Identifying potential immuno-oncology targets in salivary gland mucoepidermoid carcinoma based on inflammatory status and treatment response.

BACKGROUND: Mucoepidermoid carcinoma is a rare salivary gland malignant tumour. This study aimed to investigate inflammatory and immune signatures of mucoepidermoid carcinoma by identifying potential proteo-transcriptomic biomarkers towards the development of precision immuno-oncology treatment strategies. METHODS: A total of 30 biopsies obtained from patients diagnosed with mucoepidermoid carcinoma between 2013 and 2022 were analysed after H&E staining for scoring of histological inflammatory stroma subtypes and inflammatory hotspots with QuPath. Multiplex immunofluorescence staining and NanoString nCounter PanCancer IO 360™ panel were used to assess stroma and tumour inflammation signatures in high grade mucoepidermoid carcinoma cases in the tumour microenvironment via proteomics and transcriptomics, respectively. RESULTS: Inflammatory cells within the histological inflammatory stroma inflammatory (HIS-INF/hot) tumour neighbourhoods were greater compared to the histological inflammatory stroma-immune desert (HIS-ID/cold) (p = 0.001). A similar trend was observed between treatment non-responders and responders in stroma neighbourhoods (p = 0.0625) and in stroma-to-interface inflammatory hotspots (p = 0.0081), indicating an augmented inflammatory response in hot tumours and non-responders. Furthermore, there were striking differences in the expression of pan-immune leukocyte marker CD45 between responders and non responders particularly in the tumour neighbourhoods (p = 0.0341), but such were not robust for PD-1 and macrophage fractions. Additionally, transcriptomic analysis revealed key differences in leukocyte activation profiles between responders and non-responders. CONCLUSION: This preliminary report unveils the importance of assessing immune leukocyte cellular fractions and pathways for future prognostic biomarker discoveries in mucoepidermoid carcinoma as per the involvement of CD45-driven inflammatory and immune mediators in high grade mucoepidermoid carcinoma in non-responders to treatment. These findings will potentially contribute to the development of novel personalised immunotherapies.

A comprehensive review on Saraca asoca (Fabaceae) - Historical perspective, traditional uses, biological activities, and conservation.

ETHNOPHARMACOLOGICAL RELEVANCE: Saraca asoca (Roxb.)W.J.de Wilde, (Fabaceae) is a plant of significant medicinal value in traditional Indian medicine, with a long history of use in the treatment of gynaecological disorders and other ailments, and is held in high esteem. This plant has long existed in Indian tradition and is revered as sacred. AIM OF THE STUDY: This work aimed to explore the taxonomic revision of Saraca asoca from ancient times to the present and to evaluate the ethnobotanical, phytochemical and pharmacological information associated with traditional use and develop a roadmap for conservative strategies of species. MATERIALS AND METHODS: The study draws on a comprehensive range of herbal, traditional, ethnobotanical, and ethnopharmacological information, including ancient Ayurvedic textbooks and various databases, using a single keyword or a combination of multiple keywords. RESULTS: This review establishes a roadmap for understanding the traditional history of medicinal plants, particularly Saraca, and highlights the transfer of traditional knowledge from pharmacopoeias, materia medica, and classical textbooks over many centuries. The study also emphasises the importance of conservation strategies to protect Saraca as a valuable resource for healthcare and suggests that more research is needed to systematically evaluate its phytochemical, pharmacological, and clinical properties, as well as to develop safety, pharmacology, and toxicology reports for traditional formulations. CONCLUSIONS: In light of this study, S. asoca could be considered an important source of potential herbal drugs. The review concludes with a call for further research and conservation efforts to protect Saraca and other traditional medicinal plants for the benefit of current and future generations.

Effect of Pulse Electrodeposition Parameters on the Microstructure and Mechanical Properties of Ni-W/B Nanocomposite Coatings.

In this work, Ni−W/B nanocomposite coatings were successfully fabricated on low carbon steel by using pulse current (PC) electrodeposition. The effects of the frequency and duty cycle on the microstructure, wear resistance, and microhardness of the coatings were studied. The results obtained show that the distribution and content of boron particles (>4 wt.%) in the PC electrodeposition coatings are significantly better than those of direct current (DC) electrodeposition coatings (less than 4 wt.%). The hardness results reveal that the highest microhardness of 1122 HV can be obtained at a frequency of 100 Hz and duty cycle of 30%. Furthermore, the relationship between the microstructure and mechanical properties was discussed.

Development of a DNA barcode library of plants in the Thai Herbal Pharmacopoeia and Monographs for authentication of herbal products.

