Effectiveness of Ya-Samarn-Phlae in diabetic wound healing: Evidence from in vitro studies and a multicenter randomized controlled clinical trial.

ETHNOPHARMACOLOGICAL RELEVANCE: Ya-Samarn-Phlae (YaSP) has traditionally been widely used in southern Thailand for treating chronic and infected wounds, including diabetic foot ulcers. However, there are only a limited number of clinical studies supporting the use of this polyherbal formulation. Therefore, the present work aims to provide clinical evidence to support the application of YaSP, prepared according to a standardized traditional procedure (T-YaSP). Additionally, its potential chemical markers and wound healing-related biological activities were examined. MATERIALS AND METHODS: The in vitro wound healing-related biological activities of YaSP ethanol extract and T-YaSP, including antibacterial activity against Staphylococcus epidermidis, inhibition and eradication of staphylococcal biofilm, anti-inflammatory effects, and enhancement of human dermal fibroblast migration in scratch wounds, were examined using well-established protocols. The chemical profiles of the ethanol extract of YaSP and T-YaSP were compared, and with promising chemical markers, arecoline, alpha-mangostin, and curcumin were selected and quantified using the HPLC method. A prospective, multicenter, randomized, controlled, parallel-group study was conducted over 12 weeks to evaluate the efficacy of the YaSP solution as an adjunct therapy, combined with standard wound care, for diabetic ulcers compared to standard treatment. RESULTS: The YaSP extract reduces NO production and can scavenge NO radicals in LPS-induced RAW 264.7 macrophage cells. Additionally, in a scratch assay, this extract and one of its herbal components, Curcuma longa, enhance the migration of human dermal fibroblasts. T-YaSP, containing 2.412 ± 0.002 mg/g of arecoline, 2.399 ± 0.005 mg/g of curcumin, and 0.017 ± 0.000 mg/g of α-mangostin, has shown the ability to inhibit the development and eradicate the mature biofilm of S. epidermidis. The use of T-YaSP as an adjunct therapy led to a significantly higher proportion of patients achieving healing within six weeks compared to the standard treatment group (36%/9 patients vs. 4%/1 patient; p = 0.013). After 12 weeks, 19 out of 25 patients in the T-YaSP group experienced complete healing, whereas only four patients in the standard treatment group achieved complete wound healing (76% in the T-YaSP group vs. 16% in the control group; p < 0.001). CONCLUSION: The results presented here represent the first randomized controlled trial to demonstrate the effectiveness of the traditional polyherbal solution, T-YaSP, which exhibits a wide range of wound healing-related activities. Utilizing T-YaSP as an adjunctive treatment resulted in a significant improvement in the number of type 2 diabetic patients achieving complete healing. However, to explore and utilize YaSP further, conducting a double-blind, randomized controlled trial with a larger population is necessary.

Clinical Evaluation of a Novel Tablet Formulation of Traditional Thai Polyherbal Medicine Named Nawametho in Comparison with Its Decoction in the Treatment of Hyperlipidemia.

