The Standardized Wood Extract of Albizia myriophylla: Its Potential as an Active Ingredient in an Anti-inflammatory Herbal Gel Formulation.

Albizia myriophylla has been used in Thai folk medicine for treating inflammation-related diseases. The wood of this medicinal plant is traditionally used as a single herbal drug in the form of an aqueous decoction and as a component in several Thai herbal formulations for the remedy of fever, sore throat, and aphthous ulcers. This study aimed to evaluate in vivo the anti-inflammatory potential and possible mechanism of action of the standardized wood extract of A. myriophylla as well as to investigate the anti-inflammatory activity and physicochemical properties of the developed herbal gel formulation containing standardized wood extract of A. myriophylla. Results of quantitative HPLC analysis demonstrated that the standardized wood extract of A. myriophylla contained 22.95 mg/g of 8-methoxy-7,3',4'-trihydroxyflavone, a bioactive marker compound of A. myriophylla. The standardized wood extract of A. myriophylla (1% w/v) exhibited remarkable inhibition (54.4 - 80.3%) in the croton oil model of topical inflammation at all assessment times, comparable to standard indomethacin (55.3 - 63.6%). Real-time quantitative reverse transcription-polymerase chain reaction was performed to clarify the anti-inflammatory mechanism of standardized wood extract of A. myriophylla, and the result showed that this standardized extract decreased lipopolysaccharide-induced nitric oxide synthase mRNA levels in a dose-dependent manner. The developed herbal gel containing standardized wood extract of A. myriophylla (1% w/w) had good physicochemical characteristics and exhibited potent inhibition (51.4 - 77.8%) of inflammation in a rat ear edema model at all assessment times, comparable to indomethacin gel (33.3 - 40.5%). The notable anti-inflammatory activity of standardized wood extract of A. myriophylla and its developed herbal gel formulation indicates their potential application as natural anti-inflammatory agents.

α-Glucosidase inhibitory, antibacterial, and antioxidant activities of natural substances from the wood of Derris reticulata Craib.

Derris reticulata (Leguminosae-Papilionoideae) has been used for the treatment of diabetes in Thai folk remedies. The phytochemical investigation of the wood of D. reticulata revealed the isolation of two new pyranoflavanones, 4'-methoxydereticulatin (1) and 2'''-hydroxy,3'''-ethoxylupinifolin (2), along with five known compounds namely lupinifolin (3), 2''',3'''-dihydroxylupinifolin (4), genistein (5), lupeol (6), and ß-sitosterol (7). Compounds 1-4 were selected for antibacterial assay using broth microdilution method, and displayed good activity against four out of five tested pathogenic bacterial strains, with MIC values ranging from 0.78 to 128 µg/mL. The result from spectrophotometric assay of α-glucosidase inhibition showed that 5 exhibited promising α-glucosidase inhibitory activity, compared with the positive control acarbose. Additionally, it was found that compounds 4 and 5 showed moderate DPPH and NO radicals scavenging activity. Modeling studies were also performed to suggest the interaction modes of compounds 3-5 in the α-glucosidase enzyme active site.

Thai herbal formulation 'Ya-Pit-Samut-Noi': Its antibacterial activities, effects on bacterial virulence factors and in vivo acute toxicity.

