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Meta-analyses of genome-wide association studies identify
multiple novel loci associated with pulmonary function
Dana B. Hancock1,29, Mark Eijgelsheim2,29, Jemma B. Wilk3,29, Sina A. Gharib4,5,29,
Laura R. Loehr1,7, Kristin D. Marciante5,6, Nora Franceschini7, Yannick M.T.A. van
Durme2,8, Ting-hsu Chen9,10, R. Graham Barr11,12,13, Matthew B. Schabath14, David J.
Couper15, Guy G. Brusselle2,8, Bruce M. Psaty5,6,16,17,18, Cornelia M. van Duijn2,
Jerome I. Rotter19, André G. Uitterlinden2,20, Albert Hofman2, Naresh M. Punjabi21,
Fernando Rivadeneira2,20, Alanna C. Morrison22, Paul L. Enright23, Kari E. North7,24,
Susan R. Heckbert6,16,17, Thomas Lumley25,30, Bruno H.Ch. Stricker2,20,26,27,30,
George T. O'Connor9,10,30, and Stephanie J. London1,28,30
1Epidemiology Branch, National Institute of Environmental Health Sciences, National Institutes of
Health, Department of Health and Human Services, Research Triangle Park, North Carolina
2Department of Epidemiology, Erasmus Medical Center, Rotterdam, The Netherlands
3Departments of Neurology and Medicine, Boston University School of Medicine, Boston,
Massachusetts 4Center for Lung Biology, University of Washington, Seattle, Washington
5Department of Medicine, University of Washington, Seattle, Washington 6Cardiovascular Health
Research Unit, University of Washington, Seattle, Washington 7Department of Epidemiology,
University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 8Department of
Respiratory Medicine, Ghent University and Ghent University Hospital, Ghent, Belgium
9Pulmonary Center, Department of Medicine, Boston University School of Medicine, Boston,
Massachusetts 10The National Heart, Lung, and Blood Institute's Framingham Heart Study,
Framingham, Massachusetts 11Division of General Medicine, Department of Medicine, College of
Physicians and Surgeons, Columbia University, New York, New York 12Division of Pulmonary,
Allergy and Critical Care, Department of Medicine, College of Physicians and Surgeons,
Columbia University, New York, New York 13Department of Epidemiology, Mailman School of
Public Health, Columbia University, New York, New York 14Risk Assessment, Detection, and
Intervention, H. Lee Moffitt Cancer Center and Research Institute, Tampa, Florida 15Department
of Biostatistics, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina
16Department of Epidemiology, University of Washington, Seattle, Washington 17Center for Health
Studies, Group Health, Seattle, Washington 18Department of Health Services, University of
Washington, Seattle, Washington 19Medical Genetics Institute, Cedars-Sinai Medical Center, Los
Angeles, California 20Department of Internal Medicine, Eramus Medical Center, Rotterdam, The
Correspondence should be addressed to S.J.L. (firstname.lastname@example.com).
29These authors contributed equally to this work
30These authors jointly directed this work.
Author Contributions
RS: M.E., Y.M.T.A.van.D., G.G.B., C.M.van.D., A.G.U., A.H., F.R., B.H.Ch.S.
Study design: T.L., B.H.Ch.S., G.T.O., S.J.L.; Data analysis: D.B.H., M.E., J.B.W., L.R.L., K.D.M., N.F., T.C.; Drafting of
manuscript: D.B.H., M.E., J.B.W., S.A.G. Critical revision of manuscript: D.B.H., M.E., J.B.W., S.A.G., L.R.L., K.D.M., N.F.,
Y.M.T.A.van.D., T.C., R.G.B., M.B.S., D.J.C., G.G.B., B.M.P., C.M.van.D., J.I.R., A.G.U., A.H., N.M.P., F.R., A.C.M. P.L.E.,
K.E.N., S.R.H., T.L., B.H.Ch.S., G.T.O., S.J.L.
The authors declare no competing financial interests.
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Published in final edited form as:
Nat Genet. 2010 January ; 42(1): 45–52. doi:10.1038/ng.500.
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Netherlands 21Departments of Medicine and Epidemiology, Johns Hopkins University, Baltimore,
Maryland 22Human Genetics Center, Division of Epidemiology and Disease Control, University of
Texas Health Science Center at Houston, Houston, Texas 23College of Public Health, University
of Arizona, Tucson, Arizona 24Carolina Center for Genome Sciences, University of North Carolina
at Chapel Hill, Chapel Hill, NC 25Department of Biostatistics, University of Washington, Seattle,
Washington 26Netherlands Genomics Initiative-sponsored Netherlands Consortium for Healthy
Aging, Rotterdam, The Netherlands 27Inspectorate of Health Care, The Hague, The Netherlands
28Laboratory of Respiratory Biology, National Institute of Environmental Health Sciences, National
Institutes of Health, Department of Health and Human Services, Research Triangle Park, North
Measurements of lung function by spirometry are heritable traits that reflect respiratory health and
predict morbidity and mortality. We meta-analyzed genome-wide association studies for two
clinically important measures, forced expiratory volume in the first second (FEV1) and its ratio to
forced vital capacity (FEV1/FVC), an indicator of airflow obstruction. This meta-analysis included
20,890 participants of European ancestry from four CHARGE consortium studies: Atherosclerosis
Risk in Communities (ARIC), Cardiovascular Health Study (CHS), Framingham Heart Study
(FHS), and Rotterdam Study (RS). We identified eight loci associated with FEV1/FVC (HHIP,
GPR126, ADAM19, AGER-PPT2, FAM13A, PTCH1, PID1, and HTR4) and one locus associated
with FEV1 (INTS12-GSTCD-NPNT) at or near genome-wide significance (P<5×10−8) in
CHARGE; all but 3 loci (FAM13A, PTCH1, and PID1) replicated with the SpiroMeta consortium.
Our findings of novel loci influencing pulmonary function may offer insights into chronic lung
disease pathogenesis.
Pulmonary function is an easily measurable and reliable index of the physiological state of
the lungs and airways1. Pulmonary function also predicts mortality in the general
population, even among never smokers with only modestly reduced pulmonary function and
without respiratory symptoms2,3. The peak level of pulmonary function attained in early
adulthood and its subsequent decline with age are likely influenced by genetic and
environmental factors. Tobacco smoking is a major environmental cause of accelerated
decline in pulmonary function with age. Other inhaled pollutants also appear to contribute.
Familial aggregation studies suggest a genetic contribution to lung function with heritability
estimates exceeding 40%4,5, but little is known about specific genetic factors involved. A
relatively uncommon deficiency of α1-antitrypsin is the only established genetic risk factor
for accelerated decline in pulmonary function and development of chronic obstructive
pulmonary disease (COPD), especially in smokers4,6. However, α1-antitrypsin accounts for
little of the population variability in pulmonary function4. Candidate gene studies suggest
that other genetic variants may influence the time course of pulmonary function and its
decline in relation to smoking, but these putative genetic risk factors remain unknown4.
Forced expiratory volume in the first second (FEV1) and its ratio to forced vital capacity
(FEV1/FVC) are two clinically relevant pulmonary function measures. While both FEV1 and
FVC are influenced by lung size and can be reduced by restrictive lung diseases, obstructive
lung disease leads to proportionately greater reduction in FEV1 than FVC. Therefore, a
reduced FEV1/FVC, an indicator of airflow obstruction that is independent of lung size, is
the primary criterion for defining an obstructive ventilatory defect1. Whereas low FEV1/
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FVC indicates the presence of airflow obstruction, FEV1 is used to classify severity and
follow the progression of obstructive lung disease over time5,7,8.
