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Page 1
The FASEB Journal • Research Communication
Melanocortin 1 receptor genotype: an important
determinant of the damage response of melanocytes
to ultraviolet radiation
Ana Luisa Kadekaro,* Sancy Leachman,§Renny J. Kavanagh,* Viki Swope,*
Pamela Cassidy,§Dorothy Supp,†,?Maureen Sartor,‡Sandy Schwemberger,†
George Babcock,†,?Kazumasa Wakamatsu,¶Shosuke Ito,¶Amy Koshoffer,*
Raymond E. Boissy,* Prashiela Manga,#Richard A. Sturm,** and
Zalfa A. Abdel-Malek*,1
*Department of Dermatology,†Department of Surgery, and‡Department of Environmental Health,
University of Cincinnati, Cincinnati, Ohio, USA;§Department of Dermatology and Huntsman Cancer
Center, University of Utah, Salt Lake City, Utah, USA;?Shriners' Burns Hospital, Cincinnati, Ohio,
USA;¶Department of Chemistry, Fujita Health University, School of Health Sciences, Toyoake,
Japan;#Ronald O. Perelman Department of Dermatology, New York University, New York, New York,
USA; and **Institute for Molecular Biosciences, University of Queensland, Brisbane, Australia
main determinant of human pigmentation, and a
melanoma susceptibility gene, because its variants
that are strongly associated with red hair color in-
crease melanoma risk. To test experimentally the
association between melanocortin 1 receptor genotype
and melanoma susceptibility, we compared the re-
sponses of primary human melanocyte cultures nat-
urally expressing different melanocortin 1 receptor
variants to ?-melanocortin and ultraviolet radiation.
We found that expression of 2 red hair variants
abolished the response to ?-melanocortin and its
photoprotective effects, evidenced by lack of func-
tional coupling of the receptor, and absence of
reduction in ultraviolet radiation-induced hydrogen
peroxide generation or enhancement of repair of
DNA photoproducts, respectively. These variants had
different heterozygous effects on receptor function.
Microarray data confirmed the observed differences
in responses of melanocytes with functional vs. non-
functional receptor to ?-melanocortin and ultraviolet
radiation, and identified DNA repair and antioxidant
genes that are modulated by ?-melanocortin. Our find-
ings highlight the molecular mechanisms by which the
melanocortin 1 receptor genotype controls genomic stability
of and the mutagenic effect of ultraviolet radiation on
human melanocytes.—Kadekaro, A. L., Leachman, S.,
Kavanagh, R. J., Swope, V., Cassidy, P., Supp, D., Sartor,
M., Schwemberger, S., Babcock, G., Wakamatsu, K., Ito,
S., Koshoffer, A., Boissy, R. E., Manga, P., Sturm, R. A.,
Abdel-Malek, Z. A. Melanocortin 1 receptor genotype: an
important determinant of the damage response of mela-
nocytes to ultraviolet radiation. FASEB J. 24, 000–000
(2010). www.fasebj.org
The melanocortin 1 receptor gene is a
Key Words: DNA repair ? melanoma susceptibility ? MC1R
variants ? cAMP pathway
Epidermal melanocytes are the precursors for mel-
anoma, the most fatal form of skin cancer. The inci-
dence of melanoma continues to increase worldwide,
especially in young adults (1–5). The resistance of
melanoma tumors to conventional chemotherapy and
radiation and lack of a cure for metastatic disease have
made it necessary to identify genetic markers for mel-
anoma that should lead to more accurate risk assess-
ment and thus prevention.
The melanocortin 1 receptor (MC1R) has emerged as a
melanoma susceptibility gene. The MC1R is a highly
polymorphic gene and a major contributor to the
diversity of human pigmentation (6–8) Some MC1R
variants, particularly R151C, R160W, and D294H, are
strongly associated with red hair color in humans
(hence the name RHC alleles) and increased mela-
noma risk (7–10). An inverse correlation exists between
the incidence of sporadic melanoma, on one hand, and
constitutive pigmentation and tanning ability, on the
other (2). Cutaneous pigmentation is determined by
the relative rate of synthesis of the brown/black eu-
melanin and the red/yellow pheomelanin by epider-
mal melanocytes (11, 12). Eumelanin is the major
contributor to skin pigmentation and photoprotection
because of its efficiency in blocking ultraviolet rays
(UV) and scavenging reactive oxygen species (13).
The principle physiological regulators of eumelanin
synthesis in human melanocytes (HMs) are ?-melano-
cortin (?-MSH) and adrenocorticotropic hormone, the
physiological agonists of the MC1R (14–16). The
MC1R is a Gs-protein-coupled receptor with 7 trans-
membrane domains that is expressed on the cell sur-
1Correspondence: Department of Dermatology, University of
Cincinnati College of Medicine, 231 Albert Sabin Way, Cincin-
nati, OH 45267-0592, USA. E-mail: firstname.lastname@example.com
doi: 10.1096/fj.10-158485
The FASEB Journal article fj.10-158485. Published online June 2, 2010.
Page 2
face of HMs (17, 18). Binding of the human MC1R by
its ligands activates the cAMP signaling pathway, a
major regulator of HM proliferation and pigmentation
(19). We have made the seminal observation that in
addition to their important role as the physiological
regulators of pigmentation, melanocortins also modu-
late the UV response by enabling HMs to overcome the
UV-induced growth arrest and apoptosis and reducing
DNA damage (20–22). We found that ?-MSH enhances
repair of UV-induced DNA photoproducts and reduces
the induction of oxidative DNA damage (21, 22). These
effects were dependent on activation of the MC1R and
preceded the increase in melanin content in response
to ?-MSH treatment. Thus activation of MC1R is ex-
pected to reduce the genotoxic effects of UV exposure,
independently of increased melanin content.
Evidence for the effect of MC1R genotype on mela-
noma risk came from the observation that RHC alleles
increase the penetrance of mutations in cyclin-dependent
kinase inhibitor 2A (23–25) and that expression of an
MC1R variant increases the chance for the somatic
V600E BRAF-activating mutation (26). Expression of 2
RHC alleles results in loss of function of MC1R (27).
However, the interaction of these variants with other
alleles and their impact on the response of HMs to UV,
a major environmental etiological factor for melanoma,
are not understood. Unraveling the molecular mecha-
nisms by which MC1R alleles affect genomic stability of
melanocytes and the risk for melanoma will lead to
improved preventive strategies and effective treatments
for melanoma, one of the most challenging human
In this study we used primary HM cultures that
naturally express different MC1R genotypes to deter-
mine systematically the impact of RHC alleles and other
common MC1R variants on receptor function and
response to UV. Our data demonstrate that loss of
function of MC1R due to expression of 2 MC1R RHC
variants, or an RHC allele and V60L variant, result in an
aberrant UV response, and that the RHC alleles are not
equivalent in their impact on receptor function. Our
results provide compelling evidence that functional
MC1R is required for optimal photoprotection and
maintenance of genomic stability and elucidate mech-
anisms by which MC1R variants determine predisposi-
tion to melanoma.
Culturing of human melanocytes
Primary cultures of HMs were established as described previ-
ously (15) from discarded neonatal foreskins or adult skin from
patients undergoing elective plastic surgery procedures. The
Institutional Review Board at the University of Cincinnati has
deemed the protocol for obtaining these skin samples exempt
from approval. Melanocyte cultures were also established from
skin biopsies obtained from patients of the Melanoma Clinic at
the Huntsman Cancer Institute after informed consent. For all
experiments, HMs were maintained in medium devoid of bo-
vine pituitary extract (BPE) to allow for response to exogenous
?-MSH, as described previously (15).
Analysis of eumelanin and pheomelanin contents of
cultured HMs
Melanocytes (2–4?106cells) were harvested, pelleted, lyoph-
ilized, and used for quantitation of eumelanin and pheomela-
nin, respectively, as described previously (27).
