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Cyclooxygenase-2 inhibitory and antioxidant compounds from the truffle Elaphomyces granulatus

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The ethanol extract of fruiting bodies of Elaphomyces granulatus, a truffle-like fungus, was evaluated for cyclooxygenase-2 (COX-2) enzyme inhibitory and antioxidant activities. Inhibition of COX-2 activity was evaluated in mouse macrophages (RAW 264.7). The extract of E. granulatus caused a 68% inhibition of COX-2 activity at 50 microg/mL. Bioassay-guided investigation led to the isolation and identification of two active compounds, syringaldehyde and syringic acid. Syringaldehyde moderately inhibited COX-2 activity with an IC(50) of 3.5 microg/mL, while syringic acid strongly inhibited COX-2 activity with an IC(50) of 0.4 microg/mL. The antioxidant activity of the extract and isolated compounds was evaluated in HL-60 cells by the DCFH-DA method. The extract of E. granulatus showed a potent antioxidant effect, with an IC(50) of 41 microg/mL. Of the pure compounds, syringic acid displayed a strong antioxidant activity, with an IC(50) of 0.7 microg/mL, while syringaldehyde showed no activity in the assay.
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COMPOUNDS FROM THE TRUFFLE ELAPHOMYCES GRANULATUS 575
Copyright © 2008 John Wiley & Sons, Ltd. Phytother. Res. 23, 575–578 (2009)
DOI: 10.1002/ptr
Copyright © 2008 John Wiley & Sons, Ltd.
PHYTOTHERAPY RESEARCH
Phytother. Res. 23, 575–578 (2009)
Published online 9 December 2008 in Wiley InterScience
(www.interscience.wiley.com) DOI: 10.1002/ptr.2698
Cyclooxygenase-2 Inhibitory and Antioxidant
Compounds from the Truffle Elaphomyces
granulatus
Rita Stanikunaite1, Shabana I. Khan2, James M. Trappe3 and Samir A. Ross1,2*
1Department of Pharmacognosy, The University of Mississippi, University, MS 38677, USA
2National Center for Natural Products Research, The University of Mississippi, University, MS 38677, USA
3Department of Forest Science, Oregon State University, Corvallis, OR 97331-5752, USA
The ethanol extract of fruiting bodies of Elaphomyces granulatus, a truffle-like fungus, was evaluated for
cyclooxygenase-2 (COX-2) enzyme inhibitory and antioxidant activities. Inhibition of COX-2 activity was evaluated
in mouse macrophages (RAW 264.7). The extract of E. granulatus caused a 68% inhibition of COX-2 activity
at 50 μμ
μμ
μg/mL. Bioassay-guided investigation led to the isolation and identification of two active compounds,
syringaldehyde and syringic acid. Syringaldehyde moderately inhibited COX-2 activity with an IC50 of 3.5 μμ
μμ
μg/mL,
while syringic acid strongly inhibited COX-2 activity with an IC50 of 0.4 μμ
μμ
μg/mL. The antioxidant activity of the
extract and isolated compounds was evaluated in HL-60 cells by the DCFH-DA method. The extract of E.
granulatus showed a potent antioxidant effect, with an IC50 of 41 μμ
μμ
μg/mL. Of the pure compounds, syringic acid
displayed a strong antioxidant activity, with an IC50 of 0.7 μμ
μμ
μg/mL, while syringaldehyde showed no activity in
the assay. Copyright © 2008 John Wiley & Sons, Ltd.
Keywords: Elaphomyces granulatus; Ascomycota; hypogeous; truffle; COX-2; RAW 264.7 cells; antiinflammatory; antioxidants;
DCFH oxidation; HL-60 cells.
Received 4 May 2007
Accepted 29 July 2008
* Correspondence to: Samir A. Ross, National Center for Natural Prod-
ucts Research, P.O. Box 1848, University, MS 38677, USA.
E-mail: sross@olemiss.edu
Contract/grant sponsor: USDA Agricultural Research Service Specific
Cooperative Agreement; contract/grant number: 58-6408-2-0009.
INTRODUCTION
Truffles and truffle-like fungi are characterized by a hypo-
geous, i.e. underground, fruiting habit, having evolved
the use of animal mycophagy for the dispersal of their
spores. These fungi produce aromatic compounds by
which the animals locate them, dig them up and eat
them (Trappe and Claridge, 2005). The animals digest
the nutritious tissues of the fruiting bodies, but the spores
pass through the digestive tract and are defecated
unharmed (Claridge and Trappe, 2005). Because of their
underground fruiting habit, truffles are difficult to find,
hence research on their chemistry and medicinal prop-
erties has been limited and generally focused on the
volatile compounds responsible for their unique aromas
(Diaz et al., 2002, 2003).