In the traditional medical system in Thailand, medicinal plants and polyherbal medicines have been prescribed as lipid-lowering agents, including Nawametho decoction. This polyherbal formulation is described in the Worayokasan scripture. It consists of nine medicinal plants (Aegle marmelos (L.), Carthamus tinctorius L., Hibiscus sabdariffa Linn., Phyllanthus emblica L., Piper longum L., Piper nigrum L., Terminalia bellirica (Gaertn.) Roxb., Terminalia chebula Retz., and Zingiber officinale Roscoe). Apart from its utilization in Thai traditional medicine, there is a lack of evidence supporting its use. This research work thereby aims to formulate and evaluate the tablet containing Nawametho decoction. The feasibility of Nawametho decoction and NawaTab for patients with borderline hyperlipidemia was additionally examined using a prospective, open-label, randomized, parallel-group design. The dry granulation technique was employed to formulate the polyherbal tablets. The tablets were developed using the spray-dried Nawametho decoction as the active ingredient in addition to other excipients. The chosen formulation, the F B (NawaTab), consisted of 385 milligrams of the extract, 12% w/w of a diluent (lactose), 8% w/w of a lubricant (magnesium stearate), 5% w/w of a disintegrant (microcrystalline cellulose), and 5% w/w of an anti-adherent (talcum). Their hardness, friability, and disintegration time were 4.4 ± 0.32 kg, 0.05 ± 0.02%, and 4.60 ± 0.05 min, respectively. Accelerated stability study results revealed that NawaTab was stable for six months at 40°C/75% RH and 25°C/60% RH. Even though taking NawaTabs (500 mg twice daily) for eight consecutive weeks was unable to improve the lipid profile of the patients, the administration of Nawametho decoction (30 mL twice daily) was associated with a significant decrease in serum triglycerides of the patients. The results show that the dry granulation technique is suitable for the formulation of NawaTab based on the tablet evaluation. Furthermore, the triglyceride-lowering effect of Nawametho decoction was reported for the first time.

Ethnomedicinal Plants in Herbal Remedies Used for Treatment of Skin Diseases by Traditional Healers in Songkhla Province, Thailand.

Skin disorders are a worldwide health problem that normally affect human life. A traditional healer is an important expert in researching notable medicinal plants for skin disease treatment. This study aimed to determine the traditional knowledge and the use of medicinal plants for the treatment of skin diseases among traditional healers in the Songkhla province, Thailand. The ethnobotanical information was collected from experienced traditional healers by semi-structured interviews and participant observations. Plant specimens were also collected and identified using the standard taxonomic method. The data were analyzed by interpretation and descriptive statistics. Twenty-five polyherbal formulations for the treatment of skin diseases were obtained from traditional healers with at least 10 years of experience. A total of 66 plant species in 38 families were documented. Leaves and trees were the most commonly employed plant parts and plant habits included in the herbal remedies, respectively. Fabaceae, Rubiaceae, and Zingiberaceae were the majority of the cited families. Oryza sativa L. and Zingiber montanum (J. Koenig) Link ex A.Dietr. were the most preferred plants combined in the prescriptions, which had the highest use value (UV = 0.83). The highest relative frequency of citation was represented by Curcuma longa L., Eurycoma longifolia Jack, Knema globularia (Lamk.) Warb, and Senna siamea (Lam.) Irwin & Barneby. (0.55 each). This research suggests the importance of traditional healers in the healing of skin diseases with herbal remedies. A variety of medicinal plants are used in the prescriptions for the treatment of skin disorders in the Songkhla province, in the south of Thailand. Pharmacological and toxicological activities as well as phytochemical constituents of polyherbal remedies should be further investigated to scientifically verify further applications of widely practiced herbal medicines.

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.

Enhanced Oral Bioavailability and Improved Biological Activities of a Quercetin/Resveratrol Combination Using a Liquid Self-Microemulsifying Drug Delivery System.

Both quercetin and resveratrol are promising plant-derived compounds with various well-described biological activities; however, they are categorized as having low aqueous solubility and labile natural compounds. The purpose of the present study was to propose a drug delivery system to enhance the oral bioavailability of combined quercetin and resveratrol. The suitable self-microemulsifying formulation containing quercetin together with resveratrol comprised 100 mg Capryol 90, 700 mg Cremophor EL, 200 mg Labrasol, 20 mg quercetin, and 20 mg resveratrol, which gave a particle size of 16.91 ± 0.08 nm and was stable under both intermediate and accelerated storage conditions for 12 months. The percentages of release for quercetin and resveratrol in the self-microemulsifying formulation were 75.88 ± 1.44 and 86.32 ± 2.32%, respectively, at 30 min. In rats, an in vivo pharmacokinetics study revealed that the area under the curve of the self-microemulsifying formulation containing quercetin and resveratrol increased approximately ninefold for quercetin and threefold for resveratrol compared with the unformulated compounds. Moreover, the self-microemulsifying formulation containing quercetin and resveratrol slightly enhanced the in vitro antioxidant and cytotoxic effects on AGS, Caco-2, and HT-29 cells. These findings demonstrate that the self-microemulsifying formulation containing quercetin and resveratrol could successfully enhance the oral bioavailability of the combination of quercetin and resveratrol without interfering with their biological activities. These results provide valuable information for more in-depth research into the utilization of combined quercetin and resveratrol.

