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.

Gold nanoparticles exposure modulates antioxidant and innate immune gene expression in the gills of Sparus aurata.

This study aimed to assess antioxidant and immune gene transcription alterations in the gills of Sparus aurata exposed during 96 h to 4, 80, and 1600 µg/L of gold nanoparticles (AuNPs) coated with citrate or polyvinyl pyrrolidone (PVP). After 96 h of exposure, gr and cat mRNA levels decreased for all tested concentrations of AuNPs, for both coatings. Instead, gst3 mRNA increased after exposure to 1600 µg/L AuNPs (both coatings) and prdx6 increased after exposure to 1600 µg/L AuNPs-citrate. Concerning immune genes, il1ß mRNA levels increased after exposure to 80 µg/L AuNPs-citrate and 1600 µg/L AuNPs-PVP and cox2 mRNA showed increased levels in fish exposed to 1600 µg/L AuNPs-citrate. Results indicate that AuNPs with distinct coatings induced different gene expression profiles in gills, though most of the studied genes remained unaltered for the tested conditions.

Immune-related gene expression and physiological responses in rainbow trout (Oncorhynchus mykiss) after intraperitoneal administration of Rhodomyrtus tomentosa leaf extract: A potent phytoimmunostimulant.

The immunostimulatory effects of Rhodomyrtus tomentosa leaf extract were evaluated in rainbow trout through changes in expression profile of genes involved in innate immune and antioxidant response, hematology and stress indicators. The concentrations of R. tomentosa at 10 and 100 µg per fish were administrated by intraperitoneal injection, alone or in combination with LPS. After 6 h of administration, the gene expression was measured in head kidney, spleen, and intestine. Results indicated that R. tomentosa exerted immunostimulatory effects by inducing the expression of il10, saa, hepcidin, and sod in head kidney and the expression of il10, tgfß, and inos in intestine. In combination with LPS, the plant suppressed the expression of pro-inflammtory cytokine il1ß, il8 and other consisting of saa and gpx1 in head kidney and il1ß in spleen, pointing out its anti-inflammatory activities. Furthermore, the plant did not exert any impact on hematological parameters, but it was able to reduce cortisol levels when co-administered with LPS, indicating that R. tomentosa could attenuate stress response in rainbow trout. Our observations suggest that R. tomentosa induced the expression of genes involved in cytokine and innate immune response and modulated the physiological stress response as indicated by the suppressed cortisol in rainbow trout.

Immunomodulatory effects of Rhodomyrtus tomentosa leaf extract and its derivative compound, rhodomyrtone, on head kidney macrophages of rainbow trout (Oncorhynchus mykiss).

Rhodomyrtus tomentosa is a medicinal plant that shows biological effects including immunomodulatory activity on human and other mammals but not in fish. In this study, we evaluated the in vitro immunomodulatory effects of R. tomentosa leaf extract and its active compound, rhodomyrtone, on the immune responses, using rainbow trout (Oncorhynchus mykiss) head kidney (HK) macrophages as a model. The tested immune functions included the expression of genes involved in innate immune and inflammatory responses and the production of reactive oxygen species (ROS). Gene expression was evaluated after exposure to 10 µg mL-1 of R. tomentosa and 1 µg mL-1 of rhodomyrtone for 4 and 24 h. R. tomentosa and rhodomyrtone induced changes in the expression of pro-inflammatory cytokines (il1ß, il8, and tnfα), anti-inflammatory cytokines (il10 and tgfß), inducible enzymes (inos, cox2, and arginase), and an antioxidant enzyme (gpx1). Co-exposure of R. tomentosa with LPS resulted in a prominent reduction in the expression of genes related to an inflammatory process (il1ß, il8, tnfα, inos, saa, hepcidin, and gpx1), suggesting anti-inflammatory effects. Similarly, co-exposure of rhodomyrtone with LPS led to a downregulation of inflammation-related genes (il1ß, inos, saa, and hepcidin). In addition, exposure to both natural plant products caused a reduction in cellular ROS levels by HK macrophages. The present results indicate that R. tomentosa and rhodomyrtone exerted immunostimulatory and anti-inflammatory effects on fish macrophages, thus opening up the possibility of using these natural products to further develop immunostimulants for health management in aquaculture.

In vitro and in vivo assessments of Rhodomyrtus tomentosa leaf extract as an alternative anti-streptococcal agent in Nile tilapia (Oreochromis niloticus L.).

