curcumin and thymoquinone

Spices have been widely used as food flavorings and folk medicines for thousands of years. Numerous studies have documented the antioxidant, anti-inflammatory and immunomodulatory effects of spices, which might be related to prevention and treatment of several cancers, including lung, liver, breast, stomach, colorectum, cervix, and prostate cancers. Several spices are potential sources for prevention and treatment of cancers, such as Curcuma longa (tumeric), Nigella sativa (black cumin), Zingiber officinale (ginger), Allium sativum (garlic), Crocus sativus (saffron), Piper nigrum (black pepper) and Capsicum annum (chili pepper), which contained several important bioactive compounds, such as curcumin, thymoquinone, piperine and capsaicin. The main mechanisms of action include inducing apoptosis, inhibiting proliferation, migration and invasion of tumors, and sensitizing tumors to radiotherapy and chemotherapy. This review summarized recent studies on some spices for prevention and treatment of cancers, and special attention was paid to bioactive components and mechanisms of action.

Topics: Alkaloids; Antineoplastic Agents, Phytogenic; Apoptosis; Benzodioxoles; Benzoquinones; Capsaicin; Capsicum; Cell Proliferation; Crocus; Curcuma; Curcumin; Garlic; Humans; Neoplasms; Nigella sativa; Phytotherapy; Piper nigrum; Piperidines; Polyunsaturated Alkamides; Spices; Zingiber officinale

Mitochondrial dysfunction may lead to cardiomyocyte death in trastuzumab (TZM)-induced cardiotoxicity. Accordingly, this study was designed to evaluate the mitochondrial protective effects of curcumin, chrysin and thymoquinone alone in TZM-induced cardiotoxicity in the rats. Forty-eight male adult Wistar rats were divided into eight groups: control group (normal saline), TZM group (2.5 mg/kg I.P. injection, daily), TZM + curcumin group (10 mg/kg, I.P. injection, daily), TZM + chrysin (10 mg/kg, I.P. injection, daily), TZM + thymoquinone (0.5 mg/kg, I.P. injection, daily), curcumin group (10 mg/kg, I.P. injection, daily), chrysin group (10 mg/kg, I.P. injection, daily) and thymoquinone group (10 mg/kg, I.P. injection, daily). Blood and tissue were collected on day 11 and used for assessment of creatine phosphokinase, lactate dehydrogenase (LDH), troponin, malondialdehyde (MDA) amount, glutathione levels and mitochondrial toxicity parameters. TZM increased mitochondrial impairments (reactive oxygen species formation, mitochondrial swelling, mitochondrial membrane potential collapse and decline in succinate dehydrogenase activity) and histopathological alterations (hypertrophy, enlarged cell, disarrangement, myocytes degeneration, infiltration of fat in some areas, hemorrhage and focal vascular thrombosis) in rat heart. As well as TZM produced a significant increase in the level of CK, LDH, troponin, MDA, glutathione disulfide. In most experiments, the co-injection of curcumin, chrysin and thymoquinone with TZM restored the level of CK, LDH, troponin, MDA, GSH, mitochondrial impairments and histopathological alterations. The study revealed the cardioprotective effects of curcumin, chrysin and thymoquinone against TZM-induced cardiotoxicity which could be attributed to their antioxidant and mitochondrial protection activities.

Topics: Animals; Antioxidants; Benzoquinones; Cardiotoxicity; Curcumin; Doxorubicin; Flavonoids; Glutathione; Male; Mitochondria; Oxidative Stress; Rats; Rats, Wistar; Trastuzumab; Troponin

Medical improvements are needed to prevent ischemia-reperfusion injury in thoracoabdominal aortic surgery. The aim of this study was to determine the antioxidant effects of thymoquinone, silymarin, and curcumin against ischemia-reperfusion injury associated with abdominal aorta.. Twenty-five Wistar albino rats were included in the study. Sham, control, and treatment (thymoquinone, silymarin, and curcumin) groups were set in equal numbers. Ischemia-reperfusion was applied by clamping (120 minutes) and de-clamping (60 minutes) the infrarenal aorta of all groups, except the sham group. Before reperfusion, thymoquinone, silymarin, and curcumin were given intraperitoneally to the treatment groups. After reperfusion, blood samples were taken from the right ventricle. Total antioxidant status (TAS), total oxidant status (TOS), and oxidative stress index (OSI) were studied in serum samples and histopathological examination was performed on the gastrocnemius muscle.. There was a significant difference in TOS and OSI values between the control and sham groups. Both values were found higher in the control group than in the sham group (P<0.05). OSI values were found to be lower in the thymoquinone group compared to the control group (P<0.05). All three parameters were found to be lower in the silymarin group than in the control group (P<0.05). TAS and TOS levels were found to be higher in the curcumin group than in the control group (P<0.05). There was no histopathological difference between the groups.. Silymarin and thymoquinone administration decreases oxidative stress in experimental aortic ischemia-reperfusion injury. Antioxidant effect of curcumin was lower than silymarin and thymoquinone.

