Analysis of Volatile Compounds from Different Parts of Houttuynia cordata Thunb.

Houttuynia cordata Thunb. is a medicinal and edible plant that has been commonly used in traditional Chinese medicine since ancient times. This study used headspace solid-phase microextraction (HS-SPME) and direct injection, combined with gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS), to identify the volatile compounds in H. cordata. Extraction from different parts of the plant using different extraction techniques for the identification of volatile compounds were determined. A total of 93 volatile components were analyzed in the leaves, stems, rhizomes, and whole plant samples of H. cordata. The leaves contained more (Z)-3-hexenal, ß-myrcene, (Z)-ß-ocimene, and (4E,6E)-allo-ocimene; the stems contained more geranyl acetate and nerolidol; and rhizomes contained more α-pinene, ß-pinene, limonene, 2-undecanone, and decanoyl acetaldehyde. Among them, the essential oil extracted by HS-SPME could produce more monoterpenes, while direct injection could obtain higher contents of aliphatic ketones, terpene esters, sesquiterpenes, and was more conducive to the extraction of 2-undecanone and decanoyl acetaldehyde.

Headspace Solid-Phase Microextraction Analysis of Volatile Components in Peanut Oil.

Peanut oil is favored by consumers due to its rich nutritional value and unique flavor. This study used headspace solid-phase microextraction (HS-SPME) combined with gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) to examine the differences in the peanut oil aroma on the basis of variety, roasting temperatures, and pressing components. The results revealed that the optimal conditions for extracting peanut oil were achieved through the use of 50/30 µm DVB/CAR/PDMS fibers at 60 °C for 50 min. The primary compounds present in peanut oil were pyrazines. When peanuts were roasted, the temperature raised from 120 °C to 140 °C and the content of aldehydes in peanut oil increased; however, the content of aldehydes in No. 9 oil at 160 °C decreased. The components of peanut shell oil varied depending on the peanut variety. The most marked difference was observed in terms of the main compound at the two roasting temperatures. This compound was a pyrazine, and the content increased with the roasting temperature in hekei oils. When the roasting temperature was lower, No. 9 oil contained more fatty acid oxidation products such as hexanal, heptanal, and nonanal. When the roasting temperature increased, No. 9 oil contained more furfural and 5-methylfurfural. Heren oil was easier to oxidize and produced nonanal that possessed a fatty aroma.

Phenylpropanoids and lignoids from the whole plant of Vaccinium emarginatum and their cytotoxicity against prostate cancer cells.

One new naturally occurring quinone, 3',4'-dihydroxy-1,2,6-trimethoxy-[1,1'-biphenyl]-4(1H)-one (1), one new diarylpropane, emarginone A (2), and one new neolignan, emarginone B (3), along with eighteen known compounds have been isolated from the chemical investigation of the EtOAc-soluble fraction of the Vaccinium emarginatum whole plant methanolic extract. The new structures were elucidated by combined analysis of spectroscopic analytical methods and comparison with the literature data obtained from known analogues. In addition, the cytotoxicity of compounds 2, 4, and 14-20 against Du145 and PC-3 prostate cancer cell lines using MTT cell proliferation assay was evaluated. Compounds 2 and 19 showed most potent cytotoxicity against Du145 with IC50 values of 7.53 and 6.63 µg/mL, respectively. Furthermore, compounds 2, 17, and 19 also exhibited significant cytotoxicity against PC-3 with IC50 values ranging from 3.44-6.64 µg/mL.

Analysis of Volatile Constituents in Platostoma palustre (Blume) Using Headspace Solid-Phase Microextraction and Simultaneous Distillation-Extraction.

