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Gun MachinePaperback
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- Publisher: Little, Brown & Company
- Format: Paperback | 352 pages
- Dimensions: 137mm x 206mm x 25mm | 272g
- Publication date: 14 January 2014
- Sales rank: 788,289
Product description
The bestselling thriller from "a seriously good writer with a seriously wicked imagination" (New York Times Book Review). After a shootout claims the life of his partner in a condemned tenement building on Pearl Street, Detective John Tallow unwittingly stumbles across an apartment stacked high with guns. When examined, each weapon leads to a different, previously unsolved murder. Confronted with the sudden emergence of hundreds of unsolved homicides, Tallow discovers that he's walked into a veritable deal with the devil. An unholy bargain that has made possible the rise of some of Manhattan's most prominent captains of industry. A hunter who performs his deadly acts as a sacrifice to the old gods of Manhattan and who may, quite simply, be the most prolific murderer in New York City's history.
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Author information
Warren Ellis is the award-winning creator of graphic novels such as "Fell," "Ministry of Space," "Planetary," and "Transmetropolitan" and the author of the novel "Crooked Little Vein." His graphic novel "RED" was adapted into the #1 hit film of the same name starring Bruce Willis and Helen Mirren. He lives in London.
Review quote
"From the wrenching violence of its first pages to its bone-jarring conclusion, "Gun Machine" never lets go of the reader and never flags in its relentless pace. In the course of 300 tightly wound pages, Ellis unloads a full clip of ideas, black humor, character, and copper-sheathed action scenes. Every sentence is a bullseye." "Joe Hill, author of A Heart-Shaped Box and Horns""


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Samsung Hires Steve 'Cyanogen' Kondik 177
from the get-the-best dept.
We've improved Slashdot's video section; now you can view our video interviews, product close-ups and site visits with all the usual Slashdot options to comment, share, etc. No more walled garden! It's a work in progress -- we hope you'll check it out (Learn more about the recent updates).
This is just another move on the chess board. Google bought Motorola which will invariably give Motorola the inside track on Android to some extent. Samsung realized that while their hardware has been quite good their software has been severely lacking in both quality and updates. This hire makes perfect sense, it allows them to produce higher quality software (the goal being to improve upon Google's not just dress it up pretty) with a better update policy. If they actually allow their software to be run like CM has been (and force it through the providers) then it puts LG and HTC in poor positions long term.
You are false data.


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Free Canby Raspberry starts.
If anyone wants some starts I have plenty. Just come and dig em up and they are yours. I'd normally dig them up for you, but I am moving half of my patch and still have to get my garden in.
send me a message at
spyfferoni at yahoo dot com.
how do i prune the canby raspberries. most things i read say only prune 2 year old canes but someone said prune them all clear down to the ground and that they bear on this years canes. what is correct? how do i prune and when
I'm not sure of the type of raspberry that I have (was here when we moved in, label says "Raspberry") but here's my limited experience.
The plant was probably planted right before we moved in, and there was very limited growth the first year. The second year, I hardly remember seeing it. It didn't grow much, maybe two or three canes and it did not produce fruit. This year, there are 20+ starts in probably a 1x1.5 foot square area. Although I noticed some canes that were completely bare (no leaves, etc) I am guessing that they were from prior year's growth, and I've read that the canes don't live after they bear fruit (2 years). I've notice that there are maybe 3 or 4 blossoms where fruit is now developing (either incredibly small or early in development). My guess would be that they are from last year's growth. The new starts have no blooms at this point.
The only thing that you would do right now (June) is to cut out the dead canes (no leaves) or any damaged, diseased, or weak looking canes. You may also want to thin them out, if they are too thick, it will limit air flow, sunlight, and cause problems like disease or weak growth.
The point you made about pruning to the ground is one approach I've read about for ever bearing varieties. That type can be cut within two inches of the ground in the fall and just produce a summer harvest instead of a light spring harvest and a summer harvest. I think Canby is a mid-season bearer, so I wouldn't do that in your situation.
After you pick fruit in summer/fall, cut the canes that you picked from. I've read that older canes appear more "woody", but again, this is something to do in the fall.
My problem is that our current raspberry plants are overwhelming a grape vine, so I'm looking to transplant everything to a different location, although that should have happened already this year, I'm hoping that the dramatic increase in starts this year means the roots are good and strong. Besides, it doesn't look like much fruit is coming this year, so a move now will help get the plants established for next year's crop.
My big question is about the type of barrier to put into the new bed to contain the starts and keep them out of the lawn as much as possible. I've read that 8-12 inches down is a good distance, but what material should be used? Anyone have ideas or experience?
I've been growing Canby raspberries for the past six years, and taking care of them is pretty simple if you just follow these rules:
1. After you collect the fruit (which is usually late June or early July), cut all the canes that bore fruit down to the ground.
2. All around should be younger, greener canes that started growing this year. Leave them alone, and they will bear fruit next year in the summer.
If you have a new planting, just let them grow until you see some canes start fruiting, which would probably be in the summer the year after you plant them.
These are great raspberries, partly because the canes are nearly thornless (there's just a tiny bit of prickliness on very young canes). They also taste great. These are also called Bear Lake Raspberries.
There are other varieties of raspberry that fruit twice a year -- once in the spring on canes from last year, I think, and then in the fall on new canes (which will also bear in the following spring). I haven't tried them, but if you have one of those, the pruning requirements are different.
I think that spring would be the best time to plant raspberries, or maybe fall, but the small starts come up in the Spring. I'm sure I'll have a bunch in the Spring and I could send some down with my husband, we live in Springville, but he works in Salt Lake.
Stevation---How long do your Canbys bear fruit? Mine are about done. I've had problems the last couple of years with cholorsis. They'll start out great, and as soon as they start bearing fruit they get the yellowing and such. I called the home extension here and talked with someone who recommended adding coco fiber which has a ph of 5 to my raspberry beds in addition to compost and iron/iron sulfate.
I also need to thin out my patch. I am going to try moving a
large portion of the patch and try adding the amendments to the soil to see if I get better luck.
Something funny that my 3 yr old daughter said when I told her we were going to make some jam, "Don't squish em, just eat em!"
Hey Tyff,
I started harvesting on June 20th, and it got better and better for a few weeks after that. It's been tapering off for the past week or so, and they're pretty much done now. We picked a small bowl of them tonight, and there will still be a few small pickings left this week, but most of what remains on the canes is drying up now.
Last year, I noticed some chlorosis on mine, and I sprayed the leaves with an iron solution after the fruiting was all done. I haven't seen it happening again this year, but it might start again soon.
I have noticed one improvement in my patch this year -- I installed a strong rotating shrub head sprinkler on a three foot riser a few months ago, so they're getting more water than in the past. My new shoots this year are really getting tall already and they're much thicker than last year. So, I think we're headed for an even better crop next summer with all this vigorous new growth!
One more thing -- I was at a friend's house today and he had me pick some of his raspberries, but I was surprised how bland the flavor was! Maybe I'm spoiled with these Canbys, but mine sure have a lot of sweetness in them. These are great berries!


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Joints are the places in your body where bones meet. You have big joints - like the ones in your hips, shoulders, and legs; and itty, bitty ones - like the ones in your hands and feet.
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Particle Physics Informs the Ultimate Questions
October 1st, 2014 | 4
Editor's Note: Author and Fermilab Senior Scientist Don Lincoln is set to teach "Mysteries of the Universe" from October 13 – 24 for Scientific American's Professional Learning Program. We recently talked with Dr. Lincoln about why he became a physicist and his motivations to share what he discovers. When I was a young boy, I [...]Keep reading »
U.S. Particle Physics Program Aims for the Future
November 25th, 2014 | 1
In the last few years, stories have abounded in the press of the successes of the Large Hadron Collider, most notably the discovery of the Higgs boson. This has led some to speculate that European research is ascendant while U.S. research is falling behind. While there is no argument that U.S. particle physics budgets have [...]Keep reading »
Particles and the People Who Love Them: Documentary Shows Human Side of Large Hadron Collider
October 3rd, 2013 | 4
Full disclosure: I cried at a movie about particle physics. And I wasn't alone. As the film showed footage of the July 4, 2012 announcement of the Higgs boson discovery, I noticed the woman next to me wiping her eyes just as I was doing the same. I was at a screening of the new [...]Keep reading »
5 Sigma—What's That?
July 17th, 2012 | 10
Chances are, you heard this month about the discovery of a tiny fundamental physics particle that may be the long-sought Higgs boson. The phrase five-sigma was tossed about by scientists to describe the strength of the discovery. So, what does five-sigma mean? In short, five-sigma corresponds to a p-value, or probability, of 3×10-7, or about [...]Keep reading »
What It Means to Find "a Higgs": Lindau Nobel Laureate Meeting, Day 3
July 4th, 2012 | 5
Felicitas Pauss, head of international relations at CERN in Geneva, asked for a show of hands from the audience of young scientists: Who worked on the ATLAS or CMS instruments at CERN's Large Hadron Collider, or LHC? Many hands went up for each. And who worked as a theorist? More hands appeared—hundreds in all. Last, [...]Keep reading »
The Higgs, Sterile Neutrinos and Spintronics: Lindau Nobel Laureate Meeting, Day 2
With excitement building about an announcement due tomorrow from scientists working at the Large Hadron Collider, today's Lindau Nobel Laureate Meeting talks kicked off with the Higgs, explored some mysterious anomalies with neutrinos and looked forward to some practical applications of spintronics coming soon in information and communication technologies. (You can read all our coverage [...]Keep reading »
What Happens If We Find the Higgs Particle–or If We Don't?