ETHNOPHARMACOLOGICAL RELEVANCE: A Thai herbal formulation 'Ya-Pit-Samut-Noi' containing Nigella sativa (seed), Piper retrofractum (fruit), Punica granatum (pericarp), and Quercus infectoria (nutgall) has long been traditionally used to treat diarrhea or bloody mucous diarrhea. Scientific information is very important to support its therapeutic effects and traditional drug development. AIM OF THE STUDY: This study aimed to evaluate the antibacterial activities of Ya-Pit-Samut-Noi against diarrhea-causing bacteria and determine its effects on bacterial virulence factors and in vivo acute toxicity. MATERIALS AND METHODS: Ethanol and water extracts of Ya-Pit-Samut-Noi and its plant components were prepared. The agar diffusion method was used for preliminary screening of antibacterial activity of the extracts against diarrhea-causing bacteria including Staphylococcus aureus, Vibrio cholerae, and Vibrio parahaemolyticus. The minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) were assessed using broth microdilution method. The effects on bactericidal activity, bacterial cell wall, and cell membrane were examined by time-kill, lysis, and leakage assays, respectively. The effects on bacterial virulence factors including quorum-sensing system, biofilm production, and swarming motility were determined. Phytochemical screening was carried out to identify the group of chemical compounds present in the formulation extracts. Acute toxicity study was conducted by a single oral dose of 2000 mg/kg body weight in Wistar albino rats. RESULTS: Ethanol and water extracts of Ya-Pit-Samut-Noi and Quercus infectoria demonstrated antibacterial efficacy against all bacterial strains as revealed by zones of inhibition ranging from 7.0 to 24.5 mm. The ethanol and water extracts of Ya-Pit-Samut-Noi and Quercus infectoria produced strong bacteriostatic activity against V. parahaemolyticus (n = 11) with an MIC range of 7.81-250 µg/ml. Only the ethanol extract of Ya-Pit-Samut-Noi produced MBC values less than or equal to 1000 µg/ml against all V. parahaemolyticus. Based on time-kill study, no surviving V. parahaemolyticus (ATCC 17802 and 5268) cells were detected within 6-12 h after treatment with the ethanol extract of Ya-Pit-Samut-Noi at MBC-4MBC concentrations. Vibrioparahaemolyticus ATCC 17802 cells treated with the ethanol extract of Ya-Pit-Samut-Noi demonstrated no lysis or leakage through the bacterial membrane was not observed. At low concentrations (0.125-0.25 µg/ml) the ethanol extract of Ya-Pit-Samut-Noi inhibited violacein production by Chromobacterium violaceum DMST 21761 without affecting the bacterial growth. The ethanol (31.25-62.5 µg/ml) and water (31.25-250 µg/ml) extracts of Ya-Pit-Samut-Noi inhibited biofilm production by S. aureus. The ethanol and water extracts of Ya-Pit-Samut-Noi at 1000 µg/ml reduced the swarming motility of Escherichia coli O157: H7 by 74.98% and 52.65%, respectively. Tannins and terpenoids were detected in both the ethanol and water extracts. Flavonoids were present only in the ethanol extract. Alkaloids and antraquinones were not noticed in either extract. In the acute toxicity study, there were no significant changes in hematological and biochemical parameters nor were adverse effects on mortality, general behaviors, body weight, or organ weights detected. CONCLUSIONS: The scientific evidence from this study supported the therapeutic effects and safety of the traditional Thai herbal formulation 'Ya-Pit-Samut-Noi' which has been used as an alternative treatment for gastrointestinal infections in Thailand.

Mechanism of Action and Biofilm Inhibitory Activity of Lupinifolin Against Multidrug-Resistant Enterococcal Clinical Isolates.

The treatment of enterococcal infections is becoming more difficult because of multidrug resistance (MDR). Lupinifolin, a prenylated flavonoid isolated from Albizia myriophylla Benth., showed a potent antimicrobial activity against enterococci. The aim of this study was to investigate antibacterial activity and action of lupinifolin against MDR enterococcal clinical isolates. Antibacterial properties of lupinifolin against 21 MDR isolates were assessed using broth microdilution method and time-kill assay. To study mode of action of lupinifolin on the isolates, propidium iodide intensity, salt tolerance assay, and electron microscopic analyses were performed. Antibiofilm formation activity of lupinifolin was conducted using crytal violet assay. Minimal inhibitory concentration (MIC) and minimal bactericidal concentration values of lupinifolin against the isolates ranged between 0.5 and 2.0 µg/mL and between 2 and 16 µg/mL, respectively. Lupinifolin at 2MIC and 4MIC inhibited the bacterial growth >2 log colony-forming units (CFU)/mL at 2 hr incubation by time-kill analysis. The compound increased membrane permeability and caused loss of salt tolerance. SEM and TEM micrographs revealed pronounced morphological and ultrastructural changes in the treated bacteria. Crystal violet staining showed the antibiofilm-producing activity of lupinifolin against four MDR enterococci. This study suggested that lupinifolin is an essential antimicrobial agent that could be useful for the treatment of MDR enterococcal infections.

