Participation of Large Ca2+ Activated Potassium Channel in Antinociceptive Activity of Chalcone Derivative (3-(2, 5-dimethoxyphenyl)-1-(5-methylfuran-2-yl) prop-2-en-1-one) DMPF-1 Action in Mice Model

The role of potassium channels in nociceptive activity was proposed in the past decade. Various type of potassium channel has been found to exert different action in propogation of action potential in nervous system. As DMPF-1, a chalcone derivative possesses antinociceptive properties. The mechanism of its action has been carried out to verify the pathway involved. The present study addressed the role of potassium channel in the contribution of the antinociceptive action of DMPF-1. The involvement of potassium channel was evaluated using acetic acid-induced abdominal writhing test.The animals were pretreated with charybdotoxin (large Ca2+ activated potassium channel blocker)(0.04mg/kg, i.p) or apamin (small Ca2+ activated potassium channel blocker)(0.02mg/kg, i.p.) 15 minutes before administration of DMPF-1. It was demonstrated that the challenge of DMPF-1 treated group with charybdotoxin has reversed the antinociceptive activity of this novel chalcone, which indicates the possible participation of large Ca2+ activated potassium channel in antinociceptive effect cause by DMPF-1 but not through small Ca2+ activated potassium channel

Establishment of human neuroblastoma cell line (SK-N-SH) as an in vitro model of morphine addiction

The usage of morphine for analgesic purposes are widely known, usually for the post-operation procedure and as chronic pain management. However, addiction and overdose liabilities prior to morphine usage are also common. Morphine addiction is observed and studied from various perspectives; tolerance, dependence and withdrawal. With growing and expending research field, researches on addiction were done using in vivo and in vitro model. However, the scientific evidence of morphine addiction using human neuroblastoma cell lines is uncommon. Thus, the present study was designed and conducted to observe the liability of SK-N-SH, as a model for morphine addiction. The cells were administrated with morphine for 24 hours before being treated with methadone. The cytosolic fraction of the cell was collected and used for determining the addiction mechanism. Data showed the involvement of the µ-opioid receptor in expressing the addictive properties of morphine. Exposure to 24-hours morphine had increased the protein level responsible for addiction and reduce the protein levels expressing the endocytic machinery, desensitisation of receptor and cellular adaptation. The altered proteins level was normalised by the treatment of methadone. The study proposed the use of SK-N-SH as an addiction model, as it showed morphine addiction and methadone anti-addiction properties

Structural conformation of Bacillus stearothermophilus F1 protease and effect of modification on its thermostability

The extracellular F1 serine protease, produced by a thermophilic Bacillus stearothermophilus F1, has been isolated and characterized as one of a serine protease. F1 protease was stable in the pH range of 8.0 to 10.0, with an optimum activity at pH 9.0. The enzyme was stable for 24h at 70°C (Rahman et al., 1994)

Thermostable proteases

New proteases are constantly isolated. Proteases, mainly the alkaline proteases are widely used in the detergent formulation. Thermostable alkaline proteases are of great interest in the detergent industry because these enzymes are stable and able to retain its activities over broad range of pH and temperatures. Most thermostable alkaline proteases reported are from Bacillus spp. In this chapter, we introduce a new thermostable alkaline protease produced from Bacillus stearothermophilus strain F1. The protease production requirements by strain F1, the properties and characterization of this enzyme are discussed. The protease F1 gene was successfully cloned and expressed into E. coli XL1-Blue. The bacteriocin release protein(BRP)system was utilized; to release the recombinant F1 protease into the culture medium. The purified native and recombinant F1 protease showed a pH optimum of 9.0 and thermostabilities ranging from a half-life of 25 min at 90◦C to a half-life of 4 h at 85◦C. The optimumtemperatures were 85◦C and 80◦C respectively

In vivo toxicity study on the effects of aqueous propolis extract from Malaysian stingless bee (Geniotrigona thoracica) in mice

Geniotrigona thoracica is a stingless bee species of Trigona genus. Propolis resinous is a natural product obtained from a honeybees hive with geographical and floral specifications or exudate as such by-products resulting from a variety of botanical processes. Despite its long use for a variety of health conditions, the toxicity profile of propolis sourced from Malaysian stingless bees has not been sufficiently evaluated. For in vivo toxicity assessment, the acute oral toxicity on the effects of aqueous propolis extracts (APE) was examined. Male mice swiss strain, were subjected to acute toxicity testing for 14 days. The APE at doses of 400, 1000 and 2000 mg/kg body weight was supplemented daily to the mice through oral gavage. The clinical signs of toxicity and general behaviour, body weight, relative organ weight, and histopathology changes were investigated. In vivo study was focused on the acute toxicity testing group consisting of 4 groups including Normal (NS), 400 mg/kg (APE 400), 1000 mg/kg (APE 1000) dan 2000 mg/kg (APE 2000). Regarding the toxicity profile, it is proposed that APE supplementation did not induce any mortality and no visible signs of toxicity. No significant changes in the body and relative organ weight were recorded. All the internal organs of the mice were macroscopically healthy with no gross lesion. Likewise, histopathological examinations of the kidney showed mild to moderate histological lesions. Interestingly, the lesion was adverse with an increased dosage of the extract supplementation. This study proposed APE has considerable anti-inflammatory activities. It also demonstrated that the propolis extract is relatively safe to be consumed orally at a dose of 2000 mg/kg body weight

