Black Pepper Extract Research Articles

Graphene oxide–metal nanocomposites for cancer biomarker detection

We report a universal protocol for the in situ bioinspired green synthesis of metal nanoparticles on simultaneously reduced graphene oxide sheets using a black pepper extract for quantification of breast cancer biomarker.

EVALUATION OF CYCLIC ADENOSINE MONOPHOSPHATE AND NEUROTRANSMITTERS IN EXPERIMENTAL OBESITY: IMPACT OF BLACK PEPPER AND COFFEE AQUEOUS EXTRACTS

ABSTRACTObjective: This study aims to evaluate the effect of black pepper and coffee extracts on chronic and acute experimental-induced obesity and energyhomeostasis.Methods: Rats were divided into 10 groups including control, high-fat diet (HFD), triton, HFD+triton, black pepper+HFD, black pepper+HFD+triton,coffee+HFD, coffee+HFD+triton, mixture+HFD, and mixture+HFD+triton groups. Blood glucose, serum insulin, and insulin resistance were estimated.Body mass index, food efficiency intake, and body weight gain were calculated. Lipid profile, liver and kidney functions were measured, serum and braincyclic adenosine monophosphate (cAMP) was estimated, and brain neurotransmitters were measured by high-performance liquid chromatography.Furthermore, histopathology of liver was performed.Results: Findings showed that blood glucose, insulin resistance, lipid profile, kidney and liver functions as well as brain cAMP and neurotransmitterswere significantly increased, concomitant with a significant decrease in insulin resistance and serum cAMP in both HFD and triton-induced obesitygroups compared to control.Conclusion: Supplementation with black pepper extract, coffee extract, and a mixture of both significantly improved these findings. In conclusion,black pepper and coffee extracts are overlooked as promising weight reduction and antihyperlipidemic agents.Keywords: Energy homeostasis, Obesity, Black pepper extract, Coffee extract, Cyclic adenosine monophosphate, Neurotransmitters.

Development of chitosan based gold nanomaterial as an efficient antifilarial agent: A mechanistic approach

The gold nanoparticles (AuNPs) have been synthesized biogenically by using black pepper (Piper nigrum) extract according to the principles of green chemistry in presence and absence of a biopolymer, chitosan. A comprehensive study (up to cellular level) on the antifilarial (against Setaria cervi) activity of AuNPs has been made for the first time with a view to use it clinically. The bioactivity of biopolymer capped biogenic AuNP increases significantly compared to simple biogenic AuNP. The biopolymer plays an important role in inspiring AuNP through its inherent positive charges and hydrophobicity. The developed nanomaterial boosts the production of ROS (reactive oxygen species) and misbalances the antioxidant parameters of parasites such as GSH, GST, GPx, SOD and catalase. The produced ROS ultimately induces oxidative stress, which leads to apoptotic cell death in filarial worms. The synthesized nanomaterials exhibit negligible toxicity towards human PBMCs. The present study may serve as a fruitful platform to explore biopolymer capped gold nanoparticles as efficient antifilarial therapeutics.

Memory Enhancing Effect of Black Pepper in the AlCl3 Induced Neurotoxicity Mouse Model is Mediated Through Its Active Component Chavicine.

Black pepper (Piper nigrum Linn.) has vital pharmacological properties with profound effects on central nervous system. Neurotoxic agents like Aluminum Chloride (AlCl3) cause the oxidative stress and result in improper processing of amyloid proteins leading to accumulation of amyloid β plaques. The study aimed to explore the neuroprotective potential of black pepper (BP) extract (12.5mg/kg/day) on memory enhancement and its effect on expression of amyloid precursor protein (APP) isoforms (APP770 and APP695) in AlCl3 induced neurotoxicity (250mg/kg) mouse model. The study included the isolation and identification of pure compound from BP (chavicine) which was found pharmacologically active. Morris water maze test, elevated plus maze, fear conditioning, context and cue dependent test and social preference tests were performed to investigate the learning and memory. Gene expression (APP isoforms) and in-vitro and ex-vivo DPPH free radical scavenging activity were performed to evaluate the role of BP. BP significantly improved memory in AlCl3 induced neurotoxicity mouse model along with effectively decreasing the expression of APP770 (amyloidogenic) isoform and improved level of APP695 (non-amyloidogenic) in hippocampus, amygdala and cortex. Fear extinction learning was considerably improved in BP treated group (7.83±2.03) than AlCl3 induced neurotoxicity group (39.75±4.25). In the hippocampus, BP significantly reduced the expression of APP770 (0.37±0.05) as compared to AlCl3 induced neurotoxicity group (0.72±0.06), and effectively increased (34.80±1.39) the percentage inhibition of DPPH free radicals as compared to AlCl3 induced neurotoxicity group (14±2.68). The study revealed that BP improves memory and chavicine is a lead compound producing pharmacological effects of BP.