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.

Traditional tonifying polyherbal infusion, Jatu-Phala-Tiga, exerts antioxidant activities and extends lifespan of Caenorhabditis elegans.

BACKGROUND: The imbalance between the generation of free radicals and natural cellular antioxidant defenses, known as oxidative stress, can cause oxidation of biomolecules and further contribute to aging-associated diseases. The purpose of this study was to evaluate the antioxidant capacities of Thai traditional tonifying preparation, Jatu-Phala-Tiga (JPT) and its herbal ingredients consisting of Phyllanthus emblica, Terminalia arjuna, Terminalia chebula, and Terminalia bellirica and further assess its effect on longevity. METHOD: Antioxidant activities of various extracts obtained from JPT and its herbal components were carried out using well-established methods including metal chelating, free radical scavenging, and ferric reducing antioxidant power assays. Qualitative analysis of the chemical composition from JPT water extract was done by high-performance liquid chromatography tandem with electrospray ionisation mass spectrometry. The effect of JPT water extract on the lifespan of Caenorhabditis elegans were additionally described. RESULTS: Among the extracts, JPT water extract exerted remarkable antioxidant activities as compared to the extracts from other solvents and individual constituting plant extract. JPT water extract was found to possess the highest metal chelating activity, with an IC50 value of 1.75 ± 0.05 mg/mL. Moreover, it exhibited remarkable scavenging activities towards DPPH, ABTS, and superoxide anion radicals, with IC50 values of 0.31 ± 0.02, 0.308 ± 0.004, and 0.055 ± 0.002 mg/mL, respectively. The ORAC and FRAP values of JPT water extract were 40.338 ± 2.273 µM of Trolox/µg of extract and 23.07 ± 1.84 mM FeSO4/mg sample, respectively. Several well-known antioxidant-related compounds including amaronols, quinic acid, gallic acid, fertaric acid, kurigalin, amlaic acid, isoterchebin, chebulagic acid, ginkgolide C, chebulinic acid, ellagic acid, and rutin were found in this extract. Treatment with JPT water extract at 1 and 5 mg/mL increased C. elegans lifespan under normal growth condition (7.26 ± 0.65 vs. 10.4 0± 0.75 (p < 0.01) and 10.00 ± 0.73 (p < 0.01) days, respectively). CONCLUSIONS: The results indicated that JPT and its herbal ingredients exhibited strong antioxidant activities, in particular the water extract of the polyherbal tonic. These findings rationalize further investigation in JPT infusion as a promising agent for anti-aging and oxidative stress prevention.

Effects of a traditional Thai polyherbal medicine 'Ya-Samarn-Phlae' as a natural anti-biofilm agent against Pseudomonas aeruginosa.

