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I miss Playstation sometimes. My PS2 rarely (never) gets any love though I still have a short stack of games I want to play on the console. There are plenty of rumors of price drops and bundles for the PS3. One rumor I came across is that the new 40GB model is not going to be backwards compatible. Wha'?
Then there's the issue of well...the exclusive games just aren't there for the PS3 yet. GTAIV is going to have exclusive content for the 360. I read somewhere that Rockstar's La Noire is going to be PS3 exclusive - if it actually comes out. The game that is exclusive and looks completely awesome is Naughty Dog's Uncharted: Drake's Fortune. Really solid, top notch classic adventure games are pretty few and far between for some bizarre reason. Indiana Jones games are pretty much total disappointments. Uncharted has been getting some solid press and Naughty Dog knows what they are doing as far as making fun adventure games what with their Jak & Daxter series. Here's one of the trailers...and yes that's Nathan "Captain Tight Pants" Fillion:
Wednesday, October 10, 2007
A little love for the PS3.
Posted by Hooligan Youth Reviews at 11:49 AM
Labels: GTA: IV, LA Noire, Nathan Fillion, Naughty Dog, PS2, ps3, Uncharted: Drake's Fortune
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[hwee dzoo ng] /ˈʰwi ˈdzʊŋ/
1082–1135, emperor of China 1101–26: painter and patron of art.
Tagged: h
Read Also:
- Huitzilopochtli
[wee-tsee-loh-pohch-tlee] /ˌwi tsi loʊˈpoʊtʃ tli/ noun 1. the Aztec god of war and of the sun.
- Huizhou
[hwee-joh] /ˈʰwiˈdʒoʊ/ noun, Pinyin. 1. a city in S Guangdong province, in SE China, E of Canton.
- Huizinga
[hœi-zing-uh] /ˈhœɪ zɪŋ ə/ noun 1. Johan, 1872–1945, Dutch historian.
- Huk
[hook] /huk/ noun, plural Huks. 1. a member of the Hukbong Mapagpalaya ng Bayan (People's Liberation Army) a Communist agrarian revolutionary group in central Luzon in the Philippines. 2. a member of the Hukbalahap, an anti-Japanese resistance group in central Luzon during World War II.

subset: commoncrawl


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Marc her words . . God knows all about it
WE have just received the latest communication from Financial Astrology, an organisation which predicts world events 'based on the mathematical calculation and interpretation of planetary positions and correspondences.' Otherwise known as guesswork.
Financial Astrology is run by Rebecca Nolan, a professional mathematician who has been studying astrology for 25 years.
She says she cannot get her forecasts 100 per cent right 'otherwise I'd be God.' But she asserts she is 'getting better and better at it' which implies that God had better get used to the idea of cruising the classifieds for alternative employment.
For now, though, she claims a 71 per cent success rate for her predictions for the period March to May 1995, which may not be up to God's standards but which puts her definitely in the deity-in-waiting league.
It should be remembered that most financial experts are only right about 30 per cent of the time, which on the God scale puts them down there with the pagan worship figures of troglodytes, which seems fair enough.
Anyway, the latest newsletter informs us her predictions for the Hang Seng Index were 87 per cent correct for the month of June. There is a graph comparing her prediction with the actual performance of the index and they do, to some extent, correlate.
With an 87 per cent success record, Ms Nolan would appear to be getting dangerously close to challenging God for top spot, so it seemed in Lai See's interest to investigate her predictions for local events a little more closely.
In her predictions for the period June 1 to August 31, Ms Nolan says Mr Patten's temper will flare, causing market falls.
We learn Li Ka-shing will be 'selling property fast.' We are told that Deng Xiaoping will be 'dead or alive,' a prediction that covers most eventualities.
All pretty negative stuff. Indeed, there are going to be 'major falls in the index between July 13 and July 20', though it will 'pick up again' towards the end of the month.
The general conclusion is not good. She states confidently that 'Hong Kong is a dying market.' Basically, plenty of gloom and plenty of doom.
And then suddenly it all clicked. Rebecca Nolan is Marc Faber in drag.
Not content with one personality predicting dire and disastrous scenarios, Mr Faber, the Godfather of Glum, has invented a female alter-ego, Sister Solemnity, to match him doom for doom and gloom for gloom.
This man needs help. Consider the extent of the problem.
A financial expert given to gloomy predictions invents a rival character of the opposite sex who thinks she has the potential to be God.
Freud would have a field day.
Confirmed THE French: Another apology. We recently have implied the French have a long history of naval failure. We now accept this is completely true and only made this apology to mention it again.
Fun Times THE happy tenants of Wharf's gargantuan development, Times Square, who recently lobbied the landlord for rent reductions of up to 50 per cent alleging poor customer flow, poor signage and a host of other gripes, were thrilled to receive a letter from the management.
The letter was triggered by a breakdown in the building's automation system, which made all the alarms go off during a busy lunch hour.
The management conveyed its apologies as follows: 'We deeply appreciate the confusion caused.' Top guess OUR congratulations go out to Hobson's Property Consultants for attaining the highest standards in Making Blindingly Obvious Statements.
In a recent letter, the company announced proudly it had been appointed the leasing agent for the 28th floor of Concordia Plaza in Tsim Sha Tsui.
The company's letter says: 'The property, occupying the top floor, is not only well above other blocks in the neighbourhood, but is also higher than any other floors in the building.' How about that? The top floor is higher than the other floors. Extraordinary.
Same to you! WHILE you may be thinking twice about doing business with the above mentioned firm, you would definitely cogitate long and hard about moving into this new building in Kowloon.
Hong Kong landlords are notorious for treating their tenants with the minimum of respect, but the owners of this edifice appear to be the masters.
Tenants will have the undiluted pleasure of telling people they live in wonderfully-named Gofuku Tower.
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In this powerful, soul-searching memoir, beautifully written in the vein of A Pack of Two and Wild, animal behaviorist Dr. Patricia McConnell recounts for the first time the compelling story of her dark past, memories of which are triggered by a troubled dog named Will.
World-renowned as a source of science and soul, Patricia McConnell combines brilliant insights into canine behaviorgained from her work with aggressive and fearful dogswith heartwarming stories of her own dogs and their life on the farm. Now, she reveals that it wasnt just the dogs who had serious problems. For decades Dr. McConnell secretly grappled with her own guilt and fear, which were rooted in the harrowing traumas of her youth.
Patricia is forced to face her past by her love for a young Border Collie named Will, whose frequent, unpredictable outbreaks of fear and fury shake Patricia to her core. In order to save Will from this dangerous behavior, she must find her own will to heal, and along the way learn that will power by itself is not enough.
Interweaving enlightening stories of her clients dogs with tales of her deepening bond with Will, Patricia recounts her fight to reclaim her life. Hopeful and inspiring, the redemptive message of her journey is that, while trauma changes our brains and the past casts a long shadow, healing, for both people and dogs, is possible through hard work, compassion, and mutual devotion.
Patricia McConnell, PhD, is an internationally known Zoologist and Certified Applied Animal Behaviorist who has treated serious behavior problems in dogs for over twenty-five years. She speaks around the world about canine behavior and training, and is the author of fourteen books, including the critically acclaimed The Other End of the Leash: Why We Do What We Do around Dogs. Dr. McConnell lives with her dogs and husband on a small farm near Madison, Wisconsin.
That night I sat on the couch and worried about this bundle of behavioral problems that I had brought into the house. I turned to see Willie watching me, his face baby-soft and expectant, his body wagging from the shoulders back. I moved off the couch and lay down beside him. He nuzzled into me, the side of his head pressing against my neck. I inhaled the scent from the top of his head, as a girlfriend had told me shed breathe in the smell of her sons hair, savor it, and yearn for it when they were apart.
Even as a tiny puppy, Willie wanted nothing more than to be with me, to cuddle against me with his face pressed against my neck or chest. Willies love of people was as extreme as his fear of dogs outside of his own pack. He loved everyone on two legs and appeared to be overjoyed that the world contained an infinite number of us. When friends came over to meet Willie, hed stop for a second as he watched them get out of their cars, seemingly stunned by the appearance of yet another person. Hed quickly glance at me as if in amazementLook! Theres another one! Ive found ANOTHER ONE!and then hed charge forward, tail thumping, body soft and loose as he transported us into puppy rapture. . .
Something had sent Willie out into the world set on HIGH, like a blender with its last button pushed. Raising him was both wonderful and horrible. Underneath his crazinesshis extreme reactions to unfamiliar dogs, his phobias about noises, his disastrous digestive systemI was sure there lived the dog we all want, brimming with love and loyalty, with a face that sparkles when you come home. But Willie desperately needed to feel safe and secure.
The thing was, so did I.


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As Seen in The Hundred-Foot Journey
DESCRIPTION: A Michelin star restaurant led by Madam Mallory (Helen Mirren) gets some unexpected competition when a homey Indian restaurant opens up 100 feet away. While the impossibly chic Mallory butts heads with the new restaurant's owner, she notices amazing potential in the owner's son, Hassan Haji (Manish Dayal). Dressed in casual clothes as a chef, Hassan soon becomes bewitches by Mallory's French cuisine and kitchen. Of course, the cute Parisian sous chef is also partly to blame. Located in southwestern France, Saint-Antonin-Noble-Val is a town with a long religious history. Named for a Christian saint, the town boasts beautifully preserved architecture and urban planning that reflects this history. Scroll down to see more film locations identifed from The Hundred-Foot Journey.