Hsian-tsao (Platostoma palustre Blume) is a traditional Taiwanese food. It is admired by many consumers, especially in summer, because of its aroma and taste. This study reports the analysis of the volatile components present in eight varieties of Hsian-tsao using headspace solid-phase microextraction (HS-SPME) and simultaneous distillation-extraction (SDE) coupled with gas chromatography (GC) and gas chromatography-mass spectrometry (GC/MS). HS-SPME is a non-heating method, and the results show relatively true values of the samples during flavor isolation. However, it is a kind of headspace analysis that has the disadvantage of a lower detection ability to relatively higher molecular weight compounds; also, the data are not quantitative, but instead are used for comparison. The SDE method uses distillation 2 h for flavor isolation; therefore, it quantitatively identifies more volatile compounds in the samples while the samples withstand heating. Both methods were used in this study to investigate information about the samples. The results showed that Nongshi No. 1 had the highest total quantity of volatile components using HS-SPME, whereas SDE indicated that Taoyuan Mesona 1301 (TYM1301) had the highest volatile concentration. Using the two extraction methods, 120 volatile components were identified. Fifty-six volatile components were identified using HS-SPME, and the main volatile compounds were α-pinene, ß-pinene, and limonene. A total of 108 volatile components were identified using SDE, and the main volatile compounds were α-bisabolol, ß-caryophyllene, and caryophyllene oxide. Compared with SDE, HS-SPME sampling extracted a significantly higher amount of monoterpenes and had a poorer detection of less volatile compounds, such as sesquiterpenes, terpene alcohols, and terpene oxide.

Composition and Insecticidal Activity of Essential Oil of Bacopa caroliniana and Interactive Effects of Individual Compounds on the Activity.

Bacopa caroliniana (BC) is a perennial creeping herb and popular aquarium plant. This plant is easily cultivated; consequently, it has the potential to be a raw material which is readily available for mass production if it contains useful bioactive substances. The information about the functionality of this plant has been very limited. Therefore, the aims of this research were to analyze the composition of the essential oil (EO) of BC and to study its insecticidal effect on rice weevils. Moreover, the interactive effects of active compounds of the EO on this activity were also investigated. A total of 18 volatile compounds was identified, accounting for ca. 94% of the BC-EO in terms of quantity. Of them, α-terpinolene was the largest compound. The impact of individual volatile compounds on the inhibition of acetylcholine esterase and insecticidal activity were determined. α-Terpinolene exhibited the highest activity on these assays. Both additive and synergistic effects existed in terms of the insecticidal activity. Many compounds found in the BC-EO are widely present in other EOs. Thus, the information obtained from this study is useful for EO-related research, applications in selecting EOs or in seeking the best combination of EOs or individual compounds to achieve efficient insecticidal effects.

Identification of ß-Sitosterol as in Vitro Anti-Inflammatory Constituent in Moringa oleifera.

ß-Sitosterol is a well known phytosterol in plants, but owing to its poor solubility in typical media, determining its cellular mechanisms has been proven to be difficult. In this study, we investigated the anti-inflammatory activity of ß-sitosterol (BSS) isolated from Moringa oleifera in two cell lines. Over a dose range of 7.5 to 30 µM, BSS dispersed well in the medium as nanoparticles with diameters of 50 ± 5 nm and suppressed the secretion of inflammatory factors from keratinocytes and macrophages induced by PGN, TNF-α, or LPS, such as TNF-α, IL-1ß, IL-6, IL-8, and ROS, separately. In addition, BSS significantly reduced the expression of NLRP3, a key component of NLRP3 inflammasomes, and inhibited the activation of caspase-1. There was partial inhibition of NF-κB in macrophages. This is the first study to report an increase in the solubility of nearly water-insoluble phytosterols via the formation of nanoparticles and to delineate the formulation's capacity to inhibit the signal transduction pathways of inflammation in macrophages.

Assessment of biochemical and immunomodulatory activity of sulphated polysaccharides from Ulva intestinalis.

The biochemical characteristics and immunomodulatory activity of sulphated polysaccharides isolated from Ulva intestinalis and fractionated using a silica-silica column were investigated. The unfractionated (USP) and fractionated sulphated polysaccharides (FSP4, FSP30, and FSP32) consisted mostly of carbohydrates (4.84-26.55%) and sulphates (2.85-20.42%). Structural analyses showed that USP, FSP4, FSP30 and FSP32 had molecular weights of 300, 80, 110 and 140kDa, respectively. FSP30 exhibited the strongest DPPH radical scavenging activity. Moreover, FSP30 showed stronger immunomodulatory activities than UPS in term of stimulating the production of pro-inflammatory cytokines, including nitric oxide (NO), tumour necrosis factor-α (TNF-α), and interleukin-1ß (IL-1ß), in macrophage J774A.1 cells. USP and FSP30 were not cytotoxic to mouse macrophage at the tested concentrations (6.25-50µg/mL). The results suggested that U. intestinalis polysaccharides could be explored as potential antioxidant and immunomodulatory agents to be used as complementary medicine or functional foods.