Stakeholder involvement and preferences in landscape protection decision-making: a systematic literature review

Stakeholder involvement and preferences are pivotal in the decision-making process for landscape protection within a specific context. However, many decisions regarding landscape protection options still rely on management choices with little or no consideration of all stakeholders. Previous scholars emphasized the importance of establishing an integrated framework to gain an adequate understanding of the process of stakeholders’ decision-making in landscape protection. Therefore, a systematic literature review was conducted on the topic of stakeholders’ involvement and preferences in landscape protection decision-making. This review included research articles published from 2013 to 2023 using two databases and registers, namely, Science Direct and Google Scholar. A total of 110 research articles were identified and qualified for review based on the screening requirements, with an additional 15 documents for theories and backgrounds to provide a better understanding and outcomes for the study. The results of this study were organized based on concepts from the resulting research articles and were integrated to propose a conceptual framework for Stakeholders’ involvement and preferences in landscape protection. Additionally, this study’s findings indicate the significance of incorporating diverse stakeholders and their preferences in landscape protection processes to ensure awareness of inclusivity in decision-making and secure long-term support

Involvement of opioid receptors in Boesenbergia pandurata's esssential oil (BPEO)-induced antinociceptive activity in animal model of nociception

Boesenbergia pandurata is a folklore remedy for relieving stomach, abdominal, joint, and muscular pain. Previous study from our research group has shown that Boesenbergia pandurata’s essential oil (BPEO) possesses antinociceptive activity against chemical and thermal models of pain. The present study was conducted to evaluate participation of opioid receptors in BPEO-induced antinociceptive activity. The involvement opioid receptors were assessed using acetic acid-induced abdominal writhing test. The acetic acid-induced writhing test was conducted by administering the non-selective opioid receptor antagonists (naloxone) 15 minutes before administration of BPEO orally, and selective opioid receptor antagonists (beta-funaltrexamine, norbinaltorphimine, and naltrindole) 1 day before BPEO administration. 0.6% acetic acid was later injected intraperitoneally and 5 minutes after the injection mice was observed for writhing response in 30 minutes time span. It was demonstrated that oral administration of BPEO 300 mg/kg produced 82.19% inhibition of nociception induced by 0.6% acetic acid injection. Opioid receptor antagonists administration solely did not modify acetic acid-induced nociceptive behavior. However, administration of naloxone (non-selective opioid antagonist) significantly increases the nociceptive response of animal receiving BPEO in the acetic acid-induced writhing test. Furthermore, administration of beta-funaltrexamine (mu opioid receptor antagonist and norbinaltorphimine kappa opioid receptor antagonist) significantly reserved antinociceptive activity induced by BPEO. Together, these result suggested participation of opioid receptors in inducing antinociceptive in animal model. In conclusion, BPEO may exert its antinociceptive activity through activation of mu opioid receptor, as well as kappa opioid receptor

Antinociceptive and anti inflammatory activities of 3-(2,5 dimethoxyphenyl)-1-(5 methylfuran-2-YL) prop-2-en-1 in mice