Topics: Animals; Antioxidants; Aorta, Abdominal; Curcumin; Ischemia; Rats; Rats, Wistar; Reperfusion; Reperfusion Injury; Silymarin

Curcumin, resveratrol, and thymoquinone are the potential natural bio-actives reported with good anti-psoriatic activity. However, poor aqueous solubility and limited skin permeation of these natural bio-actives hinder their effective delivery and potential therapeutic outcome. In this regard, current research work focuses on the design and optimization of nanoemulsion (NE) gel formulation for the concurrent delivery of these three drugs. The NE system is consisting of oleic acid as oil phase, Tween 20 as surfactant, and PEG 200 as co-surfactant. The optimized formulation exhibited the droplet size 76.20 ± 1.67 nm, PDI of 0.12 ± 0.05, RI of 1.403 ± 0.007, and viscosity of 137.9 ± 4.07 mp. Carbopol 940 (0.5% w/v) was used as the gelling agent to prepare the NE gel which exhibited a good texture profile. The optimized formulation exhibited a higher % of growth inhibition on A-431 cells and demonstrated good anti-angiogenic activity in the HET-CAM test. Finally, in vivo studies in Balb/c mice model showed improved anti-psoriatic conditions which indicated that the triple natural bio-actives combination in nanoemulgel formulation is effective in the management of psoriasis.

Topics: Animals; Benzoquinones; Curcumin; Emulsions; Mice; Nanoparticles; Particle Size; Psoriasis; Resveratrol

Overall survival of high-risk neuroblastoma patients is still poor, emphasizing the need for novel therapeutic options. There is evidence for anti-cancer properties of the herbal substances thymoquinone and curcumin. These substances are isolated from Nigella sativa L. and Curcuma longa L., respectively, which are used in traditional medicine.. We investigated cytotoxic effects of thymoquinone and curcumin on neuroblastoma cell lines NLF, NB69, and SK-N-BE(2), in vitro.. Cytotoxicity of compounds was investigated by MTT cell viability assays. For analyzing effects on cell proliferation BrdU assays were employed and induction of apoptosis was detected by Cell Death ELISA assays.. Both substances showed cytotoxic effects in all three neuroblastoma cell lines, whereby primary human fibroblast cells reacted less sensitively. Overall, lower IC50 values could be calculated for curcumin (3.75-7.42 µM) than for thymoquinone (5.16-16.3 µM). Decreased proliferation and increased apoptosis rates were observed under treatment.. Both substances showed anti-tumoral properties on neuroblastoma cell lines and should be further investigated as therapeutic agents.. Einleitung: Das Gesamtüberleben bei Hochrisiko-Neuroblastom-Patienten ist immer noch sehr schlecht, was den Bedarf an neuen Therapieoptionen unterstreicht. Für die beiden pflanzlichen Substanzen Thymoquinon und Curcumin liegen Studien vor, die auf krebshemmende Eigenschaften hinweisen. Diese Substanzen werden aus den in der Traditionellen Medizin verwendeten Pflanzen Nigella sativa L. bzw. Curcuma longa L. isoliert. Ziel: Wir haben die zytotoxischen Effekte von Thymoquinon und Curcumin auf die Neuroblastomzelllinien NLF, NB69 und SK-N-BE(2) in vitro untersucht. Methoden: Die Zytotoxizität der Substanzen wurde mittels MTT Zellviabilitäts-Assays untersucht. Effekte auf die Proliferation und die Apoptose wurden durch ELISA Assays analysiert. Ergebnisse: Beide Substanzen wirkten zytotoxisch in allen drei Neuroblastomzelllinien, wobei die primäre humane Fibroblastenzelllinie am resistentesten war. Insgesamt lagen die ermittelten IC50 Werte bei Curcumin (3,75–7,42 µM) niedriger als bei Thymoquinon (5,16–16,3 µM). Unter der Behandlung kam es zu einer reduzierten Proliferations- und erhöhten Apoptoserate. Schlussfolgerung: Thymoquinon und Curcumin zeigen anti-tumorale Wirkungen auf Neuroblastomzelllinien und sollten daher weiter als mögliche Therapeutika untersucht werden.

