Diaporthe phaseolorum causing dieback disease on Melia dubia cav. in Karnataka state (India).

Melia dubia is an important tree species grown worldwide for its medicinal and timber values. It is widely used in timber and pulp industry and also as an organic pesticide, fertilisers, agro-forestry and herbal formulations. During 2019-2022, a dieback disease in plantations of M. dubia was recorded in Mysore, Mandya, Chamarajanagar, Hassan and Tumkur districts of Karnataka state (India) with disease incidence of 26.25%. The associated pathogen was isolated on PDA medium and its morpho-cultural characteristics were studied. The genomic DNA of the pathogen was isolated, and rDNA was amplified and sequenced using universal primers. Based on the microscopic, morpho-cultural, sequence data and phylogenetic analysis, the pathogen was identified as Diaporthe phaseolorum (Cooke & Ellis) Sacc. Koch's postulates were performed both in vitro and in vivo and the typical symptoms of dieback disease were recorded on post-inoculated saplings. The dieback disease is responsible for the poor growth of Melia species in the region, and hence, there is an urgent need to manage the disease in plantations using integrated management practices. This is the first report of the occurrence of D. phaseolorum on M. dubia plantations in India.

Differential gene expression analysis reveals the fast-growth mechanisms in Melia dubia at different stand ages.

BACKGROUND: Melia dubia Cav. is a fast-growing multipurpose tree suitable for agroforestry and has been widely cultivated for wood-based industries, particularly pulp and paper production. Despite its high economic value in India, there is a lack of information regarding the molecular mechanism driving its fast-growth. Therefore, this study aimed to elucidate the molecular mechanisms responsible for fast-growth by expression analysis of selective candidate genes. METHODS AND RESULTS: Initially, growth traits were assessed, including tree height and diameter at breast height (DBH), across three different ages (one-year-old, two-year-old, and three-year-old) of M. dubia plantations. Tree volume based on tree height and DBH, was also calculated. The analysis of annual tree height increment revealed that the second-year plantation exhibited the higher increment, followed by first and third years. In contrast, DBH was maximum in third-year plantation, followed by the second and first years. Similarly, annual tree volume increment showed a similar trend with DBH that maximum in the third year, followed by second and first years. Furthermore, a differential gene expression analysis was performed using qRT-PCR on four genes such as Phloem Intercalated with Xylem (PXY), Clavata3/Embryo Surrounding Region-Related 41 (CLE41), 1-aminocyclopropane-1-carboxylic acid synthase (ACS-1) and Hemoglobin1 (Hb1) for downstream analysis. The relative gene expression showed up-regulation of CLE41, ACS-1, and Hb1 genes, while the PXY gene was downregulated across the tree ages. Interestingly, a positive association was observed between tree growth and the expression of the selected candidate genes. CONCLUSION: Our results pave the way for further research on the regulatory mechanisms of genes involved in fast-growth and provide a basis for genetic improvement of Melia dubia.

Development of a medication literacy assessment instrument (MELIA) for older people receiving home care.

AIM: To develop a consensus-based instrument [MELIA] to assess the medication literacy of older home care patients to ultimately optimize medication safety. DESIGN: This study was part of the project 'Study of Medication Safety in Home Care' (doMESTIC), which took place from 2016 to 2020 in Switzerland. The development process for the medication literacy assessment instrument encompassed six steps. METHOD: First, a scoping literature search was conducted in the Pubmed, CINAHL, EMBASE and Cochrane Library databases as 2) a basis for the development of assessment items. This was followed by 3) a cognitive interview with home care patients and 4) the first round of a Delphi process. Then, 5) a focus group interview with home care experts was conducted before 6) the second Delphi round. The project took place between August 2020 and June 2021. With these different steps, perspectives of both patients and various home care and medication safety experts were included in the development of the assessment instrument. RESULTS: A detailed instrument consisting of 20 items as well as a 7-item short version were developed. The short version is intended for efficient preliminary screening to identify patients at high risk for medication management-related problems. CONCLUSION: Medication literacy in patients 65 years and older receiving professional home care is a key issue in preventing medication errors. A targeted assessment, starting with an efficient short version of MELIA, allows for prioritization of patients for interventions to optimize medication safety while ensuring their independence as much as possible. IMPACT: Systematic assessment of patients' medication literacy helps to provide them with targeted and individual support in their medication management to avoid medication errors and increase patient safety. The development of MELIA is a first step in providing an assessment instrument specifically for the home care setting. PATIENT OR PUBLIC CONTRIBUTION: Patient participation was an integral part of the instrument development. The initial 23 items were optimized based on cognitive interviews with four home care patients. The next steps of the instrument development were based on feedback of health care professionals-encompassing advance practice nurses, regular nurses, pharmacists and general practitioners-during a two-step Delphi process as well as a focus group discussion.