April 20th, 2012 | 11
With instruments offering "tantalizing hints" in support of the Higgs boson, the elementary particle thought to endow matter with mass, we stand at a singular moment in time for physics. Will we get sufficient evidence to confirm the existence of the Higgs, thus helping to complete the decades-old Standard Model? Will science have to go [...]Keep reading »
Physics Hasn't Looked This Hot Since The Big Bang
August 16th, 2013 | 3
The ATLAS detector at CERN is overwhelming to mere mortals like myself. It's one of four detectors along the Large Hadron Collider designed to detect the most fundamental particles in our universe. It sits in a cave 92 meters below ground, is over 45 meters long and weighs a mere 7000 metric tons (that's equivalent [...]Keep reading »


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Page 1
The role of sphingosine 1-phosphate in the TNF-α induction of
IL-8 gene expression in lung epithelial cells
Hemakumar Chandru, Vijayakumar Boggaram⁎
Department of Molecular Biology, University of Texas Health Center at Tyler, 11937 US Highway 271 Tyler, TX 75708-3154, United States
Received 24 August 2006; received in revised form 13 November 2006; accepted 11 December 2006
Available online 22 December 2006
Received by A.J. van Wijnen
Tumor necrosis factor-α (TNF-α) is an important cytokine involved in the pathogenesis of inflammatory diseases of the lung. Interleukin-8
(IL-8), a C-X-C chemokine, is induced by TNF-α and initiates injury by acting as a chemoattractant for neutrophils and other immune cells.
Although sphingolipids such as ceramide and sphingosine 1-phosphate (S1-P) have been shown to serve as signaling molecules in the TNF-α
inflammatory response, their role in the TNF-α induction of IL-8 gene expression in lung epithelial cells is not known. We investigated the role of
sphingolipids in the TNF-α induction of IL-8 gene expression in H441 lung epithelial cells. We found that TNF-α induced IL-8 mRNA levels by
increasing gene transcription, and the stability of IL-8 mRNA was not affected. Exogenous S1-P but not ceramide or sphingosine increased IL-8
mRNA levels and IL-8 secretion. Dimethylsphingosine, an inhibitor of sphingosine kinase, partially inhibited TNF-α induction of IL-8 mRNA
levels indicating the importance of intracellular increases in S1-P in the IL-8 induction. S1-P induction of IL-8 mRNA was due to an increase in
gene transcription, and the stability of IL-8 mRNAwas not affected. S1-P induction of IL-8 mRNAwas associated with an increase in the binding
activity of AP-1 but the activities of NF-κB and NF IL-6 were unchanged. S1-P induced the phosphorylation of ERK, p38 and JNK MAPKs.
Pharmacological inhibitors of ERK and p38 but not JNK partly inhibited S1-P induction of IL-8 mRNA levels. These data show that increases in
the intracellular S1-P partly mediate TNF-α induction of IL-8 gene expression in H441 lung epithelial cells via ERK and p38 MAPK signaling
pathways and increased AP-1 DNA binding.
© 2006 Elsevier B.V. All rights reserved.
Keywords: Sphingolipids; Transcription; Lung injury; Inflammation
1. Introduction
by maintaining alveolar stability through secretion of surfactant,
clearance of inhaled particulates by mucociliary action, and
facilitates phagocytosis of pathogens by secretory immunoglob-
ulin and surfactant production. Additionally the respiratory
epithelium plays important roles in the control of various aspects
of inflammation such as initiation, amplification and down
regulation as well as tissue repair through the elaboration of
various mediators including cytokines (Standiford et al., 1990;
necrosis factor-α (TNF-α), a cytokine expressed predominantly
by the alveolar macrophages has been implicated in the
pathophysiology of inflammatory diseases of the lung resulting
Gene 391 (2007) 150–160
Abbreviations: IL-8, interleukin-8; TNF-α, tumor necrosis factor-alpha;
ARDS, acute respiratory distress syndrome; PAF, platelet activating factor; SSC,
0.15 M NaCl/0.015 M Na3·citrate pH7.6; ELISA, enzyme-linked immunosorbent
assay; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; EMSA, electropho-
retic mobility shift assay; DMS, N,N-dimethyl-D-erythro-sphingosine; DTT,
dithiothreitol; MAP kinase, mitogen activated protein kinase; MAPP, D-erythro-
2-(N-myristoylamino)-1-phenol-1-propanol; MOPS, [3-(N-morpholino) propane-
sulfonic acid]; NOE, N-oleoyl-ethanolamine; S1-P, sphingosine 1-phosphate;
SiRNA, small inhibitor RNA; SP, surfactant protein; NF-κB, nuclear factor
kappaB; AP-1, activator protein 1; NF IL-6, nuclear factor IL-6; CCT-α, CTP:
phosphocholine cytidyltransferase.
⁎Corresponding author. Tel.: +1 903 877 7780.
E-mail address: email@example.com (V. Boggaram).
0378-1119/$ - see front matter © 2006 Elsevier B.V. All rights reserved.
Page 2
(Tracey et al., 1988; Piguet, 1990; Suter et al., 1992). TNF-α
levels are markedly elevated in the bronchoalveolar lavage fluid
from patients with acute respiratory distress syndrome (ARDS)
(Suter et al., 1992). Elevated TNF-α levels are associated
with increased IL-8 levels, and TNF-α is a major inducer of
IL-8 expression in lung epithelial cells (Standiford et al., 1990).
IL-8, a member of the C-X-C family of chemokines is a potent
immune cells (Kunkel et al., 1995). Neutralizing IL-8 antibodies
prevented lung injury in animal models of lung disease,
indicating that IL-8 is an important mediator of lung injury
(Broaddus et al., 1994; Modelska et al., 1999).
The sphingolipid ceramide has long been recognized as a
signaling molecule in the inflammatory response. It acts as a
a host of stress agents including TNF-α (Kim et al., 1991),
ionizing radiation (Haimovitz-Friedman et al., 1994) and platelet
activating factor (PAF) (Goggel et al., 2004). Ceramide can be
produced via de novo synthesis or hydrolysis of membrane
sphingomyelin by sphingomyelinase enzymes. TNF-α increases
intracellular levels of ceramide by activating lysosomal and
plasma membrane-associated sphingomyelinases that display
different pH and metal requirements. There is accumulating
pathophysiology of acute lung injury. Lung cells express high
levelsofsphingolipidenzymes andsphingolipidmetabolites,and
sphingolipid levels are elevated in acute lung injury. Intratracheal
administration of TNF-α and ceramide into rats increases lung
permeability and decreases surface tension lowering effects of
surfactant (Ryan et al., 2003). In other studies, platelet activating
2004). PAF induced edema was suppressed in acid sphingomye-
linase deficient animals and was reduced by the non-specific acid
acid sphingomyelinase-mediated production of ceramide is
responsible for the edema (Goggel et al., 2004). Ceramide once
produced can be metabolized further into sphingosine and S1-P
by the sequential actions of ceramidase and sphingosine kinase
in diverse cellular activities such as cell growth and survival, cell
motility and immunity (Spiegel and Milstien, 2003).
Although TNF-α induction of IL-8 gene expression in lung
epithelial cells has been studied previously, information is
lacking on the role of sphingolipid metabolites in the induction
process. Considering that the respiratory epithelium plays key
roles in the initiation and amplification of inflammatory
responses and that sphingolipid metabolites are recognized as
important signaling molecules in the inflammatory response, we
sought to understand the roles of sphingomyelin metabolites in
the TNF-α induction of IL-8 mRNA. In this investigation, we
studied the effects of TNF-α and sphingolipids on IL-8 gene
expression in lung H441 cells, a cell line with characteristics of
bronchiolar (Clara) epithelial cells. We found that TNF-α
induced IL-8 mRNA levels by increasing gene transcription
without altering IL-8 mRNA stability. Of the sphingolipid
metabolites tested, S1-P but not ceramide or sphingosine in-
duced IL-8 mRNA and protein levels. TNF-α induction of
IL-8 mRNA levels was partly reduced by dimethylsphingo-
sine, an inhibitor of sphingosine kinase, indicating the impor-
tance of elevated levels of intracellular S1-P in the induction.
Consistent with the effects of dimethylsphingosine, SiRNA-
mediated inhibition of sphingosine kinase partially blocked
TNF-α induction of IL-8 mRNA. S1-P induction of IL-8mRNA
was associated with an increase in AP-1 DNA binding activity,
however NF-κB binding activity was unchanged. S1-P in-
inhibitors, SB203580 and PD98059 indicating the involve-
ment of ERK and p38 MAPKs in the S1-P regulation of
IL-8 expression.
2. Methods
2.1. Cell culture
NCI-H441 cells [American Type Culture Collection (ATCC)
HTB-174], a human lung adenocarcinoma cell line of
bronchiolar (Clara) cell lineage were grown on plastic tissue
culture dishes in RPMI 1640 medium containing 10% fetal
bovine serum, penicillin (100 unit/ml), streptomycin (100 μg/
ml) and amphotericin B (0.25 μg/ml). In all experiments the
medium was changed to RPMI 1640 without serum for 24 h
before the start of the experiment. In experiments to test the
effects of S1-P on IL-8 expression, BSA (0.4% w/v) was added
to the culture medium.
2.2. Materials
TNF-α was purchased from R & D Sytems (Minneapolis,
MN). TRI Reagent was from Molecular Research Center (Cin-
cinnati, OH). Actinomycin D, bovine serum albumin [low endo-
toxin (≤0.1 ng/ml) and fatty acid-free], Staphylococcus aureus
sphigomyelinase and sphingosine were from Sigma (St. Louis,
MO). 5,6-dichloro-1-b-D-ribofuranozyl-benzimidazole (DRB)
was from Calbiochem. C2-ceramide, sphingosine 1-phosphate
and N,N-Dimethyl-D-erythro-sphingosine (DMS) were obtained
from Avanti (Alabaster, AL). S1-P was dissolved in a mixture
of methanol–water (95:5) at 0.5 mg/ml by heating at 45 °C for
10–15 min followed by sonication for 10 s each time for three
times. Solubilized S1-P was dried under nitrogen and recon-
stituted in cell culture medium containing 0.4% bovine serum
2.3. RNA isolation and Northern blot analysis
Experimental procedures for total RNA isolation and
Northern blotting analysis are as described previously (Bog-
garam and Margana, 1994). Cytosolic RNA was isolated
according to published protocol (Greenberg and Bender,
1997). IL-8 and GAPDH RNA bands were quantified with a
PhosphorImager using Quantity One Image Acquisition and
Analysis Software (Bio-Rad) and IL-8 mRNA levels were
normalized to 18S rRNA levels to correct for variations in the
quantification, loading and transfer of RNA. The expression of
151H. Chandru, V. Boggaram / Gene 391 (2007) 150–160
Page 3
GAPDH mRNA was assessed as an internal control. Plasmids
encoding human IL-8 cDNA were kindly provided by Drs.