Bioactive components, antioxidant, and anti-inflammatory activities of the wood of Albizia myriophylla

Abstract The scientific basis corresponding with the folkloric use of Albizia myriophylla Benth., Fabaceae, for the treatment of inflammation-related diseases was established by measuring antioxidant potential using 2,2-diphenyl-1-picrylhydrazyl, 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) free radicals, and ferric reducing antioxidant power assays as well as anti-inflammatory effect using nitrite assay and ethyl phenylpropiolate (EPP)-induced rat ear edema model. Both ethanol extract (DPPH, IC50 46.23 µg/ml; ABTS, IC50 57.14 µg/ml; FRAP, 950.14 mM Fe (II)/g) and dichloromethane fraction (DPPH, IC50 29.54 µg/ml; ABTS, IC50 40.36 µg/ml; FRAP, 946.69 mM Fe (II)/g) from A. myriophylla demonstrated a promising antioxidant activity. Furthermore, it was found that the ethanol extract of A. myriophylla showed significant inhibitory activity against lipopolysaccharide (LPS)-induced nitric oxide production in murine macrophage cells (IC50 13.8 µg/ml). The ethanol extract (15% w/v) also exhibited the maximum percentage inhibition (81-95%) of inflammation in the ear edema model at all assessment times comparable to indomethacin (0.5 mg/ear). Among all isolates 1-5 from the active extract of A. myriophylla, indenoic acid (1) (DPPH, IC50 8.96 µg/ml; ABTS, IC50 10.12 µg/ml) and 8-methoxy-7,3′,4′-trihydroxyflavone (2) (DPPH, IC50 5.05 µg/ml; ABTS, IC50 7.89 µg/ml) had potent free radical scavenging effects comparable to those of ascorbic acid (DPPH, IC50 2.12 µg/ml; ABTS, IC50 3.26 µg/ml). Compound 2 also displayed remarkable reducing power in FRAP test (261.81 mg QE/g) and showed a marked inhibition of the cellular nitric oxide production (IC50 27.7 µg/ml). Our results suggest that the anti-inflammatory mechanism of A. myriophylla is most probably based on its capacity to suppress nitric oxide production as well as to be free radical scavenger.

Alpha-glucosidase inhibitory activity of ethanol extract, fractions and purified compounds from the wood of Albizia myriophylla.

Albizia myriophylla Benth. is a medicinal herb which is used as a traditional remedy for various ailments including diabetes in Thailand. In our continued investigation of the biological activity of A. myriophylla, the ethanol extract, fractions and the isolated compounds from the wood of this plant were evaluated for in vitro α-glucosidase inhibition using spectrophotometric method. The plant ethanol extract and its different fractions possessed α-glucosidase inhibitory activity in a concentration-dependent manner. Dichloromethane fraction of the wood ethanol extract exhibited the highest percent inhibition against α-glucosidase (69.30%) among all fractions. Subsequent α-glucosidase inhibition assay proved that indenoic acid (1), 8-methoxy-7, 3',4'-trihydroxyflavone (2) and 3,4,7,3'-tetrahydroxyflavan (3) were partially rational for antidiabetic effect of this plant species. Among these compounds, 3 (IC50 98.59 µg/mL) exhibited potent inhibition of α-glucosidase, compared with a positive control acarbose (IC50 125 µg/mL). The inhibitory effect towards α-glucosidase of compounds 1-3 was reported herein for the first time.