Insecticidal Triterpenes in Meliaceae: Plant Species, Molecules, and Activities: Part II (Cipadessa, Melia).

Plant-originated triterpenes are important insecticidal molecules. Research on the insecticidal activity of molecules from Meliaceae plants has always been a hotspot due to the molecules from this family showing a variety of insecticidal activities with diverse mechanisms of action. In this paper, we discussed 116 triterpenoid molecules with insecticidal activity from 22 plant species of five genera (Cipadessa, Entandrophragma, Guarea, Khaya, and Melia) in Meliaceae. In these genera, the insecticidal activities of plants from Entandrophragma and Melia have attracted substantial research attention in recent years. Specifically, the insecticidal activities of plants from Melia have been systemically studied for several decades. In total, the 116 insecticidal chemicals consisted of 34 ring-intact limonoids, 31 ring-seco limonoids, 48 rearranged limonoids, and 3 tetracyclic triterpenes. Furthermore, the 34 ring-intact limonoids included 29 trichilin-class chemicals, 3 azadirone-class chemicals, and 1 cedrelone-class and 1 havanensin-class limonoid. The 31 ring-seco limonoids consisted of 16 C-seco group chemicals, 8 B,D-seco group chemicals, 4 A,B-seco group chemicals, and 3 D-seco group chemicals. Furthermore, among the 48 rearranged limonoids, 46 were 2,30-linkage group chemicals and 2 were 10,11-linkage group chemicals. Specifically, the 46 chemicals belonging to the 2,30-linkage group could be subdivided into 24 mexicanolide-class chemicals and 22 phragmalin-class chemicals. Additionally, the three tetracyclic triterpenes were three protolimonoids. To sum up, 80 chemicals isolated from 19 plant species exhibited antifeedant activity toward 14 insect species; 18 chemicals isolated from 17 plant species exhibited poisonous activity toward 10 insect species; 16 chemicals isolated from 11 plant species possessed growth-regulatory activity toward 8 insect species. In particular, toosendanin was the most effective antifeedant and insect growth-regulatory agent. The antifeedant activity of toosendanin was significant. Owing to its high effect, toosendanin has been commercially applied. Three other molecules, 1,3-dicinnamoyl-11-hydroxymeliacarpin, 1-cinnamoyl-3-methacryl-11-hydroxymeliacarpin, and 1-cinnamoyl-3-acetyl-11-hydroxymeliacarpin, isolated from Meliaazedarach, exhibited a highly poisonous effect on Spodoptera littoralis; thus, they deserve further attention.

Study protocol of a randomized, double-blind, placebo-controlled, multi-center trial to treat antipsychotic-induced weight gain: the Metformin-Lifestyle in antipsychotic users (MELIA) trial.