The first genome-wide association study (GWAS) for pulmonary function evaluating 70,987
single nucleotide polymorphisms (SNPs) in about 1,220 Framingham Heart Study (FHS)
participants revealed no genome-wide significant loci9. Recently, a GWAS of FEV1/FVC
using 2,540,223 SNPs in 7,691 FHS participants identified several chromosome 4q31 SNPs
near HHIP with genome-wide significance10. A GWAS of COPD11 also implicated the
HHIP region along with CHRNA3/5 on chromosome 15, previously associated with nicotine
We conducted meta-analyses of GWAS results for a cross-sectional analysis of pulmonary
function (FEV1/FVC and FEV1) in 20,890 individuals of European ancestry from four
Cohorts for Heart and Aging Research in Genomic Epidemiology (CHARGE) consortium14
studies: Atherosclerosis Risk in Communities (ARIC), Cardiovascular Health Study (CHS),
FHS, and Rotterdam Study (RS-I and RS-II). Given that cigarette smoking is a major risk
factor for pulmonary function decline, we conducted meta-analyses with adjustment for
smoking status and quantity, and in subgroups of ever and never smokers. Significant
findings and other selected high-signal hits were evaluated for replication with the
SpiroMeta consortium, an independent consortium having a combined sample size of 20,228
participants of European ancestry as described in the accompanying manuscript.
Meta-analyses of CHARGE genome-wide association results
Meta-analyses for FEV1/FVC and FEV1 were conducted using approximately 2,534,500
SNPs in 20,890 CHARGE participants of European ancestry (N=7,980 from ARIC,
N=3,140 from CHS, N=7,694 from FHS, N=1,224 from RS-I, and N=852 from RS-II) and
in subgroups of ever (N=11,963) and never smokers (N=8,927). Characteristics of the cohort
participants are presented in Table 1. We applied genomic control, although cohort-specific
genomic inflation factors (λgc) were low (for FEV1/FVC ranging from 1.00 (RS-I and RS-II)
to 1.05 (ARIC) and for FEV1 ranging from 1.01 (RS-II) to 1.05 (FHS)) suggesting minimal
population stratification. The meta-analysis λgc was 1.04 for FEV1/FVC and 1.03 for FEV1
in all participants. Quantile-quantile (Q-Q) plots show large deviations between observed
and expected P values for high-signal SNPs in analyses of FEV1/FVC and FEV1 in all
participants (Supplementary Fig. 1a,b), FEV1/FVC in never smokers (Supplementary Fig.
2a), and FEV1 in ever smokers (Supplementary Fig. 3c). Genome-wide significant
associations (P<5×10−8) were found for multiple SNPs in each of these analyses (Fig. 1a,b
for overall analyses and Supplementary Fig. 2b,d and Supplementary Fig. 3b,d for analyses
stratified by ever/never smoking). The top 2,000 SNPs associated with each measure, FEV1/
FVC and FEV1, beyond genome-wide significance (P>5×10−8) are presented in
Supplementary Table 1.
For FEV1/FVC, genome-wide significant associations were seen for 119 SNPs at seven loci
(Supplementary Table 2). The SNP with the smallest P value, rs1980057 (P=4.90×10−11), is
located on chromosome 4q31.22, 81 kb away from the 5'-end of HHIP. There were 27 other
genome-wide significant SNPs in the HHIP region (Fig. 2a). Additionally, 69 genome-wide
significant SNPs were located in or near the 3'-end of GPR126 on chromosome 6q24.1, with
the top SNP (rs3817928) having P=2.60×10−10 (Fig. 2b). Fifty-nine of these 69 GPR126
SNPs were associated with FEV1/FVC at genome-wide significance among never smokers
(Supplementary Table 2). Seven chromosome 5q33.3 SNPs located in ADAM19 (Fig. 2c),
two correlated chromosome 6p21.32 SNPs (r2=0.66, Fig. 2d) located in two genes (AGER
and PPT2), four chromosome 4q22.1 SNPs near the 5'-end of FAM13A (Fig. 2e), two
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chromosome 9q22.32 SNPs in PTCH1 (Fig. 2f), and six chromosome 2q36.3 SNPs near the
3'-end of PID1 (Fig. 2g) were also significantly associated with FEV1/FVC in all
participants. SNPs in AGER, PPT2, PTCH1, and PID1 had minor allele frequencies (MAFs)
between 4 and 10%, while all other significantly associated SNPs had MAFs exceeding
10%. Absolute β values (per-allele change in FEV1/FVC) ranged from 0.44 to 1.14%. The β
directions were consistent across the CHARGE cohorts for all genome-wide significant
SNPs except for the GPR126 SNPs noted in Supplementary Table 2. A borderline
significant association (P=5.37×10−8, MAF=0.42, β=−0.43) with FEV1/FVC was noted for
the chromosome 5q33.1 SNP rs11168048 in HTR4 (Fig. 2h). Cohort-specific association
results for SNPs with the smallest P value from each locus implicated at or near genome-
wide significance are shown in Supplementary Table 3.
For FEV1, genome-wide significant associations were observed for 46 chromosome 4q24
SNPs in or near four adjacent genes (Supplementary Table 4). The SNP with the smallest P
value, rs17331332 (P=4.00×10−10), is located near NPNT. The 45 other significantly
associated SNPs include four SNPs located near the 5'-end of NPNT, five SNPs located in
INTS12 or near its 3'-end, seven SNPs located in FLJ20184 or near its 3'-end, and 29 SNPs
located in GSTCD. FLJ20184 encodes a hypothetical protein according to several genome
browsers including the UCSC genome browser15, but there is no approved HUGO gene
name for this locus16. The SNP rs17331332 is correlated at r2>0.5 with most other
significantly associated SNPs in this region (Fig. 3), suggesting that the associations in the
four adjacent genes represent one independent finding. The significantly associated SNPs
had MAFs between 6 and 8%. The absolute β values (per-allele change in FEV1) ranged
from 55.92 to 71.43 mL (Supplementary Table 4), and the β directions were consistent
across the CHARGE cohorts for all 46 genome-wide significant SNPs (Supplementary
Table 3 for rs17331332). Among these 46 SNPs, 39 were associated with FEV1 at genome-
wide significance among ever smokers (Supplementary Table 4).
To evaluate whether other loci may also influence pulmonary function, we created Q-Q
plots for FEV1/FVC and FEV1 among all participants after removing SNPs (1,862 for FEV1/
FVC and 284 for FEV1) at or close to genome-wide significance and nearby SNPs
correlated at r2>0.2 with the top SNP for each locus. The resulting Q-Q plots show some
excess of small P values for FEV1/FVC (Supplementary Fig. 4a) and FEV1 (Supplementary
Fig. 4b).
Putative functional polymorphisms
Three SNPs among the 119 genome-wide significant SNPs for FEV1/FVC are non-
synonymous (missense) polymorphisms: rs11155242 (Lys to Gln) in GPR126, rs1422795
(Ser to Gly) in ADAM19, and rs2070600 (Gly to Ser) in AGER. The Polymorphism
Phenotyping (PolyPhen) program17 predicts that the amino acid substitutions resulting from
rs11155242 and rs1422795 cause benign changes but predicts that rs2070600 has a possibly
damaging impact on the structure and function of AGER.
