Nanobacterial Cellulose from Kombucha Fermentation as a Potential Protective Carrier of Lactobacillus plantarum under Simulated Gastrointestinal Tract Conditions.

Kombucha bacterial cellulose (KBC), a by-product of kombucha fermentation, can be used as a biomaterial for microbial immobilization. In this study, we investigated the properties of KBC produced from green tea kombucha fermentation on days 7, 14, and 30 and its potential as a protective carrier of Lactobacillus plantarum, a representative beneficial bacteria. The highest KBC yield (6.5%) was obtained on day 30. Scanning electron microscopy showed the development and changes in the fibrous structure of the KBC over time. They had crystallinity indices of 90-95%, crystallite sizes of 5.36-5.98 nm, and are identified as type I cellulose according to X-ray diffraction analysis. The 30-day KBC had the highest surface area of 19.91 m2/g, which was measured using the Brunauer-Emmett-Teller method. This was used to immobilize L. plantarum TISTR 541 cells using the adsorption-incubation method, by which 16.20 log CFU/g of immobilized cells was achieved. The amount of immobilized L. plantarum decreased to 7.98 log CFU/g after freeze-drying and to 2.94 log CFU/g after being exposed to simulated gastrointestinal tract conditions (HCl pH 2.0 and 0.3% bile salt), whereas the non-immobilized culture was not detected. This indicated its potential as a protective carrier to deliver beneficial bacteria to the gastrointestinal tract.

A Comprehensive Review of Andrographis paniculata (Burm. f.) Nees and Its Constituents as Potential Lead Compounds for COVID-19 Drug Discovery.

The COVID-19 pandemic has intensively disrupted global health, economics, and well-being. Andrographis paniculata (Burm. f.) Nees has been used as a complementary treatment for COVID-19 in several Asian countries. This review aimed to summarize the information available regarding A. paniculata and its constituents, to provide critical points relating to its pharmacological properties, safety, and efficacy, revealing its potential to serve as a source of lead compounds for COVID-19 drug discovery. A. paniculata and its active compounds possess favorable antiviral, anti-inflammatory, immunomodulatory, and antipyretic activities that could be beneficial for COVID-19 treatment. Interestingly, recent in silico and in vitro studies have revealed that the active ingredients in A. paniculata showed promising activities against 3CLpro and its virus-specific target protein, human hACE2 protein; they also inhibit infectious virion production. Moreover, existing publications regarding randomized controlled trials demonstrated that the use of A. paniculata alone or in combination was superior to the placebo in reducing the severity of upper respiratory tract infection (URTI) manifestations, especially as part of early treatment, without serious side effects. Taken together, its chemical and biological properties, especially its antiviral activities against SARS-CoV-2, clinical trials on URTI, and the safety of A. paniculata, as discussed in this review, support the argument that A. paniculata is a promising natural source for drug discovery regarding COVID-19 post-infectious treatment, rather than prophylaxis.

Clinical Pilot Study of Rectal Suppository Containing Combined Extract of Cissus quadrangularis Linn. and Acmella paniculata (Wall ex. DC.) R. K. Jansen in Acute Hemorrhoids.

BACKGROUND: Cissus quadrangularis Linn. (CQ) is a medicinal plant with good evidence for the treatment of hemorrhoids, listed in the Thai National List of Herbal Products in the oral dosage form. Acmella paniculata (Wall ex. DC.) R. K. Jansen. (AP) is a medicinal plant with a local anesthetic effect. OBJECTIVE: To investigate the potential of rectal suppositories containing CQ and AP extracts to alleviate symptoms of hemorrhoids compared with the commercialized rectal suppository containing hydrocortisone and cinchocaine. MATERIALS AND METHODS: Hemorrhoid outpatients (n = 105) with different severity grades (I, II, or III) from eight hospitals in northern Thailand were included in this study. Hemorrhoid severity was graded by proctoscopy associated with either anal pain or bleeding related to hemorrhoids or both. The patients were randomly allocated to two groups: CQ-AP group (n = 52) or the commercialized rectal suppository group (n = 53). One suppository was rectally administered twice daily in the morning and at bedtime for seven days. Evaluations were performed by physicians on days 1, 4, and 8 of the study. The primary endpoints were bleeding and prolapse size, while the secondary endpoint was anal pain. RESULTS: Baseline demographics, lifestyle, constipation, number of prolapses, grade of hemorrhoid severity, and duration of experiencing hemorrhoids were comparable in both groups of patients. The effects of CQ-AP and the commercialized rectal suppository on bleeding, prolapse size, and anal pain were comparable. The patients in both groups were satisfied with both products at comparable levels and stated a preference for further use in the case of hemorrhoids recurrence. In terms of safety, the patients in the commercialized rectal suppository group experienced a higher incidence of adverse events, including anal pain and bleeding. CONCLUSION: Rectal suppositories containing a combined extract of CQ and AP show potential in alleviating hemorrhoidal symptoms with a good safety profile.