In the course of our program aimed at the investiga-
tion of biological activity and chemistry of truffles from
North America, a study on Elaphomyces granulatus
Fr. (Elaphomycetaceae, Ascomycota) was initiated. The
most widely distributed and common hypogeous fungus
in the Northern Hemisphere, it occurs as a beneficial,
mycorrhizal symbiont with feeder rootlets of trees from
subarctic and subalpine forests south to the tropics
(Trappe, 1971). In Europe, E. granulatus has been used
as an aphrodisiac and a cheap but illegitimately marketed
substitute for more expensive truffles (Arora, 1986). The
analysis of the chemical composition of the spore mass
and outer layer of E. granulatus revealed the occurrence
of mixtures of higher aliphatic esters, free fatty acids,
hydrocarbons, mannitol, ergosterol, pyrocatechol, proto-
catechuic acid, salicylic acid, resorcinol, 3-hydroxy- and
4-hydroxy benzoic acids (Solberg, 1976). However, no
other studies of its biological activity and/or chemical
constituents have been reported.
Cyclooxygenase-2 (COX-2) enzyme plays an important
role in the inflammatory process. COX-2 is an induc-
ible isoform of cyclooxygenase enzyme responsible for
the production of pro-inflammatory prostaglandins in
neoplastic and inflamed tissues. COX-2 inhibitors have
a well established role in the treatment of inflamma-
tory disorders, as well as potential application for the
prevention and treatment of other diseases, such as
cancer (Flower, 2003; Amir and Agarwal, 2005). Reac-
tive oxygen species (ROS) and oxidative stress also play
an important role in the etiology and progression of
human degenerative diseases. ROS have been impli-
cated in inflammation, aging, cancer, heart disease and
other disorders (Pietta, 2000). Antioxidants act as ROS
scavengers and are important for protecting against
oxidative tissue damage in vital organs. Although,
numerous in vitro solution-based chemical assay systems,
such as 2,2-diphenyl-1-picrylhydrazyl radical (DPPH)
assay, have been used for the evaluation of antioxidants
(Cuendet et al., 1997), the use of 2,7-dichlorodihydro-
fluorescein diacetate (DCFH-DA) as a specific probe
in a cell based assay provides a better system to evalu-
ate an antioxidant effect in live cells. This method is
useful for the direct examination of ROS inhibitory
activity of natural products in live human cells
(Takamatsu et al., 2003; Choi et al., 2006).
As part of our investigation of biologically active
compounds from E. granulatus, the ethanol extract of
Copyright © 2008 John Wiley & Sons, Ltd. Phytother. Res. 23, 575–578 (2009)
DOI: 10.1002/ptr
576 R. STANIKUNAITE ET AL.
fruiting bodies of E. granulatus was evaluated for COX-
2 enzyme inhibitory and antioxidant activities in cellu-
lar assays. The bioassay-guided isolation and biological
characterization of compounds from E. granulatus with
antioxidant and COX-2 inhibitory activities were also
performed.
MATERIALS AND METHODS
General experimental procedures. The 1H-NMR, 13C-
NMR, HMBC and HMQC spectra were recorded on a
Bruker DRX 400 MHz. The Bruker NMR spectrometer
operated at 400 MHz for 1H and 100 MHz for 13C. The
NMR spectra were recorded in ppm using the residual
solvent peak as an internal standard. The HRESIMS
data were acquired on a Bruker BioAPEX 30es mass
spectrometer.
Fungal material. Elaphomyces granulatus was collected
by Mr Adrian Beyerle of the North American Truffling
Society (NATS) in Oregon, Wasco Co., Mt Hood
National Forest, near 15-Mile Creek on 16 August 2002.
All specimens were identified by Dr James M. Trappe.
A voucher specimen is deposited in the USDA National
Fungus Collections, Beltsville, MD (BPI 864222). Mr
Adrian Beyerle subsequently provided additional speci-
mens from the area to supplement the original collection.
Additional collections were contributed by other NATS
members.
Extraction and isolation. The fruiting bodies of E.
granulatus were dried for 24 h in a forced air dehydrator
at 35 °C. Powdered material (482 g) was extracted
exhaustively by maceration with 95% EtOH at room
temperature, and the combined extracts were concen-
trated under reduced pressure to yield 12.5 g of residue.
The crude extract was divided into EtOH soluble and
insoluble fractions. The EtOH soluble fraction (9 g) was
subjected to a silica gel gravity column (230 g, 457 mm
× 51 mm) and eluted with chloroform, and chloroform–
methanol (2%–100%) to yield 16 fractions. The active
fractions 4–7 were combined (800 mg) and subjected
to another silica gel column (50 g, 450 mm × 17 mm)
with hexane–chloroform–ethyl acetate (3:3:1–1:1:1–0:1:1)
to give 108 fractions. Fractions 52–74 were combined
(61 mg) and separated on preparative TLC (Si gel
CF254, 250 μm, Uniplate) with hexane–chloroform–ethyl
acetate (1:1:1) to yield syringaldehyde (7 mg). Frac-
tions 98–102 were combined (63 mg) and separated
on preparative TLC (Si gel CF254, 250 μm, Uniplate)
with chloroform–methanol (2%) to yield syringic acid
(10 mg).