BACKGROUND: Antipsychotic-induced Weight Gain (AiWG) is a debilitating and common adverse effect of antipsychotics. AiWG negatively impacts life expectancy, quality of life, treatment adherence, likelihood of developing type-2 diabetes and readmission. Treatment of AiWG is currently challenging, and there is no consensus on the optimal management strategy. In this study, we aim to evaluate the use of metformin for the treatment of AiWG by comparing metformin with placebo in those receiving treatment as usual, which includes a lifestyle intervention. METHODS: In this randomized, double-blind, multicenter, placebo-controlled, pragmatic trial with a follow-up of 52 weeks, we aim to include 256 overweight participants (Body Mass Index (BMI) > 25 kg/m2) of at least 16 years of age. Patients are eligible if they have been diagnosed with schizophrenia spectrum disorder and if they have been using an antipsychotic for at least three months. Participants will be randomized with a 1:1 allocation to placebo or metformin, and will be treated for a total of 26 weeks. Metformin will be started at 500 mg b.i.d. and escalated to 1000 mg b.i.d. 2 weeks thereafter (up to a maximum of 2000 mg daily). In addition, all participants will undergo a lifestyle intervention as part of the usual treatment consisting of a combination of an exercise program and dietary consultations. The primary outcome measure is difference in body weight as a continuous trait between the two arms from treatment inception until 26 weeks of treatment, compared to baseline. Secondary outcome measures include: 1) Any element of metabolic syndrome (MetS); 2) Response, defined as ≥5% body weight loss at 26 weeks relative to treatment inception; 3) Quality of life; 4) General mental and physical health; and 5) Cost-effectiveness. Finally, we aim to assess whether genetic liability to BMI and MetS may help estimate the amount of weight reduction following initiation of metformin treatment. DISCUSSION: The pragmatic design of the current trial allows for a comparison of the efficacy and safety of metformin in combination with a lifestyle intervention in the treatment of AiWG, facilitating the development of guidelines on the interventions for this major health problem. TRIAL REGISTRATION: This trial was registered in the Netherlands Trial Register (NTR) at  https://www.trialregister.nl/trial/8440 as NTR NL8840 on March 8, 2020.

MS diagnostic model and rapid distinguishing of bioactive limonoids in fruits of Melia toosendan using solid-phase extraction coupled with LC-MS/MS.

INTRODUCTION: Melia toosendan Sieb. et Zucc. has been used as a Chinese folk medicine for roundworm treatment since ancient times. Many diverse limonoids have been isolated from Meliaceae plants, but it remains difficult to isolate and identify other limonoids because of their small natural concentrations. OBJECTIVE: This study was performed to overcome the difficulties associated with fast and accurate identification of limonoids and establish a reliable and sensitive method for the analysis of minor limonoids in M. toosendan fruits. METHODS: An efficient strategy for enrichment, detection, and identification of minor limonoids from M. toosendan fruits using solid-phase extraction with high-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (SPE-HPLC-Q-TOF-MS/MS) was developed herein. RESULTS: Characteristic fragmentations and fragmentation ions containing trichilin-, nimbin-, and vilasinin-class limonoid skeletons were initially studied, and characteristic diagnostic ions involved retro Diels-Alder (RDA) reactions or homolytic cleavages, which were used to identify minor limonoids. In total, 13 limonoids, including four new ones, were identified. CONCLUSION: This is the first report on the analysis of M. toosendan fruits to identify limonoids. This novel analysis method may stimulate further research regarding the identification of limonoids in other plant species.

Aeromicrobium endophyticum sp. nov., a novel endophytic actinobacterium isolated from bark of Melia azedaeach L.

A Gram-staining-positive, aerobic, rod-shaped, non-spore-forming actinobacterium, designated strain M2KJ-4T, was isolated from a surface-sterilized bark of Melia azedaeach L. collected from Xinpu in Guizhou, PR China and characterized using a polyphasic approach to determine its taxonomic position. M2KJ-4T grew optimally with 1 % (w/v) NaCl at 25 °C and pH 8.0. Substrate mycelia and aerial mycelia were not formed, and no diffusible pigments were observed on the media tested. Phylogenetic analysis based on 16S rRNA gene sequence indicated that M2KJ-4Trepresented a member of the genus Aeromicrobium and shared the highest 16S rRNA gene sequence similarity with Aeromicrobium fastidiosum DSM 10552T (Z78209) (98.95 %). The DNA G+C content of M2KJ-4T was 70.6 mol%. The average nucleotide identity value and estimated DDH value between M2KJ-4T and the type strain of A. fastidiosum were 86.1 % and 30.2 %, respectively. The cell-wall peptidoglycan contained ll-diaminopimelic acid and MK-9(H4) was the predominant menaquinone. The predominant polar lipids were diphosphatidylglycerol, phosphatidylglycerol, unidentified phospholipids and unidentified lipids. The major fatty acids were C16 : 0, 10-methyl C18 : 0, C16 : 0 2-OH and C18 : 1ω9c. On the basis of the results from phylogenetic, phenotypic and chemotaxonomic analysis, strain M2KJ-4T represents a novel species of the genus Aeromicrobium, for which the name Aeromicrobium endophyticum sp. nov. is proposed. The type strain is M2KJ-4T (=KCTC 49174T=CGMCC 1.13666T).