Sequencing of the MC1R gene
The entire coding region of the MC1R was amplified using
reverse transcriptase and nested PCR amplification, followed
by sequencing as described previously (27).
Determination of cyclic AMP (cAMP) levels and tyrosinase
Cyclic AMP levels were determined using125I-labeled radio-
immunoassay kit (Perkin Elmer, Waltham, MA, USA), as
described previously (18, 27), in HMs that were treated with
0 (control), 1, or 10 nM ?-MSH, or 1 ?M forskolin for 45 min
in the presence of 0.1 mM isobutyl methylxanthine. Tyrosi-
nase activity was measured after treatment of melanocytes
with 1 or 10 nM ?-MSH or 1 ?M forskolin for 6 d, as described
previously (15, 18, 27).
Irradiation of HMs with UV
Melanocytes were irradiated with a bank of FS 20 lamps, with
75% emission in the UVB, and 25% emission in the UVA
spectra, with peak emission at 313 nm wavelength. Any
emission of UVC rays from the UV source (FS 20 lamps) was
blocked by a Kodacel filter (Eastman Kodak, Rochester, NY,
USA). Prior to irradiation, the culture medium was replaced
by PBS, which was removed after UV exposure, and fresh
medium was added.
Measurement of UV-induced generation of hydrogen
Melanocytes were irradiated with 105 mJ/cm2UV and treated
immediately while in PBS with 0, 1 nM ?-MSH or 1 ?M
forskolin. Generation of hydrogen peroxide was measured at
45 min after UV exposure, as described previously (21).
Quantitation of cyclobutane pyrimidine dimers (CPDs)
CPDs were detected using flow cytometry analysis. Melano-
cytes were pretreated with 0 (control), 10 nM ?-MSH, or 1 ?M
forskolin for 4 d prior to, and 2 d after, exposure to 105
mJ/cm2UV, harvested 48 h post-irradiation, and fixed with
70% ethanol; the DNA was denatured with 1N HCl, then
renatured with sodium tetraborate. Melanocytes were incu-
bated overnight at 4°C with the anti-CPD antibody (diluted
1:500; TDM-2 clone), then with goat anti-mouse IgG Alexa
Fluor 488 (Invitrogen, Carlsbad, CA, USA) for 1 h, and finally
resuspended in PBS containing RNase A and propidium
iodide. Melanocytes were analyzed on a Coulter EPICS XL
2 Vol. 24 October 2010 KADEKARO ET AL.The FASEB Journal ? www.fasebj.org
Page 3
flow cytometer (Beckman Coulter, Miami, FL, USA) using a
488-nm argon ion laser.
Measurement of UV-induced apoptosis by Annexin Va
Melanocytes were maintained for the duration of the exper-
iment in medium lacking BPE, and phorbol 12-myristate
13-acetate (PMA; TPA), to determine the survival effect of
?-MSH in the absence of the antiapoptotic effect of TPA.
Melanocytes were treated with 0, 1 nM ?-MSH or 1 ?M
forskolin for 4 d prior to, and 24 h after, exposure to a dose
of 105 mJ/cm2UV, then stained for Annexin Va, as described
previously (21).
Transfection of HMs expressing loss-of-function MC1R with
the wild-type MC1R
Melanocytes naturally expressing nonfunctional MC1R (2n;
R160W/D294H) were transfected with pcDNA3 plasmid con-
taining the wild-type MC1R gene (pMC1R-WT) and pcDNA3
vector only, using the electroporation method from Amaxa
Transfection System (Lonza Walkersville, Walkerville, MD,
USA), then selected for stable transfection in medium con-
taining geneticin.
Ultrastructural visualization of DOPA-stained HMs
Parental HMs expressing loss-of-function MC1R and the
same HM strain stably transfected with wild-type MC1R
were plated in 4-well chamber slides and treated for 4 d
with 0 (control) or 1 nM ?-MSH. Cells were processed for
electron microscopy as described previously (28). Mic-
rotome sections were analyzed in a Jeol JEM-1230 transmis-
sion electron microscope (Jeol Ltd., Akishima, Japan),
and photomicrographs (?15,000) of the perinuclear area
and dendrites were taken for each experimental group.
The number (n?200) and stage of the melanosomes in the
perinuclear area and dendrites were determined in the
control and ?-MSH-treated cells from parental and trans-
fected HM.
Gene expression profiling and data analysis
Total RNA was isolated from HMs treated with 1 nM ?-MSH
as described for experiments to measure CPD, 8 and 24 h
after UV exposure, using RNeasy columns following the
manufacturer's protocol (Qiagen, Valencia, CA, USA).
After quality checks were performed with the Agilent 2100
bioanalyzer (Agilent Technologies, Santa Clara, CA, USA),
RNA was used to generate the cDNA that was hybridized
with in-house (Genomics and Microarray Core of the
University of Cincinnati) glass slide arrays (OmniGrid,
multiaxis microarrayer). The slides were prepared using
the DNA source from the Research Genetics Human
UNIGENE Library (31,742 clones) and scanned, and dif-
ferential gene expression levels were determined by the
calculated ratio of Cy-3 to Cy-5 emission, using the Axon
GenePixPro 3.0 software (Axon Instruments, Foster City,
CA). For data analysis, only genes with an expected false
positive less than one and more than 2-fold change in
expression were considered. A complete list of genes
included in the microarray analysis can be found online
(http://microarray.uc.edu). The arrays were normalized
and tested for differential expression using the methods
described in Borchers et al. (29).
Detection of MC1R cell surface expression
Melanocytes were treated with 0 (control) or 10 nM ?-MSH
and/or irradiated with 105 mJ/cm2UV. At 14 h after ?-MSH
treatment or 24 h after UV exposure, the culture dishes were
placed at 4°C for 2 h, and HMs were harvested with cold 5
mM EDTA. The cells were incubated with 4 ?g/ml rabbit
anti-MC1R (H-60; Santa Cruz Biotechnology, Santa Cruz, CA,
USA) for 1 h at 4°C, then incubated with 20 ?M goat
anti-rabbit Quantum Dot 655 for 1 h (Invitrogen). The cells
were postfixed in 1% paraformaldehyde and analyzed using a
BD LSRII flow cytometer (Becton-Dickinson, San Jose, CA,
USA). Ten thousand events were collected for each sample.
Western blot analysis
Melanocytes were treated as described for the microarray
experiments and cell extracts were obtained 24 or 48 h after
UV irradiation. Western blotting was carried out to validate
some of the microarray data, using the following antibodies
for Mitf (a gift from David Fisher, Harvard University, Cam-
bridge, MA, USA), Cdk2 (M2), PCNA (F-2), (Santa Cruz
Biotechnology), Peroxiredoxin 1 (Prx1; AbCam, Cambridge,
MA, USA), Bcl2 (Ab-1) (EMD, San Diego, CA, USA), and
DDB1 (Zymed Laboratories, San Francisco, CA, USA). As
control for loading, antibodies against histone H1 (AE-4) and
actin (C-11) were used (Santa Cruz Biotechnology).
MC1R genotype and activation of MC1R
A total of 21 different primary HM cultures, 14 derived
from neonatal foreskins (n) and 7 from individual adult
(a) skin samples, were established as described previously
(15) and utilized for the experiments described below.
MC1R genotypes, as well as eumelanin and pheomelanin
contents, were determined (Table 1), as described previ-
ously (27). Responsiveness to ?-MSH was first determined
by measuring the increase in cAMP levels and tyrosinase
activity. Strains highly responsive to ?-MSH were those
homozygous for the wild-type MC1R (?/?; 17n-19n, 20a,
and 21a), or heterozygous for the pseudoalleles (p/?)
