pallida (Trad-MCN). T. pallida inflorescences were treated with different concentrations of ethanolic extracts from the selected plant species. For C. mandioccana C. moschata and P. nitens, Trad-MCN assays were carried out simultaneoulsly, followed by positive control (formaldehyde 10000 ppm), negative control (Hoagland's solution), and vehicle control (Tween 20 20% or DMSO 3%). MCN present in tetrads were quantified in 300 tetrads/inflorescence and the mean (%) and standard error (SE) were established for at least 10 inflorescences per treatment. The extracts demonstrated dose response mutagenicity (clastogenic/aneugenic effects), respectively, C. mandioccana (0.5, 1.0 and 2.0 mg/mL) and P. nitens (1.0 and 2.0 mg/mL) However, no mutagenic effect was observed to C. moschata at the concentrations evaluated in the present study. We can conclude that the C. mandioccana and P. nitens extracts demonstrated clastogenic/aneugenic effects in highest concentrations whereas C. moschata extract did not demonstrate the same effect. © 2006 Sociedade Brasileira de Toxicologia.

Modulatory effect of Byrsonima basiloba extracts on the mutagenicity of certain direct and indirect-acting mutagens in Salmonella typhimurium assays

Byrsonima basiloba A. Juss. species is a native arboreal type from the Brazilian cerrado (tropical American savanna), and the local population uses it to treat diseases, such as diarrhea and gastric ulcer. It belongs to the Malpighiaceae family, and it is commonly known as murici. Considering the popular use of B. basiloba derivatives and the lack of pharmacological potential studies regarding this vegetal species, the mutagenic and antimutagenic effect of methanol (MeOH) and chloroform extracts were evaluated by the Ames test, using strains TA97a, TA98, TA100, and TA102 of Salmonella typhimurium. No mutagenic activity was observed in any of the extracts. To evaluate the antimutagenic potential, direct and indirect mutagenic agents were used: 4 nitro-o-phenylenediamine, sodium azide, mitomycin C, aflatoxin B1, benzo[a]pyrene, and hydrogen peroxide. Both the extracts evaluated showed antimutagenic activity, but the highest value of inhibition level (89%) was obtained with the MeOH extract and strain TA100 in the presence of aflatoxin B1. Phytochemical analysis of the extracts revealed the presence of n-alkanes, lupeol, ursolic and oleanolic acid, (+)-catechin, quercetin-3-O-α-L-arabinopyranoside, gallic acid, methyl gallate, amentoflavone, quercetin, quercetin-3-O-(2″-O-galloyl)-β-D- galactopyranoside, and quercetin-3-O-(2″-O-galloyl)-α-L- arabinopyranoside. © 2008 Mary Ann Liebert, Inc.

Determination of calcium and magnesium in hydroethanolic extracts of propolis by atomic absorption flame spectrophotometry

Propolis is a natural product collected by honeybees and has a large range of pharmacological activity, including antimicrobial, antitumoral, antioxidant and anti-inflammatory. Its use as a popular medicine is increasing all over the world, creating a need for quality control of the commercial products. In this study the levels of calcium and magnesium in commercial hydroalcoholic propolis extracts from varies states of Brazil were determined by atomic absorption flame spectrophotometry and different values were obtained for northern and southern states. This study can be extended to the analysis of metals that are harmful to health. The results showed that the calibration curves were linear over a wide concentration range (0.5-4.0 μg.mL -1 for calcium and 0.05-0.4 μg.mL -1 for magnesium) with good correlation coefficients (0.999 and 0.988, respectively). Good analytical recovery (94%) was obtained. The proposed method showed adequate precision and relative standard deviation lower than 2 %. The method is accurate and precise as well as having advantages such as simplicity and speed.

BMP implant associated with platelet-rich plasma in orbit fracture repair

Purpose: To evaluate a bone morphogenetic protein (BMP) implant with and without platelet-rich plasma (PRP), which is supposed to accelerate fracture consolidation in the orbit fracture treatment. Methods: Thirty-six white rabbits were subjected to orbital fracture and treated in three groups: BMP implant fracture repair (G1), BMP plus PRP implant fracture repair (G2), and fracture and spontaneous repair (G3). The animals were sacrificed at 7, 30, 90, and 180 days after surgery. A radiology evaluation was carried out on the 7th day after the fracture and at the sacrifice moments. After the animals' death, the orbital content material was removed and prepared for morphological and morphometric analysis. Results: Radiology suggested intramembranous and progressive cavitation and ossification without a reduction in implant size and with signs of calcium deposition; these events were confirmed by histological analysis, which showed a lymphomononuclear inflammatory reaction in G1 and G2, more intense 7 days after surgery and reducing after 30 days. Associating PRP with BMP did not accelerate bone induction. Conclusion: BMP implant promotes bone induction, integration at fracture site, scarce inflammatory reaction, and may be a good alternative in orbit fracture reconstruction. The addition of PRP to the BMP plate did not accelerate the resolution, and its use is not necessary. Copyright © Informa Healthcare USA, Inc.

