Physicochemical Characteristics of Brazilian Green Propolis Evaluated During a Six-Year Period.

BACKGROUND: Propolis has been used as a natural health product mainly due to the presence of polyphenols, flavonoids, phenolic aldehydes, amino acids, vitamins and others bioactive constituents. To this natural substance are attributed different biological and pharmacological properties which are influenced by its chemical composition and organoleptic properties. The aim of this work was to evaluate the physicochemical properties and parameters of green propolis collected during a period of six years (2008-2013) in the state of Minas Gerais, located at the southeastern region of Brazil. METHODS: The methodology were in accordance with Brazilian legislation on the identity and quality standards of propolis. The evaluated parameters of hydroalcoholic from green propolis were total flavonoids, antioxidant activity - DPPH method, oxidation index, wax content, humidity and insoluble impurities. RESULTS: Propolis samples collected in different seasons during the years 2008 to 2013 presented mean values of total flavonoids (3.4 ± 0.11 mg/g), antioxidant activity DPPH (4.76 ± 0.16 µg/mL), oxidation index (3, 4 ± 0.33 seconds) and wax (15.14 ± 0.78% m/m), which are in accordance with Brazilian legislation. CONCLUSION: Green propolis did not show abrupt seasonal changes during the six years of investigation, and may be considered as an adequate functional ingredient.

Inactivation of NKX6.3 in the stomach leads to abnormal expression of CDX2 and SOX2 required for gastric-to-intestinal transdifferentiation.

Intestinal metaplasia in gastric mucosa is considered a preneoplastic lesion that progresses to gastric cancer. However, the molecular networks underlying this lesion formation are largely unknown. NKX6.3 is known to be an important regulator in gastric mucosal epithelial differentiation. In this study, we characterized the effects of NKX6.3 that may contribute to gastric intestinal metaplasia. NKX6.3 expression was significantly reduced in gastric mucosae with intestinal metaplasia. The mRNA expression levels of both NKX6.3 and CDX2 predicted the intestinal metaplasia risk, with an area under the receiver operating characteristic curve value of 0.9414 and 0.9971, respectively. Notably, the NKX6.3 expression level was positively and inversely correlated with SOX2 and CDX2, respectively. In stable AGS(NKX6.3) and MKN1(NKX6.3) cells, NKX6.3 regulated the expression of CDX2 and SOX2 by directly binding to the promoter regions of both genes. Nuclear NKX6.3 expression was detected only in gastric epithelial cells without intestinal metaplasia. Furthermore, NKX6.3-induced TWSG1 bound to BMP4 and inhibited BMP4-binding activity to BMPR-II. These data suggest that NKX6.3 might function as a master regulator of gastric differentiation by affecting SOX2 and CDX2 expression and the NKX6.3 inactivation may result in intestinal metaplasia in gastric epithelial cells.

Comparison of effects of the ethanolic extracts of brazilian propolis on human leukemic cells as assessed with the MTT assay.

Propolis is a resinous product collected by honey bees. It was also reported that propolis has a wide variety of biological actions, including antimicrobial activity and antioxidant, anti-inflammatory, and suppressive effects of dioxin toxicity activities. The aim of this study was to compare the in vitro cytotoxic activities of green propolis (G12) and red propolis (G13) in human leukemia cells. These cells were incubated with different concentrations of propolis and 48 hours after the IC(50) was calculated for each cell. The results showed that the red propolis has cytotoxic effect in vitro higher than green propolis. Red propolis was showed to be cytostatic in K562 cells and caused the same amount of apoptosis as its control Gleevec. In conclusion, these results showed that red propolis is more cytotoxic than the green propolis in a variety of human cell lines of leukemia. Red propolis may contain drugs capable of inhibiting cancer cell growth. Therefore, further isolation of respective chemical ingredients from the red propolis (G13) for identification of the activities is necessary.

Brazilian red propolis--chemical composition and botanical origin.

