Centella asiatica (L.) Urb. and Hydrocotyle umbellata L. identification and quality assessment: A methodology comparison.

INTRODUCTION: Centella is an important genus in the Apiaceae family. It includes Centella asiatica, which has significant edible and medicinal values. However, this species is easily confused due to its similar morphological traits to Hydrocotyle umbellata, hindering its utilization in the consumer and pharmacological industries. OBJECTIVE: The study aims to differentiate these two closely related plant species using reliable methods of confirming the authenticity of natural herbal medicines. METHODS: Our work mainly focuses on the basic morphological characteristics, chemical markers, genetic fingerprints, and their biological responses. RESULTS: The plants can be clearly differentiated using their leaf shapes, stipules, petioles, inflorescences, and fruit structures. Although the phytochemical compositions of the C. asiatica extract were similar to that of H. umbellata which included flavonoids, tannins, and saponins important to the plant's ability to reduce inflammation and promote healing of wounds, the H. umbellata extract showed significantly higher toxicity than that of C. asiatica. High-performance liquid chromatography analysis was used to identify chemical fingerprints. The result revealed that C. asiatica had major triterpene glycoside constituents including asiaticoside, asiatic acid, madecassoside, and madecassic acid, which have a wide range of medicinal values. In contrast, triterpenoid saponins were not identified in H. umbellata. Furthermore, using SCoT1-6 primers was possible to effectively and sufficiently created a dendrogram which successfully identified the closeness of the plants and confirmed the differences between the two plant species. CONCLUSION: Therefore, differentiation can be achieved through the combination of morphometrics, molecular bioactivity, and chemical analysis.

Galectin, another lectin from Fenneropenaeus merguiensis, contributed in shrimp immune defense.

Numerous lectins act as pattern recognition receptors (PRRs) in the innate immune system of invertebrates. Here, a galectin (FmGal) was isolated from hemocytes of Fenneropenaeus merguiensis. FmGal contained one open reading frame encoding a peptide of 338 amino acids. The primary sequence of FmGal comprised a carbohydrate recognition domain with a specific galactose binding site. The FmGal transcripts were found mostly in hemocytes of healthy shrimp. The expression of FmGal was up-regulated upon challenge with Vibrio parahaemolyticus and white spot syndrome virus (WSSV). Gene-silencing with FmGal double-stranded RNA resulted in extreme down-regulation of FmGal. Knockdown with a co-injection of pathogens reduced the survival rate of shrimp. The recombinantr protein of FmGal (rFmGal) required Ca2+ to agglutinate pathogenic bacteria and exhibited sugar-specificity to galactose, lactose, lipopolysaccharide (LPS) and lipoteichoic acid (LTA). The ELISA-validated binding of rFmGal revealed higher affinity to LTA than LPS. rFmGal did not exhibit antibacterial activity but could enhance the phagocytosis and encapsulation of pathogenic invaders by hemocytes. Encapsulation was suppressed by galactose and lactose. Moreover, rFmGal also promoted the in vivo clearance of V. parahaemolyticus. FmGal, a galectin in F. merguiensis, participated in shrimp immunity, functioning as a PRR which might be involved in certain cellular responses.

Flavonoids and Phenols, the Potential Anti-Diabetic Compounds from Bauhinia strychnifolia Craib. Stem.

Bioactive compounds from medicinal plants are good alternative treatments for T2DM. They are also sources of lead molecules that could lead to new drug discoveries. In this study, Bauhinia strychnifolia Craib. stem, a traditional Thai medicinal plant for detoxification, was extracted into five fractions, including crude extract, BsH, BsD, BsE, and BsW, by ethanolic maceration and sequential partition with hexane, dichloromethane, ethyl acetate, and water, respectively. Among these fractions, BsE contained the highest amounts of phenolics (620.67 mg GAE/g extract) and flavonoids (131.35 mg QE/g extract). BsE exhibited the maximum inhibitory activity against α-glucosidase (IC50 1.51 ± 0.01 µg/mL) and DPP-IV (IC50 2.62 ± 0.03 µg/mL), as well as dominantly promoting glucose uptake on 3T3-L1 adipocytes. Furthermore, the four compounds isolated from the BsE fraction, namely resveratrol, epicatechin, quercetin, and gallic acid, were identified. Quercetin demonstrated the highest inhibitory capacity against α-glucosidase (IC50 6.26 ± 0.36 µM) and DPP-IV (IC50 8.25 µM). In addition, quercetin prominently enhanced the glucose uptake stimulation effect on 3T3-L1 adipocytes. Altogether, we concluded that quercetin was probably the principal bioactive compound of the B. strychnifolia stem for anti-diabetic, and the flavonoid-rich fraction may be sufficiently potent to be an alternative treatment for blood sugar control.