Identification of ellagic acid derivatives in methanolic extracts from Qualea species

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Influence of diltiazem in combination with a sucrose-rich diet on gingival alterations in rats

The aim of this study was to evaluate the influence of diltiazem in combination with a sucrose-rich diet on gingival alterations in rats. One hundred and twenty male Holtzman rats were randomly assigned to 10 groups (n = 12), being 2 control groups treated with saline and 8 test groups treated with diltiazem in daily doses of 5, 25, 50 and 100 mg/kg during 40 or 60 days. Afterwards, the mandibles were removed for macroscopic, histologic and histometric analyses of the buccal gingiva of the mandibular right first molar. No macroscopic characteristic of gingival overgrowth was observed in any of the groups. The microscopic analysis showed characteristics of normality with inflammatory cells only adjacent to the crevicular epithelium in all groups for both periods. The histometric analysis showed significant differences only for the epithelial tissue area in the 40-day period (Kruskal-Wallis; P = 0.032). Comparing the periods, significant differences regarding the connective and epithelial tissue areas were observed only in the group treated with a 25 mg/kg dose (Mann-Whitney; P = 0.004 and P = 0.007, respectively). Oral administration of diltiazem in combination with a sucrose-rich diet did not induce gingival alterations in rats. © 2009 Sociedade Brasileira de Pesquisa Odontológica.

Repair of bone cavities in dog's mandible filled with inorganic bovine bone and bioactive glass associated with platelet rich plasma

The aim of this study was to evaluate the effect of platelet rich plasma (PRP) associated to bovine inorganic bone (Bio-Oss®; Geistlich) or bioactive glass (Bio-Gran®; Orthovita, Implant Innovations) on bone healing. Bone cavities were prepared in both sides of the mandible of 4 adult male dogs. The cavities were divided into 4 groups according to the filling material as follows: control, PRP, PRP/Bio-Oss, PRP/Bio-Gran. The animals were sacrificed after 120 days and histological and histomorphometrical analysis was performed. The control group showed 80.6% of bone formation in the longitudinal sections at 6 mm depth and 83.7% at 13 mm depth. The transverse sections displayed 74.2% at both 6 and 13 mm depths. The PRP group showed 21.1% of bone formation in the longitudinal sections at 6 mm depth, and 23.1% at 13 mm depth. The transverse sections presented 28.98% of bone formation at 6 mm depth and 41.2% at 13 mm depth. The PRP/Bio-Gran group showed 25.1% of bone formation in the longitudinal sections at 6 mm depth and 30.4% at 13 mm depth. In the transverse sections, the bone formation was 43.0% at 6 mm depth and 39.7% at 13 mm depth. The PRP/Bio-Oss group showed 35.5% of bone formation in the longitudinal sections at 6 mm depth and 42% at 13 mm depth. In the transversal sections, the bone formation was 26.8% and 31.2% at the depths of 6 and 13 mm, respectively. PRP alone or associated with bovine inorganic bone or bioglass had no significant effect in bone healing.

Oxidative stability of soybean oil added to Lentinus edodes and Agaricus blazei mushrooms extracts in an accelerated storage test

Purpose: The purpose of this paper is to evaluate the oxidative stability of soybean oil added by Lentinus edodes and Agaricus blezei extracts in accelerated storage test. Design/methodology/approach: The following treatments were subjected to accelerated storage test in an oven at 60°C for 15 days: Control (soybean oil without antioxidants), TBHQ (soybean oil + 100 mg/kg of TBHQ), BHT (soybean oil + 100 mg/kg of BHT), L. edodes (soybean oil + 3,500 mg/kg of L. edodes extract) and A. blazei (soybean oil + 3,500 mg/kg of A. blazei extract). The samples were taken every three days and analyzed for peroxide values and conjugated dienes. Findings: At the end of 15 days, the treatments TBHQ, A. blazei, L. edodes, Control and BHT showed 6.47, 8.81, 41.53, 71.28 and 78.40 meq/kg, respectively, for peroxide values and 0.37, 0.40, 0.67, 1.07 and 1.00 per cent, respectively, for conjugated dienes. Originality/value: The research indicates that mushrooms may be a promising source of natural antioxidants. Therefore, natural extracts of mushrooms can be applied to vegetable oils as a way to reduce the degradation caused by lipid oxidation. © Emerald Group Publishing Limited.