Corrosion inhibition and adsorption behaviour of black pepper extract on pure aluminum in hydrochloric acid medium

Purpose– The purpose of this paper is to investigate the inhibitive properties of black pepper extract (BPE) for aluminium in 1M hydrochloric acid (HCl) medium.Design/methodology/approach– Gravimetric, electrochemical impedance spectroscopy, galvanostatic polarization, scanning electron microscopy with energy dispersive X-ray examinations (SEM-EDX) techniques were used to study the corrosion inhibitive study.Findings– The gravimetric measurement indicates that inhibition efficiency shows direct proportional relation with concentration of inhibitor. The impedance results illustrates that there was a presence of protective layer of inhibitor adsorbed on the metal/solution interface. Polarization outcome showed that BPE is mixed type inhibitor. The existence of adherent layer of inhibitor on the Al surface was confirmed by SEM-EDX. Quantum chemical calculations were performed using the density functional theory at B3LYP/6-31G(d) level of theory to evaluate the activity of inhibitor molecules present in extract towards the corrosion inhibition of Al.Research limitations/implications– Due to the presence of large number of compounds in the extract, it becomes difficult to understand the most active compound responsible for inhibition. However, from gas chromatography mass spectrometry and quantum data, the approximation has been made that the major compound piperine present in the extract can be most probable component responsible for the inhibition activity. Further calculation of binding energy between Al and inhibitor molecules can be performed using Material Studio software.Practical implications– The extract can be used in cleaning and etching solutions. It can be used to limit the loss of Al metal during etching process.Originality/value– BPE can be used as a potential source of eco-friendly corrosion inhibitor for Al in HCl medium.

Pesticidal Potentials of Seed Extracts Of Black Pepper (<i>Piper Nigrum</i> L) in the Control of Maize Grain Weevil (<i>Sitophilus Zeamais</i> Mots) in Storage

This study was conducted between November 2009 – May 2010 at the Teaching and Research farm of Imo State University, Owerri, Nigeria. Five rates of ethanolic and powder extracts of black pepper, Piper nigrum L were used (0.0, 0.1, 0.2, 0.3, and 0.4) in milliliters and grams as treatments respectively, using Completely Randomized Design (CRD). The parameters determined were mortality rate, grain damage (exit holes), germination percentage, presence of phytochemicals and infrared spectroscopy. Results showed that both ethanolic and powder extracts, killed Sitophilus zeamais but the ethanolic extracts were significant, more effective and quicker in action at P ≤ 0.05. However, it was observed that the bio efficacy of the extracts increased with increase in the level of concentration. Ethanolic and powder extracts of P. nigrum L. at 0.4, 0.3 and 0.2mls/g were significantly (P ≤ 0.05) more effective in the control of maize grain weevil in storage than 0.1 mls/g within12-48hours and 12-72 intervals respectively. There were no grain damage (exit holes) on treatments with ethanolic extracts but with powder extracts, exit holes were noticed but differed significantly at (P ≤ 0.05). On seed germination tested 6 months after treatments, 0.2 and 0.4mls gave the highest number of maize grains with radicule and plumule emergence, 80% each while 0.1 and 0.3mls gave 70% each. None of the treatments was significant at (P< 0.05) except over the control. Phytochemical screening of the plant extract revealed the presence of bio-active compounds comprising alkaloids (50.13 – 8 5.67%), flavonoids (24.56 – 4.02%), Tannins (17.79 – 0.22%), Saponnins (5.51 – 7.52%) and Phenols (2.01 – 0.71) % in decreasing order of percentage. The infrared spectroscopy revealed some functional groups – OH, - COOH radicals and double bond ketones. These compounds have the implication of exerting different toxicities on organisms. Keywords: P. nigrum , S. zeamais , Maize, Phytochemical, Pesticidal Potentials