Edmund Miller and Usha Pendurthi, University of Texas Health
Center at Tyler, Tyler, TX.
2.4. Determination of IL-8
IL-8 levels in cell medium were determined by enzyme-
linked immunosorbent assay (ELISA) using a matched antibody
pair according to the manufacturer's protocol (R & D Systems,
Minneapolis, MN).
2.5. Isolation of nuclei and nuclear run-on transcription assay
Methods for the isolation of nuclei and run-on transcription
assay are as described previously (Greenberg and Bender,
1997; Boggaram and Margana, 1994). Total RNA from labeled
nuclei was isolated and equal amounts of radioactive RNAs
(10–30×106counts/min) were hybridized to nitrocellulose
membranes containing immobilized plasmid DNAs containing
human IL-8 and GAPDH cDNAs and pBluescript. After wash-
ing, radioactivity bound to the filters was quantified with a
PhosphorImager. Radioactivity bound to pBluescript was con-
sidered as background.
2.6. Plasmid DNA isolation
Luciferase reporter plasmids containing −546/+44 and
−133/+44 bp sequences of human IL-8 gene were kindly pro-
vided by Dr. Naofumi Mukaida, Cancer Research Institute,
Kanazawa University, Kanazawa, Japan. Plasmid DNAs were
amplified in Escherichia coli top 10 strain (Invitrogen,
Carlsbad, CA) and purified by anion exchange chromatography
using QIA filter plasmid purification kit (Qiagen, Valencia,
2.7. Transient transfection and reporter gene assay
Plasmid DNAs were transiently transfected into cells by lipo-
some-mediated DNA transfer with Lipofectamine 2000 (Invitro-
gen) according to the manufacturer's instructions. β-galactosidase
and luciferase reporter activities in cell extracts were measured by
chemiluminescence assays (Tropix, Bedford, MA and Promega,
Madison, WI).
2.8. Transfection of SiRNA oligonucleotides
SiRNA duplex oligonucleotides (siGENOME SMARTpool
reagent) targeting human sphingosine kinase (Human SPHK1)
and non-targeting SiRNA duplex oligonucleotides were pur-
SMARTpool SiRNA oligonucleotides comprise of four SiRNAs
negative control having no perfect matches to known human or
mouse genes. H441 cells (30–50% confluent) plated on T25
oligonucleotide and 20 μl Oligofectamine (Invitrogen) per dish
according to the manufacturer's protocol. Transfected cells were
cultured for 72 h in order to achieve maximum silencing effects
and then subjected to treatments.
2.9. Preparation of nuclear extracts and electrophoretic
mobility shift analysis (EMSA)
Nuclear extracts were prepared according to the methods
described previously (Schreiber et al., 1989; Singh and
Aggarwal, 1995). Protein concentrations of nuclear extracts
were determined by Bradford's method using Bio-Rad protein
assay reagent.
Synthetic oligonucleotides were annealed by heating equi-
molar concentrations of sense and antisense oligonucleotides in
10 mM Tris–HCl, pH 7.5 containing 10 mM MgCl2and 50 mM
NaCl at 95 °C for 5 min and then allowed to cool to room
temperature over a period of 1 h. The sense strand sequences of
the human IL-8 promoter oligonucleotides (binding sites are
underlined) used in EMSA are as follows:
NF-κB: 5′-AATCGTGGAATTTCCTCTGA-3′
(−84/−65 bp)
(−133/−114 bp)
(−94/−76 bp).
Double stranded oligonucleotides were 5′end labeled using
[γ32P] and T4 polynucleotide kinase. EMSAs were performed
as described previously (Berhane and Boggaram, 2001) by
incubating 0.5–1.0 ng (100,000 cpm) of the labeled oligonu-
cleotide with 5 μg of nuclear protein in 20 μl of binding buffer
[13 mM Hepes, pH 7.9 containing 13% glycerol, 80 mM KCl,
as non-specific competitor DNA] at 30 °C for 20 min. After
electrophoresis, the gel was dried and exposed to an X-ray film.
2.10. Immunoblotting analysis
Cells were rinsed twice with cold phosphate buffered saline
and incubated in lysis buffer (50 mM Tris–Cl, pH 7.4 containing
150 mM NaCl, 1 mM EDTA, 1% Triton X-100, 1 mM sodium
vanadate, 2.5 mM sodium pyrophosphate, 2 μg/ml leupeptin and
aprotinin, 1 mM PMSF and 15% glycerol) for 15 min on ice. The
celllysatewasclearedbycentrifugationat14,000rpmfor 10min
and the supernatant was used for Western immunoblotting
analysis. SDS-PAGE separation and transfer of proteins to
membrane were carried out with an XCell II Mini-Cell apparatus
(Novex, San Diego, CA) according to the manufacturer's
instructions. Equal amounts of cellular protein (10 μg) were
separated by SDS-PAGE on 10% Bis-Tris gels with MOPS
running buffer and electrophoretically transferred to PVDF
membranes. Membranes were successively incubated with rabbit
polyclonal antibodies against phosphorylated and total p38, p44/
42 and JNK MAPKs (Cell Signaling, Beverly, MA) at 1:1000
dilution overnight at 4 °C followed by goat anti-rabbit alkaline
phosphatase conjugated-secondary antibody (Cell Signaling) at
152 H. Chandru, V. Boggaram / Gene 391 (2007) 150–160
Page 4
1:2000 dilution for 1 h at room temperature. Protein bands were
visualized by the Enhanced Chemifluorescence (ECF) detection
method (Amersham Pharmacia Biotech, Piscataway, NJ) accord-
ing to the manufacturer's instructions. Membranes were scanned
witha fluorescencescanner for visualizationofproteinbandsand
the intensity of the bands was quantified using Quantity One
Image Acquisition and Analysis Software (Bio-Rad).
2.11. Statistics
Data are shown as means±SD/SE. In experiments where IL-
8 mRNA levels in control cells were arbitrarily set at 100%,
statistical significance was analyzed by one-sample t-test. For
other samples, unpaired t-test was used to analyze statistical
significance. One-tailed P values of b0.05 were considered
3. Results
3.1. TNF-α increases IL-8 mRNA in H441 lung epithelial cells
in a time-dependent manner
H441 cells are human lung epithelial cells that possess many
of the characteristics of Clara cells such as morphologic and
ultrastructural features as well as expression of lung-specific
a cell line representingthe distal lung epithelium.We studied the
We found that IL-8 mRNA was barely detectable in untreated
of incubation (Fig. 1A and B). Elevated IL-8 mRNA levels in
TNF-α-treated cells were sustained for extended periods of
time. The levels of GAPDH mRNAwere unaffected by TNF-α
(Fig. 1A).
3.2. TNF-α increases IL-8 gene transcription rate in H441 cells
TNF-α induction of IL-8 gene expression is subject to
regulation at the transcriptional and mRNA stabilization levels.
To determine the roles of transcriptional and posttranscriptional
mechanisms, we analyzed the effects of TNF-α on IL-8 gene
transcription rate and IL-8 mRNA levels by transcription run-on
assay and Northern blotting, respectively. We found that in cells
treated with TNF-α IL-8 transcription increased after 0.5 h and
declined thereafter (Fig. 2A and C, dot # 1). Northern
blot analysis of cytosolic RNA isolated from the same cells
showed that, in agreement with our earlier data, the steady state
IL-8 mRNA levels increased to maximum levels after 1 h of
treatment (Fig. 2B). TNF-α induction of IL-8 gene transcription
(∼10-fold relative to control at 1 h) was similar to the increase
of cytosolic IL-8 mRNA levels (∼10-fold relative to control at
1 h) indicating that the inductive effects of TNF-α are exerted
Fig. 1. TNF-α induces IL-8 mRNA levels in H441 lung epithelial cells. IL-8 and
GAPDH mRNA levels in H441 cells incubated in control (C) or TNF-α
containing medium (25 ng/ml) for the indicated periods of time were determined
by Northern blotting. A. A representative autoradiogram showing the effects of
TNF-α on IL-8, GAPDH mRNA levels. 18S rRNA levels visualized by
ethidium bromide staining are also shown. B. IL-8 mRNA levels in control cells
were arbitrarily set at 100, and the levels in treated cells relative to control cells
are shown. Data are means±SD of 2–3 independent experiments.⁎Pb0.05 and
⁎⁎Pb0.01 for TNF-α-treated samples compared to control.
Fig. 2. TNF-α increases IL-8 gene transcription rate. IL-8 and GAPDH
transcription rates and cytosolic IL-8 mRNA levels in H441 cells incubated in
control (C) or TNF-α (25 ng/ml) containing medium were determined by
transcription run-on assay and Northern blotting, respectively. A. A represen-
tative autoradiogram showing the effects of TNF-α on IL-8 and GAPDH
transcription rates. 1, IL-8; 2, GAPDH; 3, pBluescript; 4, blank. B. A
representative autoradiogram of the effects of TNF-α on cytosolic IL-8 mRNA
levels. C. Radioactivity bound to pBluescript was considered as background and
IL-8 gene transcription rate was normalized to GAPDH transcription rate. Data
shown are means±SD of 2 independent experiments.