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Charles Garnier Paris
Charles Garnier Paris continues its tradition of fine, innovative jewelry with their Sterling Silver Constellation and Couture Silver Chains Collections. Replete in a history of exquisite quality and styling since 1901, Charles Garnier Paris continues their legacy in design and technology for today's discriminating shoppers.
Here is a selection of Charles Garnier jewelry. We have an even greater assortment at Scheherazade Jewelers. Be sure and contact us for the availability of what you may be looking for from this remarkable Designer.


text: Morning guys. Anyway i have a Galaxy I5700 from Three.co.uk. i recently unlocked the phone too. last week i rooted the phone with leshaks tutorial at Root Samsung Galaxy Spica i5700 With LeshaK's Kernel after a few days i decided to put the phone back to normal as the 2.2 update is coming in november, so i downloaded the firmware from http://www.samfirmware.com/WEBPROTECT-i5700.htm, i took the EXXJH6 firmware. I flashed the firmware on and it worked fine except it was dutch, so i changed the language and plugged the phone to kies to see if their are any updates. kiesw now says this phone cannot be updated. Is this because the phone needs a specific firmware from three. is the firmware from a dutch phone and since im in the uk it wont update. Any help appreciated cheers


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Caution in Conclusion
Posted by alisond on 10 Aug 2010 at 08:23 GMT
We note that this study reports the same SNP, rs4784227, that we found to exhibit the strongest evidence in our study of >27,000 subjects of European, Asian and African ancestry. (Udler MS. et al. Fine scale mapping of the breast cancer 16q12 locus. Hum. Mol. Genet. 19, 2507-15, doi: 10.1093/hmg/ddq122). However, we are concerned by the interpretation of this present study: that rs4784227 is functionally related to breast cancer risk.
The SNPs examined here were selected from HapMap phase II (http://hapmap.ncbi.nlm.ni...), but this is an incomplete catalogue of common variants. In our study we identified 26 highly correlated, candidates for causative variant via our own re-sequencing coupled with the 1000 Genomes Project (http://www.1000genomes.or...). Only seven of these 26 are present in HapMap phase II.
Although SNP rs4784227 gives the strongest signal among those examined, this does not imply that the others can be definitively excluded. Of our 26 original candidates, we were able to exclude 12 as being at least 100 times less likely than the best. Within the remaining 14 candidates, although rs4784227 had the highest likelihood, nine others were, statistically, only marginally less likely to be causative. Four of these are almost perfectly correlated (r2 ≥ 0.96) in both Europeans and Asians and are thus unlikely to be separable by epidemiological studies alone. Notably, five of the remaining 14 candidates are tightly clustered in open chromatin in the evolutionarily conserved gap between TOX3 and LOC643714 and three of these are also predicted to affect transcription factor binding sites. Although SNP rs4784227 lies in a predicted C/EBP binding site, it falls outside this conserved region of open chromatin.
In most post-GWAS fine scale mapping studies to date, evidence clearly identifying directly causative variants has been elusive - hampered by strong linkage disequilibrium and small effect sizes. Where there are multiple, tightly linked candidates, exemplifying that any one is functional does not prove its causality. Any conclusions should be cautious until combined epidemiological and functional studies can determine the exact variant, or variants, directly responsible for breast cancer risk at this important locus.
Alison M. Dunning
Miriam S. Udler
Douglas F. Easton
We read with interest the comment by Dr. Dunning et al on our paper regarding the association of rs4784227 (16q12.1) with breast cancer risk in Asians and in vitro functional characterization of this SNP (Long J et al. Identification of a functional genetic variant at 16q12.1 for breast cancer risk: Results from the Asia Breast Cancer Consortium. PLoS Genet. 2010;6:e1001002). As described in the paper, our study consists of a multi-stage GWAS and a series of in vitro functional analyses that specifically evaluated the potential functionality of rs4784227. The DNA constructs with the T allele for the luciferase reporter assays were generated by site-directed mutagenesis from the DNA constructs with the C allele, and the sequences of oligonucleotide probes used in the electrophoretic mobility shift assay were identical except for the rs4784227 site. To minimize potential laboratory assay errors and chance findings, we used four cell lines for the luciferase reporter assays and three cell lines for the electrophoretic mobility shift assay. These in vitro experiments have shown consistently that the T allele of rs4784227 reduced luciferase activities and altered DNA-nuclear protein interactions compared to the C allele (Figure 3), suggesting that rs4784227 may be functionally significant.
As described in the paper, three SNPs at 16q12.1 (rs8051542, rs12443621, and rs3803662) were reported from previous GWAS conducted in women of European ancestry (Easton DF et al. Genome-wide association study identifies novel breast cancer susceptibility loci. Nature. 2007 Jun 28;447:1087-93). However, the SNP (rs4784227) identified in our GWAS is not in LD with rs12443621 (r2=0.07) and in weak LD with rs3803662 (r2=0.14) and rs8051542 (r2 = 0.37) in Asians (Table 3 and Figure 2). Furthermore, the association of breast cancer with rs4784227 remained highly statistically significant after adjusting for all three previously reported SNPs (Table 4). In fact, in Asian women, rs12443621 was not associated with breast cancer risk, and the association of breast cancer risk with rs8051542 and rs3803662 was weaker than the one observed for rs4784227 (Table 4). Adjusting for rs4784227 substantially attenuated the association with rs3803662 and completely eliminated the association with rs8051542. Similarly, among European-American women, rs4784227 was found to be associated with breast cancer risk even after adjusting for rs12443621 or rs8051542 at P < 0.05 (Table 5). SNPs rs3803662 and rs4784227 are highly correlated in European descendants (r2=0.86). Including these two SNPs in the same model eliminated the association with rs3803662 but did not alter the risk estimate for rs4784227, although the statistical test was no longer significant (Table 5). As reported in our paper, rs12443621 and rs8051542 were weakly associated with breast cancer risk in European-Americans. Evidently, of the SNPs evaluated in this locus, rs4784227 showed the strongest and most robust association with breast cancer risk in both Asians and European-Americans.
In an extensive search for the causal variant directly responsible for the observed associations at 16q12.1, Udler et al have identified 14 variants that are all strong candidates for being causally important for breast cancer risk (Udler MS et al. Fine scale mapping of the breast cancer 16q12 locus. Hum Mol Genet. 2010 Jun 15;19:2507-15). The authors cautioned that although rs4784227 showed the strongest signal as the potential causal variant for breast cancer, the other 13 remaining candidates could not be excluded due to the LD structure in this region. The authors called for larger epidemiological studies among women of Asian and African ancestry to further evaluate this important locus. It is possible that multiple causal variants may exist at this locus. Nevertheless, results from Udler et al's study support our finding suggesting that rs4784227 is an important candidate for future epidemiological studies and functional characterization.
Wei Zheng
Jirong Long
Qiuyin Cai