Characterization of a 2,3-oxidosqualene cyclase in the toosendanin biosynthetic pathway of Melia toosendan.

Toosendanin, bearing a furan ring, is a limonoid belonging to the group of tetranortriterpenoids. Toosendanin is a phytochemical found in the medicinal plant Melia toosendan Sieb. et Zucc. of the Meliaceae family. Toosendanin and its derivatives demonstrate high insecticidal activity and are important pesticides derived from plants. Despite intensive investigation of limonoids over several decades, the biosynthetic pathway of these triterpenoids is less understood. To identify the key enzymes involved in the toosendanin biosynthetic pathway, we analyzed the contents of toosendanin in various plant tissues and parts and found that the highest level of toosendanin was found in the developing fruit and gradually decreased as the fruit matured. More than 346 116 transcripts were assembled based on 394 million paired-end Illumina reads and 6 million PacBio reads from the pooled RNA samples of fruits, leaves and young barks. A total of 186 263 genes were predicted. Six 2,3-oxidosqualene cyclase (OSC) genes were identified by analyzing the association between gene expression and metabolite profiles. Functional analyses using the Nicotiana benthamiana transient expression assay showed that MtOSC1 catalyzed 2,3-oxidosqualene to produce a tetracyclic triterpene skeleton, tirucalla-7,24-dien-3ß-ol, which is predicted as the precursor for toosendanin biosynthesis. We identified another OSC, MtOSC6, which is a lupeol synthase. Using synthetic biology methods, these identified enzymes could be used to model a biosynthetic pathway to produce large quantities of toosendanin.

Characterization of Limonoids Isolated from the Fruits of Melia toosendan and Their Antifeedant Activity against Pieris rapae.

The fruits of Melia toosendan Sieb. et Zucc. (Meliaceae) are a source of bioactive limonoids that can be used as effective pesticides. In this study, two novel limonoids, 6-acetylsendanal and 6-ketocinamodiol, were isolated together with fourteen known compounds, namely four protolimonoids, six trichilin-class limonoids, and four C-seco limonoids. The structures of the new compounds were determined by extensive spectroscopic analyses (HR-ESI-MS, UV, IR, 1D and 2D NMR). The bioassay results revealed that eleven of the extracted limonoids exhibited interesting antifeedant activities against the larvae of Pieris rapae with AFC50 values in the range of 0.11-1.79 mm. Particularly, mesendanin H, with an AFC50 value of 0.11 mm, exhibited a higher activity than the positive control toosendanin. Information on new bioactive limonoids may provide further insight into M. toosendan as a source of bioactive components.

Effect of ethanolic extract of Melia dubia leaves on full-thickness cutaneous wounds in Wistar rats.

In recent years, the therapeutic effects of phyto-principles have been appreciated for their promising effects on wound healing. Melia dubia (Malabar neem) possesses anti-cancer, anti-diabetic, anti-tumor, anti-inflammatory, antioxidant, antibacterial, and fungicidal properties. Here, we studied the wound healing efficacy of ethanolic extract of M. dubia leaves on cutaneous wound healing for the first time. The ethanolic extract of M. dubia was applied topically on the wounds of the experimental rats until the wounds heal completely. Wounds of the control rats were treated with PBS. Granulation tissues formed on wound surfaces of the excision wound were harvested on days 4 and 8 and analyzed to determine the total collagen and hexosamine content. Total collagen and hexosamine were significantly (p < .001) higher in M. dubia treated rats compared to control. The rate of wound closure was significantly higher (p < .001) and period of epithelialization was shorter in M. dubia treated rats. Incision wound tissue was used for the tensile strength measurement. Tensile strength was improved in M. dubia treated wound tissues. Results concluded that the topical application of ethanolic extracts of M. dubia improved the rate of wound contraction and tensile strength by increased synthesis of collagen.

Fructose furoic acid ester: An effective quorum sensing inhibitor against uropathogenic Escherichia coli.