V92M (14n and 15a) or R163Q (16n), or the mild allele
V60L (r/?; 13n) (Fig. 1). These strains exhibited ?70%
increase in cAMP levels and ?2-fold increase in tyrosinase
activity above control in response to 10 nM ?-MSH. In
contrast, HM strains that were unresponsive to ?-MSH
were those homozygous or compound heterozygous for 2
RHC alleles (R/R; 1n, 2n, 3a, 4n, 5n), or compound
heterozygous for the RHC allele R160W and V60L (R/r;
6n and 7a).
Results from HMs heterozygous for RHC alleles
(R/?) were variable. Strain 12a (R151C/?) was highly
responsive to ?-MSH in both assays, whereas 10n
(R160W/?) and 11n (D294H/?) demonstrated a rel-
atively low (?50% increase above control), yet statisti-
cally significant, stimulation of cAMP and no or modest
increase in tyrosinase activity, respectively, following
?-MSH treatment (Fig. 1). Statistical analysis revealed
that 12a had a significantly higher response than 10n to
Page 4
10 or 1nM ?-MSH, and in turn 10n had a significantly
higher response than 11n to 1 nM ?-MSH in the cAMP
assay, as determined by one-way ANOVA. As for activa-
tion of tyrosinase activity, 12a had a statistically higher
than 11n in response to 1 or 10 nM ?-MSH, as
determined by Student's t test. Expression of R163Q
with R160W in 8n did not contribute further to the
disruptive effect of the latter RHC allele (10n) on
MC1R activation by ?-MSH, because there was no
statistical difference in the response of the 2 strains to
1 or 10 nM ?-MSH in the cAMP assay. Melanocytes with
this genotype exhibited a modest increase in cAMP
(50% above control) and marked (2.5-fold) stimulation
of tyrosinase activity in response to 10 nM ?-MSH. As in
12a, which is heterozygous for R151C, 9a, compound
heterozygous for R151C and V92M, responded dramat-
ically to 10 nM ?-MSH with 4-fold increase in cAMP
formation, and 2.5-fold stimulation of tyrosinase activ-
ity. Depending on the availability of HMs, forskolin, a
direct activator of adenylate cyclase, was used as a
positive control and found to increase cAMP levels and
tyrosinase activity, regardless of MC1R genotype.
MC1R genotype and response of HMs to UV
We have reported that ?-MSH inhibits the UV-in-
duced generation of hydrogen peroxide by HMs
within minutes (21, 22). Melanocyte strains 7a
(V60L/R160W), and 2n, 3a, 4n, and 5n, which
express 2 RHC alleles, with the exception of 1n,
which was homozygous for R160W, failed to respond
to ?-MSH with significant inhibition of hydrogen
peroxide generation, as measured 45 min after expo-
sure to 105 mJ/cm2UV (Fig. 2A). In contrast, all
remaining strains showed a marked decrease (25–
65%) in hydrogen peroxide generation after ?-MSH
In preliminary experiments, we compared induction
of CPD immediately after UV irradiation in untreated
vs. ?-MSH-treated HMs and found that ?-MSH did not
affect initial CPD formation (data not shown). How-
ever, treatment with ?-MSH enhanced the repair of
CPD in HMs, as measured 48 h after UV exposure,
except in those expressing 2 RHC alleles (1n–5n) or
compound heterozygous for V60L and R160W (7a)
(Fig. 2B). The latter strains maintained high levels of
CPD with no evidence of significant removal. These
results were validated using Southwestern blot analysis
of 2 HM strains, 1n (R160W/R160W) and 4n (R151C/
D294W) (data not shown).
We examined the effect of ?-MSH on UV-induced
apoptosis (Fig. 2C). With the exception of HM strains
expressing 2 RHC alleles (1n, 2n, 3a, 4n, 5n), 6n and 7a
(V60L/R160W), 14n (V92M/?) which was the most
TABLE 1.MC1R genotype, eumelanin and pheomelanin content, and response to ?-MSH in various assays of HM strains
Strain MC1R genotypeEumelanin (ng/106cells)Pheomelanin (ng/106cells)cAMP Tyrosinase activity ApoptosisH2O2
Strains: R, RHC alleles; r, V/60L; p, pseudoallele; ?, wild-type allele; a, adult; n, neonatal. Response to ?-MSH in the various assays: ?, high
response; ?, lack of response; ?, partial response; ND, not determined. ?, most resistant strain to UV-induced apoptosis even 48 h after UV
4Vol. 24 October 2010 KADEKARO ET AL. The FASEB Journal ? www.fasebj.org
Page 5
resistant to UV-induced apoptosis, and the 2 strains 9a
(R151C/V92M), and 10n (R160W/?), the remaining
HM strains responded to 1 nM ?-MSH with reduction
of the apoptotic effect of 105 mJ/cm2UV. The latter 2
HM strains (9a and 10n) were responsive to the anti-
apoptotic effect of forskolin but not ?-MSH, despite
significant reduction in hydrogen peroxide generation
and enhanced CPD repair in response to ?-MSH treat-
ment, suggesting inefficient activation of survival path-
ways by the dose of 1 nM ?-MSH (Fig. 2A, B).
Transfection with wild-type MC1R restores MC1R
function and normalizes the UV response of HMs
naturally expressing 2 RHC alleles
To establish the causality between lack of response to
?-MSH and the aberrant response to UV, we stably
transfected the HM strain 2n (R160W/D294H) with
wild-type MC1R. Electron microscopic examination of
HMs transfected with the wild-type MC1R and incu-
bated with L-DOPA, the substrate for tyrosinase,
showed that unlike the parental strain, they responded
to ?-MSH with stimulation of melanogenesis (Fig. 3A),
evidenced by enhanced maturation of melanosomes
(an increase in the number of mature stage III and IV
melanosomes) in the perinuclear area and dendrites,
due to increased deposition of melanin. In HMs trans-
fected with the wild-type MC1R, ?-MSH reduced UV
(105 mJ/cm2)-induced hydrogen peroxide production,
enhanced repair of CPD, and inhibited apoptosis (Fig.
?-MSH- and UV-altered gene expression
We compared global gene expression in 2 HM
strains, 13n (V60L/?), highly responsive to ?-MSH
(HM-F), and 2n (R160W/D294H) with loss-of-
function MC1R (HM-LOF) (Fig. 4A), as determined
by cAMP and tyrosinase activity assays (Table 1 and
Fig. 1). The groups that were directly compared
against each other are shown in Fig. 4A. The changes
in gene expression resulting from ?-MSH-treatment
were observed exclusively in 13n, HM-F (compare
lanes 1 and 2 that were almost black, indicating no
Figure 1. Effects of ?-MSH on cAMP genera-
tion (A) and tyrosinase activity (B) of HM
strains with different MC1R genotypes. Data
points represent the mean ? se percentage of
control of triplicate determinations. Data pre-
sented are representative of the results of one
experiment that was repeated at least twice with
similar findings. *P ? 0.001 vs. control; ANOVA
followed by Newman-Kuels multiple compari-
son test. R/R, homozygous or compound het-
erozygous for RHC alleles; R/r, compound
heterozygous for RHC and V60L; R/?, het-
erozygous for RHC allele; R/p, compound het-
erozygous for RHC allele and a pseudoallele
(V92M or R163Q); r/?, heterozygous for
V60L; p/?, heterozygous for a pseudoallele;
?/?, wild type.
Page 6
alteration in gene expression, and lanes 3 and 4 of
the ?-MSH treated groups in the heat map; Fig. 4C).