Syringaldehyde: pale yellow solid; HRESIMS m/z
183.0638 (calcd for C9H11O4 [M + H]+ 183.0657), 205.0457
(calcd for C9H10O4Na [M + Na]+ 205.0477); 1
H NMR
(CDCl3, 400 MHz):
δ
9.81 (1H, s, CHO), 7.14 (2H, s,
H-2, H-6), 3.95 (6H, s, 2OCH3); 13C NMR (CDCl3,
100 MHz):
δ
190.7 (CHO), 147.4 (C-3, C-5), 141.0 (C-
4), 128.4 (C-1), 106.8 (C-2, C-6), 56.5 (OCH3). Spectra
corresponded with previously reported data (Ralph
et al., 2004).
Syringic acid: pale yellow solid; HRESIMS m/z 199.0580
(calcd for C9H11O5 [M + H]+ 199.0606), 221.0398 (calcd
for C9H10O5Na [M + Na]+ 221.0426); 1H NMR (DMSO-
d6, 400 MHz):
δ
7.21 (2H, s, H-2, H-6), 3.79 (6H, s,
2OCH3); 13C NMR (DMSO-d6, 100 MHz):
δ
167.9
(COOH), 148.1 (C-3, C-5), 140.9 (C-4), 121.1 (C-1),
107.6 (C-2, C-6), 56.6 (OCH3). Spectra corresponded
with previously reported data (Ralph et al., 2004).
Cell-based assay for inhibition of COX-2 activity. Mouse
macrophages (RAW 264.7, ATCC) were cultured in
a 75 cm2 culture flask in RPMI-1640 medium (Gibco)
supplemented with 10% bovine calf serum (Hyclone)
and 60 mg/L amikacin (Sigma), at 37 °C in an environ-
ment of 95% humidity and 5% CO2. For the assay, the
cells were seeded in the wells of 96-well plates (50,000
cells/well) and incubated at 37 °C for 24 h. After wash-
ing with RPMI-1640 medium, supplemented with 3%
bovine calf serum, the cells were incubated with 5 μg/
mL LPS (Escherichia coli 055:B5, Sigma) for 16 h to
induce the production of COX-2. Induced cells were
washed thoroughly with medium to remove LPS com-
pletely, and treated with different concentrations of
test samples for 2 h. Arachidonic acid (300 μM, Sigma)
was added and the cells were further incubated for
30 min. The amount of PGE2 released in the medium
was determined with PGE2 enzyme immunoassay kit
(Cayman Chem. Co.). COX-2 activity was determined
by the conversion of exogenous arachidonic acid to
PGE2 and expressed as the percent of the vehicle con-
trol. The concentration that caused 50% inhibition
of enzyme activity (IC50) was calculated from the dose
curves generated by plotting percent COX-2 activity
against the test concentrations. NS-398 (Cayman Chem.
Co.), a specific inhibitor of COX-2, was included as a
positive control in each assay.
Assay for cytotoxicity to macrophages. RAW 264.7 cells
were cultured as described above. For the assay, cells
were seeded to wells of a 96-well plate at a density of
25 000 cells/well and incubated for 24 h. Different dilu-
tions of test compounds were added to the cells and
incubated for 48 h. Cell viability was determined by the
neutral red assay (Borenfreund et al., 1990). After in-
cubation, the medium was removed and 100 μL of fresh
medium containing 0.2 mg/mL neutral red (Sigma) was
added to each well and incubated for 90 min. The cells
were washed with saline (0.9% NaCl) to remove excess
dye. The solution of acidified isopropanol (0.33% HCl)
was then added to lyse cells. As a result, the incorpo-
rated dye was liberated from viable cells, the absorb-
ance of which was measured at 490 nm using the EL312e
plate reader (Bio-Tek instruments).