Specification and DNA Barcoding of Thai Traditional Remedy for Chronic Kidney Disease: Pikad Tri-phol-sa-mut-than.

The Pikad Tri-phol-sa-mut-than (TS) remedy, a Thai traditional medicine, is officially recorded in Tamra Paetsart Sonkrau Chabub Anurak for its capabilities in treating kidney deficiency. TS remedy is composed of three fruit species-Aegle marmelos (L.) Corrêa., Coriandrum sativum L., and Morinda citrifolia L.-in an equal part by weight. The quality of the raw material is one of the essential factors that can affect the effectiveness and safety of treatment by herbal remedy. The pharmacognostic evaluation and DNA barcode of the three fruit species and TS remedy were performed in this study to authenticate them from contamination, and to provide the scientific database for further uses. Macroscopic and microscopic examination, chemical profile by TLC, and DNA barcoding were employed to positively identify the raw materials bought from the herbal market, especially the powder form. Consequently, the outcomes of this investigation can be used to develop an essential and effective tool for the authentication of crude drugs and herbal remedies.

Antiviral medicinal plants found in Lanna traditional medicine.

Traditional medicine uses a multitude of plants to create medicinal formulations, some of which show antiviral properties that may be of benefit in treating emerging viral diseases, including Covid-19. Lanna, an ancient Kingdom in Northern Thailand, with a thriving culture that continues to this day and has a rich history of traditional medicine using local plants that is still practiced today. To find potential antiviral medicinal candidates, we examined ancient manuscripts, interviewed traditional healers practicing today, and inventoried current traditional medicines to catalogue 1400 medicinal formulations used in Lanna traditional medicine. We then narrowed this list to find those traditionally used to treat diseases that in their original use and descriptions most likely map to those we know today to be viral diseases. We identified the plants used in these formulations to create a list of 64 potential antiviral herbal candidates drawn from this ancient Lanna wisdom and matched these to the scientific literature to see which of these plants had already been shown to possess antiviral properties, generating a list of 64 potential antiviral medicinal candidates from Lanna traditional medicine worth further investigation for treating emerging viral diseases.

Injectable Thermosensitive Chitosan/Pullulan-Based Hydrogels with Improved Mechanical Properties and Swelling Capacity.

Thermosensitive chitosan/ß-glycerophosphate (CS/BGP) systems have been developed as injectable hydrogels. However, the hydrogels exhibited poor mechanical properties due to their physically crosslinked networks. In this work, CS/BGP hydrogels were reinforced by covalent crosslinking using genipin (GE) and concomitantly semi-interpenetrating networks using pullulan (PL). Based on response surface methodology, the optimized formulation was composed of CS (1.05%, w/v), PL (1%, w/v), BGP (6%, w/v), and GE (70.79 mcg/mL). The optimized hydrogels exhibited Young's modulus of 92.65 ± 4.13 kPa and a percentage of equilibrium swelling ratio of 3259.09% ± 58.90%. Scanning electron micrographs revealed a highly porous structure with nanofibrous networks in the CS/PL/BGP/GE hydrogels. The chemical interactions between the compositions were investigated by Fourier-transform infrared spectroscopy. Rheological measurements illustrated that the optimized hydrogels displayed sol-gel transition within one minute at 37 °C, a lower critical solution temperature of about 31 °C, and viscoelastic behavior with high storage modulus. Furthermore, the optimized hydrogels demonstrated higher resistance to in vitro enzymatic degradation, compared to the hydrogels without GE. Our findings could suggest that the thermosensitive CS/PL/BGP/GE hydrogels with enhanced mechanical properties and swelling capacity demonstrate the potential for use as scaffolds and carriers for cartilage tissue engineering and drug delivery applications.