In HM-F, we identified changes in expression of
genes associated with functional categories such as
cell adhesion, transcription factors, intracellular sig-
naling, protein trafficking, and biosynthesis in re-
sponse to ?-MSH (Fig. 4B). Global gene expression
in HM-F (13n) showed that a total of 3458 genes were
significantly altered in expression, with 668 genes af-
fected only by ?-MSH, 454 genes affected only by UV,
and 2336 genes influenced by both ?-MSH and UV.
Because our aim was to investigate the photoprotec-
tive mechanisms of ?-MSH, we focused our analysis of
the microarray data on gene categories that were
melanocyte-specific or involved in control of melano-
genesis, oxidative stress, DNA repair, cell cycle, and
apoptosis. Data analysis of genes belonging to these
functional categories revealed that ?-MSH had the
opposite effect of UV on the expression of 34.6% of all
UV and ?-MSH-responsive genes. Of those, 78% were
up-regulated by ?-MSH (Fig. 4D). In contrast, in HM-
LOF, comparison of the group treated with ?-MSH and
irradiated with UV to that only irradiated with UV
revealed no differences in gene expression between the
2 groups, indicating lack of modulation of the UV
response in HM-LOF by ?-MSH. Similarly, comparison
of HM-LOF treated with ?-MSH and irradiated with UV
to the same HMs treated with ?-MSH showed that the
genes that were altered were those only affected with
UV, further confirming lack of response of these HMs
to ?-MSH. Important conclusions derived from the
microarray data were that 1) ?-MSH mostly induced,
while UV mainly suppressed, gene expression; 2) ?-MSH
and UV had opposing effects on many of the analyzed
genes, and ?-MSH modulated the effects of UV on these
genes (Supplemental Data); and 3) loss of function of
MC1R compromised the UV response of HMs, due to
absence of the effects of ?-MSH on important genes that
affect the DNA damage response.
Validation of microarray data
To validate the microarray data, we selected genes
representative of the functional categories of interest.
The microarray result showed that MC1R expression
was suppressed 7-fold by UV, and induced 7-fold by
?-MSH. We confirmed these results by demonstrating
with immunostaining increased cell surface expression
of MC1R in HMs following ?-MSH treatment, and its
reduction following exposure to UV (Fig. 5A). By
Western blotting, we verified that the transcription
factor Mitf, the master regulator of melanogenesis and
melanocyte survival (30), was induced by ?-MSH and
repressed by UV (Fig. 5B). Peroxiredoxin1 (Prx1),
which reduces endogenous peroxides (31, 32), was one
Figure 2.
peroxide generation, repair of CPD, and apoptosis of HM
strains with different MC1R genotypes. A) Effects of 1 nM
?-MSH or 1 ?M forskolin on UV-induced generation of
hydrogen peroxide that was determined 45 min after UV
irradiation. ?-MSH is expected to reduce hydrogen peroxide
generation only in HMs expressing functional MC1R. Data
points represent the mean ? se percentage of UV-induced
hydrogen peroxide generation; 3 independent determina-
tions. B) Repair of DNA photoproducts was determined 48 h
after UV exposure by immunostaining of CPD and flow
cytometry analysis of cells (n?104). Data are expressed as
mean ? se percentage of median fluorescence of the UV
group; 3–5 determinations/group. C) UV-induced apoptosis
was determined by flow cytometry analysis of Annexin V-
stained cells (104cells from each of triplicate samples/
Effects of ?-MSH on the UV-induced hydrogen
group). Data are presented as mean percentage increase in apoptosis above control. All data are representative of the results
of one experiment that was repeated at least twice with similar trends.
6Vol. 24October 2010KADEKARO ET AL. The FASEB Journal ? www.fasebj.org
Page 7
of the most ?-MSH up-regulated oxidative stress regu-
latory genes that was suppressed by UV, according to
the microarray data. Western blotting showed in-
creased protein levels of Prx1 in response to ?-MSH,
and decreased levels following UV exposure (Fig. 5B).
The 2 DNA repair genes, damaged DNA binding
protein 1 (DDB1), which promotes global genomic
repair in the nucleotide excision repair pathway and
regulates DNA replication (33), and proliferating cell
nuclear antigen (PCNA), which plays important roles in
DNA replication and repair as well as DNA methylation,
chromatin assembly, and remodeling (34, 35), were
found to be increased by ?-MSH, and the latter to be
repressed by UV. Western blot analysis of DDB1 and
PCNA revealed that their levels were increased by
?-MSH and reduced by UV, and that ?-MSH partially
reversed the inhibitory effects of UV (Fig. 5B). We validated
by Western blotting the expression of cyclin-dependent
kinase 2 (CDK2), which binds to cyclin E, to allow entry of
cells into S phase, or to cyclin A, to enhance progression to
by ?-MSH and diminished by UV, in agreement with the
microarray data (Fig. 5B). The antiapoptotic Bcl2 is critical
for melanocyte survival (37); it was reduced at the protein
level by UV and increased by ?-MSH, which confirmed the
microarray results and our previous findings (21).
Figure 3. Transfection of HMs with loss-of-function MC1R with the wild-type MC1R restores responsiveness to ?-MSH and
normalizes the response to UV. HM strain 2n (R160W/D294H) was stably transfected with empty vector or wild-type MC1R.
A) Electron microscopic examination of parental HMs and HMs transfected with wild-type MC1R reveals stimulation of
melanogenesis by treatment of the latter HMs with 1 nM ?-MSH. Number of melanosomes in each maturation stage located in
the perinuclear and dendrites is expressed as percentage of the total number of melanosomes counted (n?200/group). *P ?
0.05 vs. parental control group; Student's t test. B) Expression of wild-type MC1R in 2n restored responsiveness to ?-MSH with
reduction of UV-induced hydrogen peroxide. C) Melanocytes transfected with wild-type MC1R, but not the parental strain 2n,
responded to ?-MSH with enhancement of repair of CPD. *P ? 0.001 vs. UV-irradiated group; ANOVA followed by
Newman-Kuels multiple comparison test. D) Transfection with the wild-type MC1R restored the ability of HMs to respond to
?-MSH with reduced UV-induced apoptosis.
Page 8
8 Vol. 24October 2010KADEKARO ET AL.The FASEB Journal ? www.fasebj.org
Page 9
In the present study, our goal was to elucidate the
impact of RHC MC1R alleles expressed as heterozy-
gous, homozygous, or compound heterozygous with
other commonly expressed alleles, in comparison to
pseudoalleles (V92M and R163Q) and wild-type allele,
on MC1R function and the UV response of HMs. The
MC1R is a melanoma susceptibility gene (7, 8, 10,
23–25); however, the molecular mechanisms by which
MC1R protects HMs from malignant transformation to
melanoma are not understood. By comparing a panel
of HM strains that naturally express different MC1R
genotypes (Table 1), we found that those expressing 2
RHC alleles (1n-5n) or compound heterozygous for
V60L and R160W (6n, 7a) were refractory to ?-MSH
(Table 1 and Fig. 1) (27). Expression of these geno-
types was associated with persistence of CPD and, with
the exception of R160W/R160W (1n), sustained hydro-
gen peroxide generation in UV-irradiated HM, in the
presence of ?-MSH (Fig. 2A, B). These results are
consistent with earlier genetic studies that RHC alleles
reduce MC1R function by different extents (38) and
suggest that MC1R is linked to signaling pathways other
than cAMP, such as inositol phospholipids and protein
kinase C (39, 40).
We have examined the possible dosage effect of various
MC1R variants suggested by Palmer et al. (7) (Table 1 and
Figs. 1 and 2). Melanocyte strain 10n (R160W/?) and
to ?-MSH with increased cAMP levels and tyrosinase
activity (Fig. 1), yet were efficient in reducing UV-induced
hydrogen peroxide production and apoptosis, and in
repairing CPD when treated with ?-MSH (Fig. 2). In
contrast, HMs heterozygous for the RHC allele, R151C
(12a), or for V60L (13n), V92M (14n, 15a), or R163Q
(16n), showed no evidence for reduced MC1R activity.