Propolis contains resinous substances collected by honey bees from various plant sources and has been used as a traditional folk medicine since ca 300 BC. Nowadays, the use of evidence-based complementary and alternative medicine (CAM) is increasing rapidly and so is the use of propolis in order to treat or support the treatment of various diseases. Much attention has been focused on propolis from Populus sp. (Salicaceae) and Baccharis dracunculifolia (Asteracea), but scientific information about the numerous other types of propolis is still sparse. We gathered six samples of red propolis in five states of Northeastern Brazil. The beehives were located near woody perennial shrubs along the sea and river shores. The bees were observed to collect red resinous exudates on Dalbergia ecastophyllum (L) Taub. (Leguminosae) to make propolis. The flavonoids of propolis and red resinous exudates were investigated using reversed-phase high-performance liquid chromatography and reversed-phase high-performance thin-layer chromatography. We conclude that the botanical origin of the reddish propolis is D. ecastophyllum. In areas where this source (D. ecastophyllum) was scarce or missing, bees were collecting resinous material from other plants. Propolis, which contained the chemical constituents from the main botanical origin, showed higher antimicrobial activity.

Antiproliferation of human prostate cancer cells by ethanolic extracts of Brazilian propolis and its botanical origin.

Propolis is a resinous substance collected by bees (Apis mellifera) from various tree buds which they then use to coat hive parts and to seal cracks and crevices in the hive. Propolis, a known ancient folk medicine, has been extensively used in diet to improve health and to prevent disease. In the present study, we have evaluated the effects of ethanolic extracts of Brazilian propolis group l2 and bud resins of botanical origin (B. dracunculifolia), and propolis group 3 on proliferation of metastasis (DU145 and PC-3) and primary malignant tumor (RC58T/h/SA#4)-derived human prostate cancer cells. The strongest inhibition was observed in propolis group 3 (sample #3) extracts whereas moderate growth inhibition was observed in human prostate epithelial cells. In the RC58T/h/SA#4 cells, resins of botanical origin of propolis group 12 (sample #1) and propolis group 12 (sample #2) induced growth inhibition that was associated with S phase arrest whereas propolis group 3 (sample #3) induced growth inhibition that was associated with G2 arrest. The mechanisms of cell cycle effects of propolis were investigated. The resins of botanical origin of propolis group 12 and propolis group 12 showed similar inhibition of cyclin D1, CDK4 and cyclin B1 expression. Propolis group 3 showed higher induction of p21 expression but no inhibition of cyclin D1, CDK4 and cyclin B1 expression. The results obtained here demonstrate that the Brazilian propolis extracts have significant inhibitory effect on proliferation of human prostate cancer cells. Inhibition was achieved through regulation of protein expression of cyclin D1, B1 and cyclin dependent kinase (CDK) as well as p21. Our results indicate that the Brazilian propolis extracts show promise as chemotherapeutic agents as well as preventive agents against prostate cancer.

Suppressive effects of ethanolic extracts from propolis and its main botanical origin on dioxin toxicity.

Suppressive effects of ethanolic extracts prepared from propolis group 12 and its main botanical origin (leaf bud of Baccharis dracunculifolia) on transformation of the aryl hydrocarbon receptor (AhR), the initial action of dioxin toxicity, were investigated. It was found that suppressive effects of propolis on AhR transformation were relatively higher than those of resins of its botanical origin in cell-free system and in Hepa-1c1c7 cells. When the composition of chemical ingredients was measured, propolis contained slightly higher amounts of flavonoid aglycones as compared with its botanical origin with the same characteristics. Moreover, antiradical activity, one of the typical biological activities of flavonoids, in propolis was also slightly higher than that in its botanical origin. These results indicate that not only propolis but also its botanical origin contains high amounts of flavonoid aglycones and that both of them are useful dietary sources for flavonoids with a potency to prevent dioxin toxicity.

Some chemical composition and biological activity of northern Argentine propolis.

Twenty-five samples of propolis were collected from seven different regions in northern Argentina; ethanolic extracts of propolis were prepared from all samples, and the respective samples were examined for UV absorption spectra, RPHPTLC, RPHPLC, antimicrobial activity, antiradical activity, and total phenolic content. It was found that 16 of the 25 samples showed a phenolic profile similar to that found in samples from southern Brazil and corresponding to poplar-based propolis and that the rest of the samples showed a different profile and higher antimicrobial and antiradical activities.

