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An endophytic fungus producing camptothecin (CPT) was isolated from the leaf of Nothapodytes nimmoniana (Sri Lanka), and culture conditions were optimised to enhance the yield of CPT. The TLC, HPLC-PDA, LC-MS/MS and spectroscopic data were used to identify and quantify CPT. Solvent extraction (chloroform: methanol 4:1 v/v) of submerged cultures in Sabouraud Dextrose Broth (SDB) detected CPT in the mycelial extract but not in the culture broth. The fungus was (KX212080) closely related to Diaporthe guangxiensis (MK335772) with 99% sequence similarity, thus tentatively identified as Diaporthe sp. F18. A significantly high CPT content (72.0 ± 0.2 µg/g) was produced in SDB, pH, 5.6 incubated at 30 °C under shake flask condition (150 rpm) for 14 days. Tryptophan significantly (p > 0.05) enhanced CPT production while ethanol increased it by 8-fold. This endophytic source produced higher CPT content than what has been reported hitherto in the literature, with fairly stable production up to sixth subculture generations.

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Murraya koenigii (M. koenigii), Micromelum minutum (M. minutum), and Clausena indica (C. indica) are three varieties of curry leaves in the family Rutaceae. They have been widely used in Ayurvedic medicine worldwide in the treatment and prevention of various diseases. Earlier findings provide strong evidence to support the three curry leaf species' potent pharmaceutical and biological effects, including antioxidant, antidiabetic, anti-inflammatory, and antitumor activities. Various parts of these plants, such as leaves, seeds, flowers, and fruit, contain constituents responsible for the modulation of numerous biological processes. Leading constituents of curry leaves play a crucial role in diabetic and anticancer management by regulating various molecular pathways, including Bcl-2, Bax, NF-κB, and TNFα, according to in vitro and in vivo models established. Therefore, this review summarizes the current knowledge on research achievements made in terms of phytoconstituents, their structures, biological activities, and pharmacological actions with clinical studies of curry leaves up to date. The review also emphasizes the necessity for comprehensive research studies on the pharmacological actions and the mechanisms of selected phytochemicals of M. koenigii, M. minutum, and C. indica to validate their efficacy as potent herbal remedies for many ailments.

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The use of plant materials in traditional medicine as a significant ingredient in synthesizing drugs in the form of decoctions had played a vital role due to their potential therapeutic action. The curry leaves, Murraya koenigii, and Micromelum minutum are two most common aromatic herbs widespread in Sri Lanka and many other Asian countries. They are rich in nutrition and exert several medicinal properties such as antidiabetic, antioxidant, antimicrobial, anti-inflammatory, and anti-carcinogenic due to various essential phytochemicals, minerals, and trace minerals. In this study, we determined the proximate composition, in vitro antioxidant activity, total phenol (TPC), flavonoid content (TFC), and antibacterial activity in both species using standard methods. Results showed that both species varied greatly in nutritional composition, antioxidant, and antibacterial activities. The nutrient composition in terms of ash, crude protein, and fat was significantly high in M. koenigii based on dry weight basis. In vitro antioxidant potential of M. koenigii and M. minutum was evaluated by means of the DPPH radical scavenging assay, and the IC50 values of M. koenigii and M. minutum were 107 ± 2 µg/mL and 208 ± 4 µg/mL, respectively. The TPC and TFC of M. koenigii were 101 ± 1 mg GAE/g and 9.75 ± 0.05 mg QE/g, and M. minutum were 80 ± 2 mg GAE/g and 9.16 ± 0.02 mg QE/g respectively. The antibacterial study was assessed against three bacterial strains. The study showed the highest inhibitory activity for M. koenigii and M. minutum against Staphylococcus aureus and Escherichia coli, respectively at 1.0 mg/mL concentration. The study indicated that M. koenigii possessed a significantly high TPC, and TFC correlated with higher antibacterial and antioxidant activity than M. minutum.