Storage and Microbial Evaluation of Black Pepper Pre-Treated Oven- Dried Moon Fish (Citharinus citharus Geoffery Saint-Hilaire 1809)

The storage and microbial evaluation of black pepper (Piper guineese) pre-treated oven-dried moon fish (Citharinus citharus) stored at an ambient temperature were studied. Thirty six (36) freshly caught moon fish weighing between 850-900 g were purchased, killed, eviscerated and rinsed thoroughly under tap water and were divided into 3 treatments of 12 fish in each. The first treatment was immersed in 3% brine without black pepper extract served as control, the second treatment was soaked in mixture of 3% brine and 1.5% black pepper extract while the third sample was immersed in 3% brine and 3% black pepper extract tagged MFSA, MFSB and MFSC respectively. Each treatment fish were soaked in the respective solutions for 30minutes prior to been oven- dried using gas as energy for 5hours at the temperature range of 80°C-90°C. After drying, samples were allowed to cool at room temperature in separately labelled clean trays and subsequently stored in the quality control room for 7 days to determine the storage and microbial characteristics. Processed fish samples were subjected to microbial analysis of which results revealed that the (control) MFSA had the highest microbial count of 17.2 x 105 followed by 10.8 x 105 and 9.6 x 105 for MFSB and MFSC respectively. Also, the microbial analysis showed that MFSA favour the growth of Staphylococcus aureus, while Klebsiella spp. and Bacillus spp. were identified for MFSB and MFSC respectively. These results therefore, indicated that the use of 3% brine with black pepper at 1.5% and 3% concentration could have caused reduction of microbial load of oven- dried fish and improve its shelf-life

Antiradical properties of essential oils and extracts from coriander, cardamom, white, red, and black peppers

The antiradical properties of essential oils and extracts from coriander seeds Coriandrum sativum L., cardamom fruits Elettaria cardamomum L., fruits of white and black pepper Piper nigrum L., and pods of red cayenne and green chili pepper Capsicum frutescens L. were studied in model reactions with the stable free 2,2-diphenyl-1-picrylhydrazyl radical. The essential oils consisted of monoand sesquiterpene hydrocarbons, alcohols, oxides and esters as the main components. Spice extracts contained flavonoids, diand triterpenoids, phenolic acids, alkaloids and carotenoids. The values of antiradical efficiency were low and decreased in the following order: black pepper extract > cayenne pepper extract > cardamom essential oil > chili pepper extract > cardamom extract > white pepper extract > coriander extract > black pepper essential oil > white pepper essential oil > coriander essential oil.

Antioxidant and cytotoxic effects of essential oil, water and ethanol extracts of major Indian spices

Essential oils, water and ethanol extracts of black pepper, ginger, turmeric, cinnamon, Garcinia indica, G. gummi-gutta, tamarind and curry leaves were examined for their antioxidant potential and cytotoxicity to cancer cell lines by in vitro methods. Essential oils of ginger, turmeric, cinnamon and curry leaf were highly cytotoxic, reducing cell viability to 14 to 30% of untreated control; water and ethanol extracts of G. indica, turmeric, cinnamon, tamarind and curry leaf were also cytotoxic, though to a lesser extent (27% to none); ethanol extracts displayed approximately 50% higher cytotoxicity than water extracts. Antioxidant potential of water and ethanol extracts were similar, and decreased after six months of storage in most extracts; most were superior to BHA and BHT. Antioxidant potential of ethanol was also greater than water extracts. Change in essential oil chemoprofile stored at 4°C for a year compared to fresh, most notably t-caryophyllene, is also reported here.

Inhibition of intestinal chloride secretion by piperine as a cellular basis for the anti-secretory effect of black peppers