Assay for antioxidant activity. Myelomonocytic HL-60
cells (ATCC) were grown in RPMI 1640 medium sup-
plemented with 10% fetal bovine serum (Hyclone) and
60 mg/mL amikacin at 37 °C in an environment of 95%
humidity and 5% CO2. For the assay, 125 μL of the cell
suspension (1 × 106cells/mL) was added to the wells
of a 96-well plate. After treating with different concen-
trations of the test samples for 30 min, the cells were
stimulated with 100 ng/mL phorbol 12-myristate 13-
acetate (PMA, Sigma) for 30 min. DCFH-DA (Mole-
cular Probe, 5 μg/mL) was added and the cells were
incubated for 15 min. The levels of DCF produced were
measured on a PolarStar plate reader with an excita-
tion wavelength at 485 nm and emission at 530 nm as
COMPOUNDS FROM THE TRUFFLE ELAPHOMYCES GRANULATUS 577
Copyright © 2008 John Wiley & Sons, Ltd. Phytother. Res. 23, 575–578 (2009)
DOI: 10.1002/ptr
described previously (Takamatsu et al., 2003; Choi et al.,
2006). The ability of the test materials to inhibit exo-
genous cytoplasmic ROS-catalysed oxidation of DCFH
to fluorescent DCF in HL-60 cells was measured in
comparison to PMA treated controls without the test
materials. The IC50 values were calculated from dose
curves of the % DCF production versus test concentra-
tions. Vitamin C (Sigma) was included as a positive
control.
Assay for cytotoxicity to HL-60 cells. Cytotoxicity of
the test samples to HL-60 cells was determined by the
XTT method after incubating the cells with test samples
for 48 h as described earlier (Takamatsu et al., 2003).
Briefly, 25 μL of XTT-PMS solution (1 mg/mL XTT
solution supplemented by 25 μM of PMS) was added
to each well. After incubating for 4 h at 37 °C, the
absorbance at 450 nm was measured on a plate reader
(EL312e; Bio-Tek instruments).
RESULTS AND DISCUSSION
The dried fruiting bodies of E. granulatus were exhaus-
tively extracted by maceration with 95% EtOH and
fractionated by various chromatographic techniques.
The bioassay-guided fractionation of E. granulatus led
to the isolation of two active compounds, syringaldehyde
and syringic acid (Fig. 1). Structures of syringaldehyde
and syringic acid were determined by using HRESIMS,
1H-NMR and 13C-NMR experiments.
The extract of E. granulatus and isolated compounds
were evaluated for inhibition of COX-2 activity in a
cell-based assay that utilizes the mouse macrophage cell
line (RAW 264.7). Unstimulated macrophages express
only a small amount of COX-2, while treatment with
bacterial lipopolysaccharide (LPS) leads to the induc-
tion of COX-2, which converts arachidonic acid to PGE2
(Chen et al., 2001). LPS-induced RAW 264.7 macrophages
were incubated in the presence, or absence, of test sam-
ples for 2 h, followed by the addition of arachidonic
acid. The effects of the test samples on COX-2 activity
were determined by measuring the PGE2 produced
in the culture medium. NS-398, a specific inhibitor of
COX-2, was used as a positive control (IC50: 0.2 μg/mL,
0.64 μM).
The extract of E. granulatus showed a potent COX-
2 inhibitory activity with 68% inhibition at 50 μg/mL.
Syringaldehyde inhibited COX-2 activity in a dose-
dependent manner (Fig. 2), with an IC50 of 3.5 μg/mL
(19.23 μM). Syringic acid showed a stronger inhibition
Figure 1. Structures of syringaldehyde and syringic acid.
Figure 2. Inhibition of COX-2 enzyme activity by syringaldehyde
() and syringic acid () in LPS-activated macrophages (RAW
264.7). Each data point represents the mean ± SD of triplicate
determination.
of COX-2 activity in a dose-dependent manner (Fig. 2),
with an IC50 of 0.4 μg/mL (2.02 μM).
The RAW 264.7 cell viability was determined to
exclude the possibility that the observed COX-2 inhibi-
tory effect was due to cytotoxicity. Examination of
cytotoxicity of syringic acid and syringaldehyde in RAW
264.7 macrophages by the neutral red assay indicated
that compounds did not affect the viability of RAW
264.7 cells in concentrations up to 25 μg/mL.
The results of this study demonstrate that syringal-
dehyde has a moderate COX-2 inhibitory activity, while
syringic acid is a strong inhibitor of the COX-2 enzyme.
This study is the first report on the occurrence of
syringaldehyde and syringic acid in E. granulatus, which
could account for the potent COX-2 inhibitory activity
of the mushroom extract. Although in previous studies
syringic acid has been reported to show antiinflam-
matory activity in vivo (Fernandez et al., 1998; Gamaniel
et al., 2000), this is the first report on its activity on the
COX-2 enzyme. The results of this study suggest that
the mechanism responsible for the antiinflammatory
activity of syringic acid might be related to COX-2
inhibition.
The antioxidant activity of the extract and isolated
compounds was evaluated in HL-60 cells using DCFH-
DA. This cell-based method examines directly the abil-
ity of test material to penetrate living cells and inhibit
ROS catalysed oxidation of DCFH to DCF. DCFH-
DA is a non-fluorescent probe that diffuses into cells.