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Special Field Trip Packages
are for Groups 15 - 50
Visit the Kentucky Museum! Field trips reservations must be made at least two weeks in advance and are filled on a first come, first served basis.Words in red are linked for more information.
In addition to the permanent collections on display, the 1815 Felts Log House, Instruments
of American Excellence, Civil War Exhibit, Duncan Hines, and the Quilt and textile
gallery, the Museum features new and changing exhibitions. Current Exhibits Upcoming Exhibits
Please have your students view this short Museum Rules Video.
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Each of these sheets has helpful parking and Museum information
Printable Teacher Information Sheet
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Printable Chaperone Information Sheet
For Groups 15-50*
*15 is the minimum number eligible for these packages. 50 is the maximum number eligible for these packages
Special Field Trip Packages
Group Size 15 - 50 Reservations Required
A. The Kentucky Museum Education Department offers four special field trip packages (see
below). Teachers should request a specific package, date and time at least two weeks
in advance. Click here for reservation request. Reservations are taken on a first come, first serve basis.
You will receive an email reply within 24 business hours. Special Field Trips are
offered on most Tuesdays and Thursdays September through the first week of May,following the University Calendar.
Group Size
Length of Tour
Museum Rules
- The chaperone along with the teacher must remain with the class at all times and is responsible for the discipline and conduct of the group. The minimum of 1 to 10 adults ratio is mandatory for admittance to the museum.
- Please instruct students prior to visit of proper museum conduct:
Do not touch unless given permission to touch
Do not cross any exhibit barriers
Do not talk while Museum docent is talking
No food or drink in exhibits
Flash photography is prohibited in some exhibits
- An individual or group will receive one warning. Further rule violation and goup or individual will be escorted to front desk, and admission will be refunded. Staff must ensure the safety of the exhibits and artifacts.
Special Field Trip Packages (Group Size 15-50):
B. *15 is the minimum number eligible for these packages. 50 is the maximum number eligible for these packages
Kentucky Experience
Civil War in Kentucky
Art in the Museum
Day @ WKU
Historic Christmas
Education Staff Guided Field Trip Request Form
Civil War Exhibit
Felts House hearth cooking
Felts House
U S Bank Celebration of the Arts
Mask making
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Despite the health-related benefits of exercise, many people do not engage in enough activity to realize the rewards, and little is known regarding the genetic or environmental components that account for this individual variation. We created and phenotyped a large G4 advanced intercross line originating from reciprocal crosses between mice with genetic propensity for increased voluntary exercise (HR line) and the inbred strain C57BL/6J. G4 females (compared to males) ran significantly more when provided access to a running wheel and were smaller with a greater percentage of body fat pre- and postwheel access. Change in body composition resulting from a 6-day exposure to wheels varied between the sexes with females generally regulating energy balance more precisely in the presence of exercise. We observed parent-of-origin effects on most voluntary wheel running and body composition traits, which accounted for 3–13% of the total phenotypic variance pooled across sexes. G4 individuals descended from progenitor (F0) crosses of HR♀ and C57BL/6J♂ ran greater distances, spent more time running, ran at higher maximum speeds/day, and had lower percent body fat and higher percent lean mass than mice descended from reciprocal progenitor crosses (C57BL/6J♀ × HR♂). For some traits, significant interactions between parent of origin and sex were observed. We discuss these results in the context of sex dependent activity and weight loss patterns, the contribution of parent-of-origin effects to predisposition for voluntary exercise, and the genetic (i.e., X-linked or mtDNA variations), epigenetic (i.e., genomic imprinting), and environmental (i.e., in utero environment or maternal care) phenomena potentially modulating these effects.
- advanced intercross line
- body composition
- mouse
- voluntary wheel running
Complex traits and parent-of-origin effects.
Complex traits, such as human disease and behavior, typically involve a number of genetic determinants and environmental factors that contribute jointly to the total phenotypic variation among individuals (31, 66). Potential sources of variation underlying the distribution of complex traits include the inherited genetic contributions and/or environmental stimuli (acting at any time after formation of the zygote, or in some cases even before). In particular, parent-of-origin effects are defined by cases in which a phenotype follows either a maternal or paternal line, as opposed to standard Mendelian inheritance patterns (73, 87). Parent-of-origin-dependent effects have been demonstrated to be important in modulating a variety of complex traits, including, but not limited to, growth and development (26, 38, 40, 41), cognitive abilities (48, 50), adult body composition (15, 36), cardiac response (2), such human diseases as Prader-Willi and Angelman syndromes, and some cancers (see Fig. 1 in Ref. 19, also Refs. 24, 62, 82). Furthermore, parent-of-origin effects have been demonstrated to play a role in the regulation of behavioral predispositions such as attention-deficit hyperactivity disorder (35), maternal behavior, learning and memory deficits, and altered rest/activity cycles (reviewed in Ref. 48). Effects are typically manifested through a broad range of possible genetic (e.g., sex-linkage) and epigenetic (e.g., x-chromosome inactivation, genomic imprinting) phenomena (37, 82, 86).
Physical activity.
A robust association exists between at least one environmental factor, physical inactivity, and the emergence and prevalence of modern chronic diseases. And, as Booth and colleagues (7) and others have hypothesized, physical inactivity may be at the very "environmental roots" of an ever-increasing list of chronic human health conditions. Variation in voluntary physical activity has been shown to be an important component of health, and regular physical activity has been positively correlated with the prevention and treatment of chronic human-health conditions including obesity and some cancers (6, 14, 28, 47, 59; but also see 84, 85). The benefits of voluntary exercise have been in part attributed to the regulation of energy balance, but there are few mechanistic studies that explain how exercise protects against disease pathogenesis. Regardless, there is extensive support at a descriptive level for the benefits of exercise on many aspects of human well being. Despite the health-related benefits of voluntary activity, more than 50% of U.S. adults do not engage in enough exercise to realize the rewards (80), and little is known regarding the genetic or environmental components that account for individual variation in predisposed voluntary activity levels. Broadly, in humans and rodents, it has been suggested that genetic architecture may play an important role in the regulation of voluntary activity levels, and empirical support for these suggestions is rooted in the demonstration of heritability (52, 55, 65, 70, 75).
Although the predisposition to engage in voluntary physical activity is heritable, used as a common therapeutic intervention for health-related disease, and may be a primary factor in the prevention of chronic health conditions, the location or nature of underlying genetic variation is still an emerging field in humans (9, 13, 22, 72) and rodents (49, 51, 56, 71, 79, 88). Thus, our long-term goal was to create a mapping population to detect quantitative trait loci (QTL) underlying the predisposition to engage in voluntary exercise. Toward this goal, we created a large advanced intercross line (AIL) of mice. Given the demonstration of the role of parent-of-origin effects on QTL for traits related to energy balance (e.g., 15) we utilized a reciprocal cross design to allow for detection of similar effects on voluntary exercise levels. This G4 population originated from a cross between mice with genetic propensity for increased voluntary exercise (HR line) and the inbred strain C57BL/6J.
Beginning in 1993, Garland and colleagues initiated an artificial selection experiment for voluntary wheel running in mice (reviewed in Refs. 30, 75). By generation 16, and continuing through generation 50 and beyond, the HR lines had diverged significantly from the control lines (C lines) with an approximate 2.5- to 3.0-fold increase in total revolutions/day. Selection history has caused the HR lines to diverge in a number of morphological, physiological, and behavioral traits. Examples include but are not limited to reduced body fat (25, 76), increased circulating adiponectin (81), decreased circulating leptin (33), and increased in home-cage activity (57, 58) and altered aspects of open-field behavior (10).
Here, we report on phenotypic measures of the G4 and present results in the context of strong differential sex and parent-of-origin effects on voluntary wheel running and body composition traits. We also explore the potential of the parent-of-origin effects to be differentially expressed in the two sexes. Finally, we discuss the mechanisms whereby these parent-of-origin effects could be transgenerationally inherited through direct genetic and/or epigenetic mechanisms.
Two strains of mice underwent a reciprocal intercross breeding protocol leading to a G4 AIL. The first of these two strains has been selectively bred for high voluntary wheel running for >40 generations (HR mice). Full details of this selection experiment have been provided elsewhere (75), so we will only offer a brief overview here. The original progenitors of the selection experiment were outbred, genetically variable house mice (Mus domesticus) of the Hsd:ICR strain [Harlan Sprague-Dawley (HSD), Indianapolis, IA]. After purchase from HSD, eight lines were created, four of these replicate lines have been selectively bred for high voluntary running (HR lines) on days 5 + 6 of a 6-day exposure to wheels (circumference = 1.12 m), while four others have been bred without regard to running as Controls (C). All procedures were approved by and are in accordance with guidelines set forth by the Institutional Animal Care and Use Committee at The University of North Carolina at Chapel Hill.
HR mice in the current experiment originated from one (lab designated #8) of the four HR lines in the 44th generation of artificial selection at the University of California, Riverside (UCR). At a mean age of 170 days (range = 157–174 days), HR males (n = 15) and females (n =15) were shipped from UCR to The Jackson Laboratory (TJL; Bar Harbor, ME) for rederivation. From 12 rederivations, 11 specific pathogen-free litters were successfully weaned and shipped to the University of North Carolina, Chapel Hill (UNC). These mice represented the founders (generation 0) of the UNC breeding colony of HR mice. HR mice for the current experiment originated from generation 2 of the UNC breeding colony.
The second strain was an inbred strain (C57BL/6J). Adult B6 males and females were purchased from TJL for the current experiment. The B6 strain was chosen for several reasons. First, Lightfoot et al. (55) examined variation in wheel running across inbred strains of mice and showed that B6 is a relatively average runner. Although generating a population from HR mice crossed to a poor runner would have maximized phenotypic divergence, it might also have uncovered variation that is not of interest. For example, extremely poor running performance (or no running at all) may not reflect a low predisposition to engage in exercise but instead may reflect a fear of the running wheel or mice that are generally unhealthy, both different traits than the one of current interest (see Ref. 30). Second, B6 is widely employed in many aspects of biomedical research, and its use provides access to a wealth of genomic tools and databases. These genomic tools and databases will be vital in follow-up investigations and future studies of voluntary exercise in the AIL. Third, the B6 strain has proven to be a reliable breeder, which was critical for the production of a large population. Finally, we have previously characterized wheel-running behavior in HR and B6 mice of both sexes and have demonstrated significant differences in running distance, time, average speed, and maximum speed (63).
Breeding design.
At ∼8 wk of age, 44 HR mice (22 males and 22 females) and 44 B6 mice (22 males and 22 females) underwent a reciprocal cross breeding protocol to produce a F1 population. The 44 progenitor HR mice represented brother/sister sibling pairs from 11 different families. In the F1 population, 16 (out of a possible 22) families from one cross-line population (HR♀ × B6♂) and 16 (out of a possible 22) families from a second cross-line population (B6♀ × HR♂) were chosen to propagate the following generation. In both of the reciprocal F1 populations each of the 11 progenitor families was represented at least once.
Once established, the two reciprocal cross-line populations were not mixed. The F2 and F3 generations were carried out as follows. After weaning, mice were housed in sex-specific cages in groups of four or five until 8 wk of age. At ∼8 wk of age, two males and two females were randomly chosen for breeding from within each of the 16 families in each reciprocal cross population. The males and females were bred across families with pairs of brothers and sisters each mated to a different family. In total, 64 mating pairs (32 from each reciprocal cross population) were established each generation. Each unique family was always represented by two breeding pairs to account for possible failure due to infertility of females, low litter size, or loss of pups before weaning. From these mating pairs at least one successful litter per family contributed to the next generation, which yielded no fewer than 16 unique families represented in each reciprocal cross population each generation.
To avoid inbreeding and increase the effective population size, interfamilial matings were assigned each generation utilizing a Latin square design. Pedigree files and genetic relationship matrices were maintained for all generations. All mice were reared in a viral-free facility, maintained at a temperature of 22°C, 30–55% relative humidity, and a light-dark cycle of 12 h:12 h beginning at 0700. Food (Prolab Isopro RMH 3000; calories provided by: protein 26%, fat 14%, carbohydrates 60%) and water were provided ad libitum. During pregnancy and lactation, breeding pairs were provided an enriched diet (Prolab RMH 2000; calories provided by protein 22%, fat 23%, carbohydrates 55%).