Effects of processing methods on composition and functionality of volatile components isolated from immature fruits of atemoya.

Atemoya is one of the most important commercial fruits of the family Annonaceae. The immature fruits of atemoya amply produced from a fruit-thinning process is normally regarded as waste and discarded. This research aimed at studying antimicrobial, antioxidant, and anti-inflammatory activities of the essential oil (EO) isolated from the immature fruits to explore its potential application. The fruits were subjected to different drying methods: solar drying (SD), oven drying at 30°C (OD-30), and at 50°C (OD-50). The oven drying method gave a higher EO yield than the solar drying method. Spathulenol was the largest compound in the EO after the drying process. Antimicrobial effect was not affected by the different drying methods. Antioxidant activity of the EO was measured by DPPH, nitric oxide, and reducing power methods. The EOOD-50 exhibited a stronger antioxidant activity than EOSD and EOOD-30. The EO also showed an anti-inflammatory activity in a cell model.

Citral alleviates an accelerated and severe lupus nephritis model by inhibiting the activation signal of NLRP3 inflammasome and enhancing Nrf2 activation.

INTRODUCTION: Lupus nephritis (LN) is a major complication of systemic lupus erythematosus. NLRP3 inflammasome activation, reactive oxygen species (ROS) and mononuclear leukocyte infiltration in the kidney have been shown to provoke the acceleration and deterioration of LN, such as accelerated and severe LN (ASLN). Development of a novel therapeutic remedy based on these molecular events to prevent the progression of the disease is clinically warranted. METHODS: Citral (3,7-dimethyl-2,6-octadienal), a major active compound in a Chinese herbal medicine Litsea cubeba, was used to test its renoprotective effects in a lipopolysaccharide (LPS)-induced mouse ASLN model by examining NLRP3 inflammasome activation, ROS and COX-2 production as well as Nrf2 activation. The analysis of mechanisms of action of Citral also involved its effects on IL-1ß secretion and signaling pathways of NLRP3 inflammasome in LPS-primed peritoneal macrophages or J774A macrophages. RESULTS: Attenuated proteinuria, renal function impairment, and renal histopathology, the latter including intrinsic cell proliferation, cellular crescents, neutrophil influx, fibrinoid necrosis in the glomerulus, and peri-glomerular infiltration of mononuclear leukocytes as well as glomerulonephritis activity score were observed in Citral-treated ASLN mice. In addition, Citral inhibited NLRP3 inflammasome activation and levels of ROS, NAD(P)H oxidase subunit p47(phox), or COX-2, and it enhanced the activation of nuclear factor E2-related factor 2 (Nrf2). In LPS-primed macrophages, Citral reduced ATP-induced IL-1ß secretion and caspase-1 activation, but did not affect LPS-induced NLRP3 protein expression. CONCLUSION: Our data show that Citral alleviates the mouse ASLN model by inhibition of the activation signal, but not the priming signal, of NLRP3 inflammasome and enhanced activation of Nrf2 antioxidant signaling.

Peroxyauraptenol Inhibits Inflammation and NLRP3 Inflammasome Activation by Inhibiting Reactive Oxygen Species Generation and Preserving Mitochondrial Integrity.

Peroxyauraptenol (PXT) is a peroxide-containing coumarin compound isolated from the seeds of Cnidium monnieri. PXT exerts anti-inflammatory activities, as it reduces the levels of inducible nitric oxide synthase, nitric oxide, IL-6, and NLRP3 inflammasome-derived IL-1ß in lipopolysaccharide-activated macrophages. PXT also exerts anti-inflammatory activity by reducing reactive oxygen species generation (including mitochondrial), mitogen-activated protein kinase, protein kinase C-α/δ phosphorylation, and the release of mitochondrial DNA into the cytosol. In addition, PXT suppresses the phagocytic activity of macrophages and IL-1ß secretion by Klebsiella pneumoniae-infected macrophages. The unique peroxide group is important for the anti-inflammatory activity of PXT, as this activity is reduced when the peroxide group is replaced by a hydroxyl group. These findings suggest that PXT may be a candidate for the development of anti-inflammatory agents or a healthy supplement for preventing and ameliorating inflammation-related diseases.