Cytoplasmic esterases hydrolyse DCFH-DA to DCFH
which is oxidized to DCF (2,7-dichlorofluorescin) that
fluoresces. The antioxidant activity of test samples is
determined by measuring the level of DCF produced
in treated cells compared with controls.
The extract of E. granulatus showed a potent anti-
oxidant effect, with an IC50 of 41 μg/mL. The inhibitory
effect of syringic acid on DCF production is shown in
Fig. 3. Syringic acid displayed a strong antioxidant activity
in a dose-dependent manner, with an IC50 of 0.7 μg/mL
(3.54 μM) which is comparable to the effect of vitamin
C, a naturally occurring antioxidant, that showed an
IC50 of 0.5 μg/mL (2.84 μM) in the same assay. Examina-
tion of the cytotoxicity of syringic acid and syringaldehyde
in HL-60 cells indicated that compounds were not
cytotoxic up to a concentration of 31.25 μg/mL.
The results of this study indicate that syringic acid has
a strong antioxidant activity in the cellular-based assay
while syringaldehyde was inactive. Most of the previous
reports on the antioxidant properties of syringaldehyde
Copyright © 2008 John Wiley & Sons, Ltd. Phytother. Res. 23, 575–578 (2009)
DOI: 10.1002/ptr
578 R. STANIKUNAITE ET AL.
Figure 3. Effect of syringic acid on DCF production in HL-60
cells. Each data point represents the mean ± SD of duplicate
determination.
and syringic acid have utilized solution-based chemical
assay systems that do not evaluate the antioxidant
activity within living cells. One recent study on ROS
production by human neutrophils, induced by opsonized
zymosan or phorbol 12-myristate 13-acetate (PMA),
found ROS inhibitory effects for both syringaldehyde
and syringic acid measured as luminol or lucigenin-
enhanced chemiluminescence (Worm et al., 2001).
This study is the first report on the potent antioxidant
and COX-2 enzyme inhibitory properties of the extract
of E. granulatus. In addition, COX-2 inhibitory activities
of syringaldehyde and syringic acid are reported here
for the first time. E. granulatus seems to have potential
health benefits due to its antioxidant and antiinflam-
matory effects. Consumption of E. granulatus as a dietary
supplement or as a food item could contribute to the
prevention of cancer and inflammatory disorders.
Acknowledgements
We want to thank Ms Shama Moktan and Mr John Trott for their
excellent technical help. We also want to thank Mr Adrian Beyerle
and other members of the North American Truffling Society for pro-
viding specimens of Elaphomyces granulatus. USDA Agricultural
Research Service Specific Cooperative Agreement No. 58-6408-2-0009
is acknowledged for partial support of this work.
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... As a result, anti-aging cosmetics are developed to repair and maintain the skin barrier. Many studies have highlighted the potential of bioactive compounds derived from mushrooms to serve as anti-aging ingredients in serums, topical creams, and other cosmetics, primarily due to their antioxidant and anti-wrinkle properties [10,58,[154][155][156][157][158][159][160][161][162]. These compounds can help protect the skin from oxidative damage, reduce the appearance of wrinkles, and improve overall skin health. ...
... COX-2 is associated with the production of ROS and inflammation in normal skin tissue. Therefore, COX-2 inhibitors are applied in anti-wrinkle cosmetics [161]. ...
... Notably, several bioactive compounds extracted from mushrooms have been found to effectively inhibit COX-2 activity. Stanikunaite, Khan, Trappe and Ross [161] reported that the ethanol extract of fruiting bodies of the truffle-like fungus Elaphomyces granulatus exhibited a 68% inhibition of COX-2 activity at a concentration of 50 mg/mL in mouse macrophages (RWA 264.7). Further investigation led to the identification of two bioactive compounds in E. granulatus, namely syringic acid and syringaldehyde acid, which were suggested to be responsible for the COX-2 inhibitory property [161]. ...
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... However, Villares et al. (2012) reported gallic, homogentisic, protocatechuic, oand p-coumaric, hydroxybenzoic and hydroxycinnamic acids as the main phenolic compounds in T. aestivum, with fewer coumpounds being found in T. indicum and T. melanosporum. Protocatechuic acid and catechin were also the main phenolics in Chondrogaster pachysporus and Setchelliogaster tenuipes truffles (Tel- Cayan et al., 2018), while syringic acid was identified in the truffle Elaphomyces granulatus (Stanikunaite et al., 2009). The extremely diverse phenolic profiles reported in these studies may be associated with truffle species as well as environmental conditions. ...