153H. Chandru, V. Boggaram / Gene 391 (2007) 150–160
Page 5
primarily at the transcriptional level. GAPDH transcription was
unaffected by TNF-α (Fig. 2A, dot # 2). We further evaluated the
effect of TNF-α on the posttranscriptional regulation of IL-8
mRNA by determining its effect on the stability of IL-8 mRNA.
control cells and was unaffected by TNF-α indicating that TNF-α
obtained when 5,6-dichlororibofuranosylbenzimidazole (DRB)
was used as a transcriptional inhibitor (data not shown). The
stability of GAPDH mRNA was not affected by TNF-α. Taken
together the data of transcription run-on assays and the RNA half-
life experiments showed that TNF-α increases IL-8 mRNA levels
in H441 cells primarily by increasing gene transcription.
In agreement with transcription run-on experiments, TNF-α
increased luciferase reportergene expression from IL-8 minimal
promoter (−133/+44 bp) in transient transfection assays (data
not shown). TNF-α induction of IL-8 mRNA was associated
with an induction of NF-κB DNA binding activity, however
AP-1 binding activity was not changed (data not shown).
3.3. Sphingomyelin metabolite(s) mediate TNF-α induction of
IL-8 mRNA levels
Sphingomyelin metabolites have been shown to play
important roles in mediating the inflammatory responses in
the lung. To determine the role of sphingomyelin metabolites in
the regulation of IL-8 gene expression, we investigated the
effects of exogenously added sphingomyelinase, ceramide,
sphingosine and S1-P on IL-8 mRNA and IL-8 secretion in
H441 cells. As S1-P was reconstituted in serum-free medium
containing 0.4% BSA, we determined the effects of 0.4% BSA
on IL-8 mRNA and IL-8 secretion and found that BSA did not
alter IL-8 expression (data not shown). Results (Fig. 4A)
showed that at 1 h incubation, sphingomyelinase (1 unit/ml) and
S1-P (1 μM) increased IL-8 mRNA levels by ∼3.5- and 4.8-
fold, respectively, compared to control whereas ceramide
(10 μM) and sphingosine (10 μM) did not cause any increase.
Preliminary experiments to test the effects of different
concentrations of ceramide (1–100 μM) did not show any
inductive effects on IL-8 mRNA levels after 1 h incubation. As
expected,TNF-α was highly effective inincreasing IL-8 mRNA
levels. Measurement of IL-8 levels by ELISA in the medium
cells treated with ceramide was also found (control=4.64±
0.5 pg/mg protein, ceramide=8.15±1 pg/mg protein) (Fig. 4B).
A time point of 6 h was chosen to allow sufficient time for
changes in protein synthesis and secretion to occur as S1-P
maximally stimulates IL-8 mRNA levels at 1 h of incubation.
The inductive effect of S1-P on IL-8 secretion was significantly
greater than its effect on IL-8 mRNA levels. H441 cells were
Fig. 3. TNF-α does not alter the stability of IL-8 mRNA. H441 cells were
incubated in control (♦) or TNF-α (25 ng/ml) (▪) containing medium for 1 h
and then incubation continued in the presence of actinomycin D (5 μM) for the
indicated times. The levels of IL-8 and GAPDH mRNAs were determined by
Northern blotting. A. A representative autoradiogram showing IL-8, GAPDH
and 18 S rRNA bands. B. IL-8 mRNA level at 0 h was arbitrarily set at 100 and
levels at other times are shown relative to 0 h. Data are means±SD of 2
independent experiments.
Fig. 4. Effects of sphingomyelinase and sphingolipids on IL-8 mRNA and
proteinlevels. IL-8 mRNA and IL-8 levels in mediumin H441 cells incubatedin
control medium (C) or medium containing TNF-α (25 ng/ml), sphingomyeli-
nase (Smase) (1 unit/ml), C2-ceramide (10 μM), sphingosine (10 μM) or S1-P
(1 μM) were determined by Northern blotting and ELISA, respectively. Cells
were incubated for 1 and 6 h for Northern blotting (A) and ELISA (B),
respectively. Data are means±SEM of 3–4 independent experiments. Effects on
IL-8 mRNA:⁎⁎⁎Pb0.001 for TNF-α-treated cells vs control;⁎Pb0.05 for
Smase and S1-P-treated cells vs control. Effects on IL-8 protein:⁎Pb0.05 for
ceramide and S1-P-treated cells vs control;⁎⁎⁎Pb0.001 for TNF-α-treated cells
vs control.
154H. Chandru, V. Boggaram / Gene 391 (2007) 150–160
Page 6
treated with ceramidase inhibitors, D-erythro-2-(N-myristoyla-
mino)-1-phenol-1-prop (MAPP) and N-oleoyl-ethanolamine
(NOE), for 1–24 h to increase the intracellular accumulation
of ceramide, and their effects on IL-8 mRNA levels were
determined. Results (Fig. 5) showed that treatment of H441
cells with ceramidase inhibitors for 1–24 h did not increase
IL-8 mRNA levels consistent with the lack of inductive effects
of exogenously added ceramide.
The inductive effect of S1-P on IL-8 mRNA levels was time-
dependent with a maximum effect at 1 h of incubation (Fig. 6).
The inductive effects of S1-P were concentration-dependent with
the compound at 0.5 and 1 μM causing maximum induction
after 1 h of treatment (data not shown). S1-P similarly increased
type II epithelial cells (Sparkman, L. and Boggaram, V., unpub-
lishedobservations).Todetermine ifsphingomyelinaseinduction
of IL-8 gene expression is due to the generation of intracellular
S1-P or not, the effects of inhibition of sphingosine kinase by N,
N-dimethylsphingosine (DMS) on the sphingomyelinase induc-
tion of IL-8 mRNAwere determined. Results showed that DMS
of IL-8 mRNA (control=1, sphingomyelinase=9.78±0.80,
DMS (10 μM)+sphingomyelinase=7.49±0.55, DMS (20 μM)
+sphingomyelinase=4.47±0.46, mean±SEM, n=3) indicating
the involvement of S1-P in the induction.
We further assessed the role of S1-P in the TNF-α induction
of IL-8 mRNA expression using DMS, an inhibitor of
sphingosine kinase. DMS reduced TNF-α induction of IL-
8 mRNA levels in a dose-dependent manner indicating that
intracellular increase in S1-P levels contributes to the TNF-α
induction of IL-8 mRNA levels (Fig. 7A and B). To further
prove that TNF-α induced increase in the intracellular S1-P is
involved in the induction of IL-8 mRNA, we used RNA-
mediated interference to reduce sphingosine kinase expression
and studied its effects on TNF-α induction of IL-8 mRNA
expression. RNA-mediated interference results in gene-specific
silencing providing superior advantage over chemical inhibi-
tors. SiRNA duplex oligonucleotides targeting sphingosine
kinase were transfected into H441 cells and the effects of TNF-
α on IL-8 mRNA induction was determined. Results (Fig. 7C
and D) showed that in agreement with the results of DMS,
SiRNA inhibition of sphingosine kinase partially reduced TNF-
α induction of IL-8 mRNA. The effects of silencing were
apparent in cells exposed to TNF-α for 1 h (data not shown) and
more pronounced after 24 h of treatment. We were unable to
detect sphingosine kinase levels by Western blotting in control
or SiRNA transfected cells.
3.4. S1-P induction of IL-8 gene expression is regulated at the
transcriptional level
Our data showed that S1-P induced IL-8 mRNA levels and
IL-8 secretion in H441 cells. To understand molecular mecha-
nisms that mediate S1-P induction of IL-8 mRNA levels, we
determined its effects on IL-8 gene transcription and mRNA
transcriptionrate andmRNA stabilityshowedthatS1-Pincreased
IL-8 gene transcription rate (Fig. 8A, dot # 1) and IL-8 promoter
(Fig. 8B) indicating that the inductive effects are mediated
primarily at the transcriptional level. GAPDH transcription was
not significantly increased by S1-P (Fig. 8A, dot # 2).
3.5. Effects of S1-P on NF-κB and AP-1 DNA binding activities
Our data showed that TNF-α-induced intracellular S1-P
levels play important roles in the induction of IL-8 mRNA. Our
data also showed that S1-P increased IL-8 mRNA levels by
increasing gene transcription. As NF-κB and AP-1 transcription
Fig. 5. Effects of ceramidase inhibitors on IL-8 mRNA levels. IL-8 mRNA
levels in H441 cells incubated in control medium (C) or medium containing
ceramidase inhibitors D-erythro-2-(N-myristoylamino)-1-phenol-1-propanol
(MAPP) (5 μM) or N-oleoyl-ethanolamine (NOE) (5 μM) for the indicated
times were determined by Northern blotting. Data are means±SD of 2
independent experiments.
Fig. 6. S1-P induces IL-8 mRNA levels. A. IL-8 mRNA levels in H441 cells
incubated in control medium (C) or medium containing S1-P for the indicated
times were determined by Northern blotting. B. Data are means±SEM of 3
independent experiments.⁎⁎Pb0.01 for the 30 min sample comparedto control,
⁎Pb0.05 for the 1 h sample compared to control,⁎⁎Pb0.01 for the 2 h sample
compared to control.