Carry forward advantages of traditional medicines in prevention and control of outbreak of COVID-19 pandemic.

Members of the China-ASEAN Joint Laboratory for International Cooperation in Traditional Medicine Research used the video conference platform to exchange and discuss the advantages of traditional medicine through the form of score exchange and report, and research and develop the amount and issues of the therapeutic COVID-19 products of concern. This paper mainly reviews the achievements of the implementation of the epidemic prevention and control plan, advances of scientific basic studies on SARS-CoV-2, analysis and screening of potential targets and pathways of antiviral compounds based on network pharmacology and development of antiviral food dual-use products. The authors believe that the declaration of the (10 + 3) special meeting of national leaders on epidemic prevention and control should raise the medical and pharmaceutical issues of common concern. It is the responsibility of our joint laboratory members to accelerate the development of traditional medicine research and industry. Also the authors believe that this exchange will certainly promote the development of the cause of cooperation.

Depletion of ß-sitosterol and enrichment of quercetin and rutin in Cissus quadrangularis Linn fraction enhanced osteogenic but reduced osteoclastogenic marker expression.

BACKGROUND: Cissus quadrangularis Linn. (CQ) has been used in Indian and Thai traditional medicine for healing bone fractures because of numerous active ingredients in CQ. It is still unclear which compounds are the active ingredients for bone formation. METHODS: The molecular docking technique, the ethanolic extraction along with hexane fractionation, and an in vitro experiment with a human osteoblast cell line (MG-63) were used to narrow down the active compounds, to prepare the CQ extract, and to test biological activities, respectively. RESULTS: The molecular docking technique revealed that quercetin and ß-sitosterol had highest and lowest potential to bind to estrogen receptors, respectively. Compared to the crude ethanol extract (P1), the ethanolic fraction (P2) was enriched with rutin and quercetin at 65.36 ± 0.75 and 1.06 ± 0.12 mg/g, respectively. Alkaline phosphatase (ALP) activity was significantly enhanced in osteoblasts exposed to the P2 in both tested concentrations. The amount of hydroxyproline was slightly increased in the P1 treatment, while osteocalcin was inhibited. Moreover, the P2 significantly activated osteoprotegerin (OPG) and inhibited receptor activator of nuclear factor κ ligand (RANKL) expression. CONCLUSIONS: Taken together, the enriched rutin and quercetin fraction of CQ triggered the molecules involved in bone formation and the molecules inhibiting bone resorption.

Evaluation of white Kwao Krua (Pueraria candollei Grah. ex Benth.) products sold in Thailand by molecular, chemical, and microscopic analyses.

The tuberous roots of Pueraria candollei Grah. ex Benth. (Fabaceae), commonly known as white Kwao Krua, are used to relieve menopausal symptoms in Thai traditional medicine because they contain phytoestrogens. Black and red Kwao Krua crude drugs exist as well, but they have different botanical origins and pharmacological activities. There is a high demand for white Kwao Krua products, but because of the limited availability of the plant material, it is suspected that the adulteration and misidentification of white Kwao Krua crude drugs and products occur. In this study, we authenticated white Kwao Krua products collected from Thai herbal markets by molecular, chemical, and microscopic analyses. The nucleotide sequences in the internal transcribed spacer (ITS) and trnH-psbA regions of 23 samples of authentic P. candollei were analyzed, and both regions were found to have intraspecific DNA polymorphisms. Based on the single nucleotide polymorphisms in the ITS1 region, species-specific primer sets of P. candollei were designed to authenticate white Kwao Krua and differentiate it from red and black Kwao Krua. Only the PCR products of KWP02 were not amplified by the primer sets. Isoflavonoid contents and microscopic features were used to support the results of molecular analysis to clarify the botanical origin of white Kwao Krua. Molecular, chemical and microscopic methods confirmed that all the Thai Kwao Krua products examined in this study contained authentic "white Kwao Krua" as claimed on their labels.