PURPOSE: Rhodomyrtustomentosa is a Thai medicinal plant that has been attracting attention for its remarkable antibacterial properties against Gram-positive pathogenic bacteria. The purpose of this study was to evaluate the antibacterial properties of R. tomentosa leaf extract against Streptococcus agalactiae and Streptococcus iniae isolated from infected tilapia. METHODOLOGY: The anti-streptococcal activity of R. tomentosa was determined using broth microdilution assays. RESULTS: The extract demonstrated strong antibacterial activity against the fish pathogens, with minimum inhibitory concentrations (MICs) ranging from 7.8‒62.5 µg ml-1. It was found to possess a dose-dependent bacteriostatic effect on this organism. Scanning electron microscopy revealed irregular and long chains of swollen cells, as well as corkscrew shapes andincomplete separation of cell division of S. agalactiae cells following the treatment at sub-MIC. Moreover, S. agalactiae cells pre-treated with the extract became more sensitive to oxidative stress induced by H2O2 than the untreated cells. Based on the mortality of Nile tilapia after intraperitoneal infection of S. agalactiae at median lethal dose (LD50), the pre-treated cells caused a significant (P<0.01) reduction in mortality of S. agalactiae-infected Nile tilapia. CONCLUSION: The results suggested that R. tomentosa could be further developed as a simple and effective agent for the treatment of streptococcosis in Nile tilapia.

Synergistic effects of ethnomedicinal plants of Apocynaceae family and antibiotics against clinical isolates of Acinetobacter baumannii.

OBJECTIVE: To investigate the efficacy of 17 ethnomedicinal plants belonging to Apocynaceae family used in combination with 16 conventional antibiotics against non-multidrug resistant-, multidrug resistant (MDR)-, and extensive drug resistant (XDR) Acinetobacter baumannii (A. baumannii). METHODS: Antibacterial activity and resistance modifying ability of 272 combinations were determined by growth inhibition assays and further confirmed by time-kill assay. RESULTS: Among the combinations of the antibiotics with Apocynaceae ethanol extracts on this pathogen, 15 (5%) had synergistic effects, 23 (8%) had partial synergistic effects and 234 (86%) had no effects. Synergistic activity was observed mostly when the Apocynaceae extracts were combined with rifampicin or cefazolin. Interestingly, 10 out of 17 combinations between the extracts and rifampicin displayed synergistic or partial synergistic behaviors. Holarrhena antidysenterica extract was additionally tested to restore rifampicin activity against clinical isolates of MDR and XDR A. baumannii. With respect to total or partial synergy, 70% was XDR A. baumannii isolates and 66% was MDR A. baumannii isolates. CONCLUSIONS: Holarrhena antidysenterica extract clearly demonstrated the ability to restore rifampicin activity against both A. baumannii ATCC19606 and clinically isolated A. baumannii. Additional studies examining its active principles as well as mechanisms of actions such as the effects on efflux pumps and outer membrane permeability alterations are recommended.

Holarrhena antidysenterica as a resistance modifying agent against Acinetobacter baumannii: Its effects on bacterial outer membrane permeability and efflux pumps.

Increasing rates of infections caused by multidrug resistant Acinetobacter baumannii (MDRAB) and extensively drug resistant A. baumannii (XDRAB) have caused the need for searching alternative agents. The purposed of this project was to search plant-derived natural products that act as resistant modifying agents (RMAs) against A. baumannii. In this study, we further evaluated the activity of Holarrhena antidysenterica that has been previously proposed as RMA of novobiocin for a model strain, A. baumannii ATCC 19606 on clinically isolated non-MDRAB, MDRAB, and XDRAB. Effects of H. antidysenterica on outer membrane permeability and efflux pumps of the pathogen were conducted to preliminary elucidate mechanisms of this resistant modifier. Novobiocin was selected as a model antibiotic because it is well-established as an effective agent against Gram-positive pathogens. But, it possessed low level of antibacterial activity against Gram-negative pathogens due to an effective permeability barrier of these pathogens. H. antidysenterica ethanol extract possessed weak intrinsic antibacterial activity with minimum inhibitory concentration (MIC) more than 1000 µg/mL. The extract, at concentrations of 250, 125, and 62.5 µg/mL, remarkably enhanced the inhibitory effects of novobiocin (1/4 × MIC; 1-4 µg/mL) against XDRAB isolates. Synergistic effects of novobiocin at 1/4 × MIC and 1/8 × MIC in combination with H. antidysenterica either at 31.2, 15.6, or 7.8 µg/mL against clinical isolates non-MDRAB, MDRAB, and XDRAB were evidenced for 80% of the combinations (189 out of 234 combinations). Although, no enhancement of the accumulation of ethidium bromide was observed after treated with H. antidysenterica, this plant extract weakened the outer membrane of the pathogen as indicated by an increase in the N-phenyl-1-naphthylamine uptake. Our results suggested that H. antidysenterica which primarily interrupts membrane permeability should be further investigated as a promising resistant modifier for A. baumannii.