155H. Chandru, V. Boggaram / Gene 391 (2007) 150–160
Page 7
factors play key roles in the induction of IL-8 expression, we
were interested to determine the effects of S1-P on NF-κB
and AP-1 DNA binding activities. Results (Fig. 9) showed that
NF-κB DNA binding activity was undetectable in control cells
(lane 2) and exposure of cells to S1-P (lane 3) did not increase
NF-κB DNA binding activity, however AP-1 DNA binding
activity was modestly increased (lanes 4 and 5) and NF IL-6
binding activity remained unchanged (lanes 6 and 7). Similar
results were obtained when cells were incubated with S1-P for
different periods of time ranging from 10 min to 6 h (data not
3.6. Involvement of MAPKs in the S1-P induction of IL-8
Sphingolipids are known to activate MAPK signaling path-
ways in a variety of cells. We studied the involvement of MAPK
signaling pathways in the S1-P induction of IL-8 mRNA
expression. S1-P stimulated the phosphorylation of p38, ERK
and JNK MAPKs in a time-dependent manner in H441 cells;
enhanced phosphorylation was observed as early as 10 min after
exposure to S1-P and leveled off to control levels after 2 h
(Fig. 10A). The stimulatory effect of S1-P on p46 form of JNK
that MAPK signaling pathways may be involved in the S1-P
induction of IL-8 mRNA expression. We ascertained the
involvement of MAPKs in the S1-P induction of IL-8 mRNA
by determining the effects of pharmacological inhibitors of
MAPKs on S1-P induction of IL-8 mRNA. ERK, p38 and JNK
MAPKs were inhibited with PD98059, SB203580 and JNK II
inhibitor respectively and the effects of S1-P on IL-8 mRNA
levels by approximately 50 and 40%, respectively indicating the
involvement of p38 and P44/42 MAPK signaling pathways. In
contrast JNK II inhibitor alone modestly increased IL-8 mRNA
levels and additively enhanced S1-P induction of IL-8 mRNA.
4. Discussion
IL-8 gene expression is induced by a wide variety of agents
including cytokines, growth factors, bacterial and viral products,
oxidants and others (Roebuck, 1999). Induction of IL-8 gene
expression is subject to both transcriptional and posttranscrip-
tional regulation in a cell/tissue- and stimulus-specific manner
(Roebuck, 1999; Hoffmann et al., 2002). We found that in H441
cells TNF-α induced IL-8 mRNA levels primarily by increasing
to activate IL-8 promoter activity via recruitment of NF-κB
to a TNF-α response element consistent with a role for tran-
scriptional mechanisms (Brasier et al., 1998) in the induction of
IL-8 gene expression in lung epithelial cells. A relatively short
sequence of DNA spanning −133/ +41 bp is necessary and
sufficient for the basal and TNF-α induction of IL-8 promoter
Fig. 7. Inhibition of sphingosine kinase reduces TNF-α induction of IL-8 mRNA levels. A. The effects of dimethylsphingosine (DMS), an inhibitor of sphingosine
DMS (40 μM)+TNF-α compared to TNF-α alone,⁎⁎Pb0.01 for DMS (30 μM)+TNF-α compared to TNF-α alone,⁎⁎⁎Pb0.001 for DMS (50 μM)+TNF-α
compared to TNF-α alone. C. SiRNA duplex oligonucleotides for sphingosine kinase were transfected into H441 cells by liposome-mediated transfer with
Oligofectamine. After 72 h, cells were treated with medium±TNF-α (25 ng/ml) for 24 h and IL-8 and GAPDH mRNAs were analyzed by Northern blotting. 1 and 2,
non-targeting SiRNA; 3, sphingosine kinase SiRNA. D. Data are means±SD of 3 independent experiments.⁎Pb0.05 for TNF-α compared to control;⁎Pb0.05 for
sphingosine kinase SiRNA transfected cells compared to non-targeting SiRNA transfected cells.
156H. Chandru, V. Boggaram / Gene 391 (2007) 150–160
Page 8
activity (Yasumoto et al., 1992; Brasier et al., 1998). The
NF-κB and AP-1 that act independently and synergistically
to activate IL-8 promoter in response to stimulatory agents in a
cell type-specific manner [reviewed in Roebuck, 1999]. In this
study wefound that TNF-α and S1-P induced IL-8 expression by
increasing gene transcription and without altering the stability of
IL-8 mRNA. However, regulation of IL-8 mRNA stability has
different cells. Nitric oxide, lipopolysaccharide, adenovirus and
Shiga toxin increase IL-8 mRNA expression in lung epithelial
cells, THP cells, fibroblasts and A549 cells, respectively, by
increasing the stability of IL-8 mRNA (Leland Booth and
Boggaram, 2004). The half-life of IL-8 mRNA in untreated cells
was in the range of 0.5–2 h and increased by several fold
depending on the cell type and the stimulus.
Sphingomyelin metabolites are increasingly recognized as
important mediators of inflammation in the lung, and ceramide
has emerged as a putative lipid mediator in TNF-α signaling.
Despite the important roles that sphingomyelin metabolites play
in TNF-α signaling, little is known about their involvement in
the TNF-α induction of IL-8 gene expression in lung cells. Our
data showed that among the sphingomyelin metabolites, S1-P
but not ceramide or sphingosine induced IL-8 mRNA levels and
IL-8 secretion. Consistent with the lack of significant effects of
ceramide, inhibition of ceramidase to increase intracellular
ceramide levels did not increase IL-8 mRNA levels. We found
that the inductive effect of S1-P on IL-8 level in the medium
was significantly greater than its effect on IL-8 mRNA level.
Similarly we found that although ceramide did not increase
IL-8 mRNA level it caused a small increase in IL-8 level. The
observed discrepancy between IL-8 mRNA and IL-8 levels in
S1-P- and ceramide-treated cells point to possible translational
and/or posttranslational regulation of IL-8 expression. Togeth-
er our data indicated that elevated intracellular S1-P generated
as a result of activation of sphingosine kinase partly mediates
TNF-α induction of IL-8 gene expression in H441 cells.
sphingosine-phosphate lyase that catalyzes the irreversible
cleavage of S1-P (Reiss, 2004). Whether TNF-α regulates
sphingosine-phosphate lyase expression and/or activity to
modulate intracellular S1-P levels in H441 cells is not
known. In human umbilical vein endothelial cells (HUVEC)
TNF-α was found to induce the expression of E-selectin and
vascular adhesion molecule-1 (VCAM-1) via increased
generation of S1-P by the activation of sphingosine kinase
(Xia et al., 1998). Although TNF-α induced sphingomyelin
breakdown and ceramide generation, ceramide failed to mimic
the effects of TNF-α to induce E-selectin and VCAM-1
Changes in the levels of sphingolipid metabolites can occur
via coordinate activation of the entire cascade of sphingolipid
metabolizing enzymes as in the case of oxidized low density
Fig. 8. S1-P increases IL-8 gene transcription and promoter activity. A. Effect on
IL-8 gene transcription. IL-8 and GAPDH transcription rates in H441 cells
incubated in control medium (C) or medium containing S1-P (1 μM) for 1 h
were determined by transcription run-on assay. The data is representative of 2
independent experiments. B. Effect on IL-8 mRNA stability. H441 cells were
first incubated in control medium (C) or medium containing S1-P (1 μM)
(♦, control;▪, S1-P) for 1 h and then incubation continued in the presence of
actinomycinD (2.5μM) for the indicatedtimes. IL-8 and GAPDHmRNA levels
were determined by Northern blotting. Data are means±SD of 2 independent
experiments. C. Effect on IL-8 promoter activity. H441 cells were transiently
2 independent experiments.
Fig. 9. Effects of S1-P on NF-κB, AP-1 and NF IL-6 DNA binding activities.
H441 cells were incubated in control medium (C) or medium containing S1-P
(1 μM) for 1 h and nuclear extracts prepared. Binding activities of NF-κB, AP-1
and NF IL-6 to IL-8 promoter elements were analyzed by EMSA. The arrows
indicate the mobility of protein–DNA complexes. Ns, non-specific band.
157H. Chandru, V. Boggaram / Gene 391 (2007) 150–160
Page 9
lipoprotein induced mitogenesis of smooth muscle cells (Auge
et al., 1999) or via selective activation of one of the enzymes of
the pathway as in the case of TNF-α activation of sphingosine
kinase to inhibit apoptosis in HUVEC cells (Xia et al., 1999). In
some cells the activation of ceramidase may be so robust that
the levels of sphingosine and S1-P are vastly increased in the
absence of substantial increases in ceramide levels (Kolesnick,
2002). Our experiments showed that exogenous ceramide and
inhibition of ceramidase to increase intracellular ceramide levels
failed to increase IL-8 mRNA and protein levels suggesting that
the inability of ceramide to increase IL-8 may not be due to low
intracellular ceramide levels.
TNF-α activates sphingomyelin hydrolysis to increase
intracellular ceramide levels in lung (Ryan et al., 2003) and
lung cells (Vivekananda et al., 2001). Sphingolipids generated in
cytidyltransferase (CCTα) (Vivekananda et al., 2001) the rate-
an important component of lung surfactant, leading to the
perturbation of surfactant lipid composition. These findings
suggest that perturbations in surfactant lipid synthesis contribute
to lung injury associated with inflammation and that sphingoli-
pids play important roles in mediating these effects. Our findings
of the inductive effects of S1-P on IL-8 mRNA levels reveal yet
another pathway that potentially contributes to lung injury in
inflammation. Increases in IL-8 levels can lead to increased
recruitment of neutrophils into the lung contributing to lung
The molecular mechanisms and signal transduction pathways
by which S1-P induces the expression of IL-8 in H441 lung
epithelial cells remains to be investigated. Our studies showed
that S1-P increased AP-1 DNA binding activity in H441 cells
increase of IL-8 gene transcription. S1-P is known to enhance the
DNA binding activity of AP-1 (Su et al., 1994).
MAPKs regulate IL-8 expression and secretion in a variety
of cells including lung epithelial cells (Hashimoto et al., 1999;
Matsumoto et al., 1998) and MAPK regulation of IL-8 expres-
sion occurs via transcriptional and mRNA stabilization mechan-
isms (Holtmann et al., 1999). Transcription factors NF-κB and
AP-1 play central roles in the transcriptional regulation of IL-
in the regulation of IL-8 expression appears to be dependent
on the cell type and the nature of the stimulus. Induction of
IL-8 expression in BEAS-2B bronchial epithelial cells by
S1-P (Wang et al., 2002) and lysophosphatidic acid (Saatian
et al., 2006) required activation of p44/p42 whereas induction
by Streptococcus pneumoniae (Schmeck et al., 2006) and zinc
(Kim et al., 2006) involved activation of JNK and ERK
plus JNK respectively. Our studies showed that S1-P activated
p44/42, p38 and JNK phosphorylation in H441 cells, however,
pharmacological inhibitors of ERK and p38 but not JNK
MAPKs inhibited S1-P induction of IL-8 mRNA levels in-
dicating that ERK and p38 signaling pathways are required for
S1-P induction. S1-P increased AP-1 DNA binding activity
suggesting a role for AP-1 in the induction of IL-8 mRNA
expression. It remains to be determined if ERK and p38
MAPK pathways control AP-1 DNA binding activity to in-
crease IL-8 expression. It is known that p38, p44/p42 and JNK
MAPKs regulate AP-1 activity (Whitmarsh and Davis, 1996).