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ETHNOPHARMACOLOGICAL RELEVANCE: Acronychia pedunculata (family: Rutaceae) is one of the commonly used medicinal plants in Sri Lankan traditional medicine. Different parts of this plant are used for the treatment of inflammatory conditions in the form of medicinal oils and herbal porridge. AIM OF THE STUDY: The present study aimed to evaluate the anti-nociceptive activity and anti-inflammatory activity with their mechanisms and the acute toxicity of crude extracts of the fresh leaves of A. pedunculata for scientific validation of the ethnopharmacological claims for this plant. Further, attention has been focused on the isolation of active compounds from active fractions of the crude extracts. MATERIALS AND METHODS: The acute anti-inflammatory effect of the aqueous (AELA) and 70% ethanol crude extracts (EELA) and alkaloid fraction of A. pedunculata leaves were evaluated by the determination of inhibition of hind paw oedema induced by carrageenan in Wistar rats. Evolitrine was identified as the major alkaloid with significant bioactivities by column chromatography and NMR. The anti-nociceptive and anti-histamine activities of EELA and evolitrine were evaluated by acetic acid induced writhing and wheal formation tests respectively. In addition, in-vitro (2, 2-diphenyl-1-picrylhydrazyl (DPPH) assay) and in-vivo (lipid peroxidation assay) anti-oxidant activity, nitric oxide (NO) inhibitory activity and acute toxicity of EELA were evaluated. RESULTS: Acute anti-inflammatory activity of AELA and EELA were dose-dependent. EELA was more active than AELA. The 200 mg/kg body weight (b. w.) dose of EELA was found as the minimum effective dose with maximum inhibition (78%) of oedema at 5th hour compared to the negative control (p < 0.05). Evolitrine was isolated and identified as an active anti-inflammatory and analgesic compound from active alkaloid fraction of EELA. Evolitrine showed activity enhancement when compared with crude EELA. The anti-inflammatory and analgesic activities of evolitrine (50 mg/kg b. w.) were comparable to that of reference drugs indomethacin (5 mg/kg b. w) and acetylsalicylic acid (100 mg/kg b. w.). The significant (p < 0.05) anti-histamine activity, DPPH scavenging in-vitro anti-oxidant activity, in-vivo lipid peroxidation inhibitory activity in-vivo, NO inhibitory activity of EELA as compared with relevant negative controls, were identified as probable mechanisms which mediated its anti-inflammatory action. Further, EELA showed a high safety margin in the limited dose acute toxicity study. CONCLUSION: The findings of the current study rationalize the usage of leaves of A. pedunculata in Sri Lankan traditional medicine as an analgesic and anti-inflammatory agent. Possible mechanisms mediating this activity included anti-histamine, anti-oxidant and NO inhibitory activities. Evolitrine is the major analgesic and anti-inflammatory compound isolated from the active alkaloid fraction of EELA.

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Six morphotypes of Centella asiatica designated G(1), G(2), G(3), G(7), G(8) and G(12) were analyzed for carotenoid content, proximate components and selected dietary minerals. Potassium was found to be the major element present in C. asiatica and the content in G(1) and G(3) (6,165.0 ± 125.8 mg/100 g dry weight) was significantly (P < 0.05) higher than all other morphotypes. C. asiatica is more popular for its higher contents of iron and calcium. However, no significant (P>0.05) differences in calcium content among the morphotypes were observed. The imported variety G(7) had significantly higher (P < 0.05) mean content of iron (74.3 ± 34.1 mg/100 g dry weight). The morphotypes G(8) and G(12) contained significantly higher (P < 0.05) content of ß-carotene (77.2 ± 5.1 µg/g fresh weight) and lutein (133.5 ± 10.1 µg/g, fresh weight) respectively. Lutein and ß-carotene levels of these two types were also significantly higher (P < 0.05) than those in the other varieties. The study did not provide any evidence to believe that a given morphotype is superior to another in its overall nutritional value.