Piperine is the principal alkaloid in black peppers (Piper nigrum L.), which is a commonly included spice in anti-diarrheal formulations. Piperine has antispasmodic activities, but its anti-secretory effect is not known. Therefore, this study investigated the anti-secretory effect of piperine and its underlying mechanism. Piperine inhibited cAMP-mediated Cl− secretion in human intestinal epithelial (T84) cells, similar to black pepper extract. Intraluminal administration of piperine (2μg/loop) suppressed cholera toxin-induced intestinal fluid accumulation by ∼85% in mice. The anti-secretory mechanism of piperine was investigated by evaluating its effects on the activity of transport proteins involved in cAMP-mediated Cl− secretion. Notably, piperine inhibited CFTR Cl− channel activity (IC50#8’6#10μM) without affecting intracellular cAMP levels. The mechanisms of piperine-induced CFTR inhibition did not involve MRP4-mediated cAMP efflux, AMPK or TRPV1. Piperine also inhibited cAMP-activated basolateral K+ channels, but it had no effect on Na+–K+–Cl− cotransporters or Na+–K+ ATPases. Piperine suppressed Ca2+-activated Cl− channels (CaCC) without affecting intracellular Ca2+ concentrations or Ca2+-activated basolateral K+ channels. Collectively, this study indicates that the anti-secretory effect of piperine involves the inhibition of CFTR, CaCC and cAMP-activated basolateral K+ channels. Piperine represents a novel class of drug candidates for the treatment of diarrheal diseases caused by the intestinal hypersecretion of Cl−.

Classification of Cultivation Locations of Black Pepper (Piper nigrum L.) using Gas Chromatography and Chemometrics

Gas chromatography (GC) and chemometrics were applied to classify the geographical origins of black peppers (Piper nigrum L.) from Vietnam, Brazil, Indonesia, and India; which are the most important black pepper producers in the world. A gas chromatograph with a flame ionization detector (FID) profiled the black pepper extracts. A polynomial fitting baseline correction and data transformation by different numerical roots were introduced for data preprocessing. Partial least-squares discriminant analysis (PLS-DA) was used to build classification models. The classification rates of PLS-DA obtained with a leave-one-sample-out validation were used to optimize the polynomial order for the baseline correction and the degree of the root transformation. The polynomial order baseline correction and root transformation methods significantly improved the performance of PLS-DA which resulted in improved classification rates up to 100%. Therefore, a GC method coupled with chemometric analysis was devised for determination of the growing location of black pepper. Keywords: Black pepper, chemometrics, classifications, gas chromatography, PLS-DA, root transformation.

Antibacterial mechanism and activities of black pepper chloroform extract.

Black pepper extracts reportedly inhibit food spoilage and food pathogenic bacteria. This study explored the antimicrobial activity of black pepper chloroform extract (BPCE) against Escherichia coli and Staphylococcus aureus. The antibacterial mechanism of BPCE was elucidated by analyzing the cell morphology, respiratory metabolism, pyruvic acid content, and ATP levels of the target bacteria. Scanning electron micrographs showed that the bacterial cells were destroyed and that plasmolysis was induced. BPCE inhibited the tricarboxylic acid pathway of the bacteria. The extract significantly increased pyruvic acid concentration in bacterial solutions and reduced ATP level in bacterial cells. BPCE destroyed the permeability of the cell membrane, which consequently caused metabolic dysfunction, inhibited energy synthesis, and triggered cell death.

Pest Control in Green Gram Seeds (&lt;i&gt;Vigna radiata&lt;/i&gt;) by Using Plant Extracts

A experiment was conducted to investigate the efficacy of different plant derivatives against the development of the green gram pest Callosobruchus chinensis (L). fed on green gram seeds Vigna radiate for one month period. The leaf extracts of lemon (Citrus lemon), mint (Mentha arvensis) and black pepper seed (Piper nigram) powders were evaluated for their growth, adult mortality and oviposition inhibition of Callosobruhus chinensis. The results revealed that the extracts of all the three plant products caused a considerable reduction in the number of bruchids. Among the treatments Piper nigram were found to be effective in destroying the pest.

PEST CONTROL IN GREEN GRAM SEEDS ( Vigna radiata ) BY USING PLANT EXTRACTS

A experiment was conducted to investigate the efficacy of different plant derivatives against the development of the green gram pest Callosobruchus chinensis (L). fed on green gram seeds Vigna radiate for one month period. The leaf extracts of lemon ( Citrus lemon ), mint ( Mentha arvensis ) and black pepper seed ( Piper nigram ) powders were evaluated for their growth, adult mortality and oviposition inhibition of Callosobruhus chinensis . The results revealed that the extracts of all the three plant products caused a considerable reduction in the number of bruchids. Among the treatments Piper nigram were found to be effective in destroying the pest.