We conclude that RHC alleles differ in their heterozygous
effects, and that signaling pathways other than cAMP
possibly mediate the effects of ?-MSH and/or that mild
activation of cAMP is sufficient to activate antioxidant and
nucleotide excision repair pathways. Others have re-
ported that expression of any MC1R variant allele
with an RHC allele further increases the risk for
melanoma (8). In the HM strains 8n and 9a (R/p),
we found no evidence for further disruption of
MC1R by coexpression of a pseudoallele R163Q or
V92M with RHC allele. These novel results define the
interactions of these allelic variants based on multi-
ple biochemical assays that determine MC1R func-
tion and its role in the UV response of HMs.
Figure 4. Microarray experimental design, heat map, and data analysis. Microarray experiments were carried out on 13n that
expresses functional MC1R (HM-F) and 2n that expresses loss-of-function MC1R (HM-LOF). A) Schematic representation of
microarray experimental design. B) Heat map representing the global genes that were up-regulated (red) or down-regulated
(green) by the different treatments, 8 or 24 h after UV irradiation. C) Graphic representation of global gene analysis based on
various functional categories in HM-F treated with ?-MSH. D) Genes significantly altered in expression in HM-F in the listed
categories. For ?-MSH altered groups, up-regulated genes are blue. For UV-affected genes, down-regulated genes are green. For
the ?-MSH and UV-affected group, genes regulated in the opposite direction by ?-MSH and UV are yellow; of those, genes
up-regulated by ?-MSH and down-regulated by UV are pink.
Figure 5. Validation of altered expression of selected genes belonging to the melanocyte-specific, oxidative stress, DNA damage,
cell cycle, and apoptosis functional categories. A) Altered cell surface expression of MC1R in HM-F, as determined by
immunostaining and flow cytometry analysis. B) Western blot analysis for Mitf, Prx1, DDB1, PCNA, Cdk2, and Bcl2. Histone is
used as loading control for PCNA immunoblot; actin is used as a loading control for the other proteins.
Page 10
To confirm that lack of response of HMs expressing
2 RHC alleles to ?-MSH and UV is due to their MC1R
genotype, we stably transfected HM strain 2n (R160W/
D294H) with the wild-type MC1R and found that this
restored the response to ?-MSH and reduced the burden
of UV-induced oxidative stress and DNA damage (Fig. 3).
Interestingly, in the absence of ?-MSH, the extent of
UV-induced apoptosis of HMs transfected with wild-type
MC1R was markedly less than that of the LOF parental
HMs, possibly due to increased constitutive activity of the
receptor encoded for by the wild-type MC1R.
The results of the above biochemical assays were
consistent with microarrray data, which showed that in
HM-F, but not in HM-LOF, ?-MSH altered gene expres-
sion (Fig. 4B), and antagonized the effects of UV on
many genes, such as those regulating melanogenesis,
oxidative stress, DNA repair, cell cycle, and apoptosis
(Fig. 4D). Generally, ?-MSH up-regulated, while UV
down-regulated, global gene expression (Fig. 4C, D).
The suppressive effects of UV on gene expression was
also observed by others (41–43) and is thought to be
due to UV-induced DNA damage that inhibits tran-
scription by stalling RNA polymerases (42). Because of
the stochastic nature of UV-induced DNA damage, the
inhibition of a particular gene is proportional to its size
(43). We propose that by increasing CPD repair,
?-MSH reduces the number of these offending lesions
and alleviates the repressive effect of UV on gene
The effect of ?-MSH on the induction and repair of
UV-induced DNA damage was independent of increased
eumelanin synthesis, because eumelanin content was not
changed by any of our experimental protocols (data not
shown). In addition to pigmentation, inefficient DNA
repair is a risk factor for melanoma (44, 45). Our results
underscore the significance of functional MC1R in reduc-
DNA photoproducts and reducing the extent of oxidative
damage, in addition to its classic role as regulator of
melanogenesis. Reduced DNA photoproducts and in-
creased expression of DNA repair genes by melanocortins
were reported by others (46); our recent findings re-
vealed reduction of UV-induced oxidative DNA damage
by ?-MSH (22). Elucidating the impact of different MC1R
genotypes on the responses of HMs to ?-MSH and UV
sheds light on the molecular mechanisms by which MC1R
functions as a melanoma susceptibility gene, and explains
the differential susceptibility to melanoma.
The authors thank Silva Terzieva for technical assis-
tance, Toshio Mori (Nara Medical University, Kashihara,
Japan) for donating TDM-2 antibody, Jose Carlos Garcia
Borron (University of Murcia, Murcia, Spain) for his gift of
the pMC1R-WT plasmid, and David Fisher (Harvard Uni-
versity, Cambridge, MA, USA) for providing the Mitf
antibody. This work was supported in part by U.S. National
Institutes of Health grants R01ES009110 (Z.A.M.),
R011ES017561 (Z.A.M. and S.L.), and P30 ES06096
(Z.A.M. and D.S.); the University of Cincinnati Cancer
Center Pilot Project (Z.A.M.); Procter and Gamble Co.
(Cincinnati, OH, USA) (Z.A.M.), a Dermatology Founda-
tion research grant (S.L.); Ohio Cancer Research Associ-
ates (A.L.K.); and the Japan Society for the Promotion of
Sciences, KAKENHI grant 20591357 (K.W. and S.I.).
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Received for publication March 9, 2010.
Accepted for publication May 20, 2010.

subset: commoncrawl


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June 20, 2013
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* This offer can not be combined with other special offers or discounts.
Freight Free – 48 Hour Sale!
Buy any 12 bottles of wine until midnight tomorrow (Thursday) at www.winesale.co.nz and pay no delivery costs!
Whitehaven Wines
Stunning Whitehaven Wines on Special
Whitehaven have been hitting it out of the park lately scooping some major awards a long the way.
Today we have a beautiful Whitehaven Oak Aged Hidden Barrel Sauvignon Blanc which we are selling for just $12.99. Whitehaven would normally sell this wine for much more so these are big savings on the RRP. This is gorgeous sauvignon blanc which will match with a wide range of foods and will continue to improve in the bottle over the next 3 – 4 years. This is for the connoisseurs!
Also on special from Whitehaven:
Whitehaven 2011 & 2012 Pinot Grigio – Now only $12.99 a bottle. This is a delicate food winewhich show the real class you would expect from Whitehaven – not your average Pinot Gris
Whitehaven 2013 Chardonnay – has just won a Gold Medal at the Royal Easter Show. This wine is just getting better and better. Full bodied and beautifully integrated, sublime fruit, acid and oak balance this is chardonnay to die for $22.99 per bottle of just $20.99 on case buys.
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Bannock Brae
Bannock Brae 2009 Barrel Selection – End of Vintage!
If you managed to grab a copy of the NZ Herald on Saturday and read John Hawkesby's wine column you would have read how highly he rates the Bannock Brae Barrel Selection 2009 Pinot Noir dubbing it one of the best Pinots in the country. Read article here. What he didn't say is that the 2009 vintage has rolled over and this will be a very hard wine to find soon. We have only 100 bottles in store and this is the very last of this vintage we will be able to get our hands on. So be quick! Just $55.99 a bottle.
Bannock Brae Goldfields 2012 Pinot Noir – $31.99
At the Bannock Brae winery the Goldfieds Pinot Noir gets exactly the same treatment as its Barrel Selection big brother. Unlike many other wineries the Goldfields is not a second label where shortcuts are taken to meet the market. The Goldfields is made from the same grapes and undergoes the exact same winemaking process as the Barrel Selection. The only difference being the Barrel Selection is made only from select barrels of the vintage. Make no mistake this is a first class wine in its own right.


