Diaporthe sp. F18; a new source of camptothecin-producing endophytic fungus from Nothapodytes nimmoniana growing in Sri Lanka.

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.

Acute and Subacute Toxicity Studies of the Ethyl Acetate Soluble Proanthocyanidins of the Immature Inflorescence of Cocos nucifera L. in Female Wistar Rats.

Ayurvedic and traditional medical practitioners of Sri Lanka use the decoction of the immature inflorescence of Cocos nucifera L. (IC) variety aurantiaca for the treatment of menorrhagia. The progestogenic effect of the ethyl acetate soluble proanthocyanidins (EASPA) of the IC in female rats at a dose of 3.5 mg/kg body weight has been reported. Acute and subacute toxicity studies of EASPA of the IC carried out using female Wistar rats according to Organization for Economic Co-operation and Development (OECD) guidelines 423 and 407, respectively, are reported herein. In the acute toxicity study, a single dose of EASPA (2000 mg/kg body weight) was orally administered to rats, which were monitored for 14 days. In the subacute toxicity study, rats were orally administered with EASPA daily for 28 days at doses of 1.75, 3.5, 7, and 14 mg/kg body weight. No rat in either the acute or subacute toxicity study exhibited mortality or clinical signs of toxicity. Further, these rats did not show any significant change in their mean body weight, food, and water intake, haematological and biochemical parameters as well as in the results of their histopathological examinations compared to those of control group rats. According to results of the acute toxicity, the LD50 of EASPA is estimated to be greater than 2000 mg/kg body weight. Considering the results of the subacute toxicity study, the oral administration of EASPA daily for 28 days was well tolerated up to the dose, 14 mg/kg by rats. These results will be useful in the development of a novel therapeutic agent from EASPA of the IC for the treatment of menorrhagia, which incapacitates a considerable proportion of women worldwide.

Acute anti-inflammatory and anti-nociceptive activities of crude extracts, alkaloid fraction and evolitrine from Acronychia pedunculata leaves.

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.

An analysis of 1505 consecutive patients receiving continuous interscalene analgesia at home: a multicentre prospective safety study.

Continuous interscalene brachial plexus block has been shown to be the most effective analgesic technique following shoulder surgery; however, its use is uncommon due to logistical and safety concerns related to ambulatory administration. We prospectively studied 1505 consecutive patients undergoing shoulder surgery who received continuous interscalene analgesia at home. Catheter removal was by the patient between postoperative days two and five. There were no major complications although 27% of patients reported mild dyspnoea, 13% hoarseness and 7% dysphagia. Twelve percent sought medical advice and 2% reported technical issues with the pump or tubing. Complications and technical issues were associated with patient age; weight; use of ultrasound or concomitant nerve stimulation as the endpoint for final needle tip position; local anaesthetic placement via the catheter or needle; whether a catheter-related intervention for pain relief was required in the recovery area; and the type of ambulatory pump. We conclude that this study supports the safety of this underused analgesic technique.

Patient-initiated mandatory boluses for ambulatory continuous interscalene analgesia: an effective strategy for optimizing analgesia and minimizing side-effects.

BACKGROUND: This prospective, randomized study tested the hypothesis that a reduced dose continuous interscalene regimen incorporating a low background infusion with mandatory boluses would provide similar shoulder surgery analgesia compared with a dose regimen incorporating a conventional higher background infusion. METHODS: After rotator cuff surgery, patients received via an interscalene catheter, one of two elastomeric pumps, each having a 5 ml per 60 min bolus function and a 2 ml h⁻¹ (n=38) or 5 ml h⁻¹ (n=43) ropivacaine 2 mg ml⁻¹ infusion. Boluses commenced from the onset of pain and continued for >48 h as required (pro re nata, PRN) up to every hour for a numerical rating pain score (NRPS, 0-10) >2. Group 2 ml h⁻¹ received mandatory 6 hourly boluses irrespective of the NRPS. Rescue tramadol was available. Patients were questioned on postoperative days 1 and 2 for treatment effectiveness and side-effects. RESULTS: Postoperative pain was similar between the groups [Group 2 ml h⁻¹ day 2 median (IQR) (95% confidence interval of the mean) worst movement pain=4 (1-5) (2.8-4.7) vs 4 (2-5) (3.1-4.6), P=0.99], as were night awakenings and tramadol consumption. Numerically rated numbness and weakness were similar between the groups; however, nine patients (21%) in the 5 ml h⁻¹ group vs one (3%) in the 2 ml h⁻¹ group required a temporary infusion cessation due to side-effects (predominantly hand numbness) (P=0.02). CONCLUSIONS: Continuous interscalene ropivacaine 0.2% 2 ml h⁻¹ with mandatory 6 hourly (and PRN) boluses provides similar analgesia after rotator cuff repair but with reduced side-effects compared with 5 ml h⁻¹ with PRN only boluses.

Antioxidant and antibacterial activities on foodborne pathogens of Artocarpus heterophyllus Lam. (Moraceae) leaves extracts.