Evaluation of the toxicity of Virola sebifera crude extracts, fractions and isolated compounds on the nest of leaf-cutting ants

The phytochemical study of Virola sebifera leaves led to the isolation of three lignans: (+)-sesamin, (-)-hinokinin, and (-)-kusunokinin and three flavonoids: quercetin-3-O - L-rhamnoside, quercetin-3-O - D-glucoside, and quercetin-3-methoxy-7-O - D-glucoside by using techniques as high-speed counter-current chromatography and high-performance liquid chromatography. The crude extracts, fractions, and isolated compounds were evaluated for their insecticidal and fungicidal potential against Atta sexdens rubropilosa and its symbiotic fungus Leucoagaricus gongylophorus. The bioassay results showed a high insecticidal activity for the methanol crude extract of the leaves of V. sebifera and its n-hexane, dichloromethane and ethyl acetate fractions. The fungicidal bioassay revealed high toxicity of the lignans against L. gongylophorus. © 2012 Keylla Utherdyany Bicalho et al.

Antibacterial activity of gels with pomegranate, apricot and green tea glycolic extracts

Pomegranate (PGE) and green tea (GTGE) glycolic extracts are being employed in formulations because of their antiseptic and astringent effects. Apricot (AGE) glycolic extract possesses function cooling and antibacterial. The aim was to verify the antibacterial activity of these extracts incorporated in gel base. The antibacterial activity was verified by diffusion in agar method, using cylinder in plate. Plates containing Staphylococcus aureus (ATCC 6538p), Pseudomonas aeruginosa (ATCC 27853), Escherichia coli (ATCC 10536) and Salmonella sp. (ATCC 19196) were incubated at 37°C for 24 hours. After incubation, the results were analysed with a pachymeter, observing the bacterial growth inhibition halo diameter and the statistical significance level was determined. PGE presented activity only against P. aeruginosa; GTGE presented activity against S. aureus, P. aeruginosa and E. coli; and AGE presented activity against P. aeruginosa and Salmonella sp. According to the experimental conditions, it is possible to conclude that GTGE presented the greater growth inhibition halo diameter when compared with other extracts, suggesting higher antibacterial action of this extract.

Characterization of flavonoids and naphthopyranones in methanol extracts of Paepalanthus chiquitensis herzog by HPLC-ESI-IT-MSn and their mutagenic activity

A HPLC-ESI-IT-MSn method, based on high-performance liquid chromatography coupled to electrospray negative ionization multistage ion trap mass spectrometry, was developed for rapid identification of 24 flavonoid and naphthopyranone compounds. The methanol extracts of the capitulae and scapes of P. chiquitensis exhibited mutagenic activity in the Salmonella/microsome assay, against strain TA97a. © 2013 by the authors.

Inoculation of salivary gland extracts obtained from female of Rhipicephalus sanguineus (Latreille, 1806) (Acari, Ixodidae) with 2, 4, and 6 days of feeding in rabbit: I - Histopathology of the feeding lesion

This study analyzed the histopathology of rabbit skin, previously immunized with SGE2, SGE4, and SGE6 gland extracts prepared from salivary glands of Rhipicephalus sanguineus female with 2, 4, and 6 days of feeding, at the region of the R. sanguineus female feeding lesion 2, 4, and 6 days after tick attachment. In this work, infestation-naïve New Zealand White rabbits were inoculated either with the extracts (test group (TG)) or with phosphate buffer and complete Freund's adjuvant mixture (control group 2 (CG2)). Each extract-inoculated- (TG and CG2) and non-inoculated (CG1) rabbit was subsequently infested with R. sanguineus. Skin biopsies were collected from the rabbit at the tick feeding lesion at 2, 4, and 6 days of feeding. Results revealed that rabbit immunization with gland extracts induced acquisition of resistance against this species. It should be stated that the SGE4 extract was the most effective in developing an immune-inflammatory response against ectoparasites, being this process characterized by the presence of an early and intense inflammatory cell infiltrate. On the other hand, SGE6 extract caused a later appearance of resistance with less infiltrate occurrence and intense edema at the feeding lesion site. As to the inflammatory process deriving from SGE2 extract inoculation, it was the less intense. It was concluded that immunization with different extracts from R. sanguineus female salivary glands did not change microscope features of the inflammatory process, although an earlier or more intense and later response, which was also dependent on the inoculate extract, was noticed. © 2012 Springer-Verlag Berlin Heidelberg.