A large G4 was produced following the F3 generation. The production of the G4 population was created in a similar fashion to the preceding generations, with the following exceptions. Again, individuals representing the reciprocal cross-line populations (HR♀ × B6♂ and B6♀ × HR♂), established in the F1 generation, were not mixed and were equally represented in the G4. Two males and two females from each of 30 families were chosen from the F3 generation for breeding. The 30 (an additional 30 were created as "spare," see above) breeding pairs were randomly assigned to groups of five and given the opportunity to mate beginning at ∼8, 9, or 10 wk of age. Mating pairs remained together for the entirety of the breeding process such that each of the 30 original pairs produced three sequential litters. Throughout the breeding process a repeatable synthetic diet (Research Diet D10012G; 20 kcal% protein, 16 kcal% fat, 64 kcal% carbohydrate) and water were provided ad libitum. This staggered mating design, and subsequent availability of adult animals, was done to accommodate space constraints associated with the phenotyping process while reducing any substantial intergroup age differences that may contribute to phenotypic variation.
Prior to and throughout phenotyping, G4 individuals (n = 815) were housed in sex-specific cages in groups of four and provided a repeatable synthetic control diet (Research Diet D10001; 21 kcal% protein, 68 kcal% carbohydrate, 13 kcal% fat) and water ad libitum. G4 individuals were divided into 19 cohorts with ∼45 individuals in each cohort. Cohorts 1–9 (n = 402 in total; 196 females and 206 males) represented one cross-line population (HR♀ × B6♂) established in the F1 generation. Cohorts 10–19 (n = 413 in total; 211 females and 202 males) represented the second cross-line population (B6♀ × HR♂) established in the F1 generation. Cohorts were phenotyped in sequential order spanning a period of 21 wk.
Phenotypic measures.
G4 mice were weighed ( ± 0.1 g) at 4 (mean = 30 days, range 27–32), 6 (mean = 44 days, range 41–46), and 8 (mean = 57 days, range 54–59) wk of age. Immediately following mass measurements, body composition (% fat tissue and % lean tissue) was assessed utilizing an EchoMRI-100 quantitative magnetic resonance whole body composition analyzer (Echo Medical Systems, Houston, TX). Percent body fat (and lean) was calculated as (fat mass/body mass)*100. Following body composition measurements at 8 wk of age, mice were housed individually with access to running wheels (model 80850, circumference = 1.1 m; Lafayette Instruments, Lafayette, IA) for 6 days. A circumference of 1.1 m was chosen to match the conditions under which the HR mice were selectively bred. Daily wheel-running activity was monitored with Running Wheel Activity Software (AWM V9.2, Lafayette Instruments) via Activity Wheel Counters (model 86061, Lafayette Instruments) interfaced with computers. Wheel-running activity was recorded in 1-min intervals for 23–24 h of each of the 6 days of wheel access. From this information, the following daily traits were calculated: total daily revolutions, time spent running (i.e., cumulative 1-min intervals in which at least one revolution was recorded), average speed (total revolutions/time spent running), and maximum speed (highest number of revolutions in any 1-min interval within a 24 h period). Here, we chose to analyze mean values on days 5 and 6 of the 6-day exposure to wheels, as these were the focal days used in the selection experiment that generated the HR line. Finally, food consumption was calculated by weighing (± 0.1 g) food prior to and following the 6 days of wheel access. To minimize any variation in food consumption due to food wasting, bedding was examined following day 6 and any visible pieces of food were accounted for.
Following the 6th day of wheel access, mice were weighed (± 0.1 g) and body composition assessed (as described above). Percent change (following wheel access) in percent body fat (and lean) was calculated as [(% following wheel access − % prior to wheel access)/% prior to wheel access]*100. Following decapitation, a trunk blood sample was taken and blood glucose was measured using a portable glucose monitoring system (Contour model; Bayer HealthCare, Tarrytown, NY), and tissues [tail, brain, epididymal (males) and perimetrial (females) fat depots, triceps surae muscles, liver, lungs, spleen] harvested. Wet masses (± 0.001 g) of fat and muscle were recorded on an analytical balance (model XS204; Mettler-Toledo, Columbus, OH). Following dissection (and mass measurements for fat and muscle) tissues were immediately snap frozen in liquid nitrogen.
Statistical analysis.
The Mixed procedure in SAS (version 9.1; SAS Institute, Cary, NC) was used to apply two-way nested analysis of covariance (ANCOVA) models with REML estimation and type 3 tests of fixed effects. The primary grouping factors were parent-of-origin type (whether a G4 individual was descended from a progenitor cross of HR♀ × B6♂ or B6♀ × HR♂, coded as 1 or 0, respectively) and sex (male vs. female), both fixed effects. As each of the 19 phenotyped cohorts contained only one parent-of-origin type, replicate cohort (n = 19 total) was nested within parent-of-origin type, and was always considered a random effect. Degrees of freedom (DF) for testing the parent-of-origin and sex effects were always 1 and 17. The sex effect and the sex × parent-of-origin type interaction were tested relative to the sex × cohort within parent-of-origin type interaction, again with 1 and 17 DF. Family was included as additional random factor nested within cohort. And, because each dam contributed multiple litters, litter was also treated as a random factor and was nested within family. Due to the slight variation in age, it was included as a covariate in all analyses. Additionally, where applicable, body mass, wheel freeness, time of day, and z-transformed (squared) term for time of death were included in the model as covariates. Wheel freeness was calculated as the number of wheel revolutions following acceleration to a given velocity. The z-transformed (squared) term for time of death was included, as it allows for possible nonlinear relationships between time of day and the dependent variable of interest. Traits were transformed as needed to stabilize variances among groups and improve normality of residuals. Statistical significance was judged at P < 0.05, and all P values presented are two-tailed.
Partial Pearson correlations among wheel-running variables and separately among body composition traits revealed several significant pair-wise associations (P < 0.05) (results not shown). Therefore, we first analyzed both groups of related traits with a MANCOVA, utilizing models in SAS Proc Mixed similar to those presented above. Overall contrasts yielded significant results (P < 0.001) for both the wheel running and body composition traits. Therefore, we proceeded by analyzing each trait separately as described in the Statistical analysis section and present these results below.
Since multiple tests were performed on the same set of animals it is appropriate to control the type I error rate. Given the relatively low number of hypotheses tested and the distribution of P values, we adjusted for multiple comparisons using the false discovery rate (FDR) technique in R (v. 2.8.1; R Development Core Team, 2008). Results of the FDR analysis indicated a more conservative alpha level of 0.03 for judging statistical significance (corresponding to an FDR of 5%). Since this correction does not substantially alter the overall pattern of our results we have chosen to present nominal P values for two-tailed statistical tests and emphasize general patterns in the results.
Wheel running.
Among G4 individuals, after adjusting for variation in age and wheel freeness, analysis of wheel-running data revealed that females ran significantly greater distances (revolutions/day, P < 0.0001), ran at higher average speeds (rpm, P = 0.0002), and spent significantly more time running (1-min intervals/day, P < 0.0001) than males (Table 1). With regard to maximum speed (highest number of revolutions in any 1-min interval within a 24 h period), the difference between the sexes was not significant (P = 0.1046, Table 1). As indicated by statistically significant parent-of-origin effects, G4 individuals descended from progenitor (F0) crosses of HR females and C57BL/6J males (HR♀ × B6♂) ran greater distances (P = 0.0004), spent more time running (P = 0.0006), and ran at higher maximum speeds/day (P = 0.0052) than mice descended from the reciprocal B6♀ × HR♂ progenitor crosses (Table 1 and Fig. 1). Additionally, while not statistically significant, mice descended from HR♀ × B6♂ parents tended to run at higher daily average speeds (P = 0.0529, Table 1).
Two of the wheel-running traits [average and maximum speed/day (rpm)] showed significant sex by parent-of-origin interactions (P = 0.0175 and 0.0155, respectively). Thus, for these traits, the parent-of-origin effect was dependent on sex and the phenotypic differences between the sexes were dependent on parent of origin. Inspection of the least-squares means (Fig. 1) along with separate ANCOVAs of male and female mice (Table 2 and Supplemental Table S1)1 indicated the following patterns. For males, there were significant parent-of-origin effects for average (P = 0.0014, Table 2) and maximum (P < 0.0001, Table 2) daily running speed with G4 individuals descended from HR♀ × B6♂ progenitor crosses showing higher values. Conversely, for females, there were no significant parent-of-origin effects for average (P = 0.7414, Table 2) or maximum (P = 0.3016, Table 2) daily running speed. Thus, for running speed (average and maximum), the statistically significant parent-of-origin effects seem to be exclusive to males.
While parent-of-origin effects on running distance and time spent running did not show statistically significant interactions with sex (P = 0.1014 and 0.4442, respectively; Table 1), they tended to be more pronounced in males than females (especially for running distance; Fig. 1, A and B). G4 males descended from HR♀ × B6♂ progenitor crosses ran 11% more revolutions per day than individuals descended from B6♀ × HR♂ crosses, while females descended from HR♀ × B6♂ ran only 5% more than their counterparts (Table 1 and Fig. 1A). Additionally, separate-sex analyses indicated the parent-of-origin effect for running distance was more robust for males (P < 0.0001) than for females (P = 0.0395) (Table 2). With regard to time spent running, G4 males descended from HR♀ × B6♂ progenitor crosses ran 14% more per day than their counterparts while the same parent-of-origin effect accounted for a 9% increase in females (Table 1 and Fig. 1B). Again, separate-sex analyses (Table 2 and Supplemental Table S1) indicated that the parent-of-origin effect for time spent running was more robust in males (P < 0.0001) than females (P = 0.0200). Hence, while the statistically significant parent-of-origin effects for running distance and time do not appear to be exclusive to males, as seen for running speed (average and maximum), the percent increase and critical values from separate-sex analyses indicate a larger effect in males.
Body composition.
Pre-exercise body composition was assessed at 4, 6, and 8 wk of age (Table 3 and Supplemental Table S2). At 4 and 6 wk of age, female mice weighed significantly less (P < 0.0001) and had a greater percentage of body fat (P < 0.0001), with a lower percentage of lean mass (P ≤ 0.003), compared with males (Table 3). Additionally, mice descended from HR♀ × B6♀ progenitor crosses had significantly lower percentages of fat (P < 0.05) and higher percentages of lean mass (P < 0.05) compared with mice descended from B6♀ × HR♂ (Table 3), although statistical significance was only noted at 6 wk of age after correction for multiple comparisons.
At 8 wk of age, all body composition measurements were made immediately prior to and following 6 days of wheel access. Females were significantly smaller than males (P < 0.0001) prior to and following wheel access, with no parent-of-origin effect (Table 3). Wheel access reduced body mass in both males and females, with males having a larger percent change (P < 0.0001). Prior to and following wheel access, females had percent fat values that were significantly larger than males (P < 0.0001) with no difference in percent lean mass (P > 0.3). After 6 days of wheel access, males lost a significantly larger percentage of fat compared with females (on a relative basis, P < 0.0001), with no difference in percent change in percent lean mass (P = 0.6792). With regard to parent-of-origin effects, mice descended from HR♀ × B6♂ progenitor crosses had significantly lower percentages of fat (P = 0.0003) and significantly higher percentages of lean mass (P < 0.0001), both pre- and postwheel access (Table 3).
Significant interactions between sex and parent-of-origin were observed at 6 wk of age for percent fat (P = 0.0137) and lean mass (P = 0.0054). Examination of least-squares means (Supplemental Table S2) and assessment of the sexes utilizing separate ANCOVAs (results not shown) revealed that parent-of-origin effects were more pronounced in females than males. That is, while both males and females descended from HR♀ × B6♂ progenitor crosses had significantly less fat and more lean mass (compared to B6♀ × HR♂) the quantitative effect was greater in females. At 8 wk of age significant interactions between sex and parent-of-origin type were detected for percent fat (P = 0.0014) and lean mass (P = 0.0037) prior to exercise (Table 3). These significant interaction terms, and the patterns revealed by them, were similar to results at 4 wk of age. Namely, males and females descended from HR♀ × B6♂ progenitor crosses were significantly leaner, but the quantitative effect was greater in females.
Following 6 days of wheel access, there was a marginally significant sex by parent-of-origin interaction for percent change in body mass (P = 0.0396, Table 3). The mean values (± SE) for the four subgroups were as follows: −3.7% ± 0.5, −5.0% ± 0.5, −6.4% ± 0.5, and −6.2% ± 0.5 for F4 females descended from HR♀ × B6♂, females descended from B6♀ × HR♂, males descended from HR♀ × B6♂, and males descended from B6♀ × HR♂, respectively (Supplemental Table S2). Thus, the parent-of-origin effect appears to be greater in females than in males, with female progeny from HR♀ × B6♂ parental origin having the smallest percent change in body mass following 6 days of wheel access. Additionally, significant interaction terms were observed for relative changes in percent fat (P = 0.0090) and percent lean mass (P = 0.0305) following the 6-day wheel trial (Table 3). Males descended from the two progenitor crosses (HR♀ × B6♂ and B6♀ × HR♂) had similar percent fat losses resulting from wheel exposure (−37% ± 2 and −35% ± 2, respectively), while female percent fat losses were more varied between the parent-of-origin types (−26% ± 2 and −32% ± 2, respectively; Supplemental Table S2). Results for relative change in percent lean mass were similar.
Parent-of-origin effects.