Traditional herbal medicine has long been practiced as a method of health care in many countries worldwide. The usage of herbal products has been increasing and is expected to continue to do so in the future. However, admixture and adulteration are concerns regarding the quality of herbal medicine, including its safety and efficacy. We aimed to develop a reference DNA barcode library of plants listed in the Thai Herbal Pharmacopoeia (THP) and Monographs of Selected Thai Materia Medica (TMM) (n = 101 plant species) using four core barcode regions, namely, the ITS2, matK, rbcL and trnH-psbA intergenic spacer regions, for authentication of the plant origin of raw materials and herbal products. Checking sequences from samples obtained from local markets and the Thai Food and Drug Administration (Thai FDA) against our digital reference DNA barcode system revealed the authenticity of eighteen out of twenty tested samples as claimed on their labels. Two samples, no. 3 and 13, were not Cyanthillium cinereum (L.) H.Rob. and Pueraria candollei Wall. ex Benth. as claimed, respectively. They were recognized as Emilia sonchifolia (L.) DC. and Butea superba (Roxb.), respectively. Hence, it is important for the Thai FDA or regulatory agencies to immediately initiate strict enforcement for the development of pharmacopoeial standards as well as revisions or modifications of available regulatory guidelines and to implement close monitoring for the quality control of herbal products in terms of authentication before they enter the herbal market. The centralized digital reference DNA barcode database developed here could play a very important role in monitoring or checking the authenticity of medicinal plants.

Antiproliferative Effects of Artabotrys odoratissimus Fruit Extract and its Bioactive Fraction through Upregulation of p53/γH2AX Signals and G2/M Phase Arrest in MIA PaCa-2 Cells.

BACKGROUND: Artabotrys odoratissimus (Annonaceae) is a medicinal and ornamental plant widely cultivated in Southeast Asia for its famous ylang ylang essential oil. The fruits of this plant are used for health benefits, but very little is studied about the bioactive principles, their role in regulating oxidative stress and tumour progression. OBJECTIVE: The study aimed to evaluate the antiproliferative effects of fruit extract of Artabotrys odoratissimus and its bioactive fraction using cell-based assays. METHODS: The free radical scavenging and antiproliferative effects of Artabotrys odoratissimus fruit ethyl acetate (FEA) extract and its bioactive fraction were evaluated using cell viability assays, colony formation assay, double staining assay, reactive oxygen species (ROS) assay, comet assay, cell cycle analysis, and western blotting. RESULTS: The extract showed phenolic content of 149.8±0.11µg/mg Gallic acid equivalents and flavonoid content of 214.47±4.18 µg/mg Quercetin. FEA showed an IC50 value of 76.35 µg/ml in the ABTS assay and an IC50 value of 134.3±7.8 µg/ml on MIA PaCa-2 cells. The cells treated with 125 µg/ml and 250 µg/ml FEA showed increased apoptotic cells in Double staining assay, DNA damage during comet assay, enhanced ROS, and cell cycle arrest at G2M phase at 125 µg/ml and 250 µg/ml. The active fraction AF5 showed an IC50 value of 67±1.26 µg/ml on MIA PaCa-2 cells during MTT assay, displayed potential antiproliferative effects, and showed a marked increase in the expression of γH2AX and p53. CONCLUSION: These results prove that the fruit extract and the bioactive fraction demonstrate oxidative stress-mediated DNA damage, leading to apoptosis in the MIA PaCa-2 cell line.

Differentiation of Mitragyna speciosa, a narcotic plant, from allied Mitragyna species using DNA barcoding-high-resolution melting (Bar-HRM) analysis.

Mitragyna speciosa (Korth.) Havil. [MS], or "kratom" in Thai, is the only narcotic species among the four species of Mitragyna in Thailand, which also include Mitragyna diversifolia (Wall. ex G. Don) Havil. [MD], Mitragyna hirsuta Havil. [MH], and Mitragyna rotundifolia (Roxb.) O. Kuntze [MR]. M. speciosa is a tropical tree belonging to the Rubiaceae family and has been prohibited by law in Thailand. However, it has been extensively covered in national and international news, as its abuse has become more popular. M. speciosa is a narcotic plant and has been used as an opium substitute and traditionally used for the treatment of chronic pain and various illnesses. Due to morphological disparities in the genus, the identification of plants in various forms, including fresh leaves, dried leaf powder, and finished products, is difficult. In this study, DNA barcoding combined with high-resolution melting (Bar-HRM) analysis was performed to differentiate M. speciosa from allied Mitragyna and to assess the capability of Bar-HRM assays to identify M. speciosa in suspected kratom or M. speciosa-containing samples. Bar-HRM analysis of PCR amplicons was based on the ITS2, rbcL, trnH-psbA, and matK DNA barcode regions. The melting profiles of ITS2 amplicons were clearly distinct, which enabled the authentication and differentiation of Mitragyna species from allied species. This study reveals that DNA barcoding coupled with HRM is an efficient tool with which to identify M. speciosa and M. speciosa-containing samples and ensure the safety and quality of traditional Thai herbal medicines.

DNA metabarcoding to unravel plant species composition in selected herbal medicines on the National List of Essential Medicines (NLEM) of Thailand.