The combination of ITS2 and psbA-trnH region is powerful DNA barcode markers for authentication of medicinal Terminalia plants from Thailand.

The dried fruits of Terminalia plant (Combretaceae) called "Samo" have been used as herbal medicine in Thai traditional medicine. Four "Samo" crude drugs, namely, Samo thai, Samo thed, Samo dee-ngu, and Samo phiphek, are used as the main ingredients in Triphala and Trisamo recipes. Their commercial products are available in processed and powdered form, but are difficult to authenticate by conventional methods. In this study, we aimed to discriminate species of genus Terminalia for the identification of their crude drugs by a DNA barcoding technique. A total of 208 closely related nucleotide sequences were obtained from nine Terminalia species collected from Thailand and the DDBJ/EMBL/GenBank database. An effective DNA barcode marker was selected from six DNA loci (matK, rbcL, psbA-trnH, ITS, ITS1, and ITS2) and their two-locus combination. All sequences were analyzed by three major methods: (1) BLAST search; (2) the genetic divergence method using Kimura 2-parameter (K2P) distance matrices; and (3) tree topology analysis based on the neighbor-joining method. Comparison of the six candidate DNA loci indicated that ITS identified Terminalia with 100% accuracy at the species and genus levels in the BLAST1 method. ITS2 showed the highest K2P variability. The data from the single markers and the two-locus combinations revealed that only the two-locus combinations, namely, the combinations of rbcL, ITS, ITS1, and ITS2 with psbA-trnH, clearly discriminated all the species. From the results of DNA sequence analysis and the three methods, ITS2 is recommended for the identification of Terminalia species to supplement psbA-trnH.

Influence of water-soluble channeling agents on the release of diclofenac sodium from Irvingia malayana wax matrix tablets.

Irvingia malayana wax (IW) is majorly composed of esters of medium chain fatty acids. Its melting point is low and closed to the body temperature. This study aimed at investigating the potential of IW as a matrix-forming agent and evaluate the effect of soluble channeling agents on the release of diclofenac sodium (DS) from IW matrix tablets. The preformulation study by infrared spectroscopy and differential scanning calorimetry showed no incompatibility between IW and DS or soluble channeling agents, namely PEG 4000, PEG 6000 and lactose. IW retarded the release of DS from the matrix tablets more efficiently than carnauba wax due to its greater hydrophobicity and its ability to become partial molten wax at 37° C. Factors affecting the release of DS from IW matrix were drug concentrations, and types and concentrations of channeling agents. The release of DS significantly improved when DS concentration reached approximately 33%. The fast dissolving channeling agent, lactose, could enhance the drug release rate more effectively than PEG 4000 and PEG 6000, respectively. The linear relationship between the DS release rate and the concentration of the chosen channeling agent, PEG 6000, was found (r2=0.9866).

Molecular Analysis of Terminalia spp. Distributed in Thailand and Authentication of Crude Drugs from Terminalia Plants.

Terminalia, a large genus of Combretaceae, is distributed in Tropical Asia, Africa, and America. Some Terminalia plants are used in folk medicine because they possess powerful medicinal properties. Dried fruits of Terminalia bellirica and Terminalia chebula are used as the main ingredient in Triphala, a famous polyherbal formulation in Ayurvedic medicine and Thai folk medicine, because of their laxative, detoxifying, and rejuvenating effects. To clarify the phylogenetic relationships of medicinal Terminalia species (T. bellirica, T. chebula, and T. catappa) and authenticate their crude drugs, "Samo" and Triphala, nucleotide sequencing alignments in the internal transcribed spacer one-two (ITS 1-2) regions of Terminalia plants collected in Thailand were performed. The amplified fragments of Terminalia species were approximately 800 bp in length. To compare these sequences and DDBJ registered data, a molecular phylogenetic tree was constructed. Phylogenetic analysis clearly separated the sequences into two groups: Asian Terminalia and African Terminalia with some exceptions. In the analyzed sequences, the length of the ITS1-5.8S-ITS2 region was 674 bp in T. chebula, and 677 bp in T. bellirica and T. catappa. Eighty-one single nucleotide polymorphisms (SNPs) and nine insertion-deletions (indels) were observed, and the nucleotide sequences of this region showed species-specific sequences. Based on these differences, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and amplification refractory mutation system (ARMS) were applied to identify medicinal Terminalia species. Moreover, the ARMS method was chosen for fingerprinting analysis of Samo crude drugs and Triphala formulations because it was a fast, cost-effective, and reproducible approach.