Evaluation of the bone healing process utilizing platelet-rich plasma activated by thrombin and calcium chloride: A histologic study in rabbit calvaria

To evaluate the bone healing of defects filled with particulate bone graft in combination with platelet-rich plasma (PRP), added with a mixture of calcium chloride and thrombin or just calcium chloride. Two 5-mm bone defects were created in the calvaria of 24 rabbits. Each defect was filled with particulate bone graft and PRP. In one defect the PRP was activated by a mixture of calcium chloride and thrombin; in the other, PRP was activated by calcium chloride only. The animals were euthanized 1, 2, 4, and 8 weeks after the surgeries, and the calvaria was submitted to histologic processing for histomorphometric analysis. The qualitative analysis has shown that both defects presented the same histologic characteristics so that a better organized, more mature, and well-vascularized bone tissue was noticed in the eighth week. A good bone repair was achieved using either the mixture of calcium chloride and thrombin or the calcium chloride alone as a restarting agent of the coagulation process.

In vitro antimicrobial activity of auxiliary chemical substances and natural extracts on Candida albicans and Enterococcus faecalis in root canals

Objective: The aim of this study was to evaluate the antimicrobial activity of auxiliary chemical substances and natural extracts on Candida albicans and Enterococcus faecali's inoculated in root canals. Material and Methods: Seventy-two human tooth roots were contaminated with C. albicans and E. faecalis for 21 days. The groups were divided according to the auxiliary chemical substance into: G1) 2.5% sodium hypochlorite (NaOCl), G2) 2% chlorhexidine gel (CHX), G3) castor oil, G4) glycolic Aloe vera extract, g5) glycolic ginger extract, and G6) sterile saline (control). The samples of the root canal were collected at different intervals: confirmation collection, at 21 days after contamination; 1st collection, after instrumentation; and 2nd collection, seven days after instrumentation. Microbiological samples were grown in culture medium and incubated at 37°C for 48 hours. Results: The results were submitted to the Kruskal-Wallis and Dunn (5%) statistical tests. NaOCl and CHX completely eliminated the microorganisms of the root canals. Castor oil and ginger significantly reduced the number of CFU of the tested bacteria. Reduction of CFU/mL at the 1st and 2nd collections for groups G1, G2, G3 and G4 was greater in comparison to groups G5 and G6. Conclusion: It was concluded that 2.5% sodium hypochlorite and 2% chlorhexidine gel were more effective in eliminating C. albicans and E. faecalis, followed by the castor oil and glycolic ginger extract. The Aloe vera extract showed no antimicrobial activity.

Microbiological control of moisturizing mask formulation added of hibiscus flowers, assai palm, black mulberry and papaw glycolic extracts

The microbiological control of moisturizing mask formulation added of hibiscus flowers, assai palm, black mulberry and papaw glycolic extracts, determining the number of viable microorganisms and possible presence of pathogenic. The moisturizing mask formulation was composed of zinc oxide (5. 0%) and moisturizing cream constituted of triceteareth-4 phosphate (and) cetyl alcohol (and) stearyl alcohol (and) sodium cetearyl sulfate (and) oleth-10 (qs 50g). To this formulation was added hibiscus flowers glycolic extract (2. 5%), assai palm glycolic extract (1. 5%), black mulberry glycolic extract (1. 5%) and papaw glycolic extract (2. 0%). The formulation was stored in aseptically clean recipients, away from humidity and light, in fresh and airy places. The results of the microbiological analysis on the counting of aerobic mesophilic microorganisms (bacteria and fungi), of the above mentioned formulation, revealed a bioburden < 10 CFU/mL in all samples. Such data indicate adequate microbiological quality of the tested products, according to official recommendations. Furthermore, it was not detected the presence of pathogenic microorganisms, assuring the harmlessness of the formulation. The results lead us to conclude that the formulation and raw materials analyzed did not present microbial contamination, evidenced for estimating the number of viable microorganisms (<10 UFC/g) and for researching pathogens.