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The molecular and biofunctional properties of protein and phenolic fractions in edible truffles remain largely unknown. This study examined the effect of ultrasonication on the contents, profiles, and bioactive properties of free and bound phenolics (FP and BP) from desert truffle (Tirmania nivea) and its protein fractions. Protein fractions from the Osborne extraction scheme were biochemically and structurally characterized. The albumin fraction showed the highest abundance (16.8%) and yield (35.8%). Total phenolic contents were the highest in non-sonicated samples (3.5-34.1 mg/g), particularly in the albumin fraction and in whole truffle. FP extracted at 30°C (FP-30°C) accounted for the largest proportion of total phenolics in all protein fractions, whereas BP-30°C and FP-60°C were predominant in non-sonicated and sonicated truffle, respectively. The highest antioxidant activity was obtained with FP-30°C extracts from non-sonicated albumins, globulins and truffle (91.9, 72.7 and 30.0%), followed by BP-30°C from non-sonicated albumins (25.4%) and FP-60°C from sonicated glutelins-1 (24.2%). High inhibition of α-amylase was evidenced in several extracts, including FP-30°C from non-sonicated glutelins-1 (99.2%) and FP-30°C from sonicated globulins (72.4%). Several extracts also displayed high inhibition of angiotensin I-converting enzyme (ACE), including FP-60°C from non-sonicated glutelins-1 (65.1%) and sonicated glutelins-1 (71.1%) and globulins (64.7%). Most extracts were rich in epicatechin, gallic acid, chlorogenic acid and catechin. Correlations between phenolic content, antioxidant activity, anti-α-amylase and anti-ACE activities were influenced by sonication. Sonication reduced the particle size of the proteins and modified their structural characteristics. These findings demonstrate that white desert truffle proteins co-occur with bioactive phenolics whose functionalities can be tailored by protein fractionation and sonication. KEYWORDS Truffles, proteins, phenolic profile, antioxidant activity, enzyme inhibition, sonication
... This study also shows that sinapic acid is one of the active substances responsible for the beneficial effect of radish seeds on the LPS-dependent inflammatory reaction [53]. Moreover, rasathiol isolated from R. sativus L., containing syringic acid, exhibits a number of biological activities, such as inhibition of COX-2 activity, antioxidant potential, and hepatoprotective effects [54]. The beneficial biological effects of radish extract are often attributed to phenolic compounds, glucosinolates, and flavonoids [18]. ...
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In the cosmetics industry, the extract from Raphanus sativus L. is fermented using specific starter cultures. These cosmetic ingredients act as preservatives and skin conditioners. Kombucha is traditionally made by fermenting sweetened tea using symbiotic cultures of bacteria and yeast and is used in cosmetic products. The aim of this study was to evaluate the cosmetic properties of radish leaf and root extract fermented with the SCOBY. Both unfermented water extracts and extracts after 7, 14, and 21 days of fermentation were evaluated. The analysis of secondary plant metabolites by UPLC-MS showed higher values for ferments than for extracts. A similar relationship was noted when examining the antioxidant properties using DPPH and ABTS radicals and the protective effect against H2O2-induced oxidative stress in fibroblasts and keratinocytes using the fluorogenic dye H2DCFDA. The results also showed no cytotoxicity to skin cells using Alamar Blue and Neutral Red tests. The ability of the samples to inhibit IL-1β and COX-2 activity in LPS-treated fibroblasts was also demonstrated using ELISA assays. The influence of extracts and ferments on bacterial strains involved in inflammatory processes of skin diseases was also assessed. Additionally, application tests were carried out, which showed a positive effect of extracts and ferments on TEWL and skin hydration using a TEWAmeter and corneometer probe. The results obtained depended on the concentration used and the fermentation time.
... Syringaldehyde is an aromatic aldehyde that is found in spruce, maple, [7,8] and oak woods and is an important flavor component of whiskey [9,10]. Recently, syringaldehyde has been shown to have biological activity [11], including antihyperglycemic activity [12,13], antioxidant activity [14,15], and anti-inflammatory activity [16]. In fact, phenol-containing Schiff bases often show biological activity. ...
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A series of syringaldehyde imines with para-substituted anilines have been synthesized in a good yield, and their crystal structures have been analyzed. The orientation of the syringaldehyde hydroxyl group plays in important role in the intermolecular hydrogen-bonding pattern of the molecules. The O–H…N hydrogen bonding interactions primarily determine the three-dimensional packing of the molecules, even though they make up a relatively small percentage of intermolecular interactions in the molecules. The three structures with the p-hydroxy group cis to the imine group give hydrogen-bonded zigzag chains in the monoclinic crystals, while the structure with a trans hydroxy group crystallize in a hexagonal space group (R3¯) and form hydrogen-bonded hexamers. The hexagonal structure also displays Br…Br interactions, forming additional hexameric clusters. The analysis of published p-hydroxyphenyl imine crystal structures from the Cambridge Crystallographic Database revealed patterns in the length of the hydrogen bonding interactions based on steric congestion around the hydroxyl group.
