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Clausena excavata, a shrub in the Rutaceae family, has been used in traditional medicine to alleviate cold, malaria, abdominal pain, bacterial infection, and HIV infection. The major biologically active constituents in the rhizomes and roots of C. excavata are the three pyranocoumarins; clausenidin, dentatin, and nordentatin. Due to the medicinal values of this plant, we have developed a quick and convenient qHNMR method to quantify clausenidin, dentatin, and nordentatin from the rhizomes and roots of C. excavata collected from six geographical locations throughout Thailand. The dried and powdered samples were extracted with deuterated methanol. The parameters for 1H NMR measurements were optimized according to the measured T1 relaxation time of the protons from the bioactive pyranocoumarins and of 2,3,5,6-tetrachloronitrobenzene (TCNB), the internal standard. HPLC assays were also conducted to verify the results obtained from the qHNMR method. The amount of clausenidin, dentatin, and nordentatin varied from different geographical locations, and the results from the qHNMR method were comparable to those obtained from the HPLC assays. The reported qHNMR method was quick, more convenient, and more applicable to large-scale quality control or screening than conventional HPLC methodology. The method developed herein is beneficial to the pharmaceutical and health industries.

Background. Clausena excavata Burum. f. has long been applied in ethnomedicine for the treatment of various disorders like rhinitis, headache, cough, wound healing, fever, and detoxification. This study is aimed at investigating the antibacterial activity against Enterococcus faecalis ATCC 49532 using AlamarBlue assay and atomic force microscopy (AFM) as well as the cytotoxicity, anticancer, and phytotoxicity of C. excavata. Method. Bacterial cell viability was performed by using microplate AlamarBlue assay. Atomic force microscopy was used to determine morphological changes in the surface of bacterial cells. Cytotoxicity and phytotoxicity were determined by brine shrimp lethality and Lemna minor bioassay. Caco-2 (colorectal adenocarcinoma) cell line was used for the evaluation of the anticancer effects. Result. Among the fractions tested, ethyl acetate (EA) fraction was found to be active with minimum inhibitory concentration (MIC) of 750 μg/mL against E. faecalis, but other fractions were found to be insensitive to bacterial growth. Microscopically, the EA fraction-treated bacteria showed highly damaged cells with their cytoplasmic content scattered all over. The L C 50 value of the EA fraction against brine shrimp was more than 1000 μg/mL showing the nontoxic nature of this fraction. Chloroform (CH), EA, and methanol (MOH) fractions of C. excavata were highly herbicidal at the concentration of 1000 μg/mL. EA inhibited Caco-2 cell line with an I C 50 of 20 μg/mL. Conclusions. This study is the first to reveal anti-E. faecalis property of EA fraction of C. excavata leaves, natural herbicidal, and anticancer agents thus highlight the potential compound present in its leaf which needs to be isolated and tested against multidrug-resistant E. faecalis.

New derivatives were obtained from natural nordentatin (1) previously isolated from the methanol fraction of Clausena excavata by an acylation method. Herein, we report ten new pyranocoumarin derivatives 1a–1j. Their structures were elucidated based on UV-vis, FT-IR, NMR, and DART-MS data. The α-glucosidase inhibition and anticancer activities of nordentatin (1) and its derivatives were also evaluated. The α-glucosidase inhibition assay exhibited that the derivatives 1b, 1d, 1e, 1f, 1h, 1i, and 1j possess higher inhibitory activity for α-glucosidase with IC50 values of 1.54, 9.05, 4.87, 20.25, 12.34, 5.67, and 2.43 mM, whereas acarbose was used as the positive control, IC50 = 7.57 mM. All derivatives exhibited a weak cytotoxicity against a cervical cancer (HeLa) cell line with the IC50 between 0.25 and 1.25 mM. They also showed moderate to low growth inhibition of a breast cancer (T47D) cell line with IC50 values between 0.043 and 1.5 mM, but their activity was lower than that of the parent compound, nordentatin (1) (IC50 = 0.041 mM).

Objective: Clausena excavata is known to possess anti-oxidant property. However, this property through which mechanism it affects the immune cells and suppresses the production of reactive oxygen species (ROS) has not been explored. Material and Method: This study evaluated the immunomodulatory activities of ethyl acetate, petroleum ether, chloroform, and methanol C. excavata leaf extracts by decreasing the production of ROS from whole blood, polymorphonuclears (PMNs) cells and macrophages. Result and Discussion: Among the fractions tested, ethyl acetate C. excavate extract (EACE) showed potent anti-oxidant property and significantly (p < 0.001) suppressed intracellular and extracellular phagocytic oxidative ROS burst produced by the zymosan and PMA-activated whole blood, PMNs, and macrophages cells with 50% inhibitory concentration (IC50) values of 5.7 ± 0.01, 1.3 ± 0.01, and 0.7 ± 0.03 µg/mL respectively. This study provides information regarding the mechanism behind its anti-oxidant property and its herbal use in treating various higher oxidative stress associated diseases., Amaç: Clausena excavate’ın anti-enflamatuvar etkinlik gösterdiği bilinmektedir. Bununla birlikte, bu özelliği, bağışıklık hücrelerine hangi mekanizma ile etki ettiği ve reaktif oksijen türlerinin (ROS) üretimini baskıladığı araştırılmamıştır. Gereç ve Yöntem: Bu çalışmada, C. excavate yapraklarının etil asetat, petrol eteri, kloroform ve methanol ekstrelerinin tam kan, polimorfonkleer (PMN) hücreler ve makrofajlardan ROS üretimini azaltarak gösterdikleri immunomodülatör etki değerlendirilmiştir. Sonuç ve Tartışma: Test edilen fraksiyonlar arasından, C. excavate etil asetat ekstrresi en güçlü antienflamatuvar etkiyi göstermiş ve zimosan ve PMA tarafından active edilmiş tam kan, PMN ve makrofaj hücrelerinde, intraselüler ve ekstraselüler fagositik oksidatif ROS üretimini anlamlı oranda (p < 0.001) baskılamış ve %50 inhibisyon konsantrasyonları (IC50) sırasıyla 5.7 ± 0.01, 1.3 ± 0.01, and 0.7 ± 0.03 µg/mL olarak tespit edilmiştir. Bu çalışma, bitkinin antienflamatuvar aktivitesinin mekanizması ve çeşitli enflamatuvar rahatsızlıkların tedavisinde kullanımına dair bilgi sağlamaktadır.