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Monday, April 7 2008
P.O.D.: When Angels & Serpents Dance
Meet the new P.O.D. same as the old P.O.D. (minus the hip-hop, that is.) Marcos is back! Cue the power ballads and let the dance begin, again.
Sunday, January 7 2007
P.O.D.: Greatest Hits: The Atlantic Years
The Southtown boys go in blind for their first greatest hits compilation.
Tuesday, January 2 2007
P.O.D.: Testify
What went wrong with Testify?
Monday, September 10 2001
P.O.D.: Satellite
Saturday, December 31 1994
P.O.D.: Brown
P.O.D.: The Fundamental Elements of Southtown [31.Dec.94]


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TYBEE ISLAND, Ga. (AP) — After 16 hours stuck at sea, passengers stranded on a casino boat that ran aground off Georgia's coast were ferried to shore Wednesday aboard two Coast Guard cutters.
After 16 hours, an exit from stalled casino boat
About 94 of The Escapade's 96 passengers and 27 crew members arrived at the dock shortly after 4 p.m., nearly a full day after the boat left for the maiden voyage of its operator's new Savannah service Tuesday night. A second cutter carrying 20 more people arrived shortly afterward. Four people were ferried ashore by helicopter.
Passengers walked slowly off the boat and down the dock to dry land, several of them rubbing their eyes after a five-hour cruise turned into an overnight affair.
"There was a lack of sleep but they kept us entertained," said Bernard Yount of nearby Springfield, Georgia. "I don't really think anybody was scared. I didn't see anybody panic. A few people were maybe worried about kids being home or animals being in the house."
The crew gave passengers sodas and water — no alcohol — to drink and fed them hotdogs late at night. On Wednesday morning, they were served bacon and eggs, said passenger Michael Alcott of Savannah. The ship's generator kept the lights on and the air conditioning going.
"The AC was blowing cold. They had food," said Alcott, 39. He said people slept on the floor using life jackets as pillows.
Both Alcott and passenger Dina Cook of Savannah said they felt the ship strike something at 9:30 p.m., but passengers weren't notified they had run aground until after midnight. The Coast Guard said it was notified about midnight that the ship was stuck.
"It was nerve-wracking, the fact that we were not being told anything," Cook said.
Initial attempts to tow the boat failed when the tow lines broke, so those aboard were transferred first to small boats that hold about eight people, then to the two larger vessels, Petty Officer 1st Class Lauren Jorgensen said.
Tara Sinclair, waiting at the Savannah dock for her mother, 66-year-old passenger Veronica Heyward, said her mom told her passengers had to "jump from the big boat to the little boat and then climb a rope ladder" onto the cutter. "She called it a Fear Factor moment," Sinclair said.
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Sleepside: The Collected Fantasies of Greg Bear
Greg Bear has been called the "best working writer of science fiction" by The Ultimate Encyclopedia of Science Fiction. He has been awarded two Hugo and four Nebula Awards, and holds the distinction of being one of two authors to win a Nebula in every category. His novel Darwin's Radio, published by Ballantine Books, won the Nebula Award for "Best Novel of 2000," and was honored with the prestigious Endeavor Award. His novels and short stories have been translated into fifteen languages.
Now, ibooks is proud to present a fantasy-themed companion volume to the bestselling The Collected Stories of Greg Bear (published by Tor in 2003). This exclusive volume features the stories "Webster," "The White Horse Child," "Sleepside Story," "Dead Run," "Through Road No Whither," and "Petra." Also includes a new introduction by the author: "On Losing the Taint of Being a Cannibal."
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Review: Sleepside: The Collected Fantasies of Greg BearUser Review - Michael O'Donnell - Goodreads
Not too bad. Did not get into 'The Way of all Ghosts'. Probably needed to read some of the earlier 'Way' storied. Read full review


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Photo-hosting sites?
Right now I have PhotoBucket for sharing with family & close friends & love it. I used my hotmail (junk mail) address for that.
I need another photo-hosting site for sharing with the two forums I'm a member of (THS/GW & a beauty, fashion, etc., site) since I can't use my hotmail address for another PhotoBucket account & I'm not about to give them my Yahoo! (communicating w/ family & friends) address.
So what photo-hosting sites would you recommend that are similar to PhotoBucket?
there are tons of places for Free Image Hosting
another alternative for when you fins one or two that you really like and are comfortable with, is create a new free email account that is disposable and keep a list of them, then you can get unlimited accounts at any free hosting service.


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New Movies: 'Paranormal Activity — The Marked Ones'
Americans Trust Nurses, Teachers, Military Officers but Not Lobbyists or Politicians
Gallup just released the results of its annual survey investigating Americans' opinions on the trustworthiness of certain professions. In one notable surprise, members of the clergy saw their favorable rating drop below 50 percent for the first time.
Waitress Receives Massive Tip After Returning Lost Money to Wrong Guy
Massive Mega Millions Jackpot Hits $636 Million, May Reach $1 Billion
The Mega Millions lottery drawing being held Tuesday night is now for a pot of an estimated $636 million, not far off from the record $656 million Mega Millions prize awarded in March 2012. If there's no winning ticket, then Friday's drawing could top $1 billion.
Family and Friends of Newtown Victims Gather to Commemorate First Anniversary
The family and friends of the 26 people killed a year ago at Sandy Hook Elementary School in Newtown, Connecticut, held a candlelight vigil Thursday night in Washington.
New Movies: 'The Hobbit: The Desolation of Smaug,' 'A Madea Christmas'
New Movies: 'Out of the Furnace,' 'Inside Llewyn Davis'
Nelson Mandela Dies at Age 95
New Movies: 'The Hunger Games: Catching Fire,' 'Delivery Man'
50 Years Since JFK Assassination — A Look Back With Photos, Video and More
It's been 50 years since President John F. Kennedy was shot and killed while riding in a motorcade through Dallas, Texas. More words have been written about this event (and its aftermath, and what led up to it) than just about anything else, so we thought we'd take a look back through photographs and video.