Ya-Samarn-Phlae (YaSP) is a traditional Thai polyherbal formula for the treatment of chronic wounds. Although its ethanol extract has been proven to possess several wound-related biological activities, there is no scientific information available for the infused oil of YaSP, which is its traditionally prepared form. This present study therefore aimed to evaluate the efficacy of different infused oils obtained from either fresh or oven-dried herbal parts of YaSP (F-YaSP and D-YaSP) against biofilms of Pseudomonas aeruginosa, which reside in chronic wounds. Its main active herbal component, Garcinia mangostana (F-GM and D-GM), as well as α-mangostin were also tested in this study. All infused oils significantly inhibited the biofilm formation of P. aeruginosa with a percentage of reduction ranging from 50 to 90%. Visualization of the inhibition of biofilm development was confirmed using scanning electron and atomic force microscopes. All tested agents resulted in a reduction in the mean average roughness of the biofilm, whereas only treating with D-YaSP, D-GM, and α-mangostin led to a decrease in both peak height and peak-valley height. MTT reduction assays revealed that the metabolic activity of P. aeruginosa mature biofilms decreased considerably up to 50% after only 3 h of incubation and after only 9 h of exposure to D-YaSP. Confocal laser scanning micrographs illustrated that a maximum biofilm eradication was found when treated with the extracts for 3 h, whereas the biomass, the average thickness, maximum thickness, and the surface to volume ratio of the treated biofilm was reduced after up to 18 h of contact time. It can be concluded that D-YaSP can effectively inhibit biofilm formation and eradicate mature biofilms of P. aeruginosa. It should be noted that G. mangostana and α-mangostin contribute in YaSP as principle active agents for anti-biofilm efficacy.

New alkylamide from the stems of Zanthoxylum nitidum.

A new alkylamide, named (2E,6E,8E)-N-(2-methylpropyl)-10-oxo-2,6,8-decatrienamide (1), together with 22 known compounds (2-23), were isolated from the stems of Zanthoxylum nitidum. Their structures were elucidated by spectroscopic methods, including 1D and 2D NMR spectroscopy. The isolated compounds exhibited slightly antioxidant activities through DPPH and ABTS radical scavenging assays but showed no antibacterial activity against Streptococcus mutans ATCC2517, a dental caries causing bacteria.

Antioxidant capacities and total phenolic contents of 20 polyherbal remedies used as tonics by folk healers in Phatthalung and Songkhla provinces, Thailand.

BACKGROUND: Uses of polyherbal formulations have played a major role in traditional medicine. The present study is focused on the formulations used in traditional Thai folkloric medicine as tonics or bracers. Twenty documented polyherbal mixtures, used as nourishing tonics by the folk healers in Phatthalung and Songkhla provinces in southern Thailand, are targeted. Despite traditional health claims, there is no scientific evidence to support the utilization of polyherbal formulations. METHODS: The phenolic and flavonoid contents of the polyherbal formulations and a series of antioxidant tests were applied to measure their capability as preventive or chain-breaking antioxidants. In addition, the cytotoxic activity of effective formulations was assayed in Vero cells. RESULTS: Ninety-eight plant species belonging to 45 families were used to prepare the tested formulation. The preliminary results revealed that water extracts of THP-R016 and THP-R019 contain a high level of total phenolic and flavonoid contents and exhibit remarkable antioxidant activities, as tested by DPPH, ABTS, and FRAP assays. The extract of THP-R019 also showed the strongest metal chelating activities, whereas THP-R016 extract possessed notable superoxide anion and peroxyl radical scavenging abilities. CONCLUSIONS: The data provide evidence that the water extracts of folkloric polyherbal formulations, particularly THP-R016, are a potential source of natural antioxidants, which will be valuable in the pharmaceutical and nutraceutical industries. The free radical scavenging of THP-R016 may be due to the contribution of phenolic and flavonoid contents. Useful characteristics for the consumer, such as the phytochemical profiles of active ingredients, cellular based antioxidant properties and beneficial effects in vivo, are under further investigation.

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.

Inhibition of microbial adhesion to plastic surface and human buccal epithelial cells by Rhodomyrtus tomentosa leaf extract.