Objective: To investigate the anti-inflammatory properties of methanolic extract of Clausena excavata in lipopolysaccharide (LPS)-activated macrophages (J774A.1) and the effect on skin wound in a rat model through determining cytokine levels and gene expressions. Methods: The effects of methanolic extract of Clausena excavata on in vitro viability and TNF-α, IL-6, IL-10, and nitric oxide release by LPS-activated J774A.1 cells were determined. In addition, relative expressions of BAX, BCL-2 and COX-2 genes were examined in healed wounds of rats. Results: The methanolic extract of Clausena excavata was not toxic to J774A.1 cells at the highest dose of 400 μg/mL. It decreased levels of TNF-α and IL-6, while increasing IL-10 level in LPS- activated J774A.1 cells and in the healed wounds of rats. The methanolic extract of Clausena excavata also inhibited nitric oxide production in LPS-activated J774A.1 cells. The BAX and COX-2 genes were downregulated while the BCL-2 gene was upregulated in the healed wound of rats. Conclusions: The methanolic extract of Clausena excavata promotes wound healing via its anti-inflammatory and anti-apoptotic activities.

All around the world, patients with diabetes and the prevalence of its disease are currently growing. Due to these side effects of oral hypoglycemic agents and oxidative stress in complicating diabetes, there is growing interest in drugs, which possess dual function as both type II diabetes mellitus treatment and oxidative stress treatment. The objective of this research is to search effective antidiabetes and antioxidant bioactive compounds from the Myanmar medicinal plantClausena excavata. The root part ofC. excavatawas successfully extracted with 95% ethanol and followed by column chromatographic separation technique. The structure of isolated pure compounds was elucidated by using methods of spectroscopic such as UV-Vis, IR, NMR and HRFABMS spectrometry. The α-glucosidase inhibition assay was performed against baker’s yeast and rat intestine (sucrose and maltase) α-glucosidases. The activity of isolated compounds’ antioxidant was measured by using DPPH assay. Among the tested enzymes, the two isolated compounds, which were dentatin (1) and heptaphylline (2), exhibited highest inhibitory on maltase enzymes with IC50values 6.75 and 11.46 μM; as positive control, acarbose (IC50, 2.35 μM) was utilized. Moreover, scavenging activity was found to be present upon seeing the result of antioxidant activity investigation of (1) and (2) (IC50values 2.66 and 1.55 mM), where ascorbic acid (IC500.012 mM) was used as standard. Both compounds showed their antidiabetic and antioxidant activity with different fashion, especially exhibited strongest activity against on maltase α-glucosidase.

핑크왐피는 피부병, 말라리아, 복통, 이질 및 장염 치료를 위한 약용 식물로 사용되고 있다. 본 연구의 목적은 핑크왐피의 안전성을 확인하기 위하여, 수컷 ICR 마우스를 이용하여 핑크왐피를 3주 반복 경구 투여하여 최대무독성용량을 평가했다. 핑크왐피를 100, 250, 500, 1000, 및 2000 mg/kg으로 투여한 결과, 모든 시험물질 투여군에서 이상이 관찰되지 않았다. 사망률, 임상 증상, 체중 변화, 혈액 학적 검사 및 혈청 생화학 검사에서 유의적인 차이는 없었고, 경미한 변동은 핑크왐피의 투여로 인한 영향이 아닌 정상 범위 내에서의 변화로 간주 되어진다. 결과적으로, 핑크왐피는 모든 시험물질 투여군에서 독성 영향을 미치지 않았으며, 핑크왐피의 최대무독성용량은 2000 mg/kg 이상으로 간주되었다.

A new pyrano coumarin, identified as excavatin A (1) together with two known compounds nordentatin (2) and binorpocitrin (3) was isolated from the 95% EtOH extract of Clausena excavata. All structures were elucidated by using spectroscopy methods such as extensive NMR and HR-FAB-MS spectrometry. All the isolated compounds were tested on antidiabetes activity by using α-glucosidase inhibition assay and the antioxidant activity by DPPH assay. Compounds 1-3 showed antioxidant activity with IC50 values 0.286, 0.02, 0.278 mM. Among them, 2 exhibited inhibition activity against maltase (IC50 5.45 µM) and sucrase (IC50 43.57 µM). However, compounds (1) and (3) displayed inhibition on yeast α-glucosidase with IC50 values 1.92 and 5.58 mM.

In the present investigation, Clausena excavata an indegeous plant was sequentially extracted with hexane, ethyl acetate and methanol and their bio efficacy in terms of Antioxidant activity, mosquitocidal activity and antibacterial activity was tested. Among the three-extract methanol extract of C. excavata displayed significant antioxidant, larvicidal, ovicidal, pupicidal activities against Aedes aegypti and Culex quinquefasciatus. Similarly, methanol extract also showed pronounced antibacterial activity against the selected human pathogenic bacteria. Thus, the methanol extract of C. excavata was used to synthesize silver mediated nanoparticles and its (C. excavata AgNPs) bio efficacy was also tested with the selected vector mosquitoes and selected human pathogenic bacteria. This present investigation envisages the possible utilization of C. excavata as a potent candidate as green pesticides in the near future. Article DOI: https://dx.doi.org/10.20319/lijhls.2019.51.1939 This work is licensed under the Creative Commons Attribution-Non-commercial 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc/4.0/ or send a letter to Creative Commons, PO Box 1866, Mountain View, CA 94042, USA.

Background Clausena excavata Burm.f. (Rutaceae) has been used for the treatment of stomach disorders including peptic ulcer. Purpose In this study, we aimed to investigate dentatin isolated from C. excavata Burm.f., for anti-ulcer activity against ethanol ulcer model in rats. Methods Gastric acid output, ulcer index, serum profile, histological evaluation using Hematoxylin and eosin (HE), periodic acid Schiff base stainings and immunohistochemical localization for heat shock proteins 70 (HSP70) were all investigated. Possible involvement of reduced glutathione (GSH), lipid peroxidation, prostaglandin E2 (PGE2), superoxide dismutase (SOD) enzymes, radical scavenging, and anti-Helicobacter pylori activity were investigated. Results Dentatin showed anti-secretory activity against the pylorus ligature model and protected the gastric mucosa from ethanol ulceration, as revealed by the improved macroscopic and histological appearance. Dentatin significantly increased the gastric homogenate content of PGE2 GSH and SOD. Dentatin inhibited the lipid peroxidation as revealed by the reduced gastric content of malondialdehyde (MDA). Moreover, dentatin up-regulated HSP70 expression. However, dentatin showed insignificant anti-H. pylori activity. Conclusion Dentatin possesses gastro-protective activity, which could be attributed to the anti-secretory, mucus production, anti-oxidant, and HSP70 activities.

Phytochemical study on the stem bark of Clausena excavata Burm. f. led to the isolation and identification of 11 compounds, including four prenylated coumarins (1–4), three carbazole alkaloids (5–7), a limonoid (8), a sterol (9), a depside (10) and a xanthone (11). The chemical structures of the compounds were elucidated based on NMR and mass spectroscopic data, and further supported by comparison with previously literature values. This is the first report on the isolation of compounds 10 and 11 from the genus Clausena. The chemotaxonomic significance of the isolated compounds is discussed.