text: Import Export Product Description HS Code India export data and statistics of June 2011 of chapter 17-Sugars and Sugar Confectionery. Live online India Export Data Of Chapter 17 Sugars And Sugar Confectionery December 02, 2011 , New Delhi Total Export value in June of all products categories under Chapter 17 Sugars And Sugar Confectionery was USD 214.5 M, a increase of 15.4% compared to May 2011. Here is a breakup of major products categories with growth rates. Sr No. HS Code Products Value USD M Growth Compare to Prev. Month Top 5 Indian Ports Top 5 Destination Countries 1 1701 Cane or Beet Sugar and Chemically Pure Sucrose, In Solid form 200.84 14.49% JNPT, Kandla, Madras Sea, Kakinada, Calcutta Sea Sri Lanka, Sudan, United arab Emirates, Yemen, Djibouti 2 1702 Other Sugars, Including Chemically Pure Lactose, Maltose, Glucose and Fructose, In Solid form; Sugur 5.49 0.4% Mundra, JNPT, Sabarmati ICD, Ahmedabad ICD, Pipavab Sudan, Kenya, United arab Emirates, Sri Lanka, Tanzania 3 1704 Sugar Confectionery (Including White Choco-Late), Not Containing Cocoa 4.76 -0.99% JNPT, Mundra, Bangalore ICD, Madras Sea, Raxaul Togo, Benin, Senegal, Mozambique, United arab Emirates 4 1703 Molasses Resulting from The Extraction or Refining of Sugur 3.41 1894.31% JNPT, Bombay Sea, Mangalore Sea, Delhi Air Venezuela, Netherlands, Germany, Italy These findings are based on India export data through June-2011 . InfodriveIndia Trade data is the most up-to-date information on India's foreign trade , It covers 94 ports in India like JNPT, Bombay Air and Sea , Chennai Air & Sea , Delhi IGI Air, Delhi Tughlakhabad ICD, Delhi Patparganj, Kolkata Air and Sea, Bangalore Air and many more. InfodriveIndia.com trade data is available for online analysis within 3 weeks from date of export import at InfodriveIndia.com compared to Govt sources which are around 6 months old. About InfodriveIndia InfodriveIndia.com is the market leader and primary source of India Export Import Trade information in India. The India Import Export data bank, which is at the core of InfodriveIndia.com trade information services is unique. It is based on the Bills of Entry and Shipping Bills filed at various ports, InfodriveIndia reporters collect this data from every Indian port, and InfodriveIndia database yields the most timely, accurate, comprehensive information available on trade through India Ports. More specialized InfodriveIndia databases are US Import Data, UK Import Data, China Export Import Data, Russia Export Import Data, Sri Lanka Export Import Data, Latin American trade data, Chile Export Import Data, Peru Export Import Data, Argentina Export Import Data, and Uruguay Export Import Data . Latest Import Export News DateSubjectCompiled By 26-03-2015India's gold imports to explode in MarchInfodrive India Team 26-03-2015Sesa seeks Goa government help to save junior academyInfodrive India Team 26-03-2015Government to bring out strategy paper on boosting textile exportsInfodrive India Team 26-03-2015India to boost LNG imports to raise power generationInfodrive India Team 26-03-2015No role for a few senior Ranbaxy execs in merged entity, says Dilip ShanghviInfodrive India Team 25-03-2015Essar pitching to OSO Rosneft for deeper equity allianceInfodrive India Team 25-03-2015USFDA issues import alert on two plants of Aarti DrugsInfodrive India Team 25-03-2015Goa economy grows despite mining ban, GSDP at 7.71%: ReportInfodrive India Team 25-03-2015Indians lap up diamond jewellery fastInfodrive India Team 25-03-2015Russia may allow imports of Indian dairy products soonInfodrive India Team


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Posts tagged "bristol palin"
All The Deets On Bristol Palin's Memoir, "Not Afraid Of Life"
Bristol Palin's memoir is coming out this summer and it will be called Not Afraid Of Life. Kudos to you for making reference to anti-abortion propaganda, Bristol! Not Afraid Of Life, which is being published by William Morrow, will cover her 20 years on Earth, including her appearance on "Dancing With…
Jessica Wakeman
March 3, 2011
Today's Lady News: Bristol Palin's Memoir Coming This Summer
Bristol Palin's memoir is set for publication in summer of 2011. Yes, her memoir. Damn, why don't I ever hear about these kinds of ghostwriting gigs? Actually, I don't think I could ghostwrite whatever party line Bristol's going to tow about abstinence with a straight face. [Gawker] The Protect Life Act, introduced by…
Jessica Wakeman
February 7, 2011
Ooh, Burn! Bristol Palin Calls Out Her Mom On Twitter UPDATE: It's A Hoax
As Jessica said, "Baby bear isn't gettin' any freshwater salmon tonight." Wonder what the story is here... [BuzzFeed] UPDATE: Turns out this is a hoax. I guess we should have known that no one messes with Mama Grizzly. [Mediaite]…
Amelia McDonell-Parry
January 10, 2011
Bristol Palin Buys A House In Arizona
Warning! Warning! Bristol Palin is encroaching on the continental United States. Alaska's most famous single mom recently paid $172,000 for a five-bedroom house in Maricopa, Arizona, which is 30 miles from Phoenix. The house was foreclosed last January, flipped by a real estate developer and sold to Mama Grizzly's cub one month ago.
Jessica Wakeman
December 27, 2010
Kathy Griffin And Bristol Palin's Fat Feud
Kathy Griffin, who maybe sort of had a fling or flirtation or something with Bristol Palin's ex, Levi Johnston, had some unkind words about Bristol during VH1 Divas Salute the Troops monologue this weekend. "She's the only contestant in the history of the show to actually gain weight ... She gained like…
Ami Angelowicz
December 7, 2010
Bristol Palin Says She Wasn't Forced On "DWTS," Has Her Mother's Back
"I will give my friend credit for creativity, and extra points for getting so many 'facts' wrong in so few sentences. Let me be blunt: my mom did not 'force' me to go on 'DWTS.' She did not ask me either. The show approached me. I thought about it. I made the decision. It sadde…
Kate Torgovnick/Kate-Book.com
December 6, 2010
Margaret Cho Says Sarah Palin "Forced" Bristol To Compete On "Dancing With The Stars"
"I heard from someone who really should know (really should seriously know the dirt really really) ... Why did Bristol do 'Dancing with the Stars'? That the only reason Bristol was on the show was because Sarah Palin forced her to do it. Sarah supposedly blames Bristol harshly and openly (in the…
Jessica Wakeman
December 1, 2010
The "Dancing With The Stars" Finale: Bristol Palin Versus Jennifer Grey
Last night on "Dancing with the Stars," the mirror ball hopefuls got one last chance to impress America with a redemption dance, to make up for their biggest travesty of the season, and a freestyle dance of their choice. The highlight of the night was Jennifer Grey's freestyle, during which she carried a…
Kate Torgovnick/Kate-Book.com
November 23, 2010
Bristol Palin Receives "DWTS" Death Threat
Bristol Palin is turning out to be the most controversial "Dancing With the Stars" contestant ever. People are calling "conspiracy theory" on her miraculous winning streak. A Wisconsin guy even shot at his television screaming "f**king politics" after her performance last week, which earned her a spot in the finale tonight. But…
Ami Angelowicz
November 22, 2010
Man Shoots TV After Watching Bristol On "Dancing With The Stars"
More than a few people are irked by Bristol Palin's staying power on "Dancing with the Stars." And one man in Wisconsin just couldn't take it anymore. Steven Cowan, 66, was kicking back on Monday and having a few drinks while watching the show. But after this performance by Bristol, he jumped u…
Kate Torgovnick/Kate-Book.com
November 18, 2010
Willow Palin Uses Homophobic Slurs, Bad Grammar, On Facebook
It's wash-your-mouth-out-with-soap time at the Palin household. After people criticized the family's new TLC reality show, "Sarah Palin's Alaska," on Facebook, little sis Willow Palin busted out with homophobic slurs, put-downs and grammar gone rogue. "Haha your [sic] so gay. I have no idea who you are. But what I've seen pictures of,…
Jessica Wakeman
November 17, 2010
Bristol Palin And The Situation Talk About Sex, Baby
It seems as if this would be a "Saturday Night Live" skit, not an actual safe sex PSA, and yet The Candie's Foundation has managed to blow our minds with a little video clip of Bristol Palin and "Dancing With the Stars," um, coworker, Mike "The Situation" Sorrentino talking about abstinence. The Situatio…
Julie Gerstein
November 16, 2010
Oopsie! Bristol Palin Forgot To Vote
"I did not send in my absentee ballots to Alaska. I'm going to be in trouble. Sorry, mom!" -- Bristol Palin on neglecting her civic duty. "Dancing With the Stars" is waaay more important to the future of our country. I never cease to be amazed at the values that Mama Grizz imbued…
Ami Angelowicz
November 4, 2010
Bristol Palin Dances In A Monkey Suit
Last night's "Dancing with the Stars" brought a very special surprise. Bristol Palin and Mark Ballas dancing in monkey suits while doing the jive to the theme song from my favorite old TV show, "The Monkees." See the girl monkey who can't quite keep up? Yeah, that's Bristol. I wonder if Sarah…
Kate Torgovnick/Kate-Book.com
October 19, 2010
Bristol Palin And The Situation Get Sexy On "Dancing With The Stars"
So, sue us, we are watching this week's episode of "Dancing with the Stars" a little late. But we just had to bring you this clip of Bristol Palin getting sexy while rumba-ing with Mark Ballas to "Umbrella." Her dancing was totally stiff, but that hip shimmy move wasn't bad. And the faux…
Kate Torgovnick/Kate-Book.com
October 13, 2010
"Dancing With The Stars" Courted Todd Before They Asked Bristol
"They wanted Todd to be on the show. I think that would have been cool to see, too. But here Bristol is… doing something all new ... [If I were on the show], they would have to let me dribble back and forth and maybe pivot and do a jump shot or something as part…
Kate Torgovnick/Kate-Book.com
October 8, 2010
Bristol Palin Plays Mother Nature In A New Music Video
In the spirit of one-up-manship, Bristol Palin proves that her ex Levi Johnston isn't the only person with music video cred. Just days after Levi's music video debut, Bristol announces her own cameo in Alaskan rock band Static Cycle's new vid. She plays "a mother nature role" with a babushka,…
Ami Angelowicz
October 8, 2010
Sarah Palin Gets Booed On "Dancing With The Stars"
Last night, Sarah Palin dropped by to watch the live filming of "Dancing with the Stars," sitting in the front row. But apparently, several members of the audience could not contain their lack of excitement and booed her as she took her seat. The boos could be heard in the telecast—they happened before…
Kate Torgovnick/Kate-Book.com
September 28, 2010
Mark Ballas Goes Home To Alaska To Meet Bristol's Parents
Bristol Palin's "Dancing with the Stars" partner, Mark Ballas, made the trek to Alaska this weekend to meet the Palins, check out the frozen scenery, and play an acoustic guitar set at a bar called Rumrunners. He posted the pics above on his Twitter page captioned, "Me and my gorgeous partner Bristol ...