An alternative function of C-type lectin comprising low-density lipoprotein receptor domain from Fenneropenaeus merguiensis to act as a binding receptor for viral protein and vitellogenin.

A diversity of C-type lectins (CTLs) was coming reported and they are known to participate in invertebrate innate immunity by act as pattern recognition receptor (PRR). In the present study, a unique CTL containing low-density lipoprotein receptor (LDLR) domain from Fenneropenaeus merguiensis (designated as FmLdlr) was cloned. Its sequence contained a single LDLR domain and one carbohydrate recognition domain (CRD) with a QAP motif putative for galactose-specific binding. The expression of FmLdlr was detected only in hemocytes of healthy shrimp. Its expression was significantly up-regulated by Vibrio parahaemolyticus or white spot syndrome virus (WSSV) challenge. The knockdown by FmLdlr dsRNA resulted in severe gene down-regulation. The gene silencing with pathogenic co-inoculation led to reduction of the median lethal time and increasing in the cumulative mortality including the remained WSSV in WSSV co-challenge group. Recombinant proteins of FmLdlr and two domains could agglutinate various bacterial strains which LDLR domain revealed the lowest activity. Only FmLdlr and CRD could enhance phagocytosis and encapsulation by hemocytes. Both FmLdlr and CRD except LDLR domain exhibited the antibacterial activity by inhibiting the growth of pathogenic V. parahaemolyticus in cultured medium and disk diffusion assay. Only FmLdlr and CRD could bind to WSSV proteins, envelope VP28, tegument VP39A and also capsid VP15, which FmLdlr had the higher binding affinity than that of CRD. Altogether, we concluded that FmLdlr contributed in shrimp immune defense through the main action of CRD in capable of bacterial agglutination, enhancing the phagocytosis and encapsulation, antimicrobial activity and binding to viral proteins. Interestingly, ELISA approach revealed that LDLR domain displayed the highest binding affinity to vitellogenin than whole molecule and CRD. We signified a new function of FmLdlr that it might presumably act as a receptor for vitellogenin transportation in hemolymph during vitellogenesis of shrimp.

Simultaneous Delivery of Curcumin and Resveratrol via In Situ Gelling, Raft-Forming, Gastroretentive Formulations.

Curcumin and resveratrol are polyphenolic compounds that have been shown to exhibit synergistic therapeutic properties including anti-inflammatory, anticancer, and antiulcer activities, which may be exploited for the treatment of gastric diseases. However, both compounds have poor aqueous solubility and rapid metabolism, resulting in a low oral bioavailability. In situ gelling, liquid formulations were developed to produce a gastroretentive, raft-forming delivery vehicle to improve bioavailability. Solid dispersions containing a mixture of curcumin and resveratrol with Eudragit® EPO (Cur/Res-SD) were first prepared using solvent evaporation, to improve the solubility and dissolution of the compounds. Solid dispersions of a weight ratio of 1:10 curcumin/resveratrol to Eudragit® EPO were subsequently incorporated into in situ gelling, liquid formulations based on the gelling polymers, sodium alginate (low viscosity and medium viscosity), pectin, and gellan gum, respectively. Calcium carbonate and sodium bicarbonate were included to produce carbon dioxide bubbles in the gel matrix, on exposure to gastric fluid, and to achieve flotation. Moreover, the calcium ions acted as a crosslinking agent for the hydrogels. Optimized formulations floated rapidly (<60 s) in simulated gastric fluid (pH = 1.2) and remained buoyant, resulting in the gradual release of more than 80% of the curcumin and resveratrol content within 8 h. The optimized formulation based on medium-viscosity sodium alginate exhibited enhanced cytotoxic activity toward human gastric adenocarcinoma cell lines (AGS), compared with unformulated curcumin and resveratrol compounds, and increased anti-inflammatory activity against RAW 264.7 macrophage cells compared with the NSAID, indomethacin. These findings demonstrate that in situ gelling, liquid formulations, loaded with a combination of curcumin and resveratrol in the form of solid dispersions, show potential as gastroretentive delivery systems for local and systemic effects.

Development of raft-forming liquid formulations loaded with ginger extract-solid dispersion for treatment of gastric ulceration.