Lupinifolin from Albizia myriophylla wood: A study on its antibacterial mechanisms against cariogenic Streptococcus mutans.

OBJECTIVE: To determine the anti-Streptococcus mutans mechanisms of action of lupinifolin from Albizia myriophylla Benth. (Fabaceae) wood and provide scientific evidence to support the traditional use of the plant against dental caries. METHODS: The minimum inhibitory concentration (MIC) was evaluated using the broth micro-dilution method. The effects of lupinifolin on bactericidal activity, bacterial cell walls, and membranes were investigated by time-kill, lysis, and leakage assays, respectively. Electron microscopy was utilized to observe any cell morphological changes caused by the compound. Localization of lupinifolin in S. mutans was detected using the thin layer chromatography technique. RESULTS: The MIC range of lupinifolin against S. mutans (n=6) was 2-4 µg/ml. This compound displayed bactericidal effects on S. mutans ATCC 25175 by 90-99.9% killing at 4MIC-16MIC after 8-24 hours. Lupinifolin-treated cells demonstrated no lysis. However, significant cytoplasmic leakage through the bacterial membrane was observed after treatment with lupinifolin at 4MIC-16MIC. As revealed by ultrastructural analysis, lupinifolin produced some changes in bacterial cell walls and membranes. Moreover, the compound was observed in the cytoplasmic fraction of the lupinifolin-treated cells. These results suggest that lupinifolin can enter the cell of bacteria but does not accumulate in the cell envelope and subsequently disrupts the integrity of the cytoplasmic membrane, leading to cell death. CONCLUSION: The scientific evidence from this study offers valuable insights into the potential role of lupinifolin in pharmaceutical and antibiotic applications and supports the therapeutic effects of A. myriophylla, which has traditionally been used as an alternative treatment for dental caries.

Chemical constituents and biological activities of Albizia myriophylla wood.

CONTEXT: Albizia myriophylla Benth (Leguminosae) is a medicinal plant widely used in Thailand and other Asian countries as a folk medicine remedy for many ailments. OBJECTIVE: The objective of this study is to investigate the chemical compositions, antibacterial activity, and cytotoxicity of A. myriophylla wood. MATERIALS AND METHODS: The structure identification of the isolated compounds was established using spectroscopic methods. In vitro antibacterial activity against Streptococcus mutans, Staphylococcus aureus, and Bacillus cereus and the cytotoxicity against KB cells of extracts and compounds from A. myriophylla were performed using broth microdilution and resazurin microplate assays, respectively. The lupinifolin content in A. myriophylla extracts was quantified by high-performance liquid chromatography. RESULTS: A rare flavan-3,4-diol (1) together with eight known compounds (2-9) were isolated from the wood of A. myriophylla. Compounds 4-9 exhibited anti-S. mutans activity, of which lupinifolin (5) was the most potent with an MIC value of 0.98 µg/mL, followed by its dihydroxy derivative 4 with an MIC value of 62.5 µg/mL. Compounds 4 and 5 also displayed marked antibacterial activity against B. cereus and S. aureus (MIC value 15.63-125 µg/mL) and showed strong cytotoxic activity against KB cells (IC50 value 4.95-12.55 µg/mL). The lupinifolin contents in ethanol extracts from two different collections of this plant originating from central and southern Thailand were 93.85 and 0.04 mg/g, respectively. CONCLUSION: This is the first report of compounds 1-4 from A. myriophylla. Compounds 4 and 5 showed potent antibacterial and cytotoxic activities compared with other isolates. The anti-S. mutans activity of A. myriophylla extracts seems to be related to the lupinifolin content.

Chemical constituents and antimicrobial properties of the essential oil and ethanol extract from the stem of Aglaia odorata Lour.