Along with generally similar results from an F1 cross between the same replicate of the HR selection lines and a control line (Hannon RM, Kelly SA, Keeney BK, Malisch JL, Garland T Jr., unpublished data), the current findings represent the first evidence directly implicating parent-of-origin effects (and in some cases their dependence on sex) in the regulation of voluntary activity levels. Since our data are from an advanced G4 intercross, we discuss for the first time that such effects may be inherited via X-linked or mtDNA genes or may be persisting across several generations through epigenetic effects. Here, we discuss some of the potential mechanisms through which parent-of-origin effects may be modulating voluntary activity and body composition in mice in a transgenerational manner.
One potential explanation for the transgenerational parent-of-origin effect on exercise and body composition is dosage effects of X-linked QTL (see Ref. 78 for examples of X-linked gene regulation). With regard to body composition, multiple QTL have been previously mapped to the X chromosome for fatness, body weight, and weight gain (see Fig. 1 in Ref. 66 and references therein). However, among human and mouse studies that have identified specific genetic markers associated with voluntary activity levels, none have implicated the X chromosome thus far (13, 22, 51, 56, 72). It is important to note, however, that only a minority of these studies (51, 56) performed analyses with sex-linked markers, whereas the others either did not or could not examine X chromosome inheritance. In the current experiment, G4 males descended from HR♀ × C57BL/6J♂ progenitor crosses have an approximate 62% probability of inheriting an HR X allele while males descended from C57BL/6J♀ × HR♂ have only a 38% probability, while G4 females have a 69 and 31% chance of inheriting a HR X allele, respectively. Given these percentages and the strong parent-of-origin effects observed for many of the G4 phenotypes, an X-linked QTL would need to have extremely large allelic effects to be a primary regulator of the phenotypic differences observed between the HR♀ × B6♂ and B6♀ × HR♂ progenitor crosses.
Although G4 mice descended from HR♀ × B6♂ are phenotypically different than mice descended from B6♀ × HR♂, in some cases, the parent-of-origin effects are dependent on sex and the phenotypic differences between the sexes are dependent on parent-of-origin. If genes located on the X chromosome are at least partially responsible for the variation in activity and body composition between reciprocal crosses, then the source of the significant statistical interactions between parent-of-origin and sex may be somewhat explained through mechanisms associated with X chromosome inactivation (see Ref. 23 and references therein, also see Ref. 74). For females, X chromosome alleles specific to HR or B6 may be escaping inactivation (see Ref. 23 and references therein), and this could potentially dilute any parent-of-origin effect observed in females. Alternatively, sex-specific differential regulation of wheel-running activity has been hypothesized to be mediated through estrogen/testosterone pathways (54). It is possible that genes that regulate sex-specific hormones and their pathways may be colocalized on sex chromosomes with sites related to activity levels.
Mitochondrial DNA (mtDNA) variation between HR and C57BL/6J is another potential source contributing to the parent-of-origin effects observed in the current study. In mice, mtDNA variations have been previously linked to experimental autoimmune encephalomyelitis, corticosterone and neurotransmitter response to psychological stress, and anxiety-like behavior (32, 89). Currently, we do not know if HR and C57BL/6J differ in their mtDNA sequence, but if such variation did exist then it could influence exercise and body composition traits, both of which rely heavily on energy efficiency, and effects would persist transgenerationally. In response to selective breeding, HR mice exhibit several phenotypic characteristics [e.g., high maximal oxygen consumption (69); alterations in muscle metabolic capacities after wheel running, (46)] that may be explained by sequence variation in the mitochondrial genome.
We acknowledge that founder effects could potentially account for the phenotypic differences observed between parent-of-origin types in the G4 population. However, given that the 44 progenitor HR mice represented brother/sister sibling pairs from 11 different families (see materials and methods) we believe it is unlikely that founder effects play a significant role in explaining reciprocal cross phenotypic differences.
In addition to direct genetic effects, the observed parent-of-origin effects may be modulated via such epigenetic mechanisms as genomic imprinting (68). Approximately 100 imprinted genes have been identified (3) and have been shown to be important for a variety of complex traits, including development and growth (3, 12, 15, 53, 62). Furthermore, it has been demonstrated that imprinting effects can be sex dependent (37). In the current experiment, it is important to note that for imprinting to contribute the parent-of-origin effects, HR and B6 mice would have to exhibit differential imprinting (maternal or paternal). Given the hypothesized function of imprinting in evolutionary processes we cannot rule out this possibility, but view imprinting as less likely relative to the other proposed mechanisms (see Refs. 8, 64).
While direct genetic/epigenetic influences may be the most likely mechanism underlying the parent-of-origin effects, environmental influences during pregnancy and development have also been shown to be important in shaping behavior and body composition (5, 17, 18, 21, 39, 83). This is especially paramount given recent examples of the potential for parent-of-origin effects to be modulated through transgenerationally inherited epigenetic mechanisms (1, 16, 27, 43).
In the current study, we had no direct measures of differences in maternal (or paternal) behavior (e.g., activity, food consumption), or physiology (e.g., metabolic rate) in the progenitors of each generation, and thus we cannot directly evaluate whether parent-of-origin effects on activity and body composition in the G4 mice were related to early developmental perturbations. In fact, previous studies on maternal care behavior in HR lines at earlier generations have revealed only minimal differences compared with controls (34). However, Malisch et al. (57, 58) demonstrated that female and male HR mice exhibit an ∼200% increase in home-cage activity (in the absence of a wheel) compared with control lines. In recent generations, home-cage activity during pregnancy and/or lactation and maternal care and its effects on offspring phenotype have not been quantified or evaluated in HR lines (but see Ref. 34) but may now become a priority.
The absence of F1 phenotypic data in the current experiment may lead to alternative interpretations of the observed parent-of-origin effects, especially in regard to their transgenerational nature. While the F1 generation of the cross utilized in this experiment was not phenotyped, data from another reciprocal cross involving the same HR line and a control line have produced similar results (Hannon RM, Kelly SA, Keeney BK, Malisch JL, Garland T Jr., unpublished data). Wheel running phenotypes of this F1 generation revealed that F1 mice from HR dams had higher running values (revolutions/day, time, average speed, and maximum speed), mirroring results from the current G4 generation. Furthermore, the effects were generally stronger for males than for females, with significance noted for revolutions/day and time spent running. Despite the difference between the two reciprocal crosses with regard to the strain that the HR mice were crossed with and the laboratory where phenotyping was conducted, the similarity and sex-specific magnitude of the results are striking and allow us to infer that the F1 from the current experiment would have yielded similar results. Therefore, given the assumption that F1 results from HR × ICR represent F1 results from HR × B6 this significantly increases confidence in the veracity of our findings that parent-of-origin effects on voluntary exercise levels and body composition in mice are transgenerational.
As outlined in materials and methods (Statistical analysis), due to logistical constraints, cohorts were tested in sequential order and thus cohort was confounded with parent of origin. We adopted several approaches to ensure that a sequence of testing effect was not involved in the parent-of-origin effects discussed above. First, to account for any cohort-to-cohort variation we included cohort as a nested factor in all statistical models. Second, we examined the LS means and standard errors from models that replaced parent of origin with cohort as a primary grouping factor. We did not observe a discernable pattern between cohorts or among parent-of-origin types (cohorts 1–9 vs. cohorts 10–19). Lastly, since any temporal variation in sequence order is likely to be minimized between closely tested cohorts, we grouped cohorts 7–12, separated by only a 6 wk period of time, together and treated them as one large population. With this subset of data, the parent-of-origin effects remained significant.
In this study we chose to evaluate mean running values on days 5 and 6 of the 6-day exposure to wheels. We acknowledge that mice may take longer periods (up to 2 wk) to acclimate to running wheels and reach a plateau with regard to running distance. A number of factors may contribute to the acclimation process causing day-to-day variation among individuals in the trajectory of initial wheel-running behavior. We chose the current paradigm to reflect as accurately as possible the conditions under which the HR mice were selectively bred (75), to measure the same phenotype selected for and minimize any interpretational issues associated with measuring a different phenotype. While initial "learning curves" may be present in some strains of mice, this does not seem to be the case for HR mice compared with ICR control mice (see Fig. 5 in Ref. 31). However, we point out that our results are relevant to the current methods, and caution should be taken when extrapolating these findings to other related measures of voluntary exercise.
Sex effects.
Females ran significantly more, for longer periods of time, and at higher average speeds than males. These results are generally consistent with previous investigations comparing the differences in activity in female vs. male mice of a variety of strains, including the two strains used in the current study and the other replicates of HR selection lines as well as their control lines (55, 75). These sex-specific differences in voluntary activity levels have been hypothesized to be mediated by estrogen and testosterone pathways (reviewed in Ref. 54). As outlined by Lightfoot, mechanisms involved in the estrogen-α pathway are currently the most detailed and may reflect differential release of dopamine as a possible mechanism leading to the increased physical activity in females (see Fig. 4 in Ref. 54). However, while plausible mechanisms are beginning to be proposed, a plethora of alternative hypotheses regarding the mechanistic regulation of exercise via sex hormones are still largely in their infancy.
The results of sex effects on body composition are generally consistent with existing human literature examining weight loss patterns via body fat loss or oxidation in men and women as a result of exercise training (42, 44), suggesting that these sex differences in weight loss patterns may result from greater precision in energy balance among women than among men. That is, when participating in exercise-training programs, women often eat more to compensate for the negative energy balance resulting from aerobic activity while men generally do not (Ref. 42, cf. Ref. 77 on sex differences in the maintenance of body mass in the HR mice). Sex differences in weight loss patterns resulting from exercise training may be attributable to changes in circulating hormones, differences in primary substrate utilization, or a combination of these two factors along with others. Several studies (29, 45, 61) have demonstrated that females derive a relatively larger contribution from lipids to oxidative metabolism during exercise, although others (11, 60) have observed similar contributions of lipids and carbohydrates. Future analyses will focus on whether percent change differences (among individuals and between sexes) in body mass, percent fat, and percent lean varied in accordance with running distance, duration, average speed, or maximum speed.
The results of the current investigation are an important step toward uncovering the factors contributing to the variation associated with the predisposition of voluntary activity levels. The G4 advanced intercross line we have created is one of the largest and potentially most powerful experimental populations of mice created for the purpose of dissecting and understanding the genetic architecture controlling voluntary exercise. Here, we have described strong, transgenerational parent-of-origin and sex effects on exercise and outlined several possible mechanisms that may be modulating these observations. We hypothesize that several of these factors are likely making a joint contribution to the parent-of-origin effects, as opposed to a single major cause. Forthcoming genotypic and linkage analyses will be useful in determining the role of X-linked loci and genomic imprinted QTL. Further studies will be needed to elucidate the role of the mitochondrial genome and maternal environmental effects.
This work was supported by National Institutes of Health (NIH) Grant DK-076050 to D. Pomp. S. A. Kelly was supported through a National Institute of Mental Health-funded (5T32MH-075854-04) Interdisciplinary Obesity Training program. Phenotypes were collected using the Animal Metabolism Phenotyping core facility within UNC's Clinical Nutrition Research Center (funded by NIH Grant DK-056350).
No conflicts of interest are declared by the author(s).
We thank Z. Yun for assistance with animal care and data collection. We thank Chris Wiesen at UNC's Odum Institute for Research in Social Science for statistical consultation.
↵1 The online version of this article contains supplemental material.
- Copyright © 2010 the American Physiological Society
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Health and First Aid Officer
Job details
|St John's School and Sixth Form College, A Catholic Academy
|Contract type
|Part Time
|Contract term
|This job has expired
|Job starts
|as soon as possible
Health and First Aid Officer
Part time 20 hours per week - term time only
annual salary £7,433 (4 hours per day – 10 am to 2 pm)
to start as soon as possible
(also being advertised, Swim Teacher - 10 hpw for those interested and qualified in both roles)
We are looking to recruit an enthusiastic, well organised and highly motivated person to start as soon as possible.
St John's is committed to safeguarding and promoting the welfare of children and young people. The successful applicant will be subject to an enhanced Disclosure and Barring Service check.
For further details, please contact: 01388 603246, email@example.com
or visit our website: www.stjohnsrc.org.uk - Vacancies
Closing Date: 17 August 2017