OBJECTIVE: The adherence of oral pathogenic microorganisms to host tissues is the initial step for successful process of oral diseases. This study aimed to determine the effect of the Rhodomyrtus tomentosa leaf extract and rhodomyrtone, an antibacterial compound from R. tomentosa leaf, on adhesion of some oral pathogens to polystyrene plastic surface and human buccal epithelial cells. METHODS: The minimum inhibitory concentration (MIC) was evaluated using broth microdilution method. The microbial adhesion to the plastic surface and buccal cells was determined using microtiter plate method and microscopy technique. RESULTS: The ethanol extract of leaf demonstrated antibacterial activity against oral microorganisms including Staphylococcus aureus ATCC 25923, Streptococcus mutans (clinical isolate), and Candida albicans ATCC 90028 with the MIC values of 31.25, 15.62, and 1000µg/ml, respectively. Rhodomyrtone displayed activity with the MIC values of 0.78 and 0.39µg/ml against S. aureus ATCC 25923 and S. mutans, respectively. The MIC value of the compound against C. albicans ATCC 90028 was more than 100µg/ml which was the highest test concentration. All pathogenic microorganisms treated with the extract and rhodomyrtone at their subinhibitory concentrations resulted in a decrease in their adherence ability to both plastic surface and buccal cells. CONCLUSION: It is suggested that R. tomentosa extract and rhodomyrtone may be useful in therapy or as prophylaxis in infections involving oral pathogens.

Antagonistic interactions of "Ya-Sa-Marn-Phlae" ethanol extract in combination with topical antiseptics against clinical isolates of Staphylococcus aureus.

This investigation was aimed at assessing a possible interaction of a traditional Thai herbal recipe, "Ya-Sa-Marn-Phlae (YSMP)," used for wound treatments with topical antiseptics, povidone-iodine (PI) solution and hydrogen peroxide (H2O2), and effects of THR-SK010 alone and the combinations on Staphylococcus aureus. Antibacterial activities of ethanol extracts from the herbal recipe were determined against both methicillin resistant Staphylococcus aureus (MRSA) and methicillin-susceptible S. aureus (MSSA). YSMP exhibited remarkable antistaphylococcal activity with MIC values of 3.9-7.8 µg/mL. This recipe possessed bacteriostatic activity and did not reduce the tolerance of both MRSA and MSSA isolates to the high ionic strength. Interaction between THR-SK010 and the antiseptics was carried out by checkerboard testing and time-kill assay. Both indifferent and slightly antagonistic effects were observed with THR-SK010/PI and THR-SK010/H2O2 combinations against the tested isolates. In addition to commercially available antiseptics, THR-SK010 offered additional therapeutic options for the decolonization of MRSA and MSSA. Topical application of plant extracts with antioxidant activity, such as THR-SK010, should not be used immediately with PI or H2O2 and further investigation on this interaction is needed.

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.

Thai herbal formulas used for wound treatment: a study of their antibacterial potency, anti-inflammatory, antioxidant, and cytotoxicity effects.

AIM: This present work was aimed to investigate wound healing-related biologic activities of traditional herbal formulas used for wound treatment in southern Thailand. METHODS: Water and ethanol extracts of the formulas (THR-SK004, THR-SK010, and THR-SK011) were tested for their antibacterial potency against methicillin-resistant Staphylococcus aureus (MRSA) and -susceptible S. aureus. Anti-inflammatory activities of the extracts were assessed by detection of the inhibition of lipopolysaccharide-induced nitric oxide production. Anti-oxidant activities and cytotoxicity of the extracts were also measured. RESULTS: Among the tested formulas, ethanol extract of THR-SK010 consisting of four herbs: Curcuma longa L., Areca catechu L., Oryza sativa L., and Garcinia mangostana L., was found to possess promising antibacterial activities with MIC90 of 4 µg mL(-1) against MRSA isolates. This ethanol extract offered the highest anti-inflammatory activity as well as DPPH and hydroxyl radical scavenging activities. CONCLUSIONS: Remarkable antibacterial, anti-inflammatory, and antioxidant activities as well as low toxicity on Vero cells of THR-SK010 ethanol extract provide scientific information to support the topical use of the formula for wound treatment. This information proposes the potential to develop a new generation of phytopharmaceuticals based on traditional knowledge.

Antibacterial Activity of Rhodomyrtus tomentosa (Aiton) Hassk. Leaf Extract against Clinical Isolates of Streptococcus pyogenes.