Uropathogenic Escherichia coli (UPEC) are the most common cause of UTI, accounting for more than 90% infections in the normal and unobstructed urinary tracts. Multi-drug resistance (MDR) is an emerging threat to the mankind and hence, there is an urge to develop alternative therapies. Targeting quorum sensing (QS), a cell-cell communication process regulates various biofilm and virulence factors would be a most promising alternate which curbs the pathogenesis without killing the bacteria, unlike antibiotics. SdiA, a quorum regulator is well-known to control the behavioural changes of UPEC in establishing biofilm and virulence. Therefore, we have hypothesized that the SdiA-selective inhibitors derived from the plant, Melia dubia using the molecular docking would be a remarkable therapeutic candidate to down regulate the UPEC biofilm and virulence phenotypes. In this study, we have designed, synthesized and characterized the fructose-furoic acid ester by NMR and ESI-MS. In vitro studies revealed that the QSI-MD selectively inhibits UPEC adherence and confocal laser scanning microscopy (CLSM) analysis showed the effectiveness of QSI-MD to inhibit the UPEC biofilm. Genetic studies using qRT-PCR revealed the down-regulation of quorum sensing regulated genes (fimA, csgA, espA). Based on the findings, we could propose that the QSI-MD could possibly act through SdiA and show target-specific inhibition of biofilm and virulence. It is notable that more than 70 bacterial species execute their communication through the SdiA homologues (LuxIR system). Hence, the QSI-MD could be further developed as a broad-spectrum anti-infective drug.

Meliacarpinin-Type Limonoids from the Bark of Melia toosendan.

Three new meliacarpinin-type limonoids, toosendanes A⁻C (1⁻3), along with three, known meliacarpinins (4⁻6) were isolated from the bark of Melia toosendan. Their structures, along with their absolute configurations, were elucidated, based on detailed analyses. These included HRESIMS and 1D/2D-NMR, modified Mosher's method, and electronic circular dichroism (ECD). Limonoids 2 and 3 showed moderate inhibitory activity on LPS-activated, RAW 264.7 macrophages.

[Chromatographic fingerprint analysis of Toosendan Fructus by HPLC-CAD coupled with chemometrics methods].

A new method was developed for the chromatographic fingerprint analysis of Toosendan Fructus by HPLC coupled with the charged aerosol detector (CAD) in the present study. Samples were well separated on an Agilent ZOBAX SB C18 column (4.6 mm × 250 mm, 5 µm) by gradient elution using acetonitrile and water containing 0.1 % formic acid (v/v) at the flow rate of 1.0 mL·min−1. The column temperature was 30 ℃ and the injection volume was 5 µL. The nitrogen inlet pressure of the charged aerosol detector (CAD) was 35 psi, and the nebulizer chamber temperature was 35 ℃. In addition, the method of the chromatographic fingerprints combined with multivariate statistical analysis was effective and reasonable lead to an accurate classification of 20 batches of samples from different locations. The results showed that 28 common peaks were observed in the fingerprint and the samples were classified into three clusters. The established method was well validated, and showed high precision, good repeatability, and satisfactory stability. It may serve in the quality control and evaluation of Toosendan Fructus.

Detecting long-term growth trends using tree rings: a critical evaluation of methods.

Tree-ring analysis is often used to assess long-term trends in tree growth. A variety of growth-trend detection methods (GDMs) exist to disentangle age/size trends in growth from long-term growth changes. However, these detrending methods strongly differ in approach, with possible implications for their output. Here, we critically evaluate the consistency, sensitivity, reliability and accuracy of four most widely used GDMs: conservative detrending (CD) applies mathematical functions to correct for decreasing ring widths with age; basal area correction (BAC) transforms diameter into basal area growth; regional curve standardization (RCS) detrends individual tree-ring series using average age/size trends; and size class isolation (SCI) calculates growth trends within separate size classes. First, we evaluated whether these GDMs produce consistent results applied to an empirical tree-ring data set of Melia azedarach, a tropical tree species from Thailand. Three GDMs yielded similar results - a growth decline over time - but the widely used CD method did not detect any change. Second, we assessed the sensitivity (probability of correct growth-trend detection), reliability (100% minus probability of detecting false trends) and accuracy (whether the strength of imposed trends is correctly detected) of these GDMs, by applying them to simulated growth trajectories with different imposed trends: no trend, strong trends (-6% and +6% change per decade) and weak trends (-2%, +2%). All methods except CD, showed high sensitivity, reliability and accuracy to detect strong imposed trends. However, these were considerably lower in the weak or no-trend scenarios. BAC showed good sensitivity and accuracy, but low reliability, indicating uncertainty of trend detection using this method. Our study reveals that the choice of GDM influences results of growth-trend studies. We recommend applying multiple methods when analysing trends and encourage performing sensitivity and reliability analysis. Finally, we recommend SCI and RCS, as these methods showed highest reliability to detect long-term growth trends.