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text: I know I've gotten a lot of shit for never being able to make up my mind but I finally did. I signed up for ROTC and begun the paperwork for getting the ROTC scholarship so I will definitely be contracting with the Army, no more bull shit and I'm very happy about that. I plan on going active duty after college. I got to wear my ACU today and it felt badass too.


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Maria Sharapova's 15-month suspension for failing a drug test is likely to extend her career all the way to the Tokyo Olympics in 2020, according to the Russian's longtime agent Max Eisenbud.
The five-time major winner rocked the tennis world a year ago when she announced she had tested positive for meldonium, an over-the-counter heart medication commonly used in the countries formerly comprising the Soviet Union.
Sharapova said she hadn't realized the drug -- which she had taken for more than a decade for various health issues -- had been put on the 2016 prohibited list. She is scheduled to return to the women's tour on April 26 at an event in Stuttgart, Germany after organizers gave her a wildcard.
"If (the suspension) didn't happen, this probably would have been her last year," IMG's Eisenbud told CNN in a phone interview. "I think she'll play through Tokyo" if healthy.


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Tag Archives: concrete goals
Not Just an Agent – A CEO…
Good Morning! Fall has just about arrived. Are you ready to finish this year off right? Many real estate agents will wait until December to start planning for 2013. The best agents will start right now! Are you ready to … Continue reading
Posted in RE/MAX, Training Tagged concrete goals, real estate, real estate business plan Leave a comment