Resveratrol inhibits NLRP3 inflammasome activation by preserving mitochondrial integrity and augmenting autophagy.

The NLRP3 inflammasome is a caspase-1-containing multi-protein complex that controls the release of IL-1ß and plays important roles in the development of inflammatory disease. Here, we report that resveratrol, a polyphenolic compound naturally produced by plants, inhibits NLRP3 inflammasome-derived IL-1ß secretion and pyroptosis in macrophages. Resveratrol inhibits the activation step of the NLRP3 inflammasome by suppressing mitochondrial damage. Resveratrol also induces autophagy by activating p38, and macrophages treated with an autophagy inhibitor are resistant to the suppressive effects of resveratrol. In addition, resveratrol administration mitigates glomerular proliferation, glomerular sclerosis, and glomerular inflammation in a mouse model of progressive IgA nephropathy. These findings were associated with decreased renal mononuclear leukocyte infiltration, reduced renal superoxide anion levels, and inhibited renal NLRP3 inflammasome activation. Our data indicate that resveratrol suppresses NLRP3 inflammasome activation by preserving mitochondrial integrity and by augmenting autophagy.

Two new labdane-type diterpenes from the wood of Cunninghamia konishii.

Phytochemical investigation of the methanol extract of the wood of Cunninghamia konishii resulted in the isolation of two new acidic labdane-type diterpenoids, 12(S)-hydroxy-15,16-epoxylabda-8(17),13-dien-19-oic acid (1) and 12(S)-hydroxy-15,16-epoxylabda-8(17),13-dien-18-oic acid (2), along with one known labdane-type diterpene, 7,13E-labdadien-15-ol (3). Their structures were determined by analysis of spectroscopic data and comparison with the data of known analogues.

Osthole improves an accelerated focal segmental glomerulosclerosis model in the early stage by activating the Nrf2 antioxidant pathway and subsequently inhibiting NF-κB-mediated COX-2 expression and apoptosis.

Inflammatory reactions and oxidative stress are implicated in the pathogenesis of focal segmental glomerulosclerosis (FSGS), a common chronic kidney disease with relatively poor prognosis and unsatisfactory treatment regimens. Previously, we showed that osthole, a coumarin compound isolated from the seeds of Cnidium monnieri, can inhibit reactive oxygen species generation, NF-κB activation, and cyclooxygenase-2 expression in lipopolysaccharide-activated macrophages. In this study, we further evaluated its renoprotective effect in a mouse model of accelerated FSGS (acFSGS), featuring early development of proteinuria, followed by impaired renal function, glomerular epithelial cell hyperplasia lesions (a sensitive sign that precedes the development of glomerular sclerosis), periglomerular inflammation, and glomerular hyalinosis/sclerosis. The results show that osthole significantly prevented the development of the acFSGS model in the treated group of mice. The mechanisms involved in the renoprotective effects of osthole on the acFSGS model were mainly a result of an activated Nrf2-mediated antioxidant pathway in the early stage (proteinuria and ischemic collapse of the glomeruli) of acFSGS, followed by a decrease in: (1) NF-κB activation and COX-2 expression as well as PGE2 production, (2) podocyte injury, and (3) apoptosis. Our data support that targeting the Nrf2 antioxidant pathway may justify osthole being established as a candidate renoprotective compound for FSGS.

Antimicrobial activity of the essential oil of Glossogyne tenuifolia against selected pathogens.

BACKGROUND: Glossogyne tenuifolia (GT) is a perennial herb widely distributed in the areas from south Asia to Australia. Many biological effects of G. tenuifolia have been reported; however, the information about antimicrobial activity of the essential oil (EO) of the herb remains unavailable. Therefore, the aims of this study were to investigate the antimicrobial activity of the GT-EO in vitro and food systems, the antimicrobial impact (AI) of its individual compounds, and interactive effects of major active compounds (linalool, 4-terpineol, α-terpineol, ρ-cymene) on selected Gram-positive (S. aureus, L. monocytogenes, S. mutans and S. sanguinis) and Gram-negative (E. coli O157:H7, V. parahaemolyticus and S. enterica) pathogens. RESULTS: The minimal microbicidal concentration (MMC) of the GT-EO ranged from 0.75 to 12 mg mL(-1) against the test bacteria in vitro. Except for L. monocytogenes, the GT-EO exhibited more inhibitory effect on the selected Gram positive than against the Gram negative bacteria at the GT-EO concentrations ≤ 12 mg mL(-1) . The interactive effects of major active compounds (linalool, 4-terpineol, α-terpineol, ρ-cymene) are additive instead of synergistic via the checkerboard analysis. The bacteria with a microbial load of ca. 10(2) CFU mL(-1) in the milk tea could be completely inactivated by the GT-EO with the MMC of 1.5 mg mL(-1) . CONCLUSION: ρ-Cymene is the largest component in the GT-EO; however, it is not the compound predominantly affecting the entire antimicrobial activity of the EO. Instead, 4-terpineol is the most influential among the test compounds that contribute to the antimicrobial activity of the GT-EO.