In summary, our studies have shown that TNF-α induces
IL-8 gene expression in H441 lung epithelial cells by increa-
sing gene transcription and that intracellular increases in S1-P
levels play important roles in mediating TNF-α induction.
S1-P induced IL-8 mRNA expression via activation of p38
and p44/42 MAPK signaling pathways and increase in AP-1
Fig. 10. A. Effects of S1-P on MAPK activation. H441 cells were serum-starved
for 24 h and then treated with S1-P (1 μM) for the indicated periods of time. The
levels of phosphorylated and total p38, p44/42, and JNK MAP kinases were
analyzed by Western immunoblotting. Similar results were obtained in 2 other
independent experiments. B and C. Effects of pharmacological inhibitors of
MAPK activation on S1-P induction of IL-8 mRNA levels. H441 cells were first
incubated in medium±inhibitor (SB 203850, 10 μM; PD98059, 50 μM; JNK
inhibitor II, 10 μM) for 1 h and then treated with S1-P (1 μM) for an additional
1 h. IL-8, and GAPDH mRNA levels were determined by Northern blotting
analysis. A representative Northern blot is shown. Similar results were obtained
in 2 other independent experiments.
158H. Chandru, V. Boggaram / Gene 391 (2007) 150–160
Page 10
DNA binding activity. Thus TNF-α induces IL-8 gene ex-
pression in H441 lung epithelial cells via two pathways —
one involving the activation of NF-κB and the other via
intracellular elevation of S1-P that results in an increase in
AP-1 but not NF-κB binding.
This work was supported partly by the National Heart, Lung,
and Blood Institute Grant HL48048. We thank Dr. Anna
Kurdowska for the measurement of IL-8 levels by ELISA.
Human IL-8 promoter plasmids were kindly provided by Dr.
Naofumi Mukaida, Kanazawa University, Kanazawa, Japan.
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Our family of 4, including 2 teenagers found this to be one of our fondest memories. When asked what to do in Italy we tell everyone to hire Gianni for a great trip. We had a great day seeing the surrounding area of Siena. Gianni is extremely knowledgeable and takes you to not only great sites but off the beaten path quaint villages. We had a fantastic lunch at a family run spot and a visit to a wonderful vineyard. We can't say enough about this tour, guide and service.
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The Relationship between Social Custom and English Idiom Ⅰ.Introduction Idioms as a special form of language that carry a large amount of cultural information, such as history, geography, religion, custom, thinking pattern and so on. Therefore, they are closely related to the culture, especially idioms have an inseparable relationship of social customs. From a dynamic view, language and culture interact with each other and shape each other. Idiomatic expressions are even difficult for native English speakers because their meanings cannot be determined through an analysis of their individual words. For instance, "to fall out" means "to quarrel", but neither "fall" nor "out" has a sense of quarrel. Another example is "a feather in one"s cap", which means "an honor or a success, of which one can be proud". The meaning of this idiom is a metaphorical one. "Under the rose" is also very difficult to understand from the literal meanings of its component words, its true meaning is "secretly". "To have a bee in one's bonnet" means "to have a strange fixed idea about something". "To kick the bucket" means "to die". The figurative interpretation of "shoot the breeze" is "to talk without purpose". Unlike literal language, it is impossible to explain the idiom from the literal meaning of the individual words. Thus, English idioms are difficult to understand, because they are a combination of two or more words, which function as a unit of meaning. For our Chinese students, English idioms are a stumbling block. It is very possible that they often not only feel at a loss in understanding conversations because of a key idiom but also misuse idioms. In this paper, I am going to make a study of the relationship between idioms and social custom cultures and throw some light on the learning of English idioms. As we all know, language is closely related to culture and can be said as a part of culture. They are the heritage of history and product of cultural evolvement. Consequently, we can know much about social custom culture through studying idioms and in turn get better understanding of idioms by learning the cultural background behind them. Ⅱ.The Definition of English Idioms To research idiom, first of all, we must understand what is idiom. This is the old theme that we discuss it. Because to define the idiom exactly and popular accepted definition is a difficult question. In the definition of English idioms, some scholar emphasis on the words quantity of structure in idioms. That is to say, how many language units to constitute the English idioms? Is it except the phrases, words group, and words or sentences also can make up to the idioms? Others emphasize the single meanings of English idioms, it refers that the English idiom"s meaning is arbitrary. The English idiom"s meaning can not be synthesized or cut apart. Different people hold the different opinions on the definition of English idioms; they have different focal point on the definition of English idioms. So we must understand the definition of English idioms exactly through the research. Although we are unlikely to give an 1 ideal definition to the English idioms; but we should give a better definition of English idioms before the research. In the book A Research on English Idiom, the author thinks before we decide what expression is idiom, we should consider the following three points: first, the English idiom should have two words to constitute at least; second, the structure of English idioms are relatively fixed, the word-formation of idioms can not be change; third, the meaning of English idioms can not be interpreted by the meaning of the words which constitute it. (骆世平,2006:13) It is not easily to define what English idiom is. On one hand, the scope, content, structure and formation of English idioms are very complex; on the other hand, what formations of language belong to the English idioms? How to classify the idioms from the narrow sense and broad sense? Different scholars hold the different opinions, so it brings the difficult to define the English idioms. The English idiom is a special kind of English language form which come from the phrases, words groups, structure or custom expressions, the meaning of English idiom can not be deduce from the single meaning of the constitute-words. (张安德,杨元刚,2003:14) From the above definition of English idiom, we can define that English idioms are some special language structure units—phrases, words groups and expressions, their meaning always can not be inferred from the literal meaning of the constitute-words of idioms, their meaning should be understood from the conventional form of whole idioms. From the broad sense, the English idioms include the phrases, common sayings, proverbs, idiomatic phrases, slang, a two-part allegorical saying and allusion and so on. The English idioms mainly include the set phrases and idiomatic phrases. For instance, "to have one"s head in clouds" means "to meet the difficult problems that can not think the way to solve it". Another example is "the man in the street", which means "normal people, citizen". "No man is born wise or learned" means "the man is learned from the studying but not learned from the inheritance". The idiomatic phrase of "to charge someone an arm and a leg" means "the price is very expensive". The allusion of "skeleton in the cupboard" means "the disgraceful affair of a person", and the slang of "slag off" means "to criticize". The definitions of English idioms are many in our society. In my opinion, the English idioms consist of set phrases and short sentences, which are peculiar to the language in question and steeped in the national and regional culture and ideas, thus being colorful, forcible and thought-provoking. Strictly speaking, idioms are expressions that are not readily understandable from their literal meanings of individual constituents. In a broad sense, idioms may include colloquialisms, catchphrases, slang expressions, proverbs and so on. Ⅲ.The Characters of English Idioms We have been discussed the definition of English idiom in the above; we know an English idiom has its different characters from other words and expressions. I think these characters can be sum up as single unit, institutionalization and irreplaceability. But this conclusion of English idioms is more abstract so that we can 2 not easily understand. From my view of the characters of English idioms, they mainly have two aspect characters: First of all, being phrases or sentences, idioms each consist of at least two or more constituents, but each is a semantic unity. Thought the various words which make up the idiom have their respective literal meanings, they have lost their individual identities in the idiom. That is to say, their meanings are not often recognizable in the meaning of the whole idiom. Likewise, the part of speech of each element is no longer important, for quite often the idiom functions as one word. For instance, "till the cows come down" which comprises a conjunction "till", an article "the", a noun "cow", a verb "come" and an adverb "home" means "forever" and function as an adverb, thus replaceable by the word "forever".(张维友,2004:228— 229) The semantic unity of idioms is also reflected in the illogical relationship between the literal meaning of each constituent and the meaning of the idiom. Many idioms are semantically inexplicable. For example, "how do you do" is a common form of greeting used by people when they first meet. Literally it means "in what way do you do things", which is far from its idiomatic usage. Second of all, the idioms have the character of structural stability. The structure of an idiom is to a large extent invariable, unlike free phrases. Firstly, the constituents of idioms cannot be changed or replaced. For example, "kick the bucket" can not be changed as "kick the pail" or "strike the bucket", it means "to die". Secondly, the word order cannot be inverted or changed. For instance, "by twos and threes" and "tit for tat" are not being turned into "by threes and twos" and "tat for tit". Thirdly, the constituents of an idiom cannot be deleted or added to, not even an article. Take the "out of the question" for example, it means "impossible". If the article "the" is deleted, the idiomaticity will be lost and it will signify "no question" instead. Finally, "diamond cut diamond" (two parties are equally matched) is grammatically incorrect, for normally the verb "cut" should take the third person singular "-s" as the subject diamond is singular. (张维友,2004:229—230) Ⅳ.The Social Customs in English Idioms We have been discussed the definition and characters of English idioms. Now, we discuss the relationship between social customs and English idioms. We know the English idioms carry large amount cultural information such as history, geography, religion, custom, nationality psychology, thinking pattern and so on. They are closely related to the culture and social customs. But how are they related to the social customs? In what way are they related? This is the question we discuss. As far as I am concerned, the English idioms are mainly related to the religious belief, Greek and Roman mythologies, social fashions, food customs, literary masterpieces, ancient fables and so on. A. The Relationship between Religious Beliefs and English Idioms. American and Britain are the countries that have Christianity and Hebraism