All other SNPs implicated for FEV1/FVC or FEV1 are intergenic, intronic, or located in 3'
untranslated regions. Of these, three intronic GPR126 SNPs (rs9496346, rs1040525, and
rs6929442) and one intergenic SNP near NPNT (rs10516529) are located in transcription
factor binding sites, according to the UCSC genome browser15.
Replication with the SpiroMeta consortium
Thirty high-signal SNPs associated with FEV1/FVC (18 SNPs from eight loci) or FEV1 (12
SNPs from three loci) at or close to genome-wide significance were tested in the SpiroMeta
consortium. We evaluated these SNPs in 16,178 SpiroMeta participants of European
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ancestry with complete quantitative smoking data using the CHARGE analytic method,
which included adjustment for smoking status and pack-years, and performed joint meta-
analyses of CHARGE GWAS and SpiroMeta replication results (Table 2 and Table 3). P
values that exceeded the significance threshold in SpiroMeta (P<8.33×10−4 based on 60
tests) or the genome-wide significance threshold in joint meta-analyses (P<5×10−8) were
considered significant evidence for replication.
For FEV1/FVC, among 18 SNPs tested for replication, six SNPs in three loci were
significantly associated with this measure in SpiroMeta: rs1980057 and rs1032295 near
HHIP (r2=0.72), rs2070600 in AGER and rs10947233 in PPT2 (r2=0.66), and rs11168048
and rs7735184 in HTR4 (r2=0.93) (Table 2). Their joint meta-analysis P values ranged from
3.21×10−20 to 6.23×10−11 (Table 2). Five additional SNPs in GPR126 (rs3817928,
rs7776375, and rs6937121) and ADAM19 (rs2277027 and rs1422795) were not significantly
associated with FEV1/FVC at the stringent threshold in SpiroMeta, but these SNPs were
associated at genome-wide significance in the joint meta-analysis with P values ranging
from 9.93×10−11 to 1.25×10−8 (Table 2). For replicated SNPs, the allele frequencies and the
direction and magnitude of the associations with FEV1/FVC were similar between consortia
(Table 2). Further, the HHIP, ADAM19, and HTR4 SNPs were significantly associated with
FEV1 in SpiroMeta (Supplementary Table 5). The HHIP SNP rs1980057 and HTR4 SNPs
rs11168048 and rs7735184 were also associated with FEV1 at genome-wide significance in
the joint meta-analysis (P ranging from 5.86×10−9 to 1.58×10−8, Supplementary Table 5).
SNPs in FAM13A, PTCH1, and PID1 that gave genome-wide significance in CHARGE
were not confirmed in analyses with SpiroMeta.
For FEV1, among the 12 SNPs tested for replication, eight SNPs from one locus with four
adjacent genes were significantly associated with this measure in SpiroMeta, including
rs17331332 and rs17036341 near NPNT, rs11727189 and rs17036090 in or near INTS12,
rs17036052 and rs17035960 in or near FLJ20184, and rs11097901 and rs11728716 in
GSTCD (Table 3). For replicated SNPs, the allele frequencies and the direction and
magnitude of the associations with FEV1 were similar between consortia, and P values from
joint meta-analysis ranged from 4.66×10−17 to 9.42×10−14 (Table 2). None of these SNPs
were significantly associated with FEV1/FVC in CHARGE or SpiroMeta (Supplementary
Table 5).
Associations in individuals with normal pulmonary function
To address whether the genetic associations hold even among people with normal
pulmonary function, we repeated the meta-analyses after excluding individuals with asthma
or COPD, leaving 17,855 individuals (N=6,912 from ARIC, N=2,634 from CHS, N=6,371
from FHS, N=1,126 from RS-I, and N=812 from RS-II). Asthma was defined by self-report
of ever having asthma or self-report of ever having physician-diagnosed asthma. COPD was
defined spirometrically as having both FEV1/FVC and FEV1 less than the lower limit of
normal values using NHANES III prediction equations18,19. Comparing the original meta-
analyses to the meta-analyses with exclusions for asthma and COPD, β estimates were
highly correlated for the high-signal SNPs tested for replication (Pearson's r>0.99 for 18
FEV1/FVC SNPs and 12 FEV1 SNPs). β estimates remained highly correlated for SNPs with
P values as high as 0.01 in the original meta-analyses (r=0.92 for FEV1/FVC and r=0.96 for
FEV1). As expected, there was some attenuation in P values for many of the SNPs in our
implicated loci given the substantial power loss due to both reduced sample size and the
truncation of the FEV1/FVC and FEV1 distributions, but there was substantial overlap in the
top-ranking SNPs between the two meta-analyses (results not shown). The P values for
some top-ranking SNPs became smaller, including several ADAM19, FAM13A, and HTR4
SNPs associated with FEV1/FVC. Of note, 12 SNPs in HTR4, a locus with one SNP
rs11168048 showing borderline genome-wide significance in the original meta-analysis,
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gave genome-wide significance in the subset of individuals without asthma or COPD
(P=6.93×10−9 for rs11168048).
In meta-analyses of GWAS results in 20,890 CHARGE participants of European ancestry,
we identified genome-wide significant associations with FEV1/FVC for SNPs in seven
novel independent loci (GPR126, ADAM19, AGER-PPT2, FAM13A, PTCH1, PID1, and
HTR4) and with FEV1 for one novel independent locus annotated by at least three genes
(INTS12-GSTCD-NPNT). The SpiroMeta consortium independently reported genome-wide
significant associations of GSTCD, HTR4, AGER, TNS1, and THSD4 with FEV1/FVC and
FEV1 in an independent sample of 20,228 individuals of European ancestry (accompanying
manuscript). Both consortia confirm previous GWAS findings implicating the HHIP region
for FEV1/FVC10.
Several SNPs near the hedgehog interacting protein (HHIP) gene were associated with
FEV1/FVC at genome-wide significance in CHARGE and SpiroMeta, confirming earlier
GWAS findings in FHS10. The hedgehog (Hh)-signaling pathway is crucial in several
embryonic development processes, including the branching morphogenesis of the lung20,21.
Furthermore, several polymorphisms in three genes of the Hh-signaling pathway (IHH,
HHIP, and PTCH1) were significantly associated in a GWAS of adult height22. Several
PTCH1 SNPs were also significantly associated with FEV1/FVC in CHARGE, but these
associations were not confirmed in SpiroMeta. Epithelial cells produce Hh protein, which
binds to its membrane receptor (encoded by PTCH1) on mesenchymal cells and orchestrates
tissue and organ patterning. Hh pathway dysfunction during fetal life in humans is
responsible for severe lung malformations23,24. In adults, the Hh-signaling pathway may
participate in the response of the airway epithelium to injury, such as smoking and
A non-synonymous AGER SNP (rs2070600) was associated with FEV1/FVC at genome-
wide significance in our study and independently confirmed in SpiroMeta. The AGER
protein, a membrane-bound or soluble pattern recognition receptor, belongs to the
immunoglobulin superfamily of cell surface receptors. The SNP rs2070600 has functional
significance, e.g., higher ligand affinity and production of proinflammatory proteins upon
activation27. In healthy adult mice and humans, AGER is highly expressed in the lung28, and
its absence contributes to the pathogenesis of idiopathic pulmonary fibrosis29,30. AGER
signaling is involved in host defense, inflammation, and tissue remodeling, which are
relevant processes for accelerated decline in pulmonary function with age.