This study established the DNA barcoding sequences (matK and rbcL) of three plant species identified in the tribe Clauseneae, namely Clausena excavata, C. harmandiana and Murraya koenigii. The total phenolic and total flavonoid contents, together with the biological activities of the derived essential oils and methanol extracts, were also investigated. Herein, the success of obtaining sequences of these plant using two different barcode genes matK and rbcL were 62.5% and 100%, respectively. Both regions were discriminated by around 700 base pairs and these had resemblance with those of the Clausenae materials earlier deposited in Genbank at a 99&ndash, 100% degree of identity. Additionally, the use of matK DNA sequences could positively confirm the identity as monophyletic. The highest total phenolic and total flavonoid content values (p &lt, 0.05) were observed in the methanol extract of M. koenigii at 43.50 mg GAE/g extract and 66.13 mg QE/g extract, respectively. Furthermore, anethole was detected as the dominant compound in C. excavata (86.72%) and C. harmandiana (46.09%). Moreover, anethole (26.02%) and caryophyllene (21.15%) were identified as the major phytochemical compounds of M. koenigii. In terms of the biological properties, the M. koenigii methanol extract was found to display the greatest amount of antioxidant activity (DPPH, IC50 95.54 &micro, g/mL, ABTS value 118.12 mg GAE/g extract, FRAP value 48.15 mg GAE/g extract), and also revealed the highest &alpha, glucosidase and antihypertensive inhibitory activities with percent inhibition values of 84.55 and 84.95. Notably, no adverse effects on human peripheral blood mononuclear cells were observed with regard to all of the plant extracts. Furthermore, M. koenigii methanol extract exhibited promise against human lung cancer cells almost at 80% after 24 h and 90% over 48 h.

Context: Clausena excavata Burm. f is a plant used in folklore medicine for the treatment of various ailments in South East Asia. The plant parts contain chemical components that are cytotoxic to many cancer cells. Objective: The study investigated the cytotoxic effects of ethyl acetate, methanol and chloroform C. excavata leaf extracts on the non-small-lung cancer, NCI-H460, cell line. Methods: Based on the 3-(4,5-dimethylthiazol-2-yl)-2,5,-diphenyltetrazolium bromide (MTT) assay, among extracts, ethyl acetate C. excavata leaf extract (EACE) was the most potent anti-NCI-H460 cells, with IC50 value of 47.1 ± 6.1 μg/ml. The effects of EACE on NCI-H460 cells were also determined by clonogenic, 4', 6-diamidino-2-phenylindole (DAPI), and annexin-V-fluorescein isothiocyanate/propidium iodide-PI flow cytometric assays. Reactive oxygen species (ROS) production and apoptotic gene expressions was determined via flow cytometry and real-time quantitative PCR, respectively. Results: EACE-treated NCI-H460 cells after 48 h underwent apoptosis as evident by loss of cell viability, cell shrinkage, and chromatin condensation. The results also showed EACE mediated increase in ROS production by the NCI-H460 cells. After 48 h treatment, EACE increased the pro-apoptotic BAX and decreased the anti-apoptotic Bcl-2, Survivin and c-Myc gene expressions. Conclusions: EACE is a potential anti-lung cancer by increasing cancer cell ROS production and apoptosis.

Methanolic crude extracts obtained from leaves, stem bark and root bark of 57 accessions of Clausena excavata, C. harmandiana, of both varieties of C. wallichii, as well as of three unidentified Clausena samples, were analysed for the presence of coumarins and carbazoles. All accessions were collected from different locations all over Thailand. HPLC coupled with UV-DAD was applied for phytochemical profiling. Structures of known pyrano-coumarins 1–4 and of coumarins 5–8 bearing an O-geranyl moiety, as well as of carbazoles 9–12 were confirmed by NMR and MS spectroscopy, respectively. Observed chemical diversification in the studied samples is discussed in relation to geographic distribution and with respect to organ-specific accumulation. Chemotaxonomic significance is briefly addressed.

Clausena excavata (C. excavata) has been used as a traditional medicine for the treatment of abdominal pain, enteritis, dysentery, and malaria. The present study was designed to evaluate the effect of a 50% ethanol extract of C. excavata (ECE) on weight loss, adipocyte size, and obesity-related biochemical parameters in high-fat diet (HFD)-induced obese mice. After 6 weeks of HFD + ECE administration, HFD-induced total fat, subcutaneous fat, and visceral fat were evaluated by micro-computed tomography. The serum levels of triglyceride (TG), total cholesterol (TCH), high-density lipoprotein (HDL) cholesterol, and low-density lipoprotein (LDL) cholesterol were evaluated with a biochemical analyzer, and leptin and adiponectin levels in the serum were assessed via enzyme-linked immunoassay (ELISA). Moreover, adipocyte size and lipid formation in the liver were examined. We found that weight gain, epididymal fat pad weight, adipocyte size, and lipid formation were markedly attenuated in the livers of HFD-induced obese mice treated with ECE. Furthermore, TG, TCH, and leptin decreased in the serum, whereas adiponectin increased. In conclusion, our data show that ECE has potent anti-obesity activity in vivo and support the development of ECE as a potential therapeutic agent for the treatment of obesity.

Three new prenylated phenylpropenols, exotiacetals A–C (1–3), 10 new coumarin derivatives, exotimarins A–I (4–13), and 35 known analogues (14–48) were isolated from the roots of Murraya exotica. The absolute configurations of the new compounds were assigned via comparison of their specific rotations, single-crystal X-ray diffraction data, Mosher’s method, the ECD exciton coupling method, comparison of experimental and calculated ECD data, and the ECD data of the in situ formed transition metal complexes. Compounds 1–3, which possess an unprecedented hexahydro-1H-isochromen-1-ol system, are presumably biosynthesized from two prenylated p-coumaryl alcohol moieties via Diels–Alder [4+2] cycloaddition and cyclic hemiacetal formation reactions. Compounds 1, 28, 33, and 35 demonstrated inhibition against LPS-induced NO production in BV-2 microglial cells with IC50 values of 8.6 ± 0.3, 11.8 ± 0.9, 15.5 ± 0.9, and 16.9 ± 1.0 μM, respectively.

Two new flavonoid glycosides, excavaside A ( 1 ) and B ( 2 ), together with 14 known compounds, myricetin 3- O -β- d -rutinoside ( 3 ), myricetin 3,3′-di-α- l -rhamnopyranoside ( 4 ), myricetin 3′-α- l -rhamnopyranoside ( 5 ), myricitrin ( 6 ), myricetin ( 7 ), quercitrin ( 8 ), clauslactone B ( 9 ), clauslactone A ( 10 ), murrayacoumarin C ( 11 ), clauslactone E ( 12 ), triphasiol ( 13 ), clauslactone P ( 14 ), seseline ( 15 ), and 5-geranyloxy-7-hydroxycoumarin ( 16 ) have been isolated from the ethanol extract of the leaves and twigs of Clausena excavata . Among them, compounds 3 – 5 and 13 were isolated and identified from this plant for the first time. The structures of the new compounds were established by spectroscopic data interpretation, particularly HRESIMS, 1D and 2D NMR data including HSQC and HMBC. Also, those of the known compounds were identified by spectral comparison with those of the reported values.