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Total water extract, ethyl acetate, and aqueous fractions from the leaves of Artocarpus heterophyllus were evaluated for phenolic content, antioxidant, and antibacterial activities against some foodborne pathogens such as E. coli, Listeria monocytogenes, Salmonella typhimurium, Salmonella enterica, Bacillus cereus, Enterococcus faecalis, and Staphylococcus aureus. The minimum inhibitory concentration (MICs) of extract and fractions determined by the agar dilution method were ranged from 221.9 microg/mL for ethyl acetate fraction to 488.1 microg/mL for total extract. In the agar diffusion method the diameters of inhibition were 12.2 for the total extract, 10.7 and 11.5 for ethyl acetate and aqueous fractions, respectively. A. heterophyllus showed significant antioxidant activity tested in different in vitro systems (DPPH, ABTS, FRAP, and Fe(2+) chelating activity assay). In particular, in DPPH assay A. heterophyllus total extract exhibited a strong antiradical activity with an IC(50) value of 73.5 microg/mL while aqueous fraction exerted the highest activity in FRAP assay (IC(50) value of 72.0 microg/mL). The total phenols content by Folin-Ciocalteau method was determined with the purpose of testing its relationship with the antioxidant and antibacterial activities.

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The carotenoid content and in vitro accessibility of the 'Sugar baby' variety of watermelon and the 'Horana red' variety of guava from Sri Lanka was determined using high-performance liquid chromatography. The high-performance liquid chromatography chromatogram showed that the Guava 'Horana red' variety contained almost exclusively lycopene (45.3 +/- 8.0 microg/g fresh weight (FW)), with a small amount of lutein (2.1 +/- 0.6 microg/g FW), beta-carotene (2.0 +/- 0.2 microg/g FW) and beta-cryptoxanthin. As far as carotenoids in the sugar baby variety of watermelon are concerned, it contained lycopene, lutein and beta-carotene of 37.2 +/- 4.0 microg/g FW, 2.1 +/- 0.6 microg/g FW and 0.3 +/- 1 microg/g FW, respectively. The studies showed that guava contains more lycopene (45.3+/-8.0 microg/g FW) than watermelon (37.2 +/- 4.0 microg/g FW), and that the in vitro accessibility of lycopene in guava (73%) is more than that in watermelon (25.8%). Therefore it can be concluded that guava can be used as a better lycopene source than watermelon.

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Vitamin A deficiency is a public health problem in Sri Lanka, which affects especially pre-school children. Carrots (Daucus carota), pumpkins (Cucurbita maxima), squashes (Cucurbita moschata) and sweet potatoes (Ipomoea batatas) of orange, yellow-fleshed varieties are good sources of provitamin A carotenoids, but have not been studied in Sri Lanka in terms of the food as eaten. The content of carotenoids in each preparation method and the in-vitro accessibility of beta-carotene and alpha-carotene were determined by high-performance liquid chromatography. The in-vitro method simulates the conditions in the human intestinal tract. The mean contents in dry weight (DW) in different carrot preparations ranged from 406.7 to 456.3, from 183.7 to 213.5 and from 29.0 to 39.6 microg/g for beta-carotene, alpha-carotene and lutein, respectively. The content of carotenoids in different pumpkin preparations varied from 282.1 to 294.4 microg/g DW for beta-carotene, from 155.6 to 157.7 microg/g DW for alpha-carotene and from 218.0 to 228.2 microg/g DW for lutein. The squashes preparation had 44.6 and 40.0 microg/g DW for beta-carotene and alpha-carotene, respectively, and in sweet potatoes beta-carotene ranged from 204.3 to 210.3 microg/g DW. The results showed that the contribution to the recommended daily allowance is greater when these vegetables are prepared as a curry with coconut milk. The percentage contribution to recommended daily allowance from each vegetable cooked with coconut milk was 46.7, 21.8, 1.2 and 10.8 for carrots, pumpkins, squashes and sweet potatoes, respectively.

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