Polymorphisms in HTR4 were associated with FEV1/FVC at genome-wide significance in
the joint meta-analysis of CHARGE and SpiroMeta results. HTR4 encodes a G-coupled
transmembrane receptor that regulates cAMP production in response to 5-
hydroxytryptamine (serotonin). Elevated levels of free serotonin have been found in the
plasma of symptomatic asthmatics31, and serotonin signaling pathways involving HTR4
have been implicated in cholinergic and immune-mediated airway reactivity32,33. Upon
activation by serotonin, HTR4 in human airway epithelial cells regulates the release of a pro-
inflammatory cytokine, a signature characteristic of asthma34.
ADAM19 SNPs were associated with FEV1/FVC at genome-wide significance in CHARGE
and in the joint meta-analysis with SpiroMeta. ADAM19 is a member of "a disintegrin and
metalloprotease" (ADAM) family of membrane-anchored glycoproteins that control cell-
matrix interactions and help regulate growth and morphogenesis. Polymorphisms in another
ADAM family member, ADAM33, have been associated with bronchial hyperresponsiveness
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and accelerated lung function decline in asthmatics and the general population35–37.
ADAM19 has not been previously implicated in human pulmonary disorders, but it is
abundantly expressed in alveolar epithelial cells and bronchial smooth muscle tissue38.
GPR126 polymorphisms were associated with FEV1/FVC at genome-wide significance in
CHARGE and in the joint meta-analysis with SpiroMeta. GPR126 belongs to a superfamily
of G protein-coupled receptors involved in cell adhesion and signaling39. While its precise
function has not been elucidated, its expression in mice is temporally increased during
embryonic organ development and is highest in the adult lung40. In humans, recent GWA
studies have linked GPR126 variants with adult height, and more specifically, with trunk
height41–43. We adjusted all analyses for standing height. Therefore, we repeated analyses
for GPR126 SNPs adjusting for sitting height (a more reliable indicator of trunk height) in
ARIC, where both height variables were measured, and associations with FEV1/FVC
remained significant. Thus, these associations are not likely due to residual confounding by
trunk height.
Genome-wide significant associations with FEV1 were observed in CHARGE for numerous
SNPs spanning at least three genes on chromosome 4q24, and these associations were
significant for all eight SNPs tested for replication in SpiroMeta. There is moderate to strong
linkage disequilibrium among the chromosome 4q24 SNPs, and the specific genes
influencing FEV1 remain speculative. The genes are ordered INTS12-GSTCD-NPNT along
chromosome 4q24, and joint meta-analysis with SpiroMeta showed that SNPs from the
genes INTS12 and GSTCD had the most significant associations with FEV1. The product of
INTS12 is a subunit of the Integrator complex that associates with the C-terminal domain of
RNA polymerase II and mediates 3'-end processing of small nuclear RNAs44. GSTCD
(glutathione S-transferase, C-terminal domain) could influence lung function via
mechanisms involving the detoxification by glutathione S-transferases of xenobiotics that
might damage the lungs.
The most distal gene in the chromosome 4q24 region, NPNT, encodes nephronectin, which
is expressed in fetal and adult lung45,46. The NPNT SNP rs10516529 is located in a binding
site for the transcription factor POU6F1 (also known as mPOU homeobox protein), which is
known to be expressed in adult lung and hypothesized to play a role in lung development47–
49. A fourth predicted gene in the region, FLJ20184, is located proximal to the other three
genes. Although FLJ20184 encodes a hypothetical protein of unknown function, FLJ20184
contains allelic variants associated with successful smoking cessation in a GWAS of patients
in smoking cessation trials50.
The identified genetic factors gave estimated effect sizes consistent with those for well-
established risk factors for pulmonary function decline. Carrying one copy of an implicated
reference allele resulted in a FEV1 difference ranging from 50 to 70 mL. These effect sizes
correspond to approximately 2.8–3.9 years of age-related decline in pulmonary function
based on a mean decline of about 18 mL/year and to approximately 1.7–2.3 years of active
smoking-related decline based on a mean decline of about 30 mL/year51. Second-hand
smoke exposure has also been associated with decline in FEV1 (15 mL decline for a 10-year
exposure in the home and 41 mL decline for a 10-year workplace exposure)52. For FEV1/
FVC, carrying one copy of an implicated reference allele resulted in a difference ranging
from 0.30 to 1%. The lower effect size estimates are comparable with the mean FEV1/FVC
decline related to second-hand smoking (0.35 for a 10-year exposure in the home and 0.14
for a 10-year workplace exposure)52. These comparisons demonstrate that the identified
genetic factors have a moderate impact on pulmonary function. Individuals carrying these
polymorphisms will have lower pulmonary function than predicted at a given age, thus
placing them at greater risk for developing COPD and greater risk of mortality2,3.
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A GWAS of COPD identified CHRNA3/5 on chromosome 15 as a susceptibility locus11.
CHRNA3/5 has also been associated with nicotine dependence12,13. In CHARGE, one
identified SNP in this locus (rs1051730) was associated with FEV1/FVC (P=0.00070) and
FEV1 (P=0.016), while the other identified SNP in this locus (rs8034191) was not
associated with FEV1/FVC (P=0.11) or FEV1 (P=0.36). The nominal evidence for
replication may reflect differences in study design and a potential gene-environment
interaction involving smoking.
Our study has several important strengths. The CHARGE cohorts are well-phenotyped with
pulmonary function measures passing stringent quality control criteria, thus minimizing
measurement error. Our large sample size of 20,890 participants offers a powerful resource
to examine associations of common SNPs with modest to large effects14. However, we
likely have insufficient power to detect associations of polymorphisms with small effect
sizes or low frequencies. Replication in an independent consortium with similar power
offered the opportunity to confirm true genetic associations.
Population-based cohorts are subject to population stratification, and analytic steps were
taken to minimize this potential bias. Cohort-specific λgc values were low (1.00 to 1.05), and
a genomic control adjustment was made in the meta-analyses to reduce inflation in the test
statistics. The two largest cohorts, with the largest (albeit modest) λgc values (ARIC and
FHS), incorporated principal components as potential confounders in their cohort-specific
association tests. Although we cannot eliminate the possibility that some findings are subject
to residual confounding by population stratification, the Q-Q plots showing deviations
between observed and expected P values for many high- to moderate-signal SNPs and the
replication of association for multiple top loci in SpiroMeta suggest a multifactorial
influence on pulmonary function.
Our study identified several novel loci related to two clinically important pulmonary
function measures with evidence for replication, including GPR126, ADAM19, AGER-
PPT2, and HTR4 for FEV1/FVC and INTS12-GSTCD-NPNT for FEV1 and confirmed
previous reports of association with FEV1/FVC in the HHIP region. These loci include
genes with biologically plausible functions, and their identification here warrants future
investigations to elucidate the mechanisms underlying their influence on pulmonary
function. A few of the associated polymorphisms are potentially functional, but most of the
associated polymorphisms likely tag for yet unidentified functional variants. Fine mapping
in these regions might identify and characterize such variants. Understanding the genetic
determinants of pulmonary function is paramount in identifying the biological mechanisms
that lead to its decline and ultimately lessening the mortality burden associated with reduced
pulmonary function.
Pulmonary function measurements
Study design details of the participating CHARGE cohorts are described elsewhere14,53–58.
Study protocols were approved by the relevant institutional review boards, and all
participants provided written informed consent.
Pulmonary function testing was conducted by trained spirometry technicians at a single visit
for RS and at more than one visit for ARIC, CHS, and FHS. FEV1/FVC and FEV1 measures
meeting American Thoracic Society/European Respiratory Society criteria for acceptability
were tested for association with SNPs in participants of European ancestry who were
successfully genotyped and provided informed consent for genetic testing.