Propyl-cannabinol allyldimethylsilyl ether (PCAE) is one of the phytochemical compounds which were isolated from Clausena excavata Burm f. However, the cytotoxicity and selectivity properties of (PCAE) against different cancer cell lines are not elucidated yet. Hence, this study has been done to test the efficiency of (PCAE) as a new anti-cancer agent. Isolation and identification of this compound was conducted by various spectroscopic analyses including electron impact mass spectra (EI-MS), X-ray diffractometry (XRD) and previous data. In vitro, cytotoxicity of compound against different cancer cell lines was investigated by MTT assay. After exposure time, growth inhibition with significant changes in cellular morphology was clearly noticed in treated cells. The HT29 cells were most directed towards apoptosis relative to the other cells, as the result of treatment with Propyl-cannabinol allyldimethylsilyl ether (PCAE). In Conclusion Propyl-cannabinol allyldimethylsilyl ether (PCAE) has been appeared a significant effect in stopping and killing cancer cells.

Shaymaa Fadhel Abbas Albaayit,1,2 Yusuf Abba,3 Rasedee Abdullah,4 Noorlidah Abdullah1 1Faculty of Science, Institute of Biological Sciences, University of Malaya, Kuala Lumpur, Malaysia; 2Department of Biology, Collegeof Science, University of Baghdad, Baghdad, Iraq; 3Department of Veterinary Pathology and Microbiology, 4Department of Veterinary Laboratory Diagnosis, Faculty of Veterinary Medicine, Universiti Putra Malaysia, Serdang, Selangor, Malaysia Abstract: Clausena excavata is a natural herb with both antioxidant and anti-inflammatory properties. It has been used for decades in folkloric practice for the amelioration of various ailments. In this study, the gastroprotective activity of methanolic extract of C. excavata leaves (MECE) was determined in the Sprague Dawley rat ethanol-induced gastric ulcer model. Rats were pretreated with a single dose of vehicle (5% Tween 20), 20 mg/mL omeprazole, 400 and 200 mg/mL of MECE dissolved in 5% Tween 20. Ulcer was induced with 5 mL/kg of ethanol and stomach tissue was obtained after 1 hour. Histological examination was done on hematoxylin and eosin, periodic acid-Schiff, and immunochemically stained gastric mucosal tissues. Prostaglandin E2, superoxide dismutase, catalase, glutathione peroxidase, and lipid peroxidation levels of the gastric tissue homogenates were also determined. Significantly (P

To broaden and delve into the genomic information of Clausena excavata, an important medicinal plant in many Asian countries, RNA sequencing (RNA-seq) analysis was performed and a total of 16,638 non-redundant unigenes (≥ 300 bp) with an average length of 755 bp were generated by de novo assembly from 17,580,456 trimmed clear reads. The functional categorization of the identified unigenes by a gene ontology (GO) term resulted in 2305 genes in the cellular component, 5577 in the biological processes, and 8056 in the molecular functions, respectively. The top sub-category in biological processes was the metabolic process with 4374 genes. Among annotated genes, 3006 were mapped to 123 metabolic pathways by the Kyoto Encyclopedia of Genes and Genomes (KEGG) metabolic pathway analysis tool. The search for simple sequence repeats (SSRs) resulted in 845 SSRs from 749 SSR-containing unigenes and the most abundant SSR motifs was AAG/CTT with 179 occurrences. Twelve SSR markers were tested for cross transferability among five Clausena species; eight of them exhibited polymorphism. Taken together, these data provide valuable resources for genomic or genetic studies of Clausena species and other relative studies. The transcriptome shotgun assembly data have been deposited at DDBJ/EMBL/GenBank under the accession GGEM00000000.

Inhibition of the necrotizing hyaluronidase, phospholipase A2 and protease enzymes in four snake venoms by crude water and ethanol extracts of 88 plant species used against snakebites in traditional Chinese medicine was measured. High-resolution hyaluronidase inhibition profiles were constructed for the 22 plants showing highest hyaluronidase inhibition, and the results were used to guide subsequent structural analysis towards specific hyaluronidase inhibitors. Structural analysis was performed by high-performance liquid chromatography, high-resolution mass spectrometry, solid-phase extraction and nuclear magnetic resonance spectroscopy, i.e., HPLC-HRMS-SPE-NMR. This allowed identification of four non-tannin inhibitors, i.e., lansiumamide B (6) from Clausena excavata Burm.f., myricetin 3-O-β-D-glucopyranoside (7) from Androsace umbellata (Lour.) Merr., and vitexin (8) and 4',7-dihydroxy-5-methoxyflavone-8-C-β-D-glucopyranoside (9) from Oxalis corniculata L. Absolute configuration of 2,3-dihydroxy-N-methyl-3-phenyl-N-[(Z)-styryl]propanamide (1) was determined using the Mosher method, which revealed two enantiomers, i.e., (2S,3R)-2,3-dihydroxy-N-methyl-3-phenyl-N-[(Z)-styryl]propanamide and (2R,3S)-2,3-dihydroxy-N-methyl-3-phenyl-N-[(Z)-styryl]propanamide with a ratio of 7:3.

Damage caused by Citrus psorosis virus (CPsV) could be avoided obtaining resistant cultivars by breeding, but the sensitivity of most citrus hosts to CPsV is unknown. To find potential sources of resistance we inoculated 63 cultivars and hybrids of Citrus and related genera [Citrus (36), Microcitrus (5), Fortunella (6), Eremocitrus (1), Pleiospermium (1), Atalantia (2), Severinia (1), Clausena (1), Swinglea (1), Afraegle (1), Poncirus (1) and hybrids (7)] with the CPsV isolate PB-143 and monitored symptoms and CPsV infection by ELISA. Microcitrus inodora and Fortunella hindsii were symptomless but gave high ELISA values, suggesting tolerance to CPsV, whereas Citrus depresa, Clausena excavata, Cleopatra mandarin, sour orange, Carrizo citrange and CPB4475 citrumelo were ELISA negative, suggesting resistance. Further examination of CPsV infection by reverse transcription quantitative real time PCR (RT-qPCR) in Cleopatra mandarin, Poncirus trifoliata and Carrizo citrange seedlings inoculated with CPsV isolates P-121, PB-102 and PB-143, revealed different infection rates and CPsV accumulation, depending on the isolate and citrus genotype. Bud propagation of these genotypes on CPsV-inoculated sweet orange plants incited a bud union disorder that hindered flushing and scion growth, suggesting that the high viral load in the rootstock induced a hypersensitive-like reaction in the partially resistant scion.