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Free Conversation in Use for Front Liners (08 Februari 2017)
FREE Conversation in Use for ITS Front Liners
In supporting ITS recognition and internationalization, CLC opens Free Class Conversation for ITS employees (tendik ITS). Don't miss this program and rush on it. Class is conducted twice a week and will be started in 27 March, 2017 with the schedules as follow:
Monday, 14.30-16.00 and Wednesday, 14.30-16.00 (First round: Gelombang pertama)
Tuesday, 14.30-16.00 and Thursday, 14.30-16.00 (Second round: Gelombang kedua)
It is suggested that the participants choose one of the above schedules.
Letter of recommendation from the employer can be sent by email to: firstname.lastname@example.com, completed with the contact person of the participant for being followed up with courtesy call.
Thank you.
See the detail below.
Letter of Commitment Click here


text:
Latest News
Web Date: June 2, 2014
Web Date: June 2, 2014
Maker Of Rat Poison d-CON To Pull Products
Pesticides: Reckitt Benckiser to phase out 12 products targeting rodents in response to EPA safety concerns
Department: Government & Policy
Keywords: pesticides, EPA, Reckitt Benckiser, rat poison, rodent control, rodenticides
Keywords: pesticides, EPA, Reckitt Benckiser, rat poison, rodent control, rodenticides
After a long battle with federal officials, Reckitt Benckiser, maker of d-CON rat and mouse poisons, has voluntarily agreed to stop manufacturing 12 of its products by the end of this year. Distribution of the products to retailers will cease by March 31, 2015.
The company's action comes after the Environmental Protection Agency moved to ban the 12 d-CON products last year, saying they do not meet the agency's safety standards and . . .
To view the rest of this content, please log in with your ACS ID.
- Chemical & Engineering News
- Copyright © American Chemical Society


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Use the NVIDIA 3D Stereo Bracket accessory in 3D stereo applications that require active synchronization of left-eye/right-eye video frames with a projector and 3D glasses.
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text: Hydrangea Columba is the main protagonist and narrator of NOCTIS. Page in progress HistoryEdit Current EventsEdit Retrieved from "http://avenari.wikia.com/wiki/Hydrangea_Columba?oldid=4093" Ad blocker interference detected! Wikia is a free-to-use site that makes money from advertising. We have a modified experience for viewers using ad blockers Wikia is not accessible if you've made further modifications. Remove the custom ad blocker rule(s) and the page will load as expected. Categories: Add category Cancel Save