as their cultural background. Most idioms produced in this background carry rich 3 religious elements. Getting to the roots of idioms, we can find that they mainly originated from the Bible. It is firm concept of western people that God created people. They think that God is omnipotent and sacred. God, Christ, devil, Adam and some educational segments of the stories in the Bible are passed down because of people favored or even some historical and political causes. For example: Judas's kiss: a deceitful act of courtesy. (Judas betrayed his Master with a kiss.) (New Testament) Much cry and little wool: a proverbial saying expressive of contempt or derision for one who promises great things but never fulfils the promises. (Originally the proverb says,"Great cry and little wool, as the Devil said when he sheared the hogs"; and it appears in this form in the ancient mystery of David and Abigail, in which Nabal is represented as shearing his sleep, and the Devil imitates the act by "shearing a hog".) Turn the other cheek: to take no action against the person who has hurt or harmed one, especially to allow him to do it again. (Do not resist one who is evil. But if any one strikes you on the right cheek, turn to him the other also…)——(New Testament) The widow's cruse: a small supply of anything that, by good management, is made to go a long way and to be apparently inexhaustible. (New Testament) I listed four idioms from the Bible in which there are many idioms. So when we learn idioms about Bible, we can read Bible, which will increase our knowledge and help us to be familiar with the features of western culture and have a clear understanding of the hidden meanings of idioms. B. The Relationship between Greek and Roman Mythologies and English Idioms Mythologies are ancient stories that are based on popular beliefs or that explain natural or historical events because the people of the primitive society were scared of the nature and longed for the nice future. In the American and Britain, many idioms are originated from the mythologies; especially they come from the Greek and Roman mythologies, so that we can know some relationship between English idioms and mythologies. Midas touch: the excellent skill of making money. (Midas is a legendary king of Phrygia who requested of the Gods that everything he touched might be turned into gold. His request was granted, but as his food became gold the moment he touched it, he prayed the Gods to take their favor back. He was then ordered to bathe in the Pactolus, and the river ever after rolled over golden sands.) (张安德, 杨元刚, 2003: 21) Pandora's Box: a prolific source of troubles. (In Greek mythology a box containing all the evils of mankind and given by Zeus to the mythological Pandora, who opened it against the command of Zeus, thus, all the evils flew forth and they have ever since continued to afflict the world, only Hope remained in the bottom; something that produces many unforeseen difficulties) (骆世平,2006:42) Sphinx's riddle:means "the difficult riddle" (Sphinx is a monster of ancient mythology; The Grecian Sphinx was generally said to be a daughter of Typhon and 4 Chimaera; she infested Thebes, setting the inhabitants a riddle and devouring all those who could not solve it. The riddle was——What goes on four feet, on two feet, and three feet. But the more feet it goes on the weaker it be? and it was at length solved with the answer that it was a man, who as an infant crawls upon all-fours, in manhood goes erect on his two feet, and in old age supports his tottering legs with a staff. On hearing this correct answer the Sphinx killed herself and Thebes was delivered.) Here I only listed three examples of idioms from the Greek and Roman mythologies; we can see these idioms reflect the English culture. Except these idioms, there are many English idioms that originated from the Greek and Roman mythologies. C. The Relationship between Social Fashions and English Idioms The social fashion is traditional habit or is popular in a period of time in society; it concludes living habit, etiquette, tradition, behavior standards, and method of social communication and so on. Every person lives in the atmosphere of social fashion and traditional habit, and this atmosphere influences their daily life. Meanwhile, these influences reflect the language; especially they reflect the English idioms. For example: Let one's hair down: to relax after a period of tense. (Long before, in whatever occasion, the English woman"s hair were very clean and tidy that comb up in the forehead, only when they were alone which to let their hair down. Therefore, people use the idiom of "let one"s hair down" to express their relaxation after a long time of working and tense situation.) Take off one's hat to someone: pay one"s respect to someone. (Long ago, the English people wore the hat as a fashion. When they greeted with people in the social occasion and showed the respect to someone, they took off their hat. For this reason, "take off one"s hat to someone" extends the meaning to show the respect to someone.) Go down the aisle: get married, enter into matrimony. (The westerners almost believe the Christianity, their weddings always hold in the church. When the wedding starts, the father of the bride always companies with her walk down the aisle. Therefore, the idiom of "walk down the aisle" expresses "enter into matrimony". For this reason, the idiom "go down the aisle" is produced to indicate "get married".) (骆 世平,2006:49) The social fashions reflect the English idioms to the daily life of English and American. I only listed three idioms of the social fashions. When we learn the social fashions, we can understand the meaning of English idioms deeply. Meanwhile, there are many idioms that come from the social fashions. If we learn more social fashions about the English idioms, we can know the relationship between English idioms and social fashions behind the background of culture. D. The Relationship between Food Customs and English Idioms. On a large while, the food is the life of human being, the history of human being is mainly to struggle for food. Because of the differences of nature environment and geography place for each nation, they live in the different way. Even for the 5 people of one nation, they have different food customs because of their different living conditions. Therefore, nowadays, "cultural festival of food" is held in the everywhere of world. Because of the influences of sea to the Britain"s weather, the weather is not very cold or hot. So the wheat, barley and oats of main grain are adapted to this climate of Britain. The staple food of Britain people includes bread, milk, butter and cheese. If we are careful about these phenomenons, we can find many idioms of constituents including bread. For example: Bread and butter: livelihood, a way of earning money. Bread and circuses: food and amusement. Bread with someone: to have dinner with somebody. Bread line: the long queue of the unemployed people to receive the relief. Take the bread out of someone's mouth: grab somebody"s job. (骆世平, 2006:53) It is very clear that the pursuit of daily living materials for English people is mainly the bread. The bread is not very important in modern society, but the bread is the record or the reflection of history as for English idioms. The English people love to eat butter; they make the figure of speech according to the characters of butter. For instance: Butter would not melt in his mouth: very honest. Like a hot knife through butter: very easy to do something. Bread buttered on both sides: living a comfortable life. (骆世平, 2006: 53) Many idioms about the custom of food are similar to these idioms, such as: Cry in one's beer: forget worries by drinking. Big cheese: an important figure, boss. Out of a jam: live out of the desperate straits or get out of troubles. Save one's bacon: shake off one" difficulties. Live on the breadline: living in the condition of poverty line. Jam tomorrow: something you want but it does not belong to you. Cry over spilt milk: to waste time feeling sorry about something that cannot be changed for the better. Know which side your bread is buttered: to know what will be of most gain to yourself. (骆世平, 2006:53) E. The Relationship between Literary Masterpieces and English Idioms Many popular western literary masterpieces have the classic words, phrases and sentences that attract people greatly. They are shaped to evolve the hidden meaning. For instance: Screw up one's courage: to overcome one"s fear; to stop one from being afraid. Man Friday: a faithful and willing attendant, ready to turn his hand to anything. (The young savage found by Robinson Crusoe on a Friday, and kept as his servant and companion on the desert island.) (骆世平, 2006:39) To the manner born: is be born to do. (Hamlet) 6 One's pound of flesh: fair but unreasonable request. (A moneyless young Venetian, Bassanio, is making preparations to court Portia, a rich beauty of Belmont. He needs money and appeals to his friend Antonio, a merchant of Venice, for help. But Antonio"s money is invested in ships at sea. To aid Bassanio in his courtship, he borrows money from Shylock the Jewish usurer. Shylock, who has been insulted by Antonio and other Christians, agrees to lend the money on the condition that, if the loan is not paid in three months, he may cut a pound of flesh from Antonio"s body. (刘 炳善,1993:78)Later people use "one"s pound of flesh" to imply the fair but unreasonable request. The Merchant of Venice(骆世平, 2006:35) Cakes and ale: idle away one"s time in pleasure-seeking, enjoy pleasure. (Dost thou think, because thou art virtuous, there shall be no more cakes and ales) Twelfth Night (刘炳善,1993:81) (骆世平, 2006:36) Paint the lily: an unnecessary action. ( To gild refined gold, to paint the lily…Is it wasteful and ridiculous excess?) King John (骆世平, 2006:36) Man of mould: ordinary person. HenryⅤ Suffer a sea change: a person who experienced a remarkable change. The Tempest The world is someone's oyster: at one"s will, follow one"s inclinations. The Merry Wives of Windsor. Much ado about nothing: create trouble out of nothing. Much Ado About Nothing (骆世平, 2006:37) F. The Relationship between Ancient Fables and English Idioms Though ancient fable stories are short and refined, they teach people many philosophic and educational theories while being narrated. They are also parts of the kindergarten education of Chinese children. It is obvious that they are popular even among Chinese people. They, especially Aesop's Fables are the favorites of Chinese people. For example: Naked truth: the plain, unvarnished truth; the truth without trimmings. (The fable says that Truth and Falsehood went bathing; Falsehood came first out of the water, and dressed herself in Truth"s garments. Truth, unwilling to take those of Falsehood, went naked.) Sour grapes: disparage something that is beyond one"s reach. (The allusion is to Aesop"s well-known fable of the fox, which tried in vain to get at some grapes, but when he found they were beyond his reach, he went away saying, "I see they are sour.") (骆世平, 2006:44) Bell the cat: risk one"s own life to do something dangerous. (The allusion is to the fable of a cunning old mouse, who suggested that they should hang a bell on the cat"s neck to give notice to all mice of her approach.) Cat's paw: the tool of another, the medium of doing another"s dirty work. (The allusion is to the fable of the monkey who wanted to get some roasted chestnuts from the fire, and used the paw of his friend, the cat, for the purpose.) (骆世平, 2006: 44) The lion's share: the largest part; all or nearly all. (In Aesop's Fables, several 7 beasts joined the lion in a hunt; but, when the spoil was divided, the lion claimed one quarter in right of his prerogative, one for his superior courage, one for his dam and cubs, "and as for the fourth, let who will dispute it with me." Awed by his frown, the other beasts yielded and silently withdrew.) Ⅴ. Cultural Differences Reflected by Chinese and English Idioms Both China and Britain have a long history, which fertilized their brilliant cultures. The common experiences and reciprocal interaction in various aspects through long-time communication between the two countries induce a lot of common points in culture. In early British history, Britain was occupied by the Roman Empire for nearly four hundred years. The Romans left their deep marks on Britain culture. In 49 B.C. the Roman emperor Caesar sent the order of burning out boats after his army passed Lupigen River, showing the determination to take an all-out war. That is the origin of the English idiom "burn one"s boats". Incidentally, in 208 B.C. General XiangYu in China sent the similar order to smash all the boats after the army passed Yangtze River in attempt to show its determination of winning the war, which made the birth of the Chinese idiom "破釜沉舟".