... However, the existence of flavonoids in some mushroom species is uncertain, because these species lack the chalcone isomerase enzymes necessary for the biosynthesis of flavonoids [67]. Despite not being the primary component of edible and medicinal mushrooms, phenolic compounds have been shown to exhibit a number of advantageous properties, including antioxidant [68,69] and enzyme inhibition [70], which appears to be closely associated with the anti-inflammatory qualities of mushrooms [71]. ...
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A fruiting body of a basidiomycete fungus was discovered growing on chopped Ficus nitida tree trunks in the student housing on the Assiut University campus during the course of this inquiry and a normal collecting operation in the Assiut Governorate, Egypt. Following the growth of the basidioma’s inner tissue on PDA, fungal mycelial growth was achieved. Internal transcribed spacer region (ITS) sequencing has allowed for the identification of the fungus as Tomophagus colossus. On the dry weight basis, chemical analysis of T. colossus AUMC 14536 basidioma revealed that it contains 28.81% carbohydrates, 25.34% crude fats, 23.44% crude fibers, 20.64% crude proteins, and 3.02% ash, in addition to potassium, phosphorus, calcium, selenium, iron, and zinc (133.59, 114.46, 6.27, 3.08, 1.28, and 0.73 mg/100 g dry weight, respectively). The total phenolic compounds (39.26 mg/g) and total flavonoids (5.62 mg/g) were also evaluated. The basidioma extract’s antioxidant activity was assessed as %DPPH radical scavenging activity with an IC50 of 4.15 µg/mL compared with a 1.89 µg/mL IC50 of ascorbic acid. In solid-state fermentation (SSF), the fungus could ferment broad bean straw, palm leaf hay, rice husk, rice straw, sugarcane bagasse, and wheat bran to produce endoglucanase, exoglucanase, laccase, pectinase, and xylanase in substantial amounts. Specific activity exhibited the highest values for endoglucanase (81.48 U/mg), exoglucanase (114.35 U/mg), pectinase (81.94 U/mg), and xylanase (70.18 U/mg) on the rice husk, while the peak of laccase activity (94.27 U/mg) was gained on bean straw. This is the first assessment of the organism’s nutritional value, amino acid content, antioxidant activity, and enzymatic capabilities in Egypt.
Article
The bacterial glycosyltransferase YjiC1 was used to glycosylate triterpenoids from the medicinal fungus Antrodia camphorata . Eleven new compounds were obtained from enzymatic reactions. Glucosylation could increase the inhibitory activities against...
Article
Syringaldehyde (SD), a kind of flavonoid polyphenolic small molecule compound, has the antioxidant and anti-inflammatory properties. But it is unknown whether SD has properties on the treatment of rheumatoid arthritis (RA) by modulating dendritic cells (DCs). We explored the effect of SD on the maturation of DCs in vitro and in vivo. The results showed that SD significantly down-regulated the expression of CD86, CD40 and MHC II, decreased the secretion of TNF-α, IL-6, IL-12p40 and IL-23, and increased IL-10 secretion and antigen phagocytosis in vitro induced by lipopolysaccharides in a dose-dependent manner through reducing the activation of MAPK/NF-κB signaling pathways. SD also significantly inhibited the expression of CD86, CD40 and MHC II on DCs in vivo. Moreover, SD suppressed the expression of CCR7 and the in vivo migration of DCs. In arthritis mouse models induced by λ-carrageenan and complete Freund's adjuvant, SD significantly alleviated paw and joint oedema, reduced the levels of pro-inflammatory cytokines TNF-α and IL-6 and increased the level of IL-10 in serum. Interestingly, SD significantly decreased the numbers of type I helper T cells (Th1), Th2, Th17 and Th17/Th1-like (CD4+IFN-γ+IL-17A+), but increased the numbers of regulatory T cells (Tregs) in spleens of mice. Importantly, the numbers of CD11c+IL-23+ and CD11c+IL-6+ cells were negatively correlated with the numbers of Th17 and Th17/Th1-like. These results suggested that SD ameliorated mouse arthritis through inhibiting the differentiation of Th1, Th17 and Th17/Th1-like and promoting the generation of Tregs via regulation of DC maturation.
Article
Effects of 3, 5 - dimethoxy - 4 - hydroxybenzoic acid and 2, 3, 4 - trihydroxyacetophenone were studied on haemoglobin S (Hb S) polymerisation, analgesia and inflammation using Hb S solution, rats and mice. UV spectrophotometric procedure was used to monitor the polymerization of the Hb S. Acetic acid induced writhing in mice and egg albumin induced rat paw edema procedures were used to evaluate analgesic and anti-inflammatory activities of the compounds respectively. The results indicate that both drugs inhibit the process of polymerization significantly, possibly by direct action on the Hb S molecules. The drugs inhibited acetic acid induced pain and decreased egg albumin induced oedema. It is concluded that 3, 5 - dimethoxy - 4 - hydroxybenzoic acid and 2, 3, 4 - trihydroxyacetophenone may have some value in the management of sickle cell disease.