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In ARIC and CHS, pulmonary function measures and questionnaire data from the baseline
visit were analyzed. ARIC measurements were made with a Collins Survey II water-seal
spirometer (Collins Medical, Inc.) and Pulmo-Screen II software (PDS Healthcare Products,
Inc.)59. CHS measurements were made with a Collins Survey I water-seal spirometer
(Collins Medical, Inc.) and software from S&M Instruments60,61.
In three generations of families participating in FHS, data from the most recent examination
were analyzed. Eligible examinations providing spirometry and questionnaire data included
examinations 13, 16, 17, and 19 in the original cohort (in approximate two-year intervals);
examinations three, five, six, and seven in the offspring generation (in approximate four-
year intervals); and the one examination completed to date for the third generation.
Equipment used in the standard protocol evolved as technology improved over the decades
of study62. A Collins Survey water-filled spirometer (Collins Medical, Inc.) was used for
most examinations, with measurements made by Eagle II microprocessor (Collins Medical,
Inc.) or by software from the S&M Instruments. In more recent examinations, a Collins
Comprehensive Pulmonary Laboratory dry rolling-seal spirometer and Collins 2000 Plus/
SQL Software (Collins Medical, Inc.) were used.
In RS, pulmonary function was measured at the fourth center visit of participants from the
original cohort (RS-I) and the second center visit of participants from the first extension
cohort (RS-II). Spirometry was performed using a SpiroPro® portable spirometer (Erich
Jaeger GmbH)63,64.
Genotyping, imputation, and quality control
Different genotyping platforms were used across the cohorts (Table 1)14. Imputation was
conducted using either MACH65 or BIMBAM66 to generate approximately 2.5 million
autosomal SNP genotype dosages for meta-analysis. The imputation methods perform
similarly, although MACH generally produces higher accuracy rates than the imputation
process used in BIMBAM (fastPHASE)67. Differing imputation methods across cohorts is
not a source of bias for meta-analysis since all comparisons using the imputed data are
within-cohort comparisons.
ARIC—Among 8,861 self-identified white ARIC participants genotyped, 8,127 participants
remained after exclusions for call rate<95%, genotypic and phenotypic sex mismatch,
discordances with previous genotype data, suspected first-degree relative of an included
individual based on genotype data, more than eight standard deviations for any of the first
10 principal components using EIGENSTRAT68, or outlying average identity-by-state
estimates using PLINK69. Of these, 7,980 participants had pulmonary function measures and
complete covariate information.
A total of 704,588 autosomal genotyped SNPs remained after exclusions for call rate<95%,
MAF<1%, Hardy-Weinberg equilibrium (HWE) P<10−5, or lacking strand annotation.
MACH (version 1.00.16)65 was used to impute all autosomal SNPs with reference to
HapMap CEU (release 21, build 35)70 from these 704,588 SNPs. Imputed SNPs failing
additional quality control criteria (monomorphism, HWE P<10−6, or genotype frequencies
between two genotyping phases differed by P<10−6) were excluded, leaving 2,515,866
genotyped or imputed SNPs for analysis.
CHS—CHS genotyped 3,980 participants free of cardiovascular disease at baseline with
available DNA and consent to genetic testing. After exclusions for call rate <95%, sex
mismatch, or discordance with prior genotyping, 3,291 white participants remained. Of
these, 3,140 had pulmonary function measures and complete covariate information.
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A set of 306,655 autosomal genotyped SNPs remained after exclusions for call rate<97%,
HWE P<10−5, more than two duplicate errors or Mendelian inconsistency (for reference
HapMap CEU trios)70, heterozygote frequency>0, or no mapping in dbSNP. Imputation of
autosomal SNPs was based on these 306,655 SNPs using BIMBAM (version 0.99)66 with
reference to HapMap CEU (release 22, build 36)70. The analysis data set included
2,543,887 genotyped or imputed SNPs.
FHS—A total of 8,481 participants remained after exclusions for call rate<97%,
heterozygosity more than five standard deviations from the mean, or excessive non-
inheritance. The analysis data set included 7,694 participants with complete spirometry and
covariate data.
MACH (version 1.00.15)65 was used for imputation based on 378,163 autosomal SNPs
remaining after exclusions for HWE P<10−6, call rate<97%, differential missingness related
to genotype (mishap procedure in PLINK69) with P<10−9, Mendelian errors>100,
MAF<1%, or those not present in HapMap. Two hundred unrelated individuals with high
call rate were used to infer model parameters, which were subsequently applied to all 8,481
individuals. Imputation, using HapMap CEU (release 22, build 36),70 produced genotype
dosages on 2,543,887 genotyped or imputed SNPs.
RS—All RS participants with available DNA were genotyped; 5,974 RS-I participants and
2,157 RS-II participants remained after exclusion for call rate<97.5%, excess autosomal
heterozygosity, sex mismatch, or outlying identity-by-state clustering estimates. Of these,
1,224 RS-I participants and 852 RS-II participants had pulmonary function measures and
complete covariate information.
After exclusions for call rate<98%, HWE P<10−6, and MAF<1%, 512,349 autosomal SNPs
in RS-I and 466,389 autosomal SNPs in RS-II were used for imputation in MACH (version
1.00.15 for RS-I and 1.00.16 for RS-II)65 with reference to the 2,543,887 SNPs of the
HapMap CEU (release 22, build 36)70.
Statistical analysis
In cross-sectional analyses, FEV1/FVC and FEV1 were tested for association with SNP
genotypes using a one degree-of-freedom additive model of the dosage value (estimated
reference allele count with a fractional value ranging from 0 to 2.0) as a predictor in linear
regression models. Associations were examined overall and stratified into ever and never
smokers. Overall models were adjusted for age, sex, standing height, smoking status
(current/past/never), and pack-years of smoking. Current, past, or never smoking was based
on questionnaire responses, and pack-years were calculated for current and past smokers by
multiplying smoking dose (packs/day) and duration (years). Stratified models used the same
covariates as the overall models, except that the ever-smoker stratum included adjustment
for smoking status as current/past and the never-smoker stratum included no smoking-
related covariates. Additional study-specific covariates included recruitment cohort (FHS),
recruitment center (ARIC and CHS), and principal component eigenvalues for population
stratification adjustments (10 components for ARIC and statistically significant components
for FHS). Models were implemented using ProbABEL71 in ARIC, R72 in CHS, linear
mixed effects models with fixed effects for SNPs and random effects for individuals
correlated within families73 in FHS, and MACH2QTL65 in RS as implemented in
GRIMP74. In FHS, the kinship package in R generated a covariance matrix for each family
based on the kinship coefficient for each relative pair. The kinship matrix, which includes
the full set of family-specific covariance matrices, specified the covariance matrix for the
random effects.
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GWAS results from the four cohorts were combined using inverse variance weighted meta-
analysis in METAL (http://www.sph.umich.edu/csg/abecasis/metal/). Meta-analysis was
performed on approximately 2,534,500 SNPs after applying genomic control for each study
and filtering SNPs with extremely low imputation quality ratios (<0.01) and MAF (<1%).
The genome-wide significance threshold was defined a priori as P<5×10−8, the Bonferroni
adjustment for one million independent tests75. Information on SNP function and position
relative to genes, microRNA, and transcription factor binding sites was obtained using a Perl
script (J.B.W.) that queries tables of the UCSC genome browser15 (hg18, March 2006
genome build). Functional effects of non-synonymous SNPs on protein structure and
function were predicted using PolyPhen17.