(李洪涛,2002:114) The two idioms are stunningly the same in form and meaning, which reflect the same point in culture — the same concept to show inexorable determination or irrevocable decision. What"s more, some similar points in culture are resulted from the reciprocal learning and affection in the course of communication between the two countries. For example, the Chinese idiom "以眼还眼, 以牙还牙" was borrowed from the English idiom.(李洪涛, 2002:38)The English idioms "to lose one"s face" and "to save one"s face" are borrowed from the Chinese ones.(李洪涛,2002:105) Although Chinese culture and English culture have much in common as showed above, they have more in discrepancy owing to their different geographical environment, history, religion and so on. The differentiation is a common phenomenon, which appears in varied forms, one of which is idiom. Then let me analyze four cultural divergences resulting in differences between Chinese idioms and English idioms. A. Geographical Differences Geographical environment plays an important role in shaping one country"s culture and human beings have no choices or capacities to change the geography. As a result, idioms based on geographical features appear different. Britain is an island country, which does not share land border with any other countries except the Republic of Ireland. In history, its navigation industry had ever been the first one for a long time. So, British people have a special passion for water. However, the most part of China belongs to inland place where people cannot live without earth. This very geographical difference results in different idioms in the two languages. For example, to show somebody being extravagant, there is a Chinese idiom "挥金如土", but in English the idiom is "spend money like water".(李洪涛, 2002: 2) Take the other instance, in southern China with mild and moisture weather, 8 bamboo is a familiar plant for people there, so they use the idiom "雨后春笋" to describe the new things cropping up in great numbers. Correspondingly, English has the idiom "spring up like mushrooms" to tell the same meaning, since in England mushroom in spring can be seen everywhere.(张培基,2002:11-12)Besides the different idioms expressing the same notion in the two languages, there are a lot of idioms related to water in English and earth in Chinese which have no corresponding counterpart in the other language, like the English idioms "to rest on one"s oars", "to keep one"s head above water", "all at sea" and etc.(张培基,1979:14-15) B. Conventional Differences One nation"s custom is formed through a long history and firmly rooted in people"s mind. It is closely related to national psychology and acts as one important source of idioms. In different countries and nations customs drastically differ in various aspects. Due to that, a lot of idioms turn out with varied dress. Take food habit for example, cake has been a common food in English for a long history, so it is understandable that they use "a piece of cake" denoting an easy-done task.(李洪涛,2002:31) However, in China, cake cannot be seen until the last one hundred years. So it is impossible for the same idiom to appear in Chinese. On the other hand, dishes are common in China. Therefore a similar idiom "小菜一碟" turns ( out in Chinese to express the same meaning. 李洪涛, 2002: 127) Another example is related to the habitual way of working in the field. Chinese people were habituated to use the bull to plough the field before modernized agriculture was developed, while English did with a horse. Consequently, Chinese say "力大如牛" to describe someone with great strength while English do "as strong as a horse" in alternation. C. Value Orientation Differences From people"s working and living experience of one country, different values have been abstracted, accumulated and handed down from generation to generation. Because of different experience between countries, values originated from it also turned out to be largely varied and even totally opposite, which idioms are built on, therefore also differ greatly. For instance, modesty is valued in Chinese culture almost since the beginning of Chinese history. There are a great number of Chinese idioms showing the virtue to be modest and shame to be proud, such as "骄必招辱"(张耀飞,2003:30) ,"骄兵必 败"(王福祯、赵友斌,2007:282) ,"谦虚谨慎"(邓乾德、江熙泰、郑玉章, 2004:344) ,"虚心使人进步, 骄傲使人落后"(张耀飞,2003:30)and etc. On the contrary, Britain as a developed capitalist country is infested with fierce social competition so that people there emphasize individual behaviors, achievement and value. Their value orientation in this aspect appears to be in contrast with Chinese. They have idioms showing the uselessness to be modest such as "where valor true is found, true modesty will there abound",(张耀飞,2003:30) "hide not your light under a bushel",(张耀飞,2003:31) "modest dogs miss much meat", "pride feels no colds"(王福祯、赵友斌,2007:281)and etc. 9 D. Religious Differences Religion exists in every county and nation and affects much in people"s belief and life, especially before the modern civilization. Numerous idioms were generated from the fountain of religion. The people in different countries mostly believe in different schools of religion. Therefore, some differences of idioms between Chinese and English can be traced back to the origin of religion. Religion played a key role in British society especially before capitalization. The majorities of the English people believe in Christianity and regard the Bible as one of the most important classical works in their life. The Bible is comprehensive of nearly all the matters in life so that it acts not only a classic in religion but also an important supporter of British culture, or even of western culture. Innumerable idioms are originated from the Bible, such as the idiom referring to the unwise act of forcing the extremely conservative person to accept new conceptions, "to put new wine in old bottles",(李洪涛,2002:36) the idiom describing thrilling horror caused by something, "to make one"s hair stand on end",(李洪涛,2002:114) another idiom denoting revenge in the same way, "an eye for an eye, a tooth for a tooth" and etc. (李洪涛,2002:38)Besides, the idiom "as poor as a church mouse" also is somewhat influenced by religion. In Christian church, there is no devoted food, which is not the same case in Chinese temples. Therefore, the mice in the church, which have no food, can be said as poor.(万辰秀,2006:156) Different from Britain, Chinese culture evolved from Confucian School, Taoist School. There are traces of Taoist principles and Confucian principles in Buddhist doctrines. Therefore, although Buddhist culture originated from the other country, it has been assimilated in Chinese culture through a thousand years of spreading and accepting Chinese feathers at the same time. It has been a part of Chinese culture and exerted enormous influence on Chinese traditional culture and present life as well. In language, a large number of Chinese idioms can be traced back to Buddhist School. For example, the idiom"道高一尺, 魔高一丈"is originally the warning in Buddhism to warn Buddhist disciples of the outside temptations, which remind the disciples of the hard prospect to overcome all the difficulties both inside and outside in order to make the slightest development.(张培基,1979:19)Now it is used to tell there is always at least one power stronger than the other. For another instance, "一尘不染" is also a Buddhist term. Buddhist School calls sex, sound, smell, flavor, touch and rule as "六尘" and Buddhist disciples should not be affected by the above sixes. Now this idiom evolved to describe a person who is pure and honest or a thing which is clean and in order.(邓乾德、江熙泰、郑玉章,2004:537) Ⅵ.Conclusion Idioms is a part of culture, it is reflect the cultural needs. Language is not neutral codes or grammatical rules. Culture and language are intertwined and are shaping each other. It is impossible to separate the two. Therefore, to know the cultural background will help us to understanding what information of the author want to tell us is; to know the cultural background will help us to know the true meaning of idiom, especially when we know the relationship of the social custom and idiom, we 10 can better understanding the hiding meaning of idiom. Each time we select words, from sentences, and send a message, either oral or written, we make cultural choices. We all agree that language helps us in communicating with people from different backgrounds. However, we may be less aware that cultural literacy is necessary in order to understand the language being used. If we select language without being aware of the cultural implications, we may at least not communicate well and at worst send the wrong message. Therefore, we know the social customs for the sake of understanding the hiding meaning of idiom. (Linda Beamer & Iris Varner, 2003: 31-32) Today, the type of figurative language is highly used in the daily life in Britain and America. It is necessary for us to have such knowledge of how to comprehend. The ability to understand idioms is an important way to successful communication. As we all know, English idioms have close relationship in social customs and the relationship reflects the English culture. The difficult comprehension of English idioms may also lead to communication breakdown for the persons from different language communities. Failure to grasp the meanings of idioms can impinge upon an individual"s understanding of language in social, academic, and vocational settings. In order to communicate with each other fluently, the ability to know the hiding meaning of idioms is important. We study of the relationship between English idiom and social culture so as to understand the hiding meaning of idiom. To know the relationship of social customs and English idioms can help us better to understand the true meaning of English idiom. 11 References 邓乾德、江熙泰、郑玉章. 汉英常用成语词典. 昆明:云南大学出版社,2004. 刘炳善. 英国文学简史. 郑州:河南人民出版社,1993. 李洪涛. 惊讶英语习语. 吉林:吉林科学技术出版社, 2002. 骆世平. 英语习语研究. 上海: 上海外语教育出版社, 2006. 万辰秀. 英语成语趣味读. 天津:天津科技翻译出版公司,2006. 王福祯、赵友斌. 英语谚语荟萃. 北京:中国国际广播出版社,2007. 张安德、杨元刚. 英汉词语文化对比. 武汉:湖北教育出版社,2003. 张培基. 习语汉译英研究. 北京:商务印书馆,1979. 张维友. 英语词汇学教程. 武昌:华中师范大学出版社, 2004. 张耀飞. 1000 句常用英语祝福话. 北京:中国物价出版社,2003. Hymnody and Bible House, Chinese-English Bible (Old and New Testament Union-International Version). USA: Hymnody and Bible House, 1990. Linda Beamer & Iris Varner. Intercultural Communication in the Global Workplace. Beijing: Tsinghua University Press, 2003. 12
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