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Chapter 1 : the meeting
|Rating: 12+
|Chapter Reviews: 4
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Harry, Ron, and Hermione walk into the Great Hall, mumbling to themselves.
Hey, Potter! a voice calls.
The trio turns around to find a girl of 16 with bright pink hair, black nail polish and a saucy grin standing in the doorway of the Great Hall.
Well, are you just going to stand there, stupid, or are you going to give me a hug? The stranger said.
Lily? Is it really you? Harry asked.
Yep. Its me alright. Lily said.
What are you doing here? And what happened to you? You changed a lot! Harry said.
Come on Lily replied as she lead them into the Great Hall.
Her bright pink hair was so noticeable; heads turned all over the Hall.
Well, my dad brought me here, after I caused some trouble at my old school, Durmstrang. Youve probably heard of it. A really bad school it is. A lot of bad blood, Lily continued on, Oh, and you want to know what happened to me? Heh, I decided to change my style. I heard of muggles dying their hair and wearing nail polish so I decided to experiment. Then I decided I liked it so I thought Why not? and it just sort of stuck.
Wow! How did you do get your hair that colour? Hermione asked.
Oh, Im sorry. Lily, this is Ron . Harry started.
Ahem . Ron, Ron Weasley. Ron butted in.
Its a pleasure. Lily replied.
And Im Hermione Granger.
Very nice to meet you, Hermione. Oh wait! Youre the one who stole Victors heart at the Tri-wizard Tournament! Lily exclaimed.
Hermione blushed, Yeah. Hehe.
Oh yes, Victors told me all about you. Were old chums, Me and Vic. Lily said.
Oh! Here comes Dumbledore! Hermione exclaimed.
After Dumbledore speaks and the food appears, they continue their conversation.
Well. I like the new you, Lylian. Harry says.
Its Lily. Dont worry my father calls me Lylian, too. It gets on my nerves so much. Lily says.
Im sorry, Harry says.
Oh. Its okay. So hows about we start eating? Lily said.
After a while, Lily gets a visitor.
Well, well, well. What have we hear? A sly voice calls.
They all turn around.
Malfoy, Ron says, What do you want?
Just saying hello to your friend, Weasley. Go back to your pudding. Malfoy says.
Hello, my names Lily, Lily Snape.
SNAPE?! They all exclaimed.
Yes. Why? Lily said.
I never knew Snape had kids. Hermione said.
Kid. Im an only child. My mothers dead. Lily said.
So theres a potion princess. I dont think weve been properly introduced. Ahem, I am Draco Malfoy, Son of Lucious Malfoy. Malfoy sneered.
Oh really? A Malfoy? Lily replied.
Youve heard of us? Malfoy asked.
Oh, yes, but only a little. Want to enlighten me more? Lily asked.
Very well. How about tonight, 7:00? Well meet in the front hall. Malfoy said.
7:00 it is. Lily replied.
See ya then. Malfoy said.
Malfoy and his cronies, Crabbe and Goyle, walked back to their table.
What are you doing?! Harry exclaimed.
Well apparently, I have a date tonight. Lily said slyly.
Lylian, May I have a word with you? Snapes voice came from around the corner.
Father! How many times have I told you? Its Lily. And yes you may, Lily replied, Ill meet you guys upstairs.
I heard Mr. Malfoy talking about a date with you after dinner. Is this true? Snape said.
Yes, as a matter of fact, it is. Is that a problem? Lily said.
No except you cannot and will not use your powers on him. Do you understand? Snape said fiercely.
Father! Why can't I? You know I need to get to know the guys I go out with. Lily exclaimed.
You cannot use them on him, you are too powerful! And that is no way to get to know the guys you go out with! Snape said.
Oh father! This is no fair! I need practice anyway! Lily protested.
Not on Dra
Snape stopped himself just as Malfoy came around the corner.
Professor, what are you doing here? Draco said.
Lily is my daughter, Im allowed to talk to her without your permission. Snape said quickly.
Yes, youve talked to me, now you can go. Lily said.
Lily come here. Snape pulled her aside.
What is it father? Lily said quickly.
You must not use your powers. Do you understand me? Snape whispered.
Yes father. Lily replied.
Very well. Have fun you two. Snape grunted as he walked away.
We will Professor dont worry. Malfoy said.
Draco and Lily walked out of the castle onto the grounds. Lily and Draco had been talking for about an hour when the sun was beginning to set over the lake. They picked a nice patch of earth to sit on near the side of the lake. The Octopus came up and let them tickle his tentacles.
This is really nice. Lily said.
Yeah. Lily can I tell you something? Draco asked.
Of course. Lily replied.
I know this is kind of sudden but, Ive had more fun with you in an hour than any other girl Ive ever been with. Draco confessed.
Really? I feel the same way. Lily said.
Thats great. So how do you and Harry know each other? Draco asked.
Oh its a long story. It all started about a year before me and Harry started at Wizarding School. We both lived in Surrey. Except neither of us knew each other were going to Wizarding School. I went to Durmstrang and Harry went to here, to Hogwarts. I was so upset. Then, last year, I decided I should be with my father, but he didnt think it was a good idea with my powe . Lily stopped herself before she revealed that she had magical powers.
With your what? Draco asked.
Ummm, nothing, nothing. What time is it? Lily asked.
Its about 9:00. Wow! Weve been talking for at least two hours. Draco said.
Oh my gosh, we have. Im getting a little tired. Maybe we should head back to the castle now. Lily said, shivering.
Yeah its getting a bit cold out too. Lily can I ask you something? Draco said.
Yes. Lily replied.
The first quidditch match of the season is tomorrow and I was wondering if you would see me off. Draco asked.
Of course I would. Isnt Harry the seeker of Gryffindor? Lily asked.
Yes and I have to say, a good one at that. But dont tell him I said that. Draco replied.
Alright, Lily said, Now lets head back to the castle.
I agree. Draco said.
So Draco and Lily headed back to the castle shivering, but happy with what the night had brought.
Here we are, the first quidditch match of the season between Gryffindor and Slytherin. Lee Jordan commentated.
Hey Draco! Lily called.
Lily! I thought you werent going to make it. Draco exclaimed.
I wouldnt miss it. I just have to get back to Harry before Gryffindor goes onto the field. Lily said.
Oh, okay. Well, see you later. Draco said.
Bye. Good Luck. Lily said as she walked away.
Lily took out a little glass orb from her robe pocket and whispered a few words. A second later she appeared at the other end of the field.
Harry! Lily yelled.
Hey Lily. I thought you werent going to make it. Harry replied.
I . I wouldnt miss it. Lily said.
Well, I guess Ill see you after the match. Harry replied.
Yeah, I guess so, Lily said, Good luck, Harry!
Thanks Lil. Harry said.
And Gryffindor takes the win, 180 to 50. Lee Jordan said.
Lily ran down the congratulate Harry.
Good job Harry. Lily exclaimed.
Thanks Lil. I knew you always brought good luck to me. Harry said.
At that moment, Draco landed nearby to see Harry and Lily talking.
Uh, Harry. Can I have a word? Draco said.
Um, I guess Draco. Be right back Lil. Harry replied.
Okay, whats going on between you and Lily? Draco burst out.
Nothing were just really old friends. Why? Do you like her or something? Harry asked.
Yes, if you must know. I do. Draco replied.
Ok, cool. Harry said.
So, you dont care if I go out with her again? Draco asked.
Not really. Just remember one thing, If you break her heart, Ill break your neck. Got it? Harry said.
Yeah sure, Harry. I wont break her heart. Draco said.
Do I have your word on that? Harry asked.
Yeah of course. Here, lets shake on it. Draco said.
Alright. Harry said.
Ron and Hermione came up the steps just in time to see Harry and Draco shaking hands.
Harry! What are you doing? Ron exclaimed.
Um, just making a pact. Harry replied.
Ok Harry. I think you need to lie down. Ron said.
Come on Harry. You need to take your broom back to the team closet. Hermione said.
Lil, you coming? Harry asked.
Um, I think Ill hang out here for a little while. Lily replied.
Ok, see ya later. Harry said.
Harry, Ron and Hermione walked off.
So want a ride? Draco offered.
Sure. Lily said.
Soon, Lily and Draco were soaring over the treetops of the
This is amazing. Lily said.
You think this is amazing. Just you wait. Draco said.
Soon, they reached a small hill not too far from the grounds, just
beyond the Forest.
This is so beautiful. Lily said.
Like you. Draco said.
Heh, thanks, Lily said, Are you always this nice to the new
Only if theyre as nice as you are. Draco said.
Lily and Draco were talking for about half an hour.
Its getting a little chili. Maybe we should head back. Lily said.
Here take my quidditch robe. Draco said.
Thanks. Lily said.
Draco took Lily back to the castle and walked her to the
Gryffindor Tower only to meet Professor Snape at the entrance.
Lylian. Where have you been? Snape said angrily.
Its Lily dad. And Ive been with Draco, if it any of your
business. Lily said.
It is my business when it comes to my daughter. Mr. Malfoy,
you can proceed to your dormitories now. Snape said.
Okay. Goodnight Lily. Draco said.
Good night Draco. Lily said, Goodnight father.
Im not done with you yet miss. Snape said.
Oh father cant we do this tomorrow when I have the energy?
Lily said.
Very well. Goodnight Lylian. Snape replied.
Its Lily! Lily yelled as the tower door closed.
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the meeting: the meeting
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Milky Way™ Poop Emoji Soap Mold (MW 524)
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Create soaps that will be sure to bring a smile to the faces of kids and teens! This mold is shaped like a swirled pile of poo with big, adorable eyes.
Common Uses:
Melt & Pour Soap. Cold Process Soap. Lotion Bars.
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Manufacturer: Milky Way Molds
Material: Clear PVC (Not Approved for Food Contact)
Country of Origin: United States
Milky Way Soap Molds can tolerate temperatures of 135-145 degrees F. Above these temperatures, warping may occur. Soap is easiest to release 12-24 hours after cooling.
Wash molds in warm, sudsy water. Please avoid the dishwasher because that can warp your molds or make them brittle.
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Material: Plastic
Style: Bar
Avg. Bar Weight: 1.75 oz
Bar Cavities: 5
Bar Depth: 0.875
Bar Length: 2.5
Bar Width: 2.875
Max Temperature: 135ºF
Mold Capacity (oz): 8.75
Heart Eyes & Joe Cool Emoji Soap Mold (MW 521)
Melt & Pour Soap:
Pour melted soap at a temperature no hotter than 135ºF into mold. Cool until soap has hardened. Remove soap from mold. For easier unmolding, place into freezer for 30 minutes, and then immerse bottom of the mold in hot water for a few moments. Do not use cooking spray to coat mold.
Cold Process Soap:
Coat the mold with cooking spray. Pour soap at trace into mold. After 24 hours, flip mold over onto a clean surface. Gently push the back of the mold until the soap begins to release then pull the soap out of the mold with a gloved hand. Allow soap to cure for 4-8 weeks. Note that some CP soap recipes may require a week to harden enough to remove from the mold. If this is the case with your recipe, we recommend adding sodium lactate to your recipe to assist with creating a harder bar faster.
Lotion Bars:
Pour melted lotion bar mixture into mold cavities at a temperature no hotter than 135ºF. Allow mixture to solidify. Flip mold over onto clean surface. Gently push back of mold until lotion bar begins to release then pull bar out of mold.
Wash in warm, sudsy water. Please avoid the dishwasher because that can warp your molds or make them brittle.
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Jodi • Illinois
Date Submitted: 08/17/2017
I Recommend This Product!
"Too cute!"
COMMENTS: Got this mold for my daughter to use with melt and pour. I think I like it more than she does. Sturdy, unmolds well.
PRODUCT USED IN: Soap - Melt & Pour, Soap - Cold Process
• Super cute
• Sturdy
• Unmolds easily
Sandra • Colorado
Date Submitted: 05/08/2017
I Recommend This Product!
"A Little Difficult to Unmold But Worth It"
COMMENTS: Like most Milky Way plastic molds, getting the hardened soap out is tricky but it is worth the trouble. Poop Emoji soap is one of my best sellers and it's so easy to make. A little color for the eyes, let that dry, and pour the brown batter into the mold. They are easy to make and sell quickly.
PRODUCT USED IN: Soap - Melt & Pour
REBECCA • Wisconsin
Date Submitted: 05/02/2017
I Recommend This Product!
"Emoji Mold"
COMMENTS: My kids absolutely adored this mold! Easy to use and the soaps popped right out!
PRODUCT USED IN: Soap - Melt & Pour
Karen • Washington
Date Submitted: 03/28/2017
I Recommend This Product!
COMMENTS: I got this mold when my grandkids were here and they immediately insisted we make poop soaps. They turned out amazing and the kids went home very happy. This mold is just too cute! Love it!
PRODUCT USED IN: Soap - Melt & Pour
Melissa • Nevada
Date Submitted: 03/12/2017
I Recommend This Product!
"Super fun easy mold"
COMMENTS: This mold is really fun! I used it with melt and pour. I started with the eyes and then added the body color. After they were set, I popped them in the freezer for about 15 minutes. After I released them from the mold, I added the mouth color, while the soaps were still cold from the freezer. Really fun and cute!
PRODUCT USED IN: Soap - Melt & Pour
Doris • Indiana
Date Submitted: 02/17/2017
I Recommend This Product!
"Very cute"
COMMENTS: This is cute, if you can call smiling poop cute, and will be a hit at shows coming up and something to keep the kids busy while the parent shops. I fill my shop with kid attractants so the parent or grandparent won't rush out because the child is being unruly. These and rubber ducks work wonders!
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Catholic Sentinel Video: 'A Special Way of Life'
Barbara Collier, SNJM is featured in a video by the Catholic Sentinel about the Mass for Consecrated Life at Portland's St. Mary's Cathedral on January 31. "We just keep telling ourselves it is a special way of life and we try to live it out the best way we can, whatever we are doing," said Sr. Barbara. Read more at: http://www.catholicsentinel.org/main.asp?SectionID=2&SubSectionID=35&ArticleID=28058 and watch the video below.


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Cute Dog
- (Duplicated CD)
- Artist: Mark Boucher D.
- Format: CD
- Release Date: 2/20/2009
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Song(1)is about my 'cute dog' shep. Song(5)is about hanging out with my friends, looking for a little action after school. Song(6)is about the night I meet my wife, women like this song! Song(7) is what life is all about! Song(9) is too true! Song (13) is about religon, and it's true if you are a fundamentalist or undecided. Song (14) well,it's about the moon, my wife told me it's been done, and I said we'll just have to have another one. I could go on and on! I hope you like the CD, Mark.