Topics: Benzoquinones; Cell Line, Tumor; Curcuma; Curcumin; Humans; Neuroblastoma; Nigella sativa

D-galactose (D-gal) administration causes oxidative disorder and is widely utilized in aging animal models. Therefore, we subcutaneously injected D-gal at 200 mg/kg BW dose to assess the potential preventive effect of thymoquinone (TQ) and curcumin (Cur) against the oxidative alterations induced by D-gal. Other than the control, vehicle, and D-gal groups, the TQ and Cur treated groups were orally supplemented at 20 mg/kg BW of each alone or combined. TQ and Cur effectively suppressed the oxidative alterations induced by D-gal in brain and heart tissues. The TQ and Cur combination significantly decreased the elevated necrosis in the brain and heart by D-gal. It significantly reduced brain caspase 3, calbindin, and calcium-binding adapter molecule 1 (IBA1), heart caspase 3, and BCL2. Expression of mRNA of the brain and heart

Topics: Animals; Benzoquinones; Brain; Curcumin; Galactose; Immunohistochemistry; Liver; Myocardium; Organ Specificity; Oxidation-Reduction; Oxidative Stress; Rats; Structure-Activity Relationship

Fasciolosis is a neglected tropical disease caused by the liver fluke Fasciola gigantica. The absence of successful vaccine and emerging resistance in flukes against the drug of choice, triclabendazole, has necessitated the search for alternatives including phyto-therapeutic approaches. Curcumin and thymoquinone, the active ingredients of Curcuma longa and Nigella sativa plants respectively, were first screened for their binding affinity with Glutathione-S-transferase (GST) molecule through in silico molecular docking followed by in vitro treatment of worms with varying concentrations of the test compounds. The in silico molecular docking of curcumin and thymoquinone with sigma GST revealed strong hydrogen bonding as well as hydrophobic interactions with high fitness scores but showing inter-specific differences. The in vitro treatment of F. gigantica worms with both curcumin and thymoquinone resulted in a significant increase in the generation of reactive oxygen species (ROS) whereas the level of reduced glutathione, a primary redox regulator, was found to be significantly decreased (p < 0.05). The two compounds not only inhibited the GST activity, which is an important detoxification enzyme and also a key drug/vaccine target for the control of fasciolosis but also significantly inhibited the activity of antioxidant enzymes glutathione peroxidase and glutathione reductase that are vital in maintenance of redox homeostasis. The immunohistochemistry performed using anti sigma GST polyclonal antibodies revealed that both the compounds used in the present study significantly reduced immunofluorescence in the vitellaria, developing eggs present in the ovary and the intestinal caecae indicating inhibition of GST enzyme in these regions of the worms. Further, following treatment with curcumin and thymoquinone, chromatin condensation and DNA fragmentation was also observed in F. gigantica worms. In conclusion, both curcumin and thymoquinone generated oxidative stress in the worms by production of ROS and significantly inhibiting their antioxidant and detoxification ability. The oxidative stress along with induction of apoptotic like events would compromise the survival ability of worms within the host. However, further studies are required to establish their anthelmintic potential alone and in combination with the commonly used anthelmintic drugs under in vivo conditions.