Replication in the SpiroMeta consortium
We exchanged 30 SNPs for replication testing with the SpiroMeta consortium
(accompanying manuscript). No additional genotyping was required, as these SNPs were
available from the SpiroMeta GWAS. We aimed to select two SNPs from each of the top
genes implicated for FEV1/FVC or FEV1, nearly all exceeding genome-wide significance.
The SNP with the lowest P value in or near each gene was selected. A second SNP,
genotyped (instead of imputed) in at least one cohort, was selected with preference for non-
synonymous SNPs and SNPs not in strong linkage disequilibrium with the first selected
SNP. Only one SNP was available for AGER, PPT2, TSPYL4, and NT5DC1. Four SNPs
were selected from two linkage disequilibrium blocks for the largest gene, GPR126. In total,
18 SNPs from nine genes (eight independent loci) implicated for FEV1/FVC and 12 SNPs
from seven genes (three independent loci) implicated for FEV1 were tested for replication.
Unlike CHARGE, SpiroMeta used normalized residuals as phenotypes, adjusted for age2
rather than age, and did not adjust for smoking. For better comparison, SpiroMeta conducted
modified analyses following the CHARGE analytic method described above in 16,178
participants from adult cohorts with complete quantitative smoking data available. Results
from the CHARGE GWAS and SpiroMeta replication were combined in a joint meta-
analysis using inverse variance weighting with METAL. SpiroMeta results with
P<8.33×10−4, based on an overly conservative Bonferroni correction for 60 tests (30 SNPs
tested for association with two traits, FEV1/FVC and FEV1), or joint meta-analysis results
with P<5×10−8 (genome-wide significance threshold) were considered statistically
Supplementary Material
Refer to Web version on PubMed Central for supplementary material.
This work was supported in part by the Intramural Research Program of the NIH, National Institute of
Environmental Health Sciences (Z01ES043012). The Atherosclerosis Risk in Communities Study is carried out as a
collaborative study supported by National Heart, Lung, and Blood Institute contracts N01-HC-55015, N01-
R01HL59367 and R01HL086694; National Human Genome Research Institute contract U01HG004402; and
National Institutes of Health contract HHSN268200625226C. The authors thank the staff and participants of the
ARIC study for their important contributions, along with Dr. Grace Chiu, Dick Howard, and Miguel Quibrera for
their analytic contributions.
The Cardiovascular Health Study research reported in this article was supported by contract numbers N01-
HC-85079 through N01-HC-85086, N01-HC-35129, N01 HC-15103, N01 HC-55222, N01-HC-75150, N01-
HC-45133, grant numbers U01 HL080295 and R01 HL087652 from the National Heart, Lung, and Blood Institute,
with additional contribution from the National Institute of Neurological Disorders and Stroke. A full list of
principal CHS investigators and institutions can be found at http://www.chs-nhlbi.org/pi.htm. DNA handling and
genotyping was supported in part by National Center for Research Resources grant M01-RR00425 to the Cedars-
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Sinai General Clinical Research Center Genotyping core and National Institute of Diabetes and Digestive and
Kidney Diseases grant DK063491 to the Southern California Diabetes Endocrinology Research Center.
Research was conducted in part using data and resources from the Framingham Heart Study of the National Heart,
Lung, and Blood Institute of the National Institutes of Health and Boston University School of Medicine. The
analyses reflect intellectual input and resource development from the FHS investigators participating in the SNP
Health Association Resource (SHARe) project. This work was partially supported by the National Heart, Lung, and
Blood Institute's FHS (Contract No. N01-HC-25195) and its contract with Affymetrix, Inc. for genotyping services
(Contract No. N02-HL-6-4278). A portion of this research utilized the Linux Cluster for Genetic Analysis (LinGA-
II) funded by the Robert Dawson Evans Endowment of the Department of Medicine at Boston University School of
Medicine and Boston Medical Center. J.B.W. is supported by a Young Clinical Scientist Award from the Flight
Attendant Medical Research Institute (FAMRI).
The Rotterdam Study was supported from grants from the Netherlands Organisation of Scientific Research
Netherlands Organisation for Scientific Research (NOW) Investments (175.010.2005.011, 911-03-012), the
Research Institute for Diseases in the Elderly (014-93-015; RIDE2), the Netherlands Genomics Initiative (NGI)/
NWO (050-060-810), Erasmus Medical Center, Erasmus University, Rotterdam, Netherlands Organization for the
Health Research and Development (ZonMw), the Research Institute for Diseases in the Elderly (RIDE), the
Ministry of Education, Culture and Science, the Ministry for Health, Welfare and Sports, the European Commission
(DG XII), and the Municipality of Rotterdam. The authors thank Pascal Arp, Mila Jhamai, Dr. Michael Moorhouse,
Marijn Verkerk, and Sander Bervoets for their help in creating the Rotterdam GWAS database; Dr. Tobias A.
Knoch, Luc V. de Zeeuw, Anis Abuseiris, and Rob de Graaf as well as their institutions, the Erasmus Computing
Grid, Rotterdam, The Netherlands, and the national German MediGRID and Services@MediGRID part of the
German D-Grid (German Bundesministerium fur Forschung und Technology (#01 AK 803 A–H and # 01 IG 07015
G), for access to grid resources.
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genomewide association studies of nearly all common variants. Genet Epidemiol 2008;32:381–
385. [PubMed: 18348202]
Hancock et al. Page 15
Nat Genet. Author manuscript; available in PMC 2010 March 5.
NIH-PA Author Manuscript
NIH-PA Author Manuscript
NIH-PA Author Manuscript


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WA 94 Pts - According to Wine Advocate - The 2012 Cotes du Roussillon Villages Latour de France Occultum Lapidem offers dark, mineral-laced aromas and flavors of smoked earth, licorice, wild herbs and blackcurrants to go with a firm, structured and age-worthy profile on the palate. Comprised of mostly Syrah, from schist and gneiss soils, and aged in tank and demi-muid, this superb 2012 has solid mid-palate depth, ample tannin and a beautifully focused finish. It's a superb value that will be even better in another year or so, and have 12-15 years of ultimate longevity.
Winemakers Notes - Great dark garnet-red. The first nose is quite closed with a dominant of graphitic, leather, and then intense aromas of pepper, black fruits and scrubs. The attack is rich, dense. Then the wine is fleshy and solar. The final is sappy served by delicate tannins and a great freshness. Rated 90+
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L 3, looks very well.
All the best !
L 19,F 4, a fine composition !
All the best !
Delightful image,Kostia. F+L Kind regards. Jack.
Very...very nice capture!Like
Best greetings!-Sidnei
Stunning view and magnificent capture!YSL
Best greetings!-Sidnei
HBi Pradeep colous and more colours in your gallery.. I like it ! !! greetings
Good view and picture with strong colours. Friendly greetings, Thomas
Thank you very much Yuri, for your visit, nice words and LIKE.
Best wishes from Jordan.
Ich glaube auch BIG DADDY :) und besonders bei dieser Hitze täglich fast über 40C°
Thank you very much Pavel for your nice words and your YS.
Best wishes from Jordan.


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Desperate Housewives 2004 | TV Show Watchlist
Photos | Cast & Crew | Episode Guide | Video Clips
Following her suicide, Mary Alice Young keeps tabs on friends and family she left behind on bucolic suburban Wisteria Lane. But this being a prime-time soap (if a tongue-in-cheek one), they all harbor a sordid secret or two. Created by writer-producer Marc (more…)
Premiered: October 3, 2004
Rating: TV-PG
Season 7, Episode 5 Let Me Entertain You
First Aired: October 24, 2010
Susan upsets a Va-Va Broom client and Maxine notices; Renee and Gaby tell secrets about each other to the wrong people; Bree can't keep up with Keith in bed; Lynette is mad at Tom for asking his mother (Lois Smith) to be their baby's nanny without checking with her first; and housewife Emma Graham (Dana Glover) puts on a cabaret show with saxophonist Dave Koz.