Ethanol extract of Rhodomyrtus tomentosa (Aiton) Hassk. leaf was evaluated for antibacterial activity against 47 clinical isolates of Streptococcus pyogenes. The extract exhibited good anti-S. pyogenes activity against all the tested isolates with similar minimum inhibitory concentration (MIC, 3.91-62.5 µg mL(-1)) and minimum bactericidal concentration (MBC, 3.91-62.5 µg mL(-1)) ranges. No surviving cells were detected at 16 h after treatment with 8 × MIC of the extract. The extract-treated cells demonstrated no lysis and cytoplasmic leakage through the bacterial membrane. Electron micrographs further revealed that the extract did not cause any dramatic changes on the treated cells. Rhodomyrtone, an isolated compound, exhibited good anti-S. pyogenes activity (14 isolates), expressed very low MIC (0.39-1.56 µg mL(-1)) and MBC (0.39-1.56 µg mL(-1)) values. Rhodomyrtus tomentosa leaf extract and rhodomyrtone displayed promising antibacterial activity against clinical isolates of S. pyogenes.

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.

Potential antibiotic and anti-infective effects of rhodomyrtone from Rhodomyrtus tomentosa (Aiton) Hassk. on Streptococcus pyogenes as revealed by proteomics.

Rhodomyrtone from Rhodomyrtus tomentosa (Aiton) Hassk. leaf extract has a strong antibacterial activity against the bacterial pathogen Streptococcus pyogenes. Our previous studies indicated that the bactericidal activity of rhodomyrtone might involve intracellular targets. In the present studies we followed a proteomics approach to investigate the mode of action of rhodomyrtone on S. pyogenes. For this purpose, S. pyogenes was cultivated in the presence of 0.39 µg/ml rhodomyrtone, which corresponds to 50% of the minimal inhibitory concentration. The results show that the amounts of various enzymes associated with important metabolic pathways were strongly affected, which is consistent with the growth-inhibiting effect of rhodomyrtone. Additionally, cells of S. pyogenes grown in the presence of rhodomyrtone produced reduced amounts of known virulence factors, such as the glyceraldehyde-3-phosphate dehydrogenase, the CAMP factor, and the streptococcal pyrogenic exotoxin C. Taken together, these findings indicate that rhodomyrtone has both antimicrobial and anti-infective activities, which make it an interesting candidate drug.

Rhodomyrtone: a new candidate as natural antibacterial drug from Rhodomyrtus tomentosa.

Rhodomyrtone [6,8-dihydroxy-2,2,4,4-tetramethyl-7-(3-methyl-1-oxobutyl)-9-(2-methylpropyl)-4,9-dihydro-1H-xanthene-1,3(2H)-di-one] from Rhodomyrtus tomentosa (Aiton) Hassk. displayed significant antibacterial activities against gram-positive bacteria including Bacillus cereus, Bacillus subtilis, Enterococcus faecalis, Staphylococcus aureus, methicillin-resistant S. aureus (MRSA), Staphylococcus epidermidis, Streptococcus gordonii, Streptococcus mutans, Streptococcus pneumoniae, Streptococcus pyogenes, and Streptococcus salivarius. Especially noteworthy was the activity against MRSA with a minimum inhibitory concentration (MIC) and a minimum bactericidal concentration (MBC) ranging from 0.39 to 0.78 microg/ml. As shown for S. pyogenes, no surviving cells were detected within 5 and 6h after treatment with the compound at 8MBC and 4MBC concentrations, respectively. Rhodomyrtone displays no bacteriolytic activity, as determined by measurement of the optical density at 620 nm. A rhodomyrtone killing test with S. mutans using phase contrast microscopy showed that this compound caused a few morphological changes as the treated cells were slightly changed in color and bigger than the control when they were killed. Taken together, the results support the view that rhodomyrtone has a strong bactericidal activity on gram-positive bacteria, including major pathogens.