To develop natural product resources from the Clausena genus (Rutaceae), the essential oils (EOs) from four Clausena plants (Clausena excavata, C. lansium, C. emarginata, and C. dunniana) were analyzed by GC-MS. Their lethal (contact toxicity) and sublethal effects (repellency) against Liposcelis bostrychophila (LB) adults were also evaluated. Santalene sesquiterpene was the precursor substance of santalol, a valuable perfumery. It was found that plenty of α-santalol (31.7%) and α-santalane (19.5%) contained in C. lansium from Guangxi Province and α-santalene (1.5%) existed in C. emarginata. Contact toxicity of the four EOs was observed, especially C. dunniana (LD50 = 37.26 µg/cm2). Santalol (LD50 = 30.26 µg/cm2) and estragole (LD50 = 30.22 µg/cm2) were the two most toxic compounds. In repellency assays, C. excavate, C. lansium, and C. emarginata exhibited repellent effect at the dose of 63.17 nL/cm2 2 h after exposure (percentage repellencies were 100%, 98%, and 96%, respectively). Four Clausena EOs and santalol had an excellent potential for application in the management of LB. Clausena plants could be further developed to find more resources of natural products.

The aim of this paper is to evaluate the larvicidal and ovicidal activities of hexane, diethyl ether, dichloromethane, ethyl acetate, and methanol extract of Indian medicinal plant, Clausena excavata, at different concentrations against lepidopteran agricultural field pest, Spodoptera litura (Lepidoptera: Noctuidae), and Aedes aegypti, Anopheles stephensi, and Culex quinquefasciatus (Diptera: Culicidae). Twenty-five early fourth instar larvae of S. litura and A. aegypti, An. stephensi and C. quinquefasciatus were exposed to various concentrations and were assayed in the laboratory by using specific protocols. The 24 h LC50 and LC90 values of the C. excavata leaf extract was determined by probit analysis. The ovicidal activity were determined against S. litura and A. aegypti, An. stephensi, and C. quinquefasciatus and was assessed 120 hrs after treatment. Results of this study show that the selected Indian medicinal plant C. excavata may be a potential source of natural larvicidal and ovicidal activities against selected pest and vector mosquitoes.

Some selected medicinal plants used by Bentian tribe from Indonesia were evaluated for potential antimicrobial and antioxidant properties. The leaves stem bark and root of the plants were extracted with n -hexane, ethyl acetate and ethanol to give respective extracts. Antimicrobial activity against Propionibacterium acnes and Candida albicans was determined by an agar well diffusion method. Antioxidant activity was assayed by the DPPH radical scavenging activity mechanism. The results showed that n -hexane, ethyl acetate and ethanol extracts of Cananga odorata , ethanolic extract of Chromolaena odorata , and ethanolic extracts of Hyptis capitata and Ampelocissus cinnamomeae displayed good activity against P. acnes at 25–400 μg/well of the extracts tested. The n -hexane and ethanol extracts of Chromolaena odorata and H. capitata , and the n -hexane, ethyl acetate and ethanol extracts of Cananga odorata displayed more activity against C. albicans than others at 25–400 μg/well of the extracts tested. The most antioxidant activities against DPPH were displayed by the ethanol extracts of Ficus variegata stem bark, Leucosyke quadrinerva root and Clausena excavata leaves exhibiting 91%, 91% and 86% inhibition, respectively. The present results showed potential of some medicinal plants used by Bentian tribe from Indonesia as natural anti-microbial and anti-oxidant and potentially anti-inflammatory agents.

This article reports the compounds identified in the leaf oils of five Clauseana species growing in Vietnam. The hydrodistilled oils were analysed for their chemical constituents by using gas chromatography-flame ionisation detector and gas chromatography coupled with mass spectrometry. The major compounds identified in Clausena dentata (Willd.) M. Roem. were α-pinene (21.7%), sabinene (18.3%) and β-myrcene (14.3%). Clausena dimidiata Tanaka comprised mainly of safrole (56.9%) and α-terpinolene (22.1%). However, 1-menthone (70.6%) and β-phellandrene (13.0%) were the most abundant compounds of Clausena indica (Dalz.) Oliver. Sesquiterpene compounds represented mainly by β-caryophyllene (16.7%), spathulenol (11.9%) and bicyclogermacrene (7.5%) were the major constituents of Clausena excavata Burm. f., while those of Clausena engleri Tanaka include bicycloelemene (12.1%), bicyclogermacrene (11.0%) and (E)-nerolidol (6.6%). This is the first report on the chemical analysis of essential oils of C. dimidiata and C. engleri.

Continuing our exploration for dual functions antidiabetic and antioxidant agents from Myanmar medicinal plant , a new carbazole-pyranocoumarin conjugate, carbazomarin-C (1) along with a known carbazole alkaloid, mukonine (2) and a pyranocoumarin, xanthoxyletin (3), was isolated from the roots of Clausena excavata. The chemical structures of these compounds were identified using a combination of spectroscopic methods. Among isolates, there was a strong inhibition of compounds (1) and (3) on yeast α-glucosidase in a dose-dependent manner. It was shown when p-nitrophenyl-α-d-glucopyranoside was used as a substrate in vitro with IC50 values 0.22 and 4.81 mM, respectively. However, all isolated compounds displayed no inhibition against DPPH (2,2-diphenyl-1-picrylhydrazyl) radicals.

The data presented in this article are associated to the research articles, “DOI: 10.1007/s11295-019-1348-3”, [1]; and “DOI: 10.1007/s13205-018-1162-x” [2]. Clausena excavata Burm. f. and Sterculia lanceolata Cav. are medicinal tree plants [3,4] native to Southeast Asia and China, and most members of both the genus Clausena and the genus Sterculia contain various valuable secondary metabolites with a great potential for drug development. Though many phytochemical studies have been conducted using plant extracts from various parts of these plants [4,5], there are very limited genetic resources available. RNA sequencing of C. excavata and S. lanceolata was conducted using pair-end Illumina HiSeq2500 sequencing system, from which the first de novo transcriptome data were produced for both genus Clausena and Sterculia. Transcriptome shotgun assembly using three different assembly tools [2] generated a total of 16,638 non-redundant contigs (N50, 900 bp) from C. excavata and 7,857 (N50, 423 bp) from S. lanceolata. The data are accessible at NCBI BioProject: PRJNA428402 for C. excavata [2] or PRJNA435648 for S. lanceolata[1]. Keywords: Transcriptome analysis, Medicinal plant, Sterculia lanceolata, Clausena excavata