Traditional medicines are widely traded across the globe and have received considerable attention in the recent past, with expectations of heightened demand in the future. However, there are increasing global concerns over admixture, which can affect the quality, safety, and efficacy of herbal medicinal products. In this study, we aimed to use DNA metabarcoding to identify 39 Thai herbal products on the Thai National List of Essential Medicines (NLEM) and assess species composition and admixture. Among the products, 24 samples were in-house-prepared formulations, and 15 samples were registered formulations. In our study, DNA metabarcoding analysis using ITS2 and rbcL barcode regions were employed to identify herbal ingredients mentioned in the products. The nuclear region, ITS2, was able to identify herbal ingredients in the products at the genus- and family-levels in 55% and 63% of cases, respectively. The chloroplast gene, rbcL, enabled genus- and family-level identifications in 58% and 73% of cases, respectively. In addition, plant species were detected in larger numbers (Family identified, absolute %) in registered herbal products than in in-house-prepared formulations. The level of fidelity increases concerns about the reliability of the products. This study highlights that DNA metabarcoding is a useful analytical tool when combined with advanced chemical techniques for the identification of plant species in highly processed, multi-ingredient herbal products.

Differentiation of Cyanthillium cinereum, a smoking cessation herb, from its adulterant Emilia sonchifolia using macroscopic and microscopic examination, HPTLC profiles and DNA barcodes.

Cyanthillium cinereum (L.) H.Rob. is one of the most popular herbal smoking cessation aids currently used in Thailand, and its adulteration with Emilia sonchifolia (L.) DC. is often found in the herbal market. Therefore, the quality of the raw material must be considered. This work aimed to integrate macro- and microscopic, chemical and genetic authentication strategies to differentiate C. cinereum raw material from its adulterant. Different morphological features between C. cinereum and E. sonchifolia were simply recognized at the leaf base. For microscopic characteristics, trichome and pappus features were different between the two plants. HPTLC profiles showed a distinct band that could be used to unambiguously differentiate C. cinereum from E. sonchifolia. Four triterpenoid compounds, ß-amyrin, taraxasterol, lupeol, and betulin, were identified from the distinct HPTLC band of C. cinereum. The use of core DNA barcode regions; rbcL, matK, ITS and psbA-trnH provided species-level resolution to differentiate the two plants. Taken together, the integration of macroscopic and microscopic characterization, phytochemical analysis by HPTLC and DNA barcoding distinguished C. cinereum from E. sonchifolia. The signatures of C. cinereum obtained here can help manufacturers to increase the quality control of C. cinereum raw material in commercialized smoking cessation products.

Mitigating the Impact of Admixtures in Thai Herbal Products.

Medicinal plants and their products are extensively used within indigenous healthcare systems in Thailand and several other nations. The international trade of herbal products has a noteworthy impact on the worldwide economy, and the interest in herbal products is expanding in both developing and developed countries. There has been rapid growth in the medicinal plant product market and a broadening consumer base interested in herbal products from Thailand. However, in herbal industries, ingredient substitution and admixture are typical issues wherein species of lower market value are admixed with those of a higher value. The adverse consequences of consuming adulterated drugs are invariably due to the presence of an unintended herb rather than the presence of an intended herb. It has also been argued that admixtures are intentional because of the lack of regulatory policies or centralized tests for product authentication. The consequences of species admixtures can extend from the reduced efficacy of a drug to decreased trade value. This study aims to clarify the nature and extent of species admixtures reported in the Thai herbal trade market and discuss the potential reasons for such adulteration. In the broader context of species admixtures, we strongly propose the establishment of multiple herbal crude drug repositories that can be developed to facilitate the use of comparative identity tests by industry, traders, and researchers to maintain authentic natural health product (NHP) standards and to certify the authenticity of NHPs. The proposition of the establishment of centralized testing (CT) could be a promising initiative in Thailand for the development of science and technology, and the herbal medicines produced as a result of CT could be dispensed as prescription drugs based on disease consideration instead of as health foods or nutraceuticals.

Authentication of Garcinia fruits and food supplements using DNA barcoding and NMR spectroscopy.

Garcinia L. (Clusiaceae) fruits are a rich source of (-)-hydroxycitric acid, and this has gained considerable attention as an anti-obesity agent and a popular weight loss food supplement. In this study, we assessed adulteration of morphologically similar samples of Garcinia using DNA barcoding, and used NMR to quantify the content of (-)-hydroxycitric acid and (-)-hydroxycitric acid lactone in raw herbal drugs and Garcinia food supplements. DNA barcoding revealed that mostly G. gummi-gutta (previously known as G. cambogia) and G. indica were traded in Indian herbal markets, and there was no adulteration. The content of (-)-hydroxycitric acid and (-)-hydroxycitric acid lactone in the two species varied from 1.7% to 16.3%, and 3.5% to 20.7% respectively. Analysis of ten Garcinia food supplements revealed a large variation in the content of (-)-hydroxycitric acid, from 29 mg (4.6%) to 289 mg (50.6%) content per capsule or tablet. Only one product contained quantifiable amounts of (-)-hydroxycitric acid lactone. Furthermore the study demonstrates that DNA barcoding and NMR could be effectively used as a regulatory tool to authenticate Garcinia fruit rinds and food supplements.