Topics: Animals; Apoptosis; Benzoquinones; Buffaloes; Chromatin; Curcumin; DNA Damage; DNA Fragmentation; Electrophoresis, Agar Gel; Enzyme Inhibitors; Fasciola; Glutathione; Glutathione Peroxidase; Glutathione Reductase; Glutathione Transferase; Immunohistochemistry; Microscopy, Confocal; Models, Molecular; Molecular Docking Simulation; Oxidative Stress; Reactive Oxygen Species

Topics: Acute Kidney Injury; Animals; Benzoquinones; Cell Proliferation; Cisplatin; Curcumin; Cytokines; Dose-Response Relationship, Drug; Drug Combinations; HEK293 Cells; Heme Oxygenase-1; Hepatitis A Virus Cellular Receptor 1; Humans; Kidney Function Tests; Male; NF-E2-Related Factor 2; NF-kappa B; Oxidative Stress; Rats

Acrylamide, an environmental pollutant, is known to occur in food substances cooked at high temperatures. Studies on various models indicate acrylamide to cause several physiological conditions such as neuro- and reproductive toxicity, and carcinogenesis. In our study, exposure of Drosophila melanogaster (Oregon K strain) to acrylamide via their diet resulted in a concentration and time-dependent mortality, while the surviving flies exhibited significant locomotor deficits, most likely due to oxidative stress-induced neuronal damage. Also, Drosophila embryos exhibited signs of developmental toxicity as evidenced by the alteration in the migration of border cells and cluster cells during the developmental stages, concomitant to modulation in expression of gurken and oskar genes. Curcumin, a known antioxidant has been widely studied for its neuroprotective effects against acrylamide; however; very few studies focus on thymoquinone for its role against food toxicant. Our research focuses on the toxicity elicited by acrylamide and the ability of the antioxidants: thymoquinone, curcumin and combination of thereof, in reversing the same.

Topics: Acetylcholinesterase; Acrylamide; Animals; Antioxidants; Behavior, Animal; Benzoquinones; Curcumin; Drosophila melanogaster; Drosophila Proteins; Environmental Pollutants; Female; Gene Expression; Locomotion; Male; Neuroprotective Agents; Phytochemicals; Reactive Oxygen Species; Transforming Growth Factor alpha

Acetaminophen (APAP) hepatotoxicity is characterised by an extensive oxidative stress due to depletion of glutathione (GSH), which results in massive lipid peroxidation and subsequent liver injury. The current paradigm suggests that mitochondria are the main source of reactive oxygen species (ROS), which impair mitochondrial function and are responsible for cell signalling resulting in cell death. This study was designed to compare the potential impact of thymoquinone (THQ), and/or curcumin (CURC) on liver injury induced by APAP toxicity in rats.. Serum levels of alanine transaminase, aspartate transaminase, total bilirubin, and total protein were measured. In addition, liver nitric oxide (NO), malondialdehyde, reduced glutathione (GSH), and superoxide dismutase (SOD) were estimated. Moreover, these biochemical parameters were confirmed by histopathological and immunohistochemical investigations for the expression of thioredoxin, iNOS and caspase 3.. Acetaminophen toxicity elevated most of the above-mentioned parameters but decreased GSH, SOD, and total protein levels. Histologically, liver sections demonstrated liver injury characterised by hepatocellular necrosis with nuclear pyknosis, karyorrhexis and karyolysis. Immunohistochemical study revealed increased expression of iNOS and caspase 3 proteins, while the thioredoxin protein expression was decreased.. Treatment with the THQ and CURC regulated the biochemical and histopathological alterations induced by APAP toxicity. It was concluded that the combination strategy of THQ and CURC might be considered as a potential antidote in combating liver injury induced by APAP with minimal side effects.

Topics: Acetaminophen; Alanine Transaminase; Animals; Aspartate Aminotransferases; Benzoquinones; Bilirubin; Caspase 3; Curcumin; Glutathione; Immunohistochemistry; Liver; Liver Diseases; Male; Malondialdehyde; Nitric Oxide; Nitric Oxide Synthase Type II; Rats; Superoxide Dismutase; Thioredoxins

Topics: A549 Cells; Antineoplastic Agents, Phytogenic; Benzoquinones; Cell Survival; Comet Assay; Curcumin; DNA Damage; Dose-Response Relationship, Drug; Drug Carriers; Drug Compounding; Humans; Liposomes; Microscopy, Fluorescence; Oxidative Stress; Surface Properties