Suppression of dioxin mediated aryl hydrocarbon receptor transformation by ethanolic extracts of propolis.

Present study demonstrated that the ethanolic extracts of propolis containing higher concentrations of flavonoids suppressed 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-induced aryl hydrocarbon receptor transformation in a dose-dependent manner. The IC(50) values of propolis group 3 and group 12 were 1.2 and 3.6 microg/ml, respectively, indicating that propolis showed stronger antagonistic effects as compared with vegetable extracts.

Chemical constituents in Baccharis dracunculifolia as the main botanical origin of southeastern Brazilian propolis.

Previously, it was reported that one group of propolis (Group 12) was identified in southeastern Brazil, and the botanical origin of the propolis was Baccharis dracunculifolia resinous exudates. It was also observed that honeybee (Africanized Apis mellifera) mainly visited the leaf buds or unexpanded leaves of B. dracunculifolia but rarely expanded leaves. B. dracunculifolia is dioecious with male and female inflorescences, and RPHPLC of the ethanolic extracts of the respective male and female bud resinous exudates showed the same profiles. RPHPLC profiles of propolis G12 leaf buds and unexpanded and expanded leaves of B. dracunculifolia showed similarity, but unexpanded leaves quantitatively decreased in chemical constituents as compared with leaf buds. In the case of expanded leaves, all chemical constituents were severely decreased or disappeared. Artepillin C (3,5-diprenyl-4-hydroxycinnamic acid) was also identified in both propolis and resinous exudates, and both ethanolic extracts contained the highest concentrations of this compound as compared with the rest of the chemical constituents.

Effect of a mouthrinse containing selected propolis on 3-day dental plaque accumulation and polysaccharide formation.

The aim of this study was to evaluate the effect of a mouthrinse containing propolis SNB-RS on 3-day dental plaque accumulation. Six volunteers took part in a double-blind crossover study performed in two phases of 3 days. During each phase the volunteers refrained from all oral hygiene and rinsed with 20% sucrose solution 5 times a day to enhance dental plaque formation and with mouthrinse (placebo or experimental) twice a day. On the 4th day, the plaque index (PI) of the volunteers was scored and the supragingival dental plaque was analyzed for insoluble polysaccharide (IP). The PI (SD) for the experimental group was 0.78 (0.17), significantly less than for the placebo group, 1.41 (0.14). The experimental mouthrinse reduced the IP concentration in dental plaque by 61.7% compared to placebo (p < 0.05). An experimental mouthrinse containing propolis SNB-RS was thus efficient in reducing supragingival plaque formation and IP formation under conditions of high plaque accumulation.

Effect of a mouthrinse containing selected propolis on 3-day dental plaque accumulation and polysaccharide formation / Efeito de antisséptico bucal contendo própolis selecionado em 3 dias de acumulação de placa dentária e formação de Polysaccharide

The aim of this study was to evaluate the effect of a mouthrinse containing propolis SNB-RS on 3-day dental plaque accumulation. Six volunteers took part in a double-blind crossover study performed in two phases of 3 days. During each phase the volunteers refrained from all oral hygiene and rinsed with 20 percent sucrose solution 5 times a day to enhance dental plaque formation and with mouthrinse (placebo or experimental) twice a day. On the 4th day, the plaque index (PI) of the volunteers was scored and the supragingival dental plaque was analyzed for insoluble polysaccharide (IP). The PI (SD) for the experimental group was 0.78 (0.17), significantly less than for the placebo group, 1.41 (0.14). The experimental mouthrinse reduced the IP concentration in dental plaque by 61.7 percent compared to placebo (p < 0.05). An experimental mouthrinse containing propolis SNB-RS was thus efficient in reducing supragingival plaque formation and IP formation under conditions of high plaque accumulation

Effects of compounds found in propolis on Streptococcus mutans growth and on glucosyltransferase activity.