Pest Control in Green Gram Seeds (&lt;i&gt;Vigna radiata&lt;/i&gt;) by Using Plant Extracts

A experiment was conducted to investigate the efficacy of different plant derivatives against the development of the green gram pest Callosobruchus chinensis (L). fed on green gram seeds Vigna radiate for one month period. The leaf extracts of lemon (Citrus lemon), mint (Mentha arvensis) and black pepper seed (Piper nigram) powders were evaluated for their growth, adult mortality and oviposition inhibition of Callosobruhus chinensis. The results revealed that the extracts of all the three plant products caused a considerable reduction in the number of bruchids. Among the treatments Piper nigram were found to be effective in destroying the pest.

Effects of Black Pepper (Piper nigrum Linn.) Extract on Sexual Drive in Male Mice

Black pepper (Piper nigrum L.), known as the king of spices, proven to contain various types of active substances that are allegedly beneficial to the human body functions including sexual function. This study was conducted to determine whether black pepper extract can be used to improve sexual function in male subjects. Thirty six healthy male mice, four months old, weighing between 25-30 g, were used as experimental animals and grouped into four. The first one mice was given pellets that do not contain black pepper extract as a control. The second and third groups, respectively were treated with pellets containing aqueous and ethanol extracts. The last group was given pellets containing aqueous extract and ethanolic extracts with a ratio of 1:1. Pellets were given once every day for 90 days. Mating behavior test was conducted in an open round plastic tray with a diameter of 40 cm and height 25 cm. Throughout the test, videotaping was performed for the following parameters: courtship latency and mounting frequency. The results revealed that compared with control group, male mice fed on aqueous as well as ethanol extract of black pepper significantly showed a shorter courtship latency (p<0.05). On the other hand, there was no difference in the mounting frequency between treated groups. Interestingly, the mounting frequency showed a strong negative correlation with the courtship latency (r = -0.968). In conclusion, the fruit extract of black pepper potentially affect sexual drive in male mice.

In vitro antibacterial activity of seven Indian spices against high level gentamicin resistant strains of enterococci.

IntroductionThe aim of the study was to explore the in vitro antibacterial activity of seven ethanolic extracts of spices against high level gentamicin resistant (HLGR) enterococci isolated from human clinical samples.Material and methodsTwo hundred and fifteen enterococcal strains were isolated from clinical samples. High level gentamicin resistance in ethanolic extracts of cumin (Cuminum cyminum), cinnamon (Cinnamomum zeylanicum), ginger (Zingiber officinale), fenugreek (Trigonella foenum-graecum), cloves (Syzygium aromaticum), cardamom (Elettaria cardamomum Maton) and black pepper (Piper nigrum) were prepared using Soxhlet apparatus. The antibacterial effect of the extracts was studied using the well diffusion method. Statistical analysis was carried out by χ2 test using SPSS 17 software.ResultsOnly cinnamon and ginger were found to have activity against all the isolates, whereas cumin and cloves had a variable effect on the strains. Fenugreek, black pepper and cardamom did not show any effect on the isolates. The zone diameter of inhibition obtained for cinnamon, ginger, cloves and cumin was in the range 31–34 mm, 27–30 mm, 25–26 mm and 19–20 mm respectively.ConclusionsCinnamomum zeylanicum and Z. officinale showed the maximum antibacterial activity against the enterococcal isolates followed by S. aromaticum and C. cyminum. The findings of the study show that spices used in the study can contribute to the development of potential antimicrobial agents for inclusion in the anti-enterococcal treatment regimen.

Food Compounds Activating Thermosensitive TRP Channels in Asian Herbal and Medicinal Foods.