Shaymaa Fadhel Abbas Albaayit,1,2 Yusuf Abba,3 Rasedee Abdullah,4 Noorlidah Abdullah1 1Faculty of Science, Institute of Biological Sciences, University of Malaya, Kuala Lumpur, Malaysia; 2Department of Biology, Collegeof Science, University of Baghdad, Baghdad, Iraq; 3Department of Veterinary Pathology and Microbiology, 4Department of Veterinary Laboratory Diagnosis, Faculty of Veterinary Medicine, Universiti Putra Malaysia, Serdang, Selangor, Malaysia Abstract: Clausena excavata is a natural herb with both antioxidant and anti-inflammatory properties. It has been used for decades in folkloric practice for the amelioration of various ailments. In this study, the gastroprotective activity of methanolic extract of C. excavata leaves (MECE) was determined in the Sprague Dawley rat ethanol-induced gastric ulcer model. Rats were pretreated with a single dose of vehicle (5% Tween 20), 20 mg/mL omeprazole, 400 and 200 mg/mL of MECE dissolved in 5% Tween 20. Ulcer was induced with 5 mL/kg of ethanol and stomach tissue was obtained after 1 hour. Histological examination was done on hematoxylin and eosin, periodic acid-Schiff, and immunochemically stained gastric mucosal tissues. Prostaglandin E2, superoxide dismutase, catalase, glutathione peroxidase, and lipid peroxidation levels of the gastric tissue homogenates were also determined. Significantly (P

Ethnopharmacological relevance Clausena excavata Burm. f . is used locally in folk medicine for the treatment of cancer in South East Asia. Aim of the study To determine the mechanism of action of pure clausenidin crystals in the induction of hepatocellular carcinoma (hepG2) cells apoptosis. Materials and methods Pure clausenidin was isolated from Clausena excavata Burm. f . and characterized using 1 H and 13 C NMR spectra. Clausenidin-induced cytotoxicity was determined by MTT assay. The morphology of hepG2 after treatment with clausenidin was determined by fluorescence and Scanning Electron Microscopy. The effect of clausenidin on the apoptotic genes and proteins were determined by real-time qPCR and protein array profiling, respectively. The involvement of the mitochondria in clausenidin-induced apoptosis was investigated using MMP, caspase 3 and 9 assays. Results Clausenidin induced significant (p Conclusion Clausenidin extracted from Clausena excavata Burm. f . is an anti-hepG2 cell compound as shown by its ability to induce apoptosis through the mitochondrial pathway of apoptosis. Clausenidin can potentially be developed into an anticancer compound.

Clausena excavata Burm. f.(Rutaceae) has long been associated with medicinal benefits in folk medicine, particularly in the treatment of cancer and its related disorders in the eastern region of Thailand. The present study evaluates the antioxidant activity of the methanolic leaf extract of C. excavata using the lipid peroxidation model. The results indicated that the methanolic leaf extract showed marked antioxidant activity (IC 50 = 201.3 μg/ml), whereas the aqueous leaf extract showed moderate antioxidant activity (IC 50 = 450.6 μg/ml). In conclusion, the antioxidant activity of the methanolic leaf extract of C. excavata was comparable with that of propyl gallate, which is used as a standard drug, thus confirming that it might be a good source of antioxidants. The extended degree of antioxidant activity could be attributable to the presence of flavonols, gallic acid, and angelicin, which are known to be antioxidants.

Chromosomenumbersfor21Rutaceousspecies,representingfivegenera(Bergera,Clausena,Glycosmis, Micromelum, and Murraya) in the tribe Clauseneae (Aurantioideae, Rutaceae) were investigated to shed light on the relationships between these genera in the tribe in particular, and with other groups in the orange subfamily in general. Most species have the chromosome number 2n = 2x = 18, but Glycosmis shows variable chromosome numbers with ploidy levels ranging from 2x to 8x. Glycosmis parviflora is hexaploid with a chromosome number of 2n = 6x = 54 and G. longipetala octoploid with 2n = 8x = 72, and diploid with 2n = 2x = 18 was observed in other species. One species (Clausena excavata) shows intraspecies ploidy variation. The chromosome numbers of 11 species, Bergera kwangsiensis, B. microphylla, Clausena dunniana, C. emarginata, C. lenis, C. yunnanensis, Glycosmis esquirolii, G. lucida, G. oligantha, Micromelum falctum, and Murraya alata, are reported for the first time. The basic chromosome number x = 9 was inferred for all species studied. These results, together with previous chromosome counts in the tribe Citreae, reveal a cytological homogeneity, and strongly support the monophyly of the orange subfamily. The tribe Clauseneae, together with the tribe Cit- reae, were classified into the subfamily Aurantioideae ( = Citroideae) in Rutaceae according to Swingle & Reece (1967). In general, Aurantioideae can be char- acterized as small trees, shrubs, and, rarely, lianas that produce fruit with a leathery rind or hard shell and often with a juicy pulp. The leaves and fruits have schizolysigenous oil glands that release an aroma when touched, and the flowers are typically white and fra- grant. The leaves are persistent except in three mono- typic genera (Poncirus, Aegle, and Feronia), three species of Clausena, and one species of Murraya. The tribe Clauseneae consists of five genera, Micromelum, Glycosmis, Clausena, Murraya, and Merrillia, and in- cludes the more primitive genera of the orange subfam- ily. None of the species develops spines in the axils of the leaves and the odd-pinnate leaves are at once dis- tinguished from those of the tribe Citreae by having the leaflets attached alternately to the rachis. The ovary has 2-5 locules with only 1 or 2 ovules in each locule, except in Merrillia, which has 5 (rarely 6) locules and 8-10 ovules in each locule.The fruits are usually small

A new γ-lactone coumarin, named as excavarin-A, showing antifungal activity was isolated from the leaves of Clausena excavata by bioassay guided fractionation method. The structure was elucidated by spectroscopic data analysis and identified as 7((2E)-4(4,5-dihydro-3-methylene-2-oxo-5-furanyl)-3-methylbut-2-enyloxy) coumarin. Minimum inhibitory concentration (MIC) was determined against fifteen fungal strains pathogenic against plants and human. The least MIC was recorded against the human pathogen, Candida tropicalis and the plant pathogens Rhizoctonia solani and Sclerotinia sclerotiorum. Antifungal activities against the human pathogens, Aspergillus fumigatus and Mucor circinelloides and plant pathogens, Colletotrichum gloeosporioides, Lasiodiplodia theobromae, Fusarium oxysporum and Rhizopus stolonifer were stronger than that of the standard antimicrobials.