Osthole mitigates progressive IgA nephropathy by inhibiting reactive oxygen species generation and NF-κB/NLRP3 pathway.

Renal reactive oxygen species (ROS) and mononuclear leukocyte infiltration are involved in the progressive stage (exacerbation) of IgA nephropathy (IgAN), which is characterized by glomerular proliferation and renal inflammation. The identification of the mechanism responsible for this critical stage of IgAN and the development of a therapeutic strategy remain a challenge. Osthole is a pure compound isolated from Cnidiummonnieri (L.) Cusson seeds, which are used as a traditional Chinese medicine, and is anti-inflammatory, anti-apoptotic, and anti-fibrotic both in vitro and in vivo. Recently, we showed that osthole acts as an anti-inflammatory agent by reducing nuclear factor-kappa B (NF-κB) activation in and ROS release by activated macrophages. In this study, we examined whether osthole could prevent the progression of IgAN using a progressive IgAN (Prg-IgAN) model in mice. Our results showed that osthole administration resulted in prevention of albuminuria, improved renal function, and blocking of renal progressive lesions, including glomerular proliferation, glomerular sclerosis, and periglomerular mononuclear leukocyte infiltration. These findings were associated with (1) reduced renal superoxide anion levels and increased Nrf2 nuclear translocation, (2) inhibited renal activation of NF-κB and the NLRP3 inflammasome, (3) decreased renal MCP-1 expression and mononuclear leukocyte infiltration, (4) inhibited ROS production and NLRP3 inflammasome activation in cultured, activated macrophages, and (5) inhibited ROS production and MCP-1 protein levels in cultured, activated mesangial cells. The results suggest that osthole exerts its reno-protective effects on the progression of IgAN by inhibiting ROS production and activation of NF-κB and the NLRP3 inflammasome in the kidney. Our data also confirm that ROS generation and activation of NF-κB and the NLRP3 inflammasome are crucial mechanistic events involved in the progression of the renal disorder.

Anti-inflammatory lanostanoids and lactone derivatives from Antrodia camphorata.

Four new lanostanoids, ethyl lucidenate A (1), ethyl lucidenate F (2), 15-O-acetylganolucidate A (3), and 3,11,15,23-tetraoxo-27ξ-lanosta-8,16-dien-26-oic acid (4), and two new lactone derivatives, 5-hydroxy-5-(methoxymethyl)-4-methylfuran-2(5H)-one (5) and 3-(4-methoxy-2-oxo-2H-pyran-6-yl)propanoic acid (6), together with four known compounds, 11α-hydroxy-3,7-dioxolanost-8,24(E)-dien-26- oic acid (7), 3,7,11-trioxo-5α-lanosta-8,24(E)-dien-26-oic acid (8), methyl 3,7,11,12,15,23-hexaoxo-5α-lanost-8-en-26-oate (9), and ethyl 3,7,11,12,15,23-hexaoxo-5α-lanost-8-en-26-oate (10), were characterized from Antrodia camphorata. The structures of these new compounds were determined by analysis of their spectroscopic data, including 1D and 2D NMR experiments. Ten components were evaluated for anti-inflammatory activity by examining their effect on LPS-iNOS-dependent NO production in murine macrophage (RAW 264.7) cells. Among them, compounds 1, 3, 7, 8, 9, and 10 significantly suppressed the NO concentration in LPS-treated RAW 264.7 cells with IC50 values ≤ 10 µM.

New Anti-Inflammatory Aromatic Components from Antrodia camphorata.