Kate Torgovnick/Kate-Book.com
September 27, 2010
Who Celebrities Want To See Win This Season Of "Dancing With The Stars"
Former "Dancing With The Stars" contestant Kelly Osbourne knows whom she is rooting for on season 11. Kelly says that her vote is going to Bristol Palin, for her bravery. "I have a real soft spot for Bristol Palin," said Kelly. "I really do. I think she's going out there and trying,…
Kelli Bender
September 24, 2010
Simply Irresistible
All »
The 20 Hottest Sex Scenes From "Game Of Thrones"
When you play the game of thrones, you either win or you die. But you get to f*ck a lot first. So . . . it's kinda worth it. The show is known for its gratuitous amounts of nudity and raunchy sex, so what better way to celebrate the impending Game of Thrones Spring premiere…
Tres Sugar
March 29, 2015
The 10 Definitive Cult Films You Absolutely Must See
What makes a movie a "cult classic"? Film history is littered with hundreds of films that never made it big, but developed small, passionate followings that allowed them to build up a reputation over time. Sometimes a good film is poorly marketed by a studio more concerned with their bottom line; other times genuinely bad…
Ask Men
March 28, 2015
Dave Grohl Helps Out A Dying Fan
A big Foo Fighters fan with terminal cancer will get to scratch off one item from his wish list: meeting Dave Grohl and the band. The band was flying home from Australia when a flight attendant told Grohl about the wish of her friend, Australian Ken Powell, 56, who has late-stage melanoma, report…
March 28, 2015
Amanda Knox's Murder Conviction Has Been Overturned
Foxy Knoxy is off the hook.
Claire Hannum
March 27, 2015
You'll Love
If These Style Blogger Poses Could Talk...
8 More Realistic Types Of Female Orgasms
6 Terrible Date Ideas Brought To You By Celebrity Guys


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I stayed here during thanks giving vacation and the highlights are following:
1) Great staff! They helped me in easy checkout and exceeded my expectation by offering the hospitality room service even without asking.
2)Location is great , the beach is just crossing a street. The hotel is near to the diamond head so you can go for hiking easily and can save some time.
2)our room got upgraded to full ocean view.
3)helpful concierge .
4)Hertx car rental right in the building.
5)the ABC store and the crepe place near by was perfect for doing little shopping and for the breakfast.
1)hotel rooms are little old atleast our room was old. may be the other tower is newer one.
2)its at one end of the waikiki which is good and bad both. its more quieter but at the same time main maket is about 10 min walk.
- Official Description (provided by the hotel):
- WAIKIKI BEACH MARRIOTT RESORT & SPA fronts the best stretch of Waikiki Beach. Conveniently located in the heart of Waikiki, the hotel offers 1,310 refurbished guest rooms and suites with private lanai providing spectacular views of world-famous Diamond Head and Waikiki Beach. Other amenities include Kuhio Beach Grill, featuring the best in Pacific Rim cuisine; Sansei Seafood Restaurant & Sushi Bar and d.k Steak House; Arancino di Mare Italian Restaurant; newly redesigned Moana Terrace, offering live Hawaiian entertainment nightly; Seattle's Best Coffee; Paul Brown's Spa Olakino * Salon; a surf school; two freshwater, heated swimming pools; 10-person whirlpool; new 3,000 sq. ft. 24-hour fitness center; business center with 24-hour Internet access; high-speed and wireless Internet access throughout the resort's guest rooms and public spaces; shops and boutiques; and Hawaiian cultural activities. ... more less
- Reservation Options:
- TripAdvisor is proud to partner with Expedia, Booking.com, Marriott, Hotels.com, Odigeo, TripOnline SA and Tingo so you can book your Waikiki Beach Marriott Resort & Spa reservations with confidence. We help millions of travelers each month to find the perfect hotel for both vacation and business trips, always with the best discounts and special offers.
- Also Known As:
- Waikiki Beach Marriott Hotel Honolulu
- Waikiki Beach Marriott Resort And Spa
- Marriott Waikiki Beach
- Marriott Honolulu
- Honolulu Marriott
- Waikiki Beach Marriott Resort & Spa Hawaii/Honolulu


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Daihatsu Mira E:s 2012 X grade 4.5 A auction grade, Black Color, Automatic, 660cc, 46,000km Driven, Two-Tone Interior, Luxury Fabric Seats, Eco Idle function, Digital Meter, Chilled A/c, Power Steering, Power Windows, Power Mirror, Retractable Mirrors, ABS Brake, SRS Dual Airbags, and lots of features. Original Auction Report Available, Car is original from Interior and Exterior, Bring your Denter / Mechanic to Believe. Best Car for family, Fuel Efficient Car. Car is parked at Plot in Khalid Bin Waleed Road.
Auction Report Interpretation: (Code 3529) - 4.5 Grade vehicle - Full Original - remote engine starter Merchants Automobile Specialty 1) All deals as per Original Auction Report 2) No meter or condition tempering 3) Money back guarantee if Auction Report proven fake Merchants Automobile believes in relationship than salesmanship. Our company specializes in imported vehicles purchased directly from Japanese auction. Since inception Merchants Automobile has worked on the philosophy as car consultant rather than car dealer hence we get the maximum value of money for our clients. Concerned sales person Sajid Hussain Sabir Molai
Assalam o Allaikum, Want to sale my fresh import Mira E:S Echo Idle Package: G with Push start (Top of the line) Condition: 4.5 Tyre condition 8/10 (as per Auction Sheet) Original Mileage: 36000 km Package Included:- Echo Idle Push Start Original Aluminium Alloy Rims Climate Control AC Traction Control 6 Airbags Leather Stitched Steering Tinted Glass & Security Alarm Original CD Player with AUX Retractable & Winker Side Mirror Extras: -Auto Headlights sensor -Auto Viper sensor -Parking Assist Sensor on front and rear bumpers -Stylish Seats Covers *Compact Smart Car *Very Economical Fuel Consumption *100% Genuine Condition Only serious buyer can contact me Demand is final
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