Medicinal plants of the Mien (Yao) in Northern Thailand and their potential value in the primary healthcare of postpartum women.

AIM OF THE STUDY: To study the use of medicinal plants by the Mien in Nan Province and their potential value in the primary healthcare of postpartum women. Also, to survey the medicinal plant resources present there. MATERIALS AND METHODS: Free listing and interviews were used with four key informants (herbalists and collectors) to collect all qualitative and quantitative plant data. Semi-structured questionnaires were used to obtain information on the knowledge, attitude and practices of fifty-eight non-specialist informants. Transect walks of forest plots were carried out with herbalists to get more information on the status of the medicinal plants with regard to their habitat. RESULTS AND CONCLUSION: More than 168 species of medicinal plants were surveyed. These plants belonged to 80 families and 145 genera, of which 131 were wild plants and 37 species have been cultivated in home gardens. The interview data from four herbalists and fifty-eight non-specialist informants indicated that the majority of non-specialist informants who used medicinal plants were women and the most common usage categories were for birth related conditions (44 species, 26.2%). The most common method of preparation was decoction for both oral consumption and bathing uses (134 species, 79.8%). The most common species of medicinal plants were used in a postpartum herbal bath formulae and in food supplement formulas. These were Anredera cordifolia (Ten.) Steenis, Basella alba L., Ricinus communis L., Poikilospermum suaveolens (L.) Merr., Gouania leptostachya DC. Var. leptostachya, Schefflera sp. cf. Schefflera bengalensis Gamb., Blumea balsamifera (L.) DC., Chromolaena odoratum (L.) King et Robin and Cymbopogon citratus (DC.) Stapf.

Quantitative analysis of oxyresveratrol content in Artocarpus lakoocha and 'Puag-Haad'.

OBJECTIVE: To develop a thin-layer chromatography (TLC) densitometric method for the determination of oxyresveratrol content in Artocarpuslakoocha heartwood and in the traditional drug 'Puag-Haad'. MATERIALS AND METHODS: Sample solution of A. lakoocha heartwood was prepared by Soxhlet extraction of the plant material in ethanol, whereas the Puag-Haad solution was obtained by dissolving the drug in methanol. Analysis of each sample solution was performed on a Silica gel 60 F(254) TLC plate (20 x 10 cm) with methylene chloride/methanol (85:15) as the mobile phase. After development, the TLC plate was examined with a TLC scanner in the absorbance mode at 254 nm. The newly developed analytical method was validated using an authentic sample of oxyresveratrol previously isolated from A. lakoocha heartwood, and was used to analyze the oxyresveratrol content in samples of A. lakoocha heartwood and the traditional drug Puag-Haad. RESULTS: A sensitive and reliable TLC densitometric method was successfully developed. The method was validated in terms of accuracy (99.11-102.60%) and precision (1.66-4.23% coefficient of variation). The limits of detection and quantitation were 15.6 and 52 ng/spot, respectively. The amounts of oxyresveratrol in 3 samples of A. lakoocha heartwood collected from its natural habitat were 49.0-182.3 mg/g, whereas those in 11 commercial samples were in the range of 23.4-69.6 mg/g. The oxyresveratrol contents in 2 samples of traditional drug Puag-Haad were 780.1 and 837.5 mg/g. CONCLUSION: The TLC densitometric method developed in this study is a simple, convenient, sensitive and reliable procedure. It was an effective analytical tool for the evaluation of oxyresveratrol content in both A. lakoocha heartwood and the traditional drug Puag-Haad.