Lignans from the Fruits of Melia toosendan and Their Agonistic Activities on Melatonin Receptor MT1.

Investigation on the fruits of Melia toosendan afforded seven new lignans (1-7), along with seventeen known compounds (8-24). The structures of the new compounds, involving four neo-lignans (1-4), two sesquilignans (5-6), and a nor-lignan (7), were elucidated based on extensive spectroscopic analyses (high-resolution electrospray ionization mass spectra, ultraviolet, infrared, one-dimensional and two-dimensional nuclear magnetic resonance). Compound 24 exhibited activity on melatonin receptor type 1 with an agonistic rate of 57.77% at 1.02 mM according to the assay on HEK293 cell lines in vitro.

Identification of amino acid and glutathione N-conjugates of toosendanin: bioactivation of the furan ring mediated by CYP3A4.

Toosendanin (TSN) is a hepatotoxic triterpenoid extracted from Melia toosendan Sieb et Zucc. Considering that TSN contains the structural alert of the furan ring, it is believed that bioactivation of TSN may be responsible for its toxicity. Herein, the bioactivation potential and metabolism profiles of TSN were investigated. After an oral administration of 10 mg/kg TSN to rats, esterolysis and conjugation with amino acids were identified as the main metabolic pathways. The same types of conjugates were detected in liver microsomes in an NADPH-dependent manner. According to the remaining amount of the parent drug, the reactivity of trapping reagents with TSN reactive metabolites was sorted in a decreasing order of N(α)-(tert-butoxycarbonyl)-l-lysine (Boc-Lys) > alanine, lysine, taurine, phenylalanine, serine, glutamic acid, glycine, and glutathione (GSH) > cysteine. No conjugates were observed in NADPH and N-acetyl cysteine (NAC)-supplemented human liver microsomal incubations. Further phenotyping studies and the chemical synthesis of the major conjugated standards proved that TSN was bioactivated by CYP3A4 and yielded a cis-butene-1,4-dial intermediate, which was prone to undergo 1,2-addition with the amino group of amino acids and GSH to form 3-pyrroline-2-one adducts. The sulfydryl group of GSH also attacked the intermediate and yielded S-conjugates by 1,4- or 1,2-addition, which would form pyrrole conjugates by further reacting with the amino group. Compared to the well-recognized S-conjugation of the furan ring, N-conjugation with multiple amino acids and GSH played a more important part in the elimination of reactive metabolites of TSN. The significance of these conjugates requires further investigation.

1-O-tigloyl-1-O-deacetyl-nimbolinin B inhibits LPS-stimulated inflammatory responses by suppressing NF-κB and JNK activation in microglia cells.

Overactivation of microglia may contribute to the pathogenesis of neurodegenerative diseases such as Parkinson's disease, Alzheimer's disease, and HIV dementia. Thus, regulating microglial activation has been an important therapeutic strategy for treating neurodegenerative diseases. In this research, we compared three limonoids compounds extracted from Melia toosendan by a cell-based assay to investigate their anti-inflammatory effects in lipopolysaccharide (LPS)-stimulated microglia cells. Our study indicated that 1-O-tigloyl-1-O-deacetyl-nimbolinin B (TNB) markedly suppressed the production of nitric oxide (NO) and tumor necrosis factor (TNF)-α in LPS-stimulated microglia cells. TNB also inhibited the gene expression of inducible nitric oxide synthase (iNOS), TNF-α, cyclooxygenase (COX-2), and interleukin (IL)-1ß. In addition, TNB inhibited generation of intracellular reactive oxygen species (ROS). We found that TNB significantly attenuated the nuclear translocation of NF-κB, inhibiting the activation of c-jun N-terminal kinase (JNK) in LPS-stimulated BV-2 cells. Furthermore, TNB reduced cytotoxicity of activated microglia toward HT-22 hippocampal cells in a co-culture system. Taken together, our experimental results reveal, for the first time, that TNB is a potent inhibitor of microglia-mediated inflammation, and it might be a potential candidate for the treatment of neurodegenerative diseases.