Four natural pyranocoumarins clausenidin (1), nordentatin (2), clausarin (3), and xanthoxyletin (4) were isolated from the medicinal plant Clausena excavata. Recently, we found that 1 and 2 suppressed hepatitis B virus surface antigen in HepA2 cells, and in addition, 1-3 showed cytotoxic activity against four human cancer cell lines (A549, MCF7, KB, and KB-VIN). To explore the SAR of 1-4, 17 pyranocoumarin analogues (5-21) were designed and synthesized. Among these analogues, 5 and 10 were the most potent against hepatitis B virus with EC(50) values of 1.14 and 1.34microM, respectively. The most interesting result in the cytotoxicity assay was the significant activity of 1, 5, and 6 against the multi-drug resistant cell line, KB-VIN, without activity against the KB cell line. These data suggest that these three compounds could be useful hits for developing MDR-inverse drugs.

BACKGROUND: The current study investigates, for the first time, the mosquito larvicidal activities of leaf and twig essential oils from Clausena excavata Burm. f. and their individual constituents against Aedes aegypti L. and Aedes albopictus Skuse larvae. The yields of essential oils obtained from hydrodistillation were compared, and their constituents were determined by GC-MS analyses. RESULTS: The LC50 values of leaf and twig essential oils against fourth-instar larvae of Ae. aegypti and Ae. albopictus were 37.1–40.1 µg mL−1 and 41.1–41.2 µg mL−1 respectively. This study demonstrated that C. excavata leaf and twig essential oils possess mosquito larvicidal activity, inhibiting the growth of mosquito larvae for both species at a low concentration. In addition, results of larvicidal assays showed that the effective constituents in leaf and twig essential oils were limonene, γ-terpinene, terpinolene, β-myrcene, 3-carene and p-cymene. The LC50 values of these constituents against both mosquito larvae were below 50 µg mL−1. Among these effective constituents, limonene had the best mosquito larvicidal activity, with LC50 of 19.4 µg mL−1 and 15.0 µg mL−1 against Ae. aegypti and Ae. albopictus larvae respectively. CONCLUSION: The findings suggested that the essential oils from Clausena excavata leaf and twig and their effective constituents may be explored as a potential natural larvicide. Copyright © 2008 Society of Chemical Industry

A carbazole-pyranocoumarin conjugate, carbazomarin B (1), and two carbazole alkaloids, 6-methoxymukonidine (2) and 2-hydroxy-3-methoxycarbazole (3), together with 27 known compounds (4–30), were isolated from the stems of Clausena excavata. Their structures have been elucidated by spectroscopic analyses. Compound 2 showed moderate cytotoxicity to HuCCA-1, MOLT-3 and HepG2 cancer cell lines with IC50 values of 15.09–28.50 μg/mL, but none to A549 cell line. Heptaphylline (6) and nordentatin (23) were found to show moderate cytotoxic activity against HepG2 cell line with IC50 values of 12.33 and 11.33, respectively, while clausine K (27) exhibited strong cytotoxicity with IC50 value of 1.05 μg/mL, better than a standard drug (etoposide, IC50 13.40 μg/mL).

Shaymaa Fadhel Abbas Albaayit,1,2 Yusuf Abba,3 Rasedee Abdullah,3 Noorlidah Abdullah1 1Institute of Biological Sciences, Faculty of Science, University ofMalaya, Kuala Lumpur, Malaysia; 2Department of Biology, College of Science, University of Baghdad, Baghdad, Iraq; 3Department of Pathology and Microbiology, Faculty ofVeterinary Medicine, Universiti Putra Malaysia, Serdang, Selangor, Malaysia Abstract: Clausena excavata is a well-known plant used in folkloric medicine for the treatment of different ailments. This study aimed to determine the in vitro cytoxicity of its leaf solvent extracts as well as the in vivo wound healing and antioxidant activities of the methanolic extracts of C. excavata (MECE). HaCaT (keratocyte) and Vero cell lines were used for evaluation of the in vitro cytotoxic effects, while the in vivo wound healing and antioxidant activities were determined in skin wounds inflicted on rats. Twenty adult male Sprague-Dawley rats were divided into five groups of four animals each. Approximately 3.14 cm2 excisional wound was inflicted on the nape of each rat following anesthesia. The treatment groups received topical application of MECE at 50 mg/mL (MECE-LD [low dose]), 100 mg/mL (MECE-MD [medium dose]), and 200 mg/mL (MECE-HD [high dose]), while the negative control group was treated with gum acacia in normal saline and the positive control group with intrasite gel. Wound contraction was evaluated on days 5, 10, and 15 after wound infliction, and tissue from wound area was collected at day 15 post-wound infliction for antioxidant enzyme evaluation and histopathological analyses. Generally, Vero cells were more resistant to the cytotoxic effects of the solvent extracts as compared with HaCaT cells. Chloroform (CH) and ethyl acetate (EA) extracts of C. excavata were toxic to HaCaT cells at 200 and 400 µg/mL, but the same concentrations showed higher (P

Damayanti F, Helmanto H. 2015. Seed germination and seedling growth of Clausena excavata on Bioposka composting treatment. Pros Sem Nas Masy Biodiv Indon 1: 856-859. Sand is one of the most common media used for germinating the seeds. Sand has some deficiency that the surface area is relatively small, the ability to store water is so low that dry faster media. This study aimed to determine the effect of Bioposka on seed germination and seedling growth of Clausena excavata. The research method used a Completely Randomized Design (CRD). Treatment of media was sand, sand: Bioposka (1:1) and pure Bioposka. Each media was made two replications and each replication contained 10 units of seeds. The parameters measured were the number of seeds germinated and high of seedlings for 3 months. Data analysis of viability (germination total, normal germination, first day of germination, the last day of germination, germination rate coefficient, coefficient of simultaneity grow) and height of seedlings used Microsoft Excel and Analysis of Variance used STAR (Statistical Tool for Agricultural Research) software. The results showed that the viability of the media of sand: Bioposka and pure Bioposka had as high as 100% and 90% for sand media. Analysis of variance showed that seedling growth significant difference to the media in the level of 5%. Further, the test results of DMRT (Duncan's Multiple Range Test) showed that sand media produced the highest growth of C. exavata with average growth of 10.19 cm, Bioposka 7.71 cm, and sand: Bioposka 5.06 cm.

Hepatocellular carcinoma (HCC) is a primary liver cancer with high global incidence and mortality rates. Current candidate drugs to treat HCC remain lacking and those in use possess undesirable side effects. In this investigation, the antiproliferative effects of dentatin (DTN), a natural coumarin, were evaluated on HepG2 cells and DTN's probable preliminary molecular mechanisms in apoptosis induction were further investigated. DTN significantly (p0.05) suppressed proliferation of HepG2 cells with an IC50 value of 12.0 μg/mL, without affecting human normal liver cells, WRL-68 (IC5050 μg/mL) causing G0/G1 cell cycle arrest via apoptosis induction. Caspase colorimetric assays showed markedly increased levels of caspase-3 and caspase-9 activities throughout the treatment period. Western blotting of treated HepG2 cells revealed inhibition of NF-κB that triggers the mitochondrial-mediated apoptotic signaling pathway by up-regulating cytoplasmic cytochrome c and Bax, and down-regulating Bcl-2 and Bcl-xL. The current findings suggest DTN has the potential to be developed further as an anticancer compound targeting human HCC.