To investigate thymoquinone, curcumin and a combination of these two drugs were effective or not at the growth of liver.. Forty female Wistar-Albino rats distributed into five groups of eight rats each, control, thymoquinone, curcumin, and thymoquinone/curcumin groups. Pathological specimens were studied using the Ki-67 Proliferation Index(PI); and arginase(Arg), tissue plasminogen activator(tPA), ceruloplasmin(Cer) and nitric oxide(NO) were studied in biochemical analysis.. Our results showed that Ki-67 proliferation index was low in Groups 1. The proliferation coefficient was significantly higher in the Group 2 and Group 4 than in the Group 1 and Group 3.(P < 0.001 between Groups 1 and 2, 1 and 4, and 3 and 4). There was no difference between Groups 2 and 4 (P = 1). The results of the biochemical Arg, tPA and Cer test showed statistically between the Group 1 and Group 2. NO showed significant differences Group 1 and 3.. Thymoquinone and curcumin both have known positive effects on the organism. Histological and biochemical tests showed that thymoquinone is more effective than curcumin.

Topics: Animals; Antineoplastic Agents; Antioxidants; Arginase; Benzoquinones; Biomarkers; Cell Proliferation; Ceruloplasmin; Curcumin; Female; Hepatectomy; Ki-67 Antigen; Liver; Liver Neoplasms; Liver Regeneration; Liver Transplantation; Nitric Oxide; Rats; Rats, Wistar; Tissue Plasminogen Activator

Fasciolosis an economically important global disease of ruminants in the temperate and tropical regions, caused by Fasciola hepatica and F. gigantica, respectively, also poses a potential zoonotic threat. In India alone it causes huge losses to stakeholders. Anthelmintics including triclabendazole have been used to control this menace but the emerging resistance against the available compounds necessitates identification of novel and alternative therapeutic measures involving plant derived natural compounds for their anthelmintic potential. Thymoquinone (T) and curcumin (C), the active ingredients of Nigella sativa and Curcuma longa respectively have been used as antiparasitic agents but the information on their flukicidal effect is very limited. Adult flukes of F. gigantica were in vitro exposed to different concentrations of thymoquinone and curcumin separately for 3h at 37+ 1°C. A significant (p<0.05) reduction in the worm motility at 60 μM concentration of both T and C was observed though all the worms remained alive after 3h exposure, whereas the effect on egg shedding was statistically insignificant. Pronounced tegumental disruptions and erosion of spines in the posterior region and around the acetabulum was evident. A significant (p<0.05) decrease in glutathione-S-transferase and superoxide dismutase activity and reduced glutathione (GSH) level was observed, while protein carbonylation increased differentially. A significant inhibition of CathepsinL (CatL) gene expression in thymoquinone treated worms was also evident. Further, in silico molecular docking of T and C with CatL revealed a stronger interaction of curcumin with the involvement of higher number of amino acids as compared to thymoquinone that could be more effective in inhibiting the antioxidant enzymes of F. gigantica. It is concluded that both the compounds understudy will decrease the detoxification ability of F. gigantica, while inhibition of CatL will significantly affect their virulence potential. Thus, both thymoquinone and curcumin appeared to be promising anthelmintic compounds for further investigations.

Topics: Animals; Antiplatyhelmintic Agents; Benzoquinones; Curcumin; Dose-Response Relationship, Drug; Fasciola; Glutathione; Glutathione Transferase; Parasitic Sensitivity Tests

This article has been retracted: please see Elsevier Policy on Article Withdrawal (http://www.elsevier.com/locate/withdrawalpolicy).\ \ The authors retract the above paper due to: 1) conflict of interest among the authors; and 2) addition of coauthor Dr. Muhammad Younus without his knowledge or permission. The authors apologize for these two grave mistakes.

Topics: Animal Feed; Animals; Antiviral Agents; Benzoquinones; Curcumin; Diet; Dietary Supplements; Female; Immunity, Innate; Influenza A Virus, H9N2 Subtype; Influenza in Birds; Male; Poultry Diseases; Turkeys