Propolis, a resinous bee product, has been shown to inhibit the growth of oral microorganisms and the activity of bacterium-derived glucosyltransferases (GTFs). Several compounds, mainly polyphenolics, have been identified in this natural product. The present study evaluated the effects of distinct chemical groups found in propolis on the activity of GTF enzymes in solution and on the surface of saliva-coated hydroxyapatite (sHA) beads. Thirty compounds, including flavonoids, cinnamic acid derivatives, and terpenoids, were tested for the ability to inhibit GTFs B, C, and D from Streptococcus mutans and GTF from S. sanguinis (GTF Ss). Flavones and flavonols were potent inhibitors of GTF activity in solution; lesser effects were noted on insolubilized enzymes. Apigenin, a 4',5,7-trihydroxyflavone, was the most effective inhibitor of GTFs, both in solution (90.5 to 95% inhibition at a concentration of 135 microg/ml) and on the surface of sHA beads (30 to 60% at 135 microg/ml). Antibacterial activity was determined by using MICs, minimum bactericidal concentrations (MBCs), and time-kill studies. Flavanones and some dihydroflavonols, as well as the sesquiterpene tt-farnesol, inhibited the growth of S. mutans and S. sobrinus; tt-farnesol was the most effective antibacterial compound (MICs of 14 to 28 microg/ml and MBCs of 56 to 112 microg/ml). tt-Farnesol (56 to 112 microg/ml) produced a 3-log-fold reduction in the bacterial population after 4 h of incubation. Cinnamic acid derivatives had negligible biological activities. Several of the compounds identified in propolis inhibit GTF activities and bacterial growth. Apigenin is a novel and potent inhibitor of GTF activity, and tt-farnesol was found to be an effective antibacterial agent.

Botanical origin and chemical composition of Brazilian propolis.

Brazilian propolis has been classified into 12 groups based on physicochemical characteristics: five in the southern Brazil group (group 3), one in the southeastern Brazil group (group 12), and six in the northeastern Brazil group (group 6). The plant origins of these groups were investigated using reversed-phase high-performance thin-layer chromatography (RPHPTLC), reversed-phase high-performance liquid chromatography (RPHPLC), and gas chromatography-mass spectrometry (GC-MS). It was concluded that the origins of propolis group 3, group 6, and group 12 are resins of the poplar tree, Hyptis divaricata, and Baccharis dracunculifolia, respectively.

Effects of Apis mellifera propolis on the activities of Streptococcal glucosyltransferases in solution and adsorbed onto saliva-coated hydroxyapatite / Efeitos do própolis Apis mellifera nas atividades do Streptococcal Glucosyltransferases em solução e absorvido sobre hidroxyapatita coberta com saliva

Propolis, a resinous hive product collected by Apis mellifera bees, has been used for thousands of years in folk medicine. Ethanolic extracts of propolis (EEP) have been shown to inhibit the activity of a mixture of crude glucosyltransferase (Gtf) enzymes in solution. These enzymes synthesize glucans from sucrose, which are important for the formation of pathogenic dental plaque. In the present study, the effects of propolis from two different regions of Brazil on the activity of separate, purified Gtf enzymes in solution ans on the surface of saliva-coated hydroxyapatite (sHA) beads were evaluated. The EEP from Minas Gerais (MG; Southeastern Brazil) and Rio Grande do Sul (RS; Southern Brazil) were tested for their ability to inhibit the enzymes GtfB (synthesis of insoluble glucan), GtfC (insoluble/soluble glucan) and GtfD (soluble glucan). The effects of propolis on Gtf from Streptococcus sanguis (soluble glucan synthesis) was also explored...

Comparison of the flavonoid aglycone contents of Apis mellifera propolis from various regions of Brazil

Forty-six specimens of propolis which were collected by Apis mellifera were obtained from various regions of Brazil and extracted with 80 per cent aqueous ethanol(ethanolic extract of propolis, EEP). The extracts were analysed by using determination of total flavonoid concentrations using the aluminum nitrate method, UV-spectrophotometry, reversed phase-high performance thin layer chromatography (HPTLC), and reversed phase-HPLC. Six samples of EEP, which contained high total flavonoid concentrations, were selected for futher investigation. HPTLC indicated that the quality of flavonoid aglycones of EEP from the States of Minas Gerais and Säo Paulo (Southeastern Brazil), Goias and Mato Grosso do Sul (closely to Southeastern) were similar, but were different compared to those of EEP's from Paraná and Rio Grande do Sul, which are located in Southern Brazil. Reversed Phase-HPLC analysis of EEP's from Southern Brazil identified 7 flavonoid aglycones, whereas there were 9 from Säo Paulo and Minas Gerais and 7 from Goiás and Mato Grosso do Sul. These results indicate that the plant ecology of Southeastern and Southern Brazil are different. It was also observed that all samples of EEP's demonstrated inhibition of growth of Staphylococcus aureus coagulase positive, whereas Escherichia coli was not inhibited