The stem-derived essential oil of Aglaia odorata Lour. was obtained by hydrodistillation using a Clevenger-type system. Gas chromatography-mass spectrometry analysis of the oil revealed the identification of 39 compounds, representing 76.4% of the oil; germacrene D (20.3%), α-humulene (17.1%), α-himachalene (12.7%) and ß-caryophyllene (10.2%) were the major components. Ar-turmerone (1) and eichlerialactone (2) were isolated from the stem oil and ethanolic stem extract of this plant species, respectively. Antimicrobial activities of the oil and ethanol extract were tested against both Gram-positive and Gram-negative bacterial strains including Bacillus cereus ATCC 11778, Staphylococcus aureus ATCC 25923, Acinetobacter baumannii ATCC 19606 and Escherichia coli ATCC 25922, as well as three rice fungal pathogens Bipolaris oryzae, Pyricularia oryzae and Rhizoctonia solani using broth microdilution method. The oil and 1 exhibited significant antifungal activity against the three rice pathogens tested, whereas 2 exhibited good antibacterial activity against both the Gram-positive pathogens tested.

Antibacterial substances from Albizia myriophylla wood against cariogenic Streptococcus mutans.

Albizia myriophylla has been used for long by Thai traditional healers as an important ingredient herb in Thai herbal formulas for caries. In this study, three flavonoids lupinifolin (6), 8-methoxy-7,3',4'-trihydroxyflavone (7), and 7,8,3',4'-tetrahydroxyflavone (8), a triterpenoid lupeol (3) as well as four sterols ß-sitosterone (1), stigmasta-5,22-dien-3-one (2), ß-sitosterol (4), and stigmasterol (5) were isolated from A. myriophylla wood. The antibacterial activity of these compounds against Streptococcus mutans ATCC 25175 was performed using broth microdilution method. All compounds exhibited antibacterial activity against S. mutans with minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) ranging from 1-256 and 2-256 µg/ml, respectively. Among the isolated compounds, lupinifolin (6) was found to be the most potent with MIC and MBC of 1 and 2 µg/ml, respectively. Lupinifolin (6) also showed a strong activity against ten clinical isolates of S. mutans with MIC and MBC ranging from 0.25-2 and 0.5-8 µg/ml, respectively. These results reported the bioactive ingredients of A. myriophylla which support its ethnomedical claims as well. Lupinifolin (6) may have a potential to be a natural anticariogenic agent.

Anti-Streptococcus mutans efficacy of Thai herbal formula used as a remedy for dental caries.

CONTEXT: Traditional knowledge of herbal remedies plays an important role in the search for more effective alternative treatment of a variety of disorders. The ethnobotanical surveys in southern Thailand have revealed that 35 Thai herbal formulas have been used by Thai traditional healers against dental caries. However, the scientific evaluation to confirm their rational uses is scarce. OBJECTIVE: To test in vitro anti-Streptococcus mutans activity of Thai herbal formulas used against dental caries (THF-DC). MATERIALS AND METHODS: Ethanol extracts of Thai herbal formulas were evaluated for antibacterial activity against S. mutans. Agar disc diffusion was employed as a preliminary screening assay, followed by broth microdilution assay to assess minimum inhibitory concentration (MIC). Furthermore, medicinal plants contained in the most active THF-DC were investigated for their phytochemicals. RESULTS: Eleven THF-DC extracts exhibited clear inhibition zones of 7.0-22.5 mm against S. mutans. Subsequent determination of their MIC revealed that the formula containing Albizia myriophylla Benth. (Leguminosae), Alpinia galanga (L.) Willd. (Zingiberaceae), Avicennia marina (Forssk.) Vierh. (Acanthaceae), and Ocimum sanctum L. (Lamiaceae) was the most active, with MIC at 250 µg/mL. Among these medicinal plants, A. myriophylla gave the strongest activity with MIC at 3.9 µg/mL, followed by A. marina with MIC at 62.5 µg/mL. Various classes of bioactive phytochemicals including tannins, flavonoids, alkaloids, and terpenoids were found in these extracts. CONCLUSION: Anti-S. mutans activity of THF-DC extracts was established. Further investigations may be required for the isolation and chemical characterization of the active ingredients in A. myriophylla.