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We previously reported that oroxylin A, a polyphenolic compound, was a potent inhibitor of lipopolysaccharide (LPS)-induced expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). In the present study, three oroxylin A structurally related polyphenols isolated from the Chinese herb Huang Qui, namely baicalin, baicalein, and wogonin, were examined for their effects on LPS-induced nitric oxide (NO) production and iNOS and COX-2 gene expressions in RAW 264.7 macrophages. The results indicated that these three polyphenolic compounds inhibited LPS-induced NO production in a concentration-dependent manner without a notable cytotoxic effect on these cells. The decrease in NO production was in parallel with the inhibition by these polyphenolic compounds of LPS-induced iNOS gene expression. However, these three compounds did not directly affect iNOS enzyme activity. In addition, wogonin, but not baicalin or baicalein, inhibited LPS-induced prostaglandin E2 (PGE2) production and COX-2 gene expression without affecting COX-2 enzyme activity. Furthermore, N-nitro-l-arginine (NLA) and N-nitro-l-arginine methyl ester (L-NAME) pretreatment enhanced LPS-induced iNOS (but not COX-2) protein expression, which was inhibited by these three polyphenolic compounds. Wogonin, but not baicalin or baicalein, similarly inhibited PGE2 production and COX-2 protein expression in NLA/LPS or L-NAME/LPS-co-treated RAW 264.7 cells. These results indicated that co-treatment with NOS inhibitors and polyphenolic compounds such as wogonin effectively blocks acute production of NO and, at the same time, inhibits expression of iNOS and COX-2 genes.
Article
Four new iridoid glucosides 1–4, named blumeosides A–D, were isolated from the methanolic stem-bark extract of Fagraea blumei G. DON. (Loganiaceae). They were accompanied by the benzyl-alcohol derivative di-O-methylcrenatin (5) and the flavone C-glucoside swertisin (6). The structures of 1–4 were established by spectroscopic methods, including FAB-MS, and 1H- and 13C-NMR, and by alkaline hydrolysis. Blumeosides A (1) and C (3) are 10-O-(2,5-dihydroxytercphthalo) adoxosidic acid and 10-O-(2-hydroxyterephthalo)adoxosidic acid, respectively. In blumeosides B (4) and D (2), both carboxylic groups of the terephthalic-acid moiety are esterified by adoxosidic-acid units, Blumeosides A–D (1–4) inhibited bleaching of crocin induced by alkoxyl radicals. Blumeosides A (1) and D (2) also demonstrated scavenging properties towards the 2,2-diphenyl-1-picryl-hvdrazvl (CDPPH) radical in TLC autographic and spectrophotometric assays.
Article
This database was created and is administered as a cooperative effort between the US Forest Products Laboratory and the US Dairy Forage Research Center. It was designed to provide a coherent, single source of NMR data of lignin model compounds as well as compounds modeling similar structures in grasses and other forage plants. The database exists in four different formats: an interactive HyperCard© stack for the Macintosh® computer, a FileMaker Pro© database for cross-platform use, an Adobe© pdf cross-platform file for viewing and printing, and a hardcopy version derived from the FileMaker Pro database. The first three versions are available for downloading over the internet from the Dairy Forage Research Center web site: http://www.dfrc.ars.usda.gov The hardcopy is available by request from the authors at the Forest Products Laboratory. The use of trade or firm names in this publication is for reader information and does not imply endorsement by the US Dept. of Agriculture of any product or service. In general 13 C NMR data was collected in three common deuterated solvents (acetone, chloroform and dimethyl sulfoxide) for each compound. The 1 H NMR data was reported for one solvent. A standard set of acquisition parameters was used to acquire and process the spectra to keep the data as uniform and constant as possible. Those compounds with an index number less than 1000 were run on a Bruker 250 MHz spectrometer at FPL and those compounds with an index number between 1000 and 10,000 were run at the DFRC on a Bruker 360 MHz instrument. The order of the compounds in the database reflects their arrival at the spectrometer rather than a preordained plan. Search routines for the software versions allow grouping the compounds with similar traits, whereas the structure index is most useful for the hardcopy version. The inclusion of many analogous series of structures with small structural differences allows calculation of substituent effects that are invaluable for chemical shift predictions of structures not included in the database. The chemical shift assignments for most of the compounds were made by comparison with other compounds, literature values and in some cases other NMR experiments such as long and short range C-H correlations, COSY and DEPT. Every effort was made to correctly assign the chemical shifts; however, limited time and resources precluded confirming the shifts for many of the compounds. The shifts are reported to the second decimal place only to distinguish very close shifts however comparisons between spectra are practical only within ± 0.1 ppm. The authors would greatly appreciate any corrections on misassignments.