Purificaçäo e caracterizaçäo de amiloglicosidase extracelular de Candida sp. e sua utilizaçäo na produçäo de xarope de glicomaltose / Purification and characterization of extracelullar amyloglusidase from Candida sp. and its use for the production of glucomaltose syrup

A enzima amilolítica de Candida sp. ATCC 90238 foi purificada e suas características enzimáticas foram estudadas. O peso molecular da enzima purificada foi estimado em 120.000. A cromatografia em papel do hidrolizado de amido solúvel indicou que a enzima é uma amiloglicosidase. Maltotriose e amido solúvel foram eficientemente hidrolisados a glicose pela enzima, enquanto que a maltose foi menos eficiente como substrato. A enzima apresenta características diferentes quando comparada à amiloglicosidase de Aspergillus e Rhizopus sp. que hidrolisam completamente a maltose. Esta enzima é adequada para a produçäo de xarope de glicomaltose.

Purification and characterization of thermostable xylanases from thermophilic Humicola sp. and their application in pulp improvement

A linhagem termófila Humicola sp., que foi isolada de madeira em decomposiçäo, produz xilanases extracelulares termoestáveis a 50oC. As xilanases foram purificadas e foram encontradas 3 fraçöes de proteínas com atividade de xilanase. As características das 3 xilanases foram estudadas. Verificou-se que a xilanase I é uma endoxilanase; a xilanase II é uma xilosidase (exoxilanase) enquanto que a xilanase III é uma endoxilanase que também apresenta atividade de arabinosidase e CMCase. O tratamento da polpa "Kraft" branqueada obtida de eucalipto com enzima purificadas e bruta aumentou o brilho da polpa quando comparada com a polpa sem este tratamento. Xilanase I e xilanase bruta aumentaram a viscosidade da polpa enquanto xilanase II e III diminuíram a viscosiddade devido à presença de atividade CMCase

Características de uma nova linhagem de levedura "Killer" / Study of some characteristics of newly isolated killer yeas

Foi isolada uma nova linhagem de Saccharomyces cerevisuiae 337, de usina de cana de açucar, que apresentou ativudade "Killer" semelhante ao fenótipo K.A eletroforese em gel de agarose mostrou que a linhagem S.cerevisiae 337 possui dois ds-RNAs. A atividade "killer" da linhagem foi comparada com a levedura padräo"Killer" K

Produçäo e caracterizaçäo de celulases extracelular de uma linhagem termofílica do fungo Aspergillus sp / production and characterization of extracellular cellulases from a thermostable aspergillus sp

Uma linhagem termofílica do fungo Aspergillus sp., No.10-2, foi isolada de compostos.a faixa de temperatura para crescimento variou de 20 a 50ºC sendo que a temperatura ótima ficou entre 35 a 45ºC.As produçöes máximas de endo-1, 4-ß-D-glucanase EC.3.2.1.4 (CMCase), exo-1, 4-ß-D-cellobiohydrolase EC. 3.2.1.91 (Avicelase) e ß-glucosidase EC.3.2.1.21 (Celobiase)foram obtidas quando o fungo foi cultivado em meio líquido contendo papel de celulose em pó.A maior produçäo de CMCase foi obtida a 37ºC enquanto que uma extraordinária produçäo de ß-glucosidase ocorreu a 45ºC.A temperatura ótima de sacarificaçäo de celulose pela enzima foi 55ºC.Ensaios de cromatografia de papel de hidrolisados da celulose demonstraram predominantemente glicose quando comparado a hidrolisados de celulose obtidos por enzima de Trichoderma reesei QM 9414.Isto ocorreu porque a linhagem de Aspergillus sp., No.10-2, produziu maiores a enzima ß-glucosidase