Chalcone is being extensively explored due to its pharmacological properties. The search on chalcone derivatives as an analgesic agent increased as the current pain treatment caused severe side effects. This study aimed to determine the analgesic activity of new chalcone derivative, 3-(2,5-dimethoxy phenyl)-1-(5-methyl furan-2-yl) prop-2-en-1 (DMPF-1) and to identify its possible mechanism of actions. Analysis of acute and sub-acute toxicity of DMPF-1 supplementation was performed. A single oral dose (1000 mg/kg) and 28-days repeated treatment of DMPF-1 (0.1-10 mg/kg) showed no significant changes in body weight, haematological, serum biochemical, macroscopic and microscopic analysis proved the absence of changes in the treated subjects. Antinociceptive study of DMPF-1 was started using the acetic acid-induced abdominal writhing test, formalin-induced paw licking test and hot plate test. The results showed DMPF-1 significantly reduce pain in a dose-dependent manner and suggests central and peripheral antinociceptive effect. Accordingly, the study on the possible involvement of opioid receptors was done. The challenge of DMPF-1 with naloxone showed no reversion of its antinociceptive effect, suggesting no contribution to the opioid system. Further examination using capsaicin, glutamate and phorbol 12-myristate 13-acetate (PMA)-induced paw licking test showed that the systemic administration of DMPF-1 at various doses significantly reduced the nociceptive response of the mice in a dose-dependent manner. This result proposed that DMPF-1 was acted through the vanilloid, glutamatergic and protein kinase C system in mediating analgesic action. In addition, the pre-treatment of DMPF-1 with L-arginine and ODQ had reversed the DMPF-1 antinociceptive effect indicating the involvement of nitric oxide and cyclic GMP synthesis. In sequence, pre-treatment of the animals with various potassium channels blockers such as charybdotoxin, glibenclamide and tetraethylammonium significantly abolished its antinociceptive activity, which suggests the facilitation of NO-cGMP pathway, large Ca2+ activated, ATP sensitive, and voltage-dependent potassium channels in its mechanism of action implied. At present, the possible involvement of various inhibitory neuroreceptors was performed. Pre-treatment with bicuculine appears to block its antinociceptive profile in which the event was not seen in pre-treatment with phaclofen, thus suggesting the involvement of the GABAA receptor. Meanwhile, pre-treatment of the subject with haloperidol and metoclopramide was carried out to investigate the involvement of dopaminergic receptors. Marked inhibition of DMFP-1 activity by only metoclopramide was observed, which indicated the contribution of the D2 dopaminergic receptor. Further investigation using atropine, yohimbine, and caffeine attenuated its antinociceptive action, thus suggesting those receptors' participation in pain modulation. Moreover, pre-treatment of the mice with various serotonergic receptor antagonists, including WAY 100635, pindolol and kentanserin but not ondansetron, however, fail to affect the antinociceptive activity of DMPF-1. This concludes that DMPF-1 could stimulate the 5HT3 receptor in order to produce antinociception. A paw oedema test was carried out to determine its peripheral antinociceptive capability. DMPF-1 at various dosages can reduce the volume of paw oedema induced by carrageenan, bradykinin, substance P, prostaglandin E, histamine, arachidonic acid and serotonin. Results indicate that DMPF-1 exerts peripheral activity by attenuating the action of the inflammatory mediators. In conclusion, this study confirms the antinociceptive activities of DMPF-1 and elucidates the possible mechanism of action through which it exerts its antinociceptive effects

Willingness to pay for renewable energy : evidence from high wind and wave energy potential areas

The study aims to investigate consumers’ willingness to pay for the development of renewable energy to encourage renewable energy consumption in the long run. This study further identifies the factors that influence consumers’ willingness to pay for renewable energy. A face-to-face questionnaire, with a contingent valuation method was administered to 672 residential electricity users in the East Coast of Peninsular Malaysia, areas that have high wind and wave energy potentials with large hydropower. The findings show that the average consumers’ willingness to pay is about RM 4.90 or about USD 1.18 per month on electricity bills as a contribution to a Renewable Energy Fund. The results further show that the consumers’ willingness to pay for renewable energy is significantly influenced by the proposed bid price, income, confidence level and education. The findings may help the government in creating a useful framework to ensure sustainable development in the future

Acute and subacute toxicity profile of (3-(2,5-dimethoxyphenyl)-1-(5-methylfuran-2-yl) prop-2-en-1-one, chalcone derivative in experimental animal model

Evaluation of toxicity induced in laboratory animals is paramount to the screening step in the assessment of the safety profile of all compounds. 3-(2,5-dimethoxyphenyl)-1-(5-methyl furan-2-yl) prop-2-en-1-one (DMPF-1); synthetic chalcone derivative has been investigated due to its pharmacological properties, including its antinociceptive and anti-inflammatory. However, the safety profile of this compound is yet to be determined. The present study seeks to highlight the toxicity effect of this compound using acute and subacute toxicity studies in the mice model. A single highest dose (1000 mg/kg) and repeated dose (0.1, 0.5, 1, 5 and 10 mg/kg) of DMPF-1 supplementation were executed. All toxicity study performed was supported by behavioural and body weight changes, haematological, serum biochemical analysis, macroscopic and microscopic analysis of the vital organs. The present result simplifies that DMPF-1 compound supplementation for four weeks is non-toxic as it caused no significant alteration in mice body weight and behaviour. Besides, no significant changes in haematological and biochemical parameters were observed. Further evaluation of its safety profile was confirmed by the normal architecture of the tissues organs obtained. Collectively, this report showed that DMPF-1 was safe to be consumed