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.

Drug error in anaesthetic practice: a review of 896 reports from the Australian Incident Monitoring Study database.

Eight hundred and ninety-six incidents relating to drug error were reported to the Australian Incident Monitoring Study. Syringe and drug preparation errors accounted for 452 (50.4%) incidents, including 169 (18.9%) involving syringe swaps where the drug was correctly labelled but given in error, and 187 (20.8%) due to selection of the wrong ampoule or drug labelling errors. The drugs most commonly involved were neuromuscular blocking agents, followed by opioids. Equipment misuse or malfunction accounted for a further 234 (26.1%) incidents; incorrect route of administration 126 (14.1%) incidents; and communication error 35 (3.9%) incidents. The outcomes of these events included minor morbidity in 105 (11.7%), major morbidity in 42 (4.7%), death in three (0.3%) and awareness under anaesthesia in 40 (4.4%) incidents. Contributing factors included inattention, haste, drug labelling error, communication failure and fatigue. Factors minimising the events were prior experience and training, rechecking equipment and monitors capable of detecting the incident. The information gained suggests areas where improved guidelines are required to reduce the incidence of drug error. Further research is required into the effectiveness of preventive strategies.

Antiradical and antilipoperoxidative effects of some plant extracts used by Sri Lankan traditional medical practitioners for cardioprotection.

Reactive oxygen species (ROS) are implicated in many pathogenic processes including the cardiovascular system. Detoxification of ROS by antioxidants (AO) therefore affords protection against such diseases. There is a growing body of evidence suggesting that antioxidants contribute to cardioprotection. Therefore, nine plants that are components of Ayurvedic formulations used for the therapy of cardiovascular diseases were investigated to determine whether antioxidant activity is one of the mechanisms by which these plants exert cardioprotection. Initially aqueous freeze dried extracts of the plants were prepared and the antioxidant activity was measured (a) in vitro, by DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging and deoxyribose damage protection assays, and (b) in vivo, by effects on lipid peroxidation. Terminalia arjuna showed significant DPPH radical scavenging activity with EC(50) 8.3 +/- 0.3 microg/mL (similar to L-ascorbic acid). The potency of this activity was much lower in Cassia fistula (EC(50) = 59.0 +/- 2.7 microg/mL). The other seven extracts demonstrated no such activity in the concentration range tested. In the deoxyribose damage protection assay, T. arjuna> demonstrated no significant effect in the concentration range 0-20 microg/mL, but above -20 microg/mL concentration (20-125 microg/mL), a pro-oxidant activity was observed (although markedly less than demonstrated by L-ascorbic acid). A similar trend was observed with Vitex negundo. In contrast, C. fistula afforded a 30% protection against such damage at 125 microg/mL concentration. Other plant extracts did not show any activity in this assay. At a dose of 90 mg/kg (single dose) T. arjuna, cardiac lipid peroxidation in male Wistar rats was reduced by 38.8% +/- 2.6% (p<0.05) whereas the reduction was only 11.6% +/- 3.5% in the case of C. fistula even at a dose of 120 mg/kg. Of all the plants tested, T. arjuna demonstrated the highest antioxidant activity. Overall results show that only some plants used in the therapy of cardiovascular disease exert their beneficial effects via antioxidant activity.

Fermentation in traditional medicine: the impact of Woodfordia fruticosa flowers on the immunomodulatory activity, and the alcohol and sugar contents of Nimba arishta.

The impact of Woodfordia fruticosa flowers on the immunomodulatory activity, and alcohol and sugar contents of the ayurvedic drug 'Nimba arishta' was investigated by means of model preparations. The use of Woodfordia flowers in model preparations resulted in a substantial increase of the inhibition of both human complement activity and chemiluminescence generated by zymosan-stimulated human polymorphonuclear leukocytes. It was established that the increased biological activity was not due to microbial interference, but to immuno-active constituents released from the Woodfordia flowers. It was also found that the flowers themselves are not the source of alcohol-producing microorganisms. Experiments performed with yeasts isolated from commercial Nimba arishtas showed, in agreement with empirical findings, significantly raised alcohol content upon addition of Woodfordia. An invertase activity exhibited by Woodfordia flowers may be causative of this effect.

An ethnopharmacognostic approach to the search for immunomodulators of plant origin.

The search for immunomodulating plant constituents through basic and field inquiries into the literature and practices of traditional Indian medicine is treated. The strategy of data collecting proceeds through aspects of an ethnobotanical, an ethnopharmaceutical, an ethnopharmacological, and an ethnomedical nature. In the experimental immunopharmacognostic phase, immunomodulatory compounds are isolated and purified through action-guided fractionation procedures. The results described here refer to activities found on human complement activation and on PMN leucocytes activation. The immunomodulating plant compounds included in this report were isolated from Azadirachta indica bark, Woodfordia fructicosa flowers, Picrorhiza kurroa roots, and Jatropha multifida latex.