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We just finished our diving certificates, and we are so lucky that we have some really good dive sites right outside the gate of the house. We are diving several times a week off the beach here so I am sure we will have a massive underwater photo collection up soon.
With Apo island only 7 kilometers from our house we are very spoiled with the dive sites already, I would be hard pushed going back to Denmark to dive there after seeing the beautiful underwater world of the Philippines.
There an abundance of dive shops and dive centers in Dumaguete and the surrounding areas, I have compiled a short and definitely not complete list of the dive centers here:
APO ISLAND BEACH RESORT,Dumaguete City, Desk Tel #: 225-5490
Atlantis Dive Shop, Dauin, Tel # +63 (0)35 4240578
DIVER'S INN, Siliman Beach — Dumaguete City
DIVE KOREA, Bunao Dumaguete City
DIVE QUEEN, Hilaitan, Gihulngan. Mobile #: 09167084740
DIVE SOCIETY,El Dorado Beach Resort, Dauin, Tel# + 63 (0) 35 – 425-2274
JMC, STA MONICA RESORT — Dumaguete City Tel #: 255-0704
MALATAPAY COTTAGES, Maluay, Zamboanguita, Mobile #: 0918-740-0038
Mike's Dauin Beach Resort Poblacion, Dauin, Tel. 035-422-1781
NAUI WORLD WIDE, Rovina Rd. Dumaguete City. Tel #: 225-5257/422-1739
PAUL'S DIVE SHOP, Apo Island, Dauin: Tel #: 424-0888
SAN JOSE DIVE & TOURS — Tel #: 225-9648/417-0504
SCUBA WORD,COCONUT PLANTATION INN, DAUIN — Tel #: 422-2199
SEA EXPLORERS, Pura Vida Beach & Dive Resort,Dauin — Tel #: +63 (0) 35 425 2284
WHITE TIP DIVERS, Hibbard Ave. Dumaguete City — Tel #: 225-2381/225-2402, FAX: 225-7716
Make sure you talk to the instructors before signing up for a dive course, it is very important you feel comfortable and safe with the dive instructor. They are after all going to take you under water and teach you how to dive. All the mentioned dive centers are members of the PADI network, so they can all do your PADI certification.
Adventure Dive Shop
You are looking for a professional dive shop in Dumaguete with a great family-atmosphere ? Mike's Adventure Dive Shop is the place to go. Spending every day in the waters around Dumaguete and the neighborhood Islands Apo, Siquijor, Sumilon and the south-coast of Cebu many years makes Mike an expert in and around the waters […]Read More
Dauin: Diving the Car Wrecks
When people talk about a tourist destination the Philippines is definitely on their list. It is mainly because the country has a lot to offer such as the white beaches of Boracay, Pagudpud and Puerto Galera. Furthermore, the Philippines has also has a number of unexplored wonders waiting for you to explore such as the […]Read More
Dauin: Masaplod Marine Sanctuary
Dauin in the province of Negros Oriental provides tourists with exceptional dive sites. Most of them are located 5 – 10 minutes away from the Amontillado Beach & Dive Resort. In general, they are located in front of the beach and can be easily reached. Dauin boasts of several marine sanctuaries. They are home to […]Read More
Dive Shops
We just finished our diving certificates, and we are so lucky that we have some really good dive sites right outside the gate of the house. We are diving several times a week off the beach here so I am sure we will have a massive underwater photo collection up soon. With Apo island only […]Read More
Orientwind Travel & Tours
So, your flight is booked, your hotel is reserved and you have everything ready for a great trip to Dumaguete, the beautiful city on the shores of Negros Oriental in the archipelago of the Philippines. What's next? What's missing to ensure your trip is a perfect one? How can you make your vacation stress free […]Read More


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Torch – Colombia Freestyle
MMG's Torch jumps on Young Scooter's Colombia. Off Torch's Tax Season mixtape dropping March 12th.
Wale's highly-anticipated fourth studio album The Album About Nothing hits stores tomorrow! And today a new track has been leaked onto the ...
Fabolous' ongoing #FridayNightFreestyles has been a relative success thus far but with this new one right here it's either make it or ...
Bodega BAMZ' forthcoming project Sidewalk Exec is due out April 14th and having already previewed "Billy Bats" BAMZ is returning with a ...
MMG's Torch jumps on Young Scooter's Colombia. Off Torch's Tax Season mixtape dropping March 12th.
In an interview with SkeeTV, Snoop Dogg talked about squashing his beef with Suge Knight, putting Drake on his new single, working on a reggae album with Diplo, and much more.
"Time heels all wounds. And then being a bigger man. And being one who accepts responsibility, being right or wrong. I like saving lives, I don't like taking lives. If that means putting my hand out saying I'm wrong, I apologize and be cool, I'll do that. As opposed to me pulling a gun out…and me doing something vicious to you."
Watch the interview after the jump (more…)
mtvU got the premiere for this Joey Bada$$' Unorthodox produced by DJ Premier.
Producer Mathematics rounds up some elite MCs for this Rap Genius premiere. Check out the decoded lyrics here.
Download Mathematics feat. Bad Luck, Cappadonna, Eyes Low, Method Man, Raekwon & Termanology – Men of Respect
For season 2 of Iyanla: Fix My Life, she travels to DMX's hometown of Yonkers. DMX's emotions run high and his temper takes over.
DJ Premier gives the Demigodz an assist in making sure we never have pleasant dreams again.


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Abu's panel-writhing photo of Pumbaa's House and Hound Pumbaa
After you roam through Pumbaa's house, you'll come across her TV Barn in the back yard. There you can stumble on Pumbaa finishing recording a scene from her celebrated film, "The Band Bunny. Be sure to avoid confusing her with Genie!
Beware Zombie Mode
© copyright 2011–2015
A Pants Aflame production


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Fox News once again traveled to Panama Beach, Fla. for spring break to "expose the realities" of the booze and drug-fueled debauchery to be found there because "BREAKING: College kids still act like idiots on spring break!" What exactly did they find? 102 more words
Tags » Ainsley Earhardt
'WOWZA'! Are you ready for Andrea Tantaros' FNC Halloween flashback? [photo]
We're guessing a lot of people had that same reaction when they saw this:
226 more words
Flashback: Halloween 2011… with @kimguilfoyle and @ainsleyearhardt pic.twitter.com/s6km5bfMEI
— Andrea Tantaros (@AndreaTantaros) …
Special Report: College Kids 'Party' on Spring Break (Video)
WASHINGTON (CBSDC) - College students across the country have flocked to beach towns for the annual rite of passage known as "spring break."
Another way to put it is 'pour copious amounts of booze into their skulls until they pass out where they stand,' which seems to be the tone beautifully articulated in a recent special report by Ainsley Earhardt for Fox News, brought to my attention by drunk college chick connoisseur, Matt Valdez. 801 more words


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Will oil prices affect airline tickets?