There are several thermosensitive transient receptor potential (TRP) ion channels including capsaicin receptor, TRPV1. Food components activating TRPV1 inhibit body fat deposition through sympathetic nerve stimulation. TRPA1 is another pungency sensor for pungent compounds and is mainly coexpressed with TRPV1 in sensory nerve endings. Therefore, TRPA1 activation is expected to have an anti-obesity effect similar to TRPV1 activation. We have searched for agonists for TRPV1 and TRPA1 in vitro from Asian spices by the use of TRPV1- and TRPA1-expressing cells. Further, we performed food component addition tests to high-fat and high-sucrose diets in mice. We found capsiate, capsiconiate, capsainol from hot and sweet peppers, several piperine analogs from black pepper, gingeriols and shogaols from ginger, and sanshools and hydroxysanshools from sansho (Japanese pepper) to be TRPV1 agonists. We also identified several sulfides from garlic and durian, hydroxy fatty acids from royal jelly, miogadial and miogatrial from mioga (Zingiber mioga), piperine analogs from black pepper, and acetoxychavicol acetate (ACA) from galangal (Alpinia galanga) as TRPA1 agonists. Piperine addition to diets diminished visceral fats and increased the uncoupling protein 1 (UCP1) in interscapular brown adipose tissue (IBAT), and black pepper extract showed stronger effects than piperine. Cinnamaldehyde and ACA as TRPA1 agonists inhibited fat deposition and increased UCP1. We found that several agonists of TRPV1 and TRPA1 and some agonists of TRPV1 and TRPA1 inhibit visceral fat deposition in mice. The effects of such compounds on humans remain to be clarified, but we expect that they will be helpful in the prevention of obesity.

Influência de diferentes extratos aquosos de plantas medicinais no desenvolvimento de Colletotrichum gloeosporioides e de Fusarium moniliforme

O objetivo deste trabalho foi avaliar a influência dos extratos aquosos das plantas medicinais alecrim, alho, cravo-da-índia, sálvia, capim-limão, orégano ou pimenta-do-reino no desenvolvimento in vitro de Colletotrichum gloeosporioides e de Fusarium moniliforme. Os extratos foram obtidos pela infusão de 60 g de cada planta medicinal em 200 mL de água fervente. Cada extrato aquoso foi fracionado em concentrações de 0, 5, 10 e 20% (p:v) e incorporado ao meio de cultivo BDA (batata-dextrose-ágar) antes da esterilização em autoclave. Posteriormente, um disco de 8 mm de diâmetro de micélio fúngico de cada patógeno foi transferido para o centro de placas de Petri. Após 24, 48 e 96 horas de incubação em câmara de crescimento a 22 ± 2 ºC e fotoperíodo de 12 horas avaliou-se o crescimento micelial de F. moniliforme e de C. gloesporioides. No último período de incubação, também se quantificou o número de conídios de cada fungo. Para o teste de germinação adicionou-se nas cavidades de placas de teste Elisa, uma alíquota de 40 µL de cada extrato nas concentrações de 0, 5, 10 e 20%, e outra alíquota, da suspensão de conídios de cada patógeno. Após 24 horas a 22 ± 2 ºC, no escuro, a germinação dos conídios foi paralisada com a adição de 20 µL de lactofenol; avaliou-se então a porcentagem de germinação de conídios. Os experimentos foram conduzidos no delineamento inteiramente casualizado em esquema fatorial 7 x 4 (extratos de plantas medicinais x concentrações) com quatro repetições. Para ambos os patógenos o extrato aquoso de alho e cravo-da-índia apresentaram maior ou total inibição do crescimento micelial, respectivamente, quando comparado com os demais extratos. Para C. gloeosporioides, o extrato de cravo-da-índia apresentou menor número de conídios em todas as concentrações testadas, e para o extrato de alho a 20%, também não foi observada a germinação de conídios. O extrato de alho foi eficiente em reduzir o número e a germinação dos conídios de F. moniliforme na concentração de 20%. Os extratos de alecrim, cravo-da-índia, orégano e pimenta-do-reino, nas maiores concentrações, tiveram efeito positivo na redução da produção de conídios deste mesmo fungo.

Piperine, a component of black pepper, decreases eugenol-induced cAMP and calcium levels in non-chemosensory 3T3-L1 cells

This study investigated the effects of an ethanol extract of black pepper and its constituent, piperine, on odorant-induced signal transduction in non-chemosensory cells. An ethanol extract of black pepper decreased eugenol-induced cAMP and calcium levels in preadipocyte 3T3-L1 cells with no toxicity. Phosphorylation of CREB (cAMP response element-binding protein) was down-regulated by the black pepper extract. The concentration (133.8mg/g) and retention time (5.5min) of piperine in the ethanol extract were quantified using UPLC–MS/MS. Pretreatment with piperine decreased eugenol-induced cAMP and calcium levels in 3T3-L1 cells. Piperine also decreased the phosphorylation of CREB, which is up-regulated by eugenol. These results suggest that piperine inhibits the eugenol-induced signal transduction pathway through modulation of cAMP and calcium levels and phosphorylation of CREB in non-chemosensory cells.