Three new benzenoids, 3-isopropenyl-2-methoxy-6-methyl-4,5-methylenedioxy- phenol (1), 2-hydroxy-4,4'-dimethoxy-3,3'-dimethyl-5,6,5',6'-bimethylenedioxybiphenyl (2), 4,4'-dihydroxy-3,3'-dimethoxy-2,2'-dimethyl-5,6,5',6'-bimethylenedioxybiphenyl (3), together with two known benzenoids, 2,3,6-trimethoxy-5-methylphenol (4) and 2,3-methylenedioxy- 4-methoxy-5-methylphenol (5), were isolated from Antrodia camphorata. Our results support that compounds 1-5 potently inhibited LPS (lipopolysaccharide)-induced nitric oxide (NO) production in a dose-dependent manner. The IC(50) values of compounds 1, 3 and 5 were 1.8 ± 0.2, 18.8 ± 0.6 and 0.8 ± 0.3 µg/mL, respectively.

Diterpenoids with anti-inflammatory activity from the wood of Cunninghamia konishii.

Two new diterpenoids, konishone (1) and 3b-hydroxy-5,6-dehydrosugiol (2), along with three known diterpenoids--hinokiol (3), sugiol (4), and 12-hydroxy-6,7-secoabieta-8,11,13-triene-6,7-dial (5)--were isolated from the wood of Cunninghamia konishii. Compound 1 is a novel skeleton of the 7,20-dinorabietane-type diterpene. In addition, when RAW264.7 macrophages were treated with different concentrations of compounds 1, 3, and 5 together with LPS, a significant concentration-dependent inhibition of NO production was detected. The IC50 values for inhibition of nitrite production of compounds 1, 3, and 5 were about 9.8 ± 0.7, 7.9 ± 0.9, and 9.3 ± 1.3 µg/mL, respectively. This study presents the potential utilization of compounds 1, 3, and 5, as lead compounds for the development of anti-inflammatory drugs.

Lipopolysaccharide/adenosine triphosphate-mediated signal transduction in the regulation of NLRP3 protein expression and caspase-1-mediated interleukin-1ß secretion.

OBJECTIVE: Reactive oxygen species (ROS) plays a critical role in the regulation of NLRP3 inflammasome activation. However, the ROS-mediated signaling pathways controlling NLRP3 inflammasome activation are not well defined. METHODS: Using lipopolysaccharide (LPS) and adenosine triphosphate (ATP) activated murine macrophages as the testing model, cytokine release and protein expression were quantified by enzyme-linked immunosorbent assay and Western blot, respectively. ROS was scavenged by N-acetyl cysteine; NADPH oxidase, the major source of ROS, was inhibited by diphenyliodonium, apocynin or gp91-phox siRNA transfection; and protein kinase was inhibited by its specific inhibitor. RESULTS: LPS-induced NLRP3 protein expression was regulated through the NADPH oxidase/ROS/NF-κB-dependent, JAK2/PI3-kinase/AKT/NF-κB-dependent, and MAPK-dependent pathways, while ATP-induced caspase-1 activation was regulated through the NADPH oxidase/ROS-dependent pathway. CONCLUSIONS: These results demonstrate that ROS regulates not only the priming stage, but also the activation stage, of NLRP3 inflammasome activation in LPS + ATP-activated macrophages.

Improved purity and immunostimulatory activity of ß-(1→3)(1→6)-glucan from Pleurotus sajor-caju using cell wall-degrading enzymes.

The objective of this work was to improve the purity of ß-(1→3)(1→6)-glucan in the native triple helical structure from the fruiting bodies of Pleurotus sajor-caju for effective biological function using cell wall-degrading enzymes. A crude carbohydrate was extracted with hot water, then treated with crude xylanase and cellulase from Paenibacillus curdlanolyticus B-6. ß-Glucan in the extract was purified to homogeneity with a single and symmetrical peak using 650M DEAE Toyopearl and Sepharose CL-6B column chromatography. The purity of ß-glucan was confirmed by high-performance size-exclusion chromatography. Purified ß-glucan was obtained at a purity of up to 90.2%. The Congo red reaction and atomic force microscopy indicated that the purified ß-glucan exhibited a triple helix conformation. Purified ß-glucan was able to effectively up-regulate the functions of macrophages such as nitric oxide (NO) and tumor necrosis factor (TNF-α) production.