Kippreport speaks to both Etihad and Emirates to find out what the deal is...March 29, 2015 2:46
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Qatar's bourse says plans rights issue trading
Soccer-Xavi close to deal with Al Sadd, says Qatari club
Soccer-Xavi close to deal with Al Sadd, says Qatari club
Abu Dhabi Airports CEO leaving to take up U.K. defence ministry role
Kuwait February inflation edges up to 2.9 pct
MIDEAST STOCKS-Gulf markets sink after Yemen intervention; Dubai worst hit
HSBC's Islamic banking unit plans sukuk sale in Malaysia
Oil rises more than 1 pct, boosted by weaker dollar
UAE's Shah gas project seen achieving full capacity in Q2
National Bank of Fujairah issues 500 mln dirham tier 1 capital
From the managing director of Dubai Duty Free to Pulp Fiction's 'Mr Wolf", via David Beckham, in Kipp's six easy steps.
September 24, 2007 11:16 by kippreport
London-born Cannon has made a success of his career in America. He got his big break in Hollywood directing Sylvester Stallone in the 1995 film Judge Dredd, and has since gone on to produce (and direct episodes on) the money-spinning CSI franchise. With a reputation for style (if not content) his big break came in the 1993 film the Young Americans, which starred Viggo Mortensen and…
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accounting jobs near Moorhead, MN
Needed: Accounting Tutor in or near Fargo, ND. $34-$39 an hour.NEW
Tutorselect Tutoring- Fargo, ND
We have students struggling in Accounting, can you help? Work a few hours a week and earn ... goals. Our positions are rewarding and fun. Accounting tutors Fargo. $34 - $39 an...21 hours ago from TutorSelect Tutoring
GRAIN ACCOUNTING ASSISTANTCasselton, ND
GRAIN ACCOUNTING ASSISTANT MAPLE RIVER GRAIN AND AGRONOMY, LLC located in Casselton, ND, is looking for a full time grain accounting assistant. Duties include daily grain ticket...27 days ago from TheJobNetwork
Accounting Manager
Eide Bailly- Moorhead, MN
is responsible for all aspects of the accounting and financial systems of the ... ideal candidate will possess knowledge of accounting principles and practices along...30+ days ago from iHireAccounting
Internal Auditor
Titan Machinery- West Fargo, ND
This position also provides accounting and project management support to the Accounting Te ... Demonstrated experience working with accounting systems Physical Requirements...7 days ago from dieseljobs.com
Controller – Bookkeeper Premium Accounting for Retail Broker
Work at Home Vintage Employees- Fargo, ND
suited candidate must have a premium accounting background including producer ... of retail or wholesale/MGA broker premium accounting background. • Candidates must have a...30+ days ago from RetirementJobs.com
Accountant Controller
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and direction of the accounting staff. • Lead and implement accounting ... disciplines including general accounting, audit, reporting, planning and analysis,...18 days ago from ZipRecruiter
State and Local Taxation (SALT) Senior Associate
Cpa and Business Advisory Firm- Fargo, ND
compliance, preparing returns, income tax accounting, controversies, restructuring, due ... Accounting / Control, Consulting - Accounting...20 days ago from IvyExec
Accountant II
RDO Equipment- Fargo, ND
the accounting function by performing daily accounting tasks and timely account ... accounting projects * Identifies and reports accounting related issues to appropriate...30+ days ago from Professional Diversity Network
General Ledger AccountantFargo, ND
teamwork. Qualifications: 4-year degree in accounting. Exceptional written/verbal communication skills. Strong knowledge of Microsoft Office. Experience with Microsoft Dynamics GL...20 days ago from TheJobNetwork
Audit Manager
Cpa and Business Advisory Firm- Fargo, ND
business processes. Prepare and/or review accounting-related reports through a ... requirements: Bachelor's degree in Accounting Certified Public Accountant (CPA)...20 days ago from IvyExec
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Woodruff Homes for Rent
BusinessWeek "Best of the Web" Winner 3 Years Running
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Live updates
Stoke hope to tie down Arsenal and Liverpool target
Stoke City chief executive Tony Scholes says the club are hopeful of getting Asmir Begovic to sign a new deal.
The Bosnian goalkeeper, who has been linked with moves to Liverpool and Arsenal, has a contract that runs to next summer and Stoke believe they can match his ambitions.
Clearly Asmir is a terrific goalkeeper and we are very keen to keep him at the club.
I think when you've got extremely good players it is only natural that there will be speculation about their future.
These players are also very ambitious and I include Asmir in that. He is a very focused, very talented and very ambitious young man.
He wants to make sure he can play at the very best levels his abilities will enable him to and to have a very good and successful career in the game .
We hope that we can fulfil all of those things at Stoke City.
Arsenal forward makes 'silly' comment in French media
Arsenal forward Olivier Giroud has told Frence media outlet RMC Sport that he wants to score more goals than Alexis Sanchez this season in what he describes as a "silly" comment.
The Gunners pair have 13 goals apiece in the Premier League while Sanchez has bagged two more than Giroud's 17 in all competitions.
Speaking during the international break, French striker Giroud said:
I'm not going to say something silly because it'll be reported, but why not finish ahead of Alexis?
We push each other, we're on 13 goals, and I would like to score some more to help the team achieve its objectives.
Man Utd suffer blow as Brazilian joins Real Madrid
Man United have suffered an early blow to their transfer plans for next season after defender Danilo agreed to join Real Madrid.
The Brazilian full back opted for the Spanish giants despite concrete interest from United, and will leave Porto in the summer for a fee around £23m.
Madrid confirmed last night:
Real Madrid and FC Porto have reached an agreement over the transfer of Danilo, he will join the club next season and has a contract until 30 June 2021.
- ITV Report
Dortmund defender talks about Man Utd rumour on Twitter
Mats Hummels has shot down reports of a deal with Manchester United coach Louis Van Gaal to join the Premier League clubRead the full story ›
- ITV Report
Rumours: England star to swap Arsenal for Chelsea
Theo Walcott will find a new home across London at Chelsea if, as expected, he stalls on contract discussions in the summerRead the full story ›
Former Man United assistant resigns after defeat
Carlos Queiroz has stepped down as Iran coach after their 3-1 loss to Sweden in a friendly on Tuesday night.
The former Real Madrid and Portugal boss has reportedly been involved in a dispute over income tax with the Iranian Football Federation (FFIRI), and chose to make their friendlies this weekend against Austria and Sweden his last matches in charge.
Speaking after Tuesday's loss, he said on the FFIRI website: "After four years of work with the national team, I am proud of the team and the individual players...
"Now I am sad to leave the team, but that's life and I'm happy to be moving forward."
Queiroz, 62, became Iran coach in 2011 and qualified them for the 2014 World Cup.
After the tournament, he signed a new deal which was due to run until the 2018 World Cup in Russia.
- ITV Report
Spurs winger calls out Arsenal legend on Twitter
Spurs winger Andros Townsend celebrated his stunning goal against Italy by taking a swipe at Paul Merson on TwitterRead the full story ›
Rooney: Man United star Carrick was the best player on the pitch
England captain Wayne Rooney has praised the influence of Manchester United team-mate Michael Carrick in the draw with Italy.
The midfielder replaced Chris Smalling in the first-half and was instrumental in bringing England back in the game.
Plane fault forces England squad to stay in Italy
The England football team will need to stay in Italy for an extra night after their plane suffered a technical issue.
It is now hoped that the team will be able to return to England on Wednesday morning, which could annoy a few club managers.
Townsend stunner earns England draw in Italy
Spurs winger Andros Townsend came off the bench to score the England equaliser in spectacular fashion in Turin.
Graziano Pelle had given Italy the lead in the first-half, but it wasn't enough for the hosts in the end.
Watch highlights of the game on our England website.