Raft-forming liquid formulations incorporating ginger extract solid dispersion (GE-SD) were developed to achieve prolonged delivery of 6-gingerol in the stomach and thus increase the effectiveness of gastric ulcer treatment. The solubility of 6-gingerol in 0.1 N HCl (pH 1.2) was maximized (15 mg/mL) by combining ginger extract with PVP K30 at 1:3 w/w ratio to produce a solid dispersion. The nature of GE-SD was confirmed by PXRD and FT-IR analysis. PXRD pattern showed miscibility of GE and PVP K30 in amorphous solid dispersion and the FT-IR spectra confirmed the formation of hydrogen bond between GE and PVP K30. GE-SD-loaded raft-forming liquids were prepared using sodium alginate as a gel former and HPMC as a release-controlling agent. The formulations exhibited rapid floating behavior in 0.1 N HCl (<30 s) and remained afloat on the surface over 8 h. The formed raft structures provided sufficient strength (>7.5 g) and allowed sustained release of more than 70 % of the 6-gingerol content over 8 h in 0.1 N HCl. Raft-forming formulations incorporating ginger extract demonstrated anti-inflammatory activity by inhibiting nitric oxide production in LPS-stimulated RAW 264.7 macrophage cells (IC50 = 5.13 ± 0.07 µg/mL). Exposure to the formulations also had a significant cytotoxic effect on AGS human gastric adenocarcinoma cells with an IC50 of 17.45 ± 0.29 µg/mL. In addition, the raft-forming formulations enhanced the migratory behavior of L929 mouse fibroblasts in the scratch wound model. Taken together, these findings reveal the benefits of gastro-retentive, GE-SD-loaded raft-forming liquid formulations for improving the treatment of gastric ulcers.

Anti-diabetic compounds from Uvaria dulcis Dunal.

Medicinal plants have long been a source of lead compounds for drug discovery. Among these, the Annonaceae family has gained recognition for its potential to yield novel compounds, particularly those that can be used in the development of drugs targeting chronic diseases like diabetes mellitus (DM). We employed various chromatographic methods to isolate bioactive compounds from the roots, leaves, and twigs of Uvaria dulcis Dunal. We used spectroscopic methods to determine the chemical structures of these compounds. We successfully identified twelve known compounds from various parts of U. dulcis: patchoulenon, polygochalcone, 2'3'-dihydroxy-4',6'-dimethoxydihydrochalcone, 2',3'-dihydroxy-4',6'-dimethoxychalcone, chrysin, techochrysin, 8-hydroxy-5,7-dimethoxyflavanone, pinocembrin, 3-farnesylindole, onysilin, cinchonain la, and cinchonain lb. Interestingly, cinchonain la and cinchonain lb exhibited more potent anti-α-glucosidase activity than acarbose (standard drug), with IC50 values of 11.88 ± 1.41 µg/mL and 15.18 ± 1.19 µg/mL, respectively. Cinchonain la inhibited the DPP-IV enzyme, with IC50 value lower than the standard compound (diprotin A) at 81.78 ± 1.42 µg/mL. While 2',3'-dihydroxy-4',6'-dimethoxychalcone show more potent inhibitory effect than standard drug with IC50 value of 8.62 ± 1.19 µg/mL. Additionally, at a concentration of 10 µg/mL, cinchonain lb and 2',3'-dihydroxy-4',6'-dimethoxychalcone promoted glucose uptake in L6 myotubes cells to the same extent as 100 nM insulin. These findings suggest that cinchonain la, cinchonain lb, and 2',3'-dihydroxy-4',6'-dimethoxychalcone are the U. dulcis-derived bioactive compounds that hold promise as potential structures to use in the development of anti-diabetic drugs.

Development of Oral In Situ Gelling Liquid Formulations of Garcinia Extract for Treating Obesity.

Novel in situ gelling liquid formulations incorporating garcinia extract were developed to achieve prolonged delivery of hydroxycitric acid (HCA), an active compound displaying anti-obesity function, following oral administration. The optimized formulation was composed of sodium alginate (1.5% w/v), hydroxypropyl methylcellulose (HPMC K100) (0.25% w/v), calcium carbonate (1% w/v) and garcinia extract (2% w/v). The formulation displayed rapid gelation in less than a minute on exposure to 0.1 N hydrochloric acid (pH 1.2) and remained afloat for more than 24 h. The formulations were capable of gradually releasing more than 80% of HCA load over 8 h, depending on the composition. The resulting gels exhibited high values of gel strength by texture analysis, suggesting they would offer resistance to breakdown under the action of stomach content movement. The optimized formulation loaded garcinia extract significantly reduced lipid accumulation in 3T3-L1 adipocyte cells and displayed moderate anti-inflammatory activity by inhibiting the production of nitric oxide (NO) in LPS-stimulated RAW 264.7 macrophage cells. These findings demonstrate that oral in situ gelling liquid formulations based on sodium alginate and HPMC K100 offer much potential for sustained delivery of HCA and other anti-obesity compounds.