Immunomodulating effects in Balb/C mice of aqueous extract (CEHW) and the Thai folklore preparation (CEHF) of Clausena excavata Burm. f. were investigated. Haemagglutinating antibody (HA) titers at day 0, 7, 14, 21, 28 and 35 from the serum of animals fed or injected intraperitoneally with the extracts for 5 days were compared and evaluated for humoral mediated immunity (HMI). Footpad swelling test was used to determine delayed type hypersensitivity for cell mediated immunity (CMI). CEHW and CEHF injected intraperitoneally and administered orally exhibited the same maximum antibody production of 1/800. Both extracts given orally reached the maximum antibody titer at day 7, which was 2 weeks faster than by intraperitoneal administration. However, antibody titers from CEHW injected intraperitoneally diminished without retaining whereas the CEHF retained for 1 week. Moreover, CEHW gave CMI response more than the CEHF. This study suggested the potential in vivo immunomodulating activities of extracts from Clausena excavata supporting our previous in vitro studies.

Five new tetranortriterpenoids, (11beta)-21,23-dihydro-11,21-dihydroxy-23-oxoobacunone (=21,23-dihydro-21-hydroxy-231-oxozapoterin: 2), (11beta)-21,23-dihydro-11,23-dihydroxy-21-oxoobacunone (=21,23-dihydro-23-hydroxy-21-oxozapoterin; 3), (1alpha,11beta)-1,2,21,23-tetrahydro-1,11,23-trihydroxy-21-oxoobacunone (=21,23-dihydro-23-hydroxy-21-oxoclausenarin; 4), (1alpha,11beta)-23-ethoxy-1,2,21,23-tetrahydro-1,11-dihydroxy-21-oxoobacunone (=23-ethoxy-21,23-dihydro-21-oxoclausenarin; 5); (11beta)-1,2,21,23-tetrahydro-11,23-dihydroxy-21-oxoobacunoic acid; 6), were isolated from the aerial part of Clausena excavata BURM. F. (Rutaceae). All compounds possessed 3,4-seco skeletons. Their structures were established by spectroscopic studies. Tetranortriterpenoids with a 4-hydroxybut-2-eno-4-lactone moietv are rarely found in the genus Clausena.

Clausena excavata(Lour.), locally known as “Kemantu hitam,” is a common plant in Malaysian folklore medicine. This study evaluated the antioxidant properties of the solvent extracts ofC. excavataleaves and determined the acute toxicity of methanolic extractC. excavata(MECE) leaves in Sprague-Dawley rats. Harvested leaves were dried and subjected to solvent extraction using petroleum ether, chloroform, ethyl acetate and methanol in succession. The antioxidant activity of each extract was determined using the ferric-reducing antioxidant power (FRAP) and 2,2-diphenyl-1-picryl dihydrazyl (DPPH) radical scavenging activity. The total phenolic content (TPC) and total flavonoids content (TFC) were estimated by Folin-Ciocalteu and ethanolic aluminium chloride method, respectively. The chloroform extract was found to be highest in flavonoid content, while the methanolic extract showed the highest TPC and antioxidant activity. There was no mortality in rats treated with MECE leaves even at a high dose of 5000 mg/kg body weight. However, the MECE leaves produced mild to moderate pathological changes in the liver and kidneys, shown by mild degenerative changes and leucocyte infiltration. The extract did not affect the haematological parameters or relative weights of the liver or kidneys. Overall, the MECE leaves have potent antioxidant activity and are presumed safe to be used orally as health-promoting product at low to moderate doses.

Clausena excavata, a native of Southeast Asia, displays characteristics of disease tolerance and adaptability to different kinds of soils. This species contains potential parents for the improvement of Citrus fruit rootstocks. However, it is tetraploid, unlike most Citrus and related species, which are diploid. Bringing the ploidy level back to 2n=2x would allow us to consider integrating this species into the varietal creation programme by somatic hybridisation. A diplohaploidisation programme by anther culture is currently being conducted. The effects of the hormonal balance on androgenesis and on the maturation of androgenetic embryos are described. Thirty-six hormonal combinations using benzylamino purine (BAP) and dichlorophenoxyacetic acid (2,4-D) were examined. The tendency to embryogenesis varies from 0 to 9% depending on the hormonal concentration and combination. A total of 106 plants were regenerated, of which 91 were hexaploid, 11 were tetraploid, and only two were diploid after the flow cytometry evaluation. Histological study showed that the embryos were derived not from somatic embryogenesis of the mother tissues but from microspores. (Resume d'auteur)

In addition to a known alkaloid, some limonoids and coumarins, the new coumarins excavatins A–M have been isolated from Clausena excavata. Their structures have been assigned by NMR and CD investigations.

Ten new carbazole alkaloids, clausine-M, -N, -O, -P, -Q, -R, -S, -U, -V and clausenatine-A, together with 39 known compounds were isolated and identified from the acetone extract of the root bark of Clausena excavata. Their structures have been elucidated by spectroscopic analyses. This is the first report of a decarboxylated carbazole alkaloid from a natural source. The presence of a C23-skeleton carbazole alkaloid in a plant of Clausena genus is also reported for the first time.

Constituents of the roots, the stem bark, and the leaves of Clausena excavata BURM. f. (Rutaceae) grown in a greenhouse at Okitsu Branch, Fruit Tree Research Station, Shizuoka, were studied. New carbazole alkaloids named clauszoline-H (1), -I (2), and -J (4) from the roots, clauszoline-K (6) and -L (7) from the stem bark, and clauszoline-M (8) from the leaves were isolated, together with known carbazoles and coumarins, and their structures were elucidated by spectroscopic methods.

Somatic embryos were induced from immature seeds of eight Citrus relatives including Aegle marmelos, Atalantia ceylanica, Citropsis gabunensis, Clausena excavata, Glycosmis pentaphylla, Microcitrus australasica, Murraya paniculata and Severinia buxifolia on MT medium supplemented with 0.05 mg l−1 2,4-D, 0.05 mg l−1 BA and 400 mg l−1 malt extract. Approximately 20% of somatic embryos from six of the genera underwent callogenesis during subculture on the same medium. After at least six successive subcultures on MT medium containing 10 mg l−1 BA, calli became growth regulator independent. Somatic embryos from growth regulator independent calli were obtained with MT medium containing 5% lactose. Plantlets were regenerated from somatic embryos on MT medium containing 0.02 mg l−1 NAA, and established in soil.