Among noncommunicable diseases, metabolic syndrome (MS), a cluster of metabolic disorders including obesity, hyperglycemia, hyperlipidemia and hypertension, is highly prevalent in modern society. Its management requires lifestyle modifications and/or the life-long use of multiple medications, hence demanding development of safe alternative remedies. This study was aimed to establish the efficacy of combined use of black seeds and turmeric using fructose-fed rat model of MS. The high-performance liquid chromatographic fingerprints of turmeric and black seeds showed the presence of curcumin and thymoquinone, respectively, as their major constitutes. Different doses of black seeds and turmeric, individually and in combination, were administered to fructose-fed rats for up to 6 weeks representing characteristic features of MS. At 3 weeks of the treatment, black seeds and turmeric lowered (P < 0.01) high blood pressure and low-density lipoprotein cholesterol, respectively, whereas their coadministration reduced (P < 0.01) both high blood pressure and hypertriglyceridemia. At 6 weeks, the coadministration of both herbs, at half the doses of individual herbs, was the most effective (P < 0.001) in preventing hypertension, hyperglycemia, dyslipidemia, hyperinsulinemia, and endothelial dysfunction than the individual herbs. This study demonstrates the therapeutic superiority of the combination of black seeds and turmeric at low doses over individually tested herbs, in improving features of MS.

Topics: Animals; Benzoquinones; Blood Glucose; Blood Pressure; Cholesterol, LDL; Curcuma; Curcumin; Drug Therapy, Combination; Fructose; Metabolic Syndrome; Phytotherapy; Plant Preparations; Rats; Seeds; Sweetening Agents; Treatment Outcome

This study was conducted to assess the preventive effect of two plant constituents, thymoquinone and curcumin, on gentamicin-induced deleterious effect on liver function, integrity and histological architecture. The gentamicin was intraperitoneally injected to rats at dose level of 100 mg/kg b.w. (every other day) for 21 days. The thymoquinone and curcumin were concurrently and orally administered at dose level of 20 mg/kg b.w. (every other day) to gentamicin-injected rats. The present data indicated that thymoquinone and curcumin significantly prevented the gentamicin-induced elevations of serum AST, ALT and LDH activities as well as tumor necrosis factor alpha (TNF-α) and total bilirubin levels. On the other hand, both agents markedly ameliorated the gentamicin-induced decrease in serum total protein, albumin and albumin/globulin ratio. In addition, the gentamicin-induced liver histological alterations including hydropic degeneration of hepatocytes, fatty changes, inflammatory cell infiltration and congestion of portal vein were successfully amended by thymoquinone and curcumin. The elevated proapoptotic proteins caspase 3 and Bax expression in cytoplasm and nucleus of hepatocytes of gentamicin-injected rats were reduced to normal value as a result of thymoquinone and curcumin administration while the lowered expression of antiapoptotic protein Bcl-2 was increased. Based on the previous findings, it can be concluded that thymoquinone and curcumin successfully prevents the deleterious effects on liver function and histological integrity to more or less the same degree by enhancing anti-oxidant defense system, suppression of oxidative stress and attenuation of inflammation and apoptosis.

Topics: Alanine Transaminase; Animals; Anti-Inflammatory Agents; Antioxidants; Apoptosis; Aspartate Aminotransferases; bcl-2-Associated X Protein; Benzoquinones; Caspase 3; Chemical and Drug Induced Liver Injury; Curcumin; Gentamicins; Glutathione; Glutathione Peroxidase; L-Lactate Dehydrogenase; Liver; Male; Oxidative Stress; Proto-Oncogene Proteins c-bcl-2; Rats; Superoxide Dismutase; Tumor Necrosis Factor-alpha

This study was designed to compare the inhibitory effects of thymoquinone (TQ) and curcumin (CMN) on the biological changes associating asthma. TQ appeared to exhibit greater inhibitory effects on the aggregation of inflammatory cells in bronchoalveolar lavage (BAL) fluid and in lung tissues. We also measured the effects of the two agents on serum IgE and the changes in the mRNA levels of inducible nitric oxide synthase (iNOS), tumor necrosis factor-α (TNF-α) and transforming growth factor-β1 (TGF-β1). Serum IgE was significantly decreased by TQ and CMN with TQ being more potent. Also, TQ showed superior inhibitory effects on iNOS and TGF-β1. Meanwhile, CMN was more potent in inhibiting mRNA expression of TNF-α. These results suggest that TQ is more potent in inhibiting the inflammatory changes associating asthma. On the other hand, CMN was a less potent inhibitor of all measured parameters, despite its superior inhibitory effect on TNF-α mRNA levels.

Topics: Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents, Non-Steroidal; Asthma; Benzoquinones; Bronchoalveolar Lavage Fluid; Curcumin; Immunoglobulin E; Lung; Male; Mice; Nitric Oxide Synthase Type II; Transforming Growth Factor beta1; Tumor Necrosis Factor-alpha