Absolute configuration of isoeichlerialactone.

The title seco-dammarane triterpenoid, C(27)H(42)O(4) (systematic name: 3-{(3S,3aR,5aR,6S,7S,9aR,9bR)-6,9a,9b-trimethyl-3-[(R)-2-methyl-5-oxotetra-hydro-furan-2-yl]-7-(prop-1-en-2-yl)dodeca-hydro-1H-cyclo-penta-[a]naphthalen-6-yl}propanoic acid), has been isolated for the first time from the seeds of Aglaia forbesii. The mol-ecule has three fused rings and all rings are in trans-fused. The two cyclo-hexane rings are in standard chair conformations and the cyclo-pentane ring adopts an envelope conformation. Its absolute configuration was determined by the refinement of the Flack parameter to 0.26 (17). In the crystal, mol-ecules are linked into chains along [010] by O-H⋯O hydrogen bonds.

Absolute configuration of methyl isoeichlerialactone.

The title compound, C(28)H(44)O(4)·0.56H(2)O, is a co-crystal of methyl isoeichlerialactone monohydrate as the major component and methyl isoeichlerialactone as the minor component in a 0.55778 (3):0.44222 (3) ratio. The conformations of both components are identical except for that of the -COOCH(3) group of the methyl propanoate side chain on the cyclo-hexane ring which is positionally disordered over two orientations. The mol-ecule of methyl isoeichlerialactone has three fused rings and all rings are trans-fused. The two cyclo-hexane rings are in standard chair conformations and the cyclo-pentane ring adopts an envelope conformation. In the crystal, weak C-H⋯O inter-actions link methyl isoeichlerialactone mol-ecules into screw chains along [010]. The crystal structure is further stabilized by O-H⋯O hydrogen bonds and weak C-H⋯O inter-actions.

The mode of antistaphylococcal action of Eleutherine americana.

The anti-methicillin-resistant Staphylococcus aureus (anti-MRSA) activity and the possible mechanism of action of a crude extract from red bulbs of Eleutherine americana Merr. were investigated. The crude ethanolic extract from E. americana produced minimum inhibitory concentrations (MICs) of 62.5-1000 and 250 microg mL(-1) against MRSA isolates and the reference strains, respectively. Treatment of S. aureus ATCC 27664 with a crude extract at 2MIC reduced the inoculum size by 5 log at 24 h compared with the control. The combined effect of the extract and 7.5% NaCl on the enterotoxin-producing ATCC strain resulted in no detection of organisms within 24 h compared with the control. The release of cell materials after extract treatment was determined by measuring OD(260 nm), the treatment resulted in cytoplasmic leakage. Determination of OD(620 nm) showed that the extract did not cause gross cell wall damage. However, observation of S. aureus cells under an electron microscope after treatment with 2MIC and 4MIC of the crude extract revealed that the extract caused damage to membrane morphology. A knowledge of the mechanism of action of the E. americana extract may offer useful hints in the search for novel antibacterial substance.

Flavaglines and triterpenoids from the leaves of Aglaia forbesii.

Three structurally complex flavaglines of the cyclopenta[bc]benzopyran type, named desacetylpyramidaglains A, C, D (1-3), and the triterpene 23, 24, 25-trihydroxycycloartan-3-one (4) were isolated from the leaves of Aglaia forbesii together with the two rare pregnane steroids 2beta,3beta-dihydroxy-5alpha-pregn-17(Z)-en-16-one and 2beta,3beta-dihydroxy-5alpha-pregn-17(E)-en-16-one, as well as the bisamide pyramidatine, the sesquiterpene spathulenol, and the widespread triterpenoids lupeol, lupenone, and a mixture of beta-sitosterol and stigmasterol. Their structures were elucidated by 1D and 2D NMR spectroscopy and mass spectrometry. Compounds 3, 4, 5, and 6 were tested for antituberculosis and antiviral activity.