Potential anti-osteoporotic effects of herbal extracts on osteoclasts, osteoblasts and chondrocytes in vitro.

BACKGROUND: Osteoporosis (OP) is one of the most serious diseases in the modern world, and OP patients frequently suffer from fragility fractures in the hip, spine and wrist, resulting in a limited quality of life. Although bisphosphonates (BPs) are the most effective class of anti-bone-resorptive drugs currently available and the most commonly prescribed for the clinical treatment of OP, they are known to cause serious side effects such as bisphosphonate-related osteonecrosis of the jaw. Novel therapeutic materials that can replace the use of BPs have therefore been developed. METHODS: We commenced an institutional collaborative project in which candidates of herbal extracts were selected from more than 400 bioactive herbal products for their potential therapeutic effects not only in OP, but also in oral and skeletal diseases. In the present study, we report on 3 Chinese medical herbal extracts from the root barks of Melia azedarach, Corydalis turtschaninovii, and Cynanchum atratum. RESULTS: All of these extracts inhibited osteoclast proliferation and induced apoptosis by up-regulation of caspase activity and increase of mitochondrial pro-apoptotic proteins expression. Furthermore, the extracts enhanced differentiation, but did not affect proliferation of both osteoblasts and chondrocytes. The osteo-inducible effect was also observed in cultured primary bone marrow cells. CONCLUSIONS: Although these extracts have been utilized in traditional Chinese medicine for hundreds of years, there are no reports to our knowledge, on their therapeutic effects in OP. In this study, we elucidate the potency of these herbal extracts as novel candidates for OP therapy.

Exploring research trends and priorities of genus Melia.

The genus Melia is known for its secondary metabolites and recently, this genus is being explored for its timber. There are vast differences among its species. For instance, Melia azedarach is reported to be invasive and while another species, M. dubia, has diverse utility with complex germination and regeneration characteristics. Researchers globally have been working on various aspects of this genus; In this study, using topic modelling and science mapping approach, we attempted to understand research facets on this genus. The literature corpus of the Web of Science database was explored using a single keyword-"Melia" which yielded 1523 publications (1946-2022) and after scrutiny metadata of 1263 publications were used in the study. Although nine individual species were cited in the publications, only three species are accepted viz., M. dubia, M. azedarach, and M. volkensii. This implies taxonomic uncertainty, with potential confusion in assigning scientific findings to particular species. Thus, a taxonomic relook on this genus is warranted for a better assessment of the economic utility in many countries. More importantly, our results indicate that the research interests have recently shifted from the secondary metabolite constituents towards growth, biomass, wood properties, germination, plantation, and green synthesis. The shift in research focus toward wood properties of Melia sp. can impact the wood demand-supply at a global scale owing to its fast growth and the possibility of cultivation over a wider geographical range.

Meliasanines A-L, tirucallane-type triterpenoids from Melia toosendan with anti-inflammatory properties via NF-κB signaling pathway.

Meliasanines A-L, twelve previously unreported tirucallane-type triterpenoids, together with fifteen known ones, have been isolated from the stem bark of Melia toosendan. Their structures and absolute configurations were determined based on HRESIMS, and NMR, combined with calculated ECD and single-crystal X-ray diffraction analyses. Subsequently, all compounds except 10 were evaluated for their inhibitory effect on the production of nitric oxide induced by lipopolysaccharide in RAW264.7 macrophage cells. The results indicated that seven compounds (1, 13, 14, 16, 20, 22, and 23) exhibited significant NO inhibitory effects, with IC50 values ranging from 1.35 to 5.93 µM, which were more effective than the positive control indomethacin (IC50 = 13.18 µM). Moreover, the corresponding results of Western blot analysis revealed that meliasanine A (1) can significantly suppress the protein expression of inducible nitric oxide synthase and cyclooxygenase 2 in a concentration-dependent manner. The mechanism study suggested that meliasanine A exerts an anti-inflammatory effect via the nuclear factor-κB signaling pathway by suppressing phosphorylation of P65 and IκBα.