忍者ブログ
Informative and Instructive Science News
[3]  [4]  [5]  [6]  [7]  [8]  [9]  [10]  [11]  [12]  [13
×

[PR]上記の広告は3ヶ月以上新規記事投稿のないブログに表示されています。新しい記事を書く事で広告が消えます。

Navigating the Genome for Autism Clues

Two new studies connect structural variations to 1 percent of autism cases, a finding that may help unlock the enigmatic disorder's genetic footprint

A pair of research teams recently linked large-scale mutations on one of the body's 23 pairs of chromosomes (which carry cells' genetic code) to autism, a finding that helps shed light on a disorder whose genetic underpinnings have confounded scientists for decades. The revelation represents the most concrete evidence to date that structural variations in the genome play a crucial role in the condition's development, marked by symptoms that include a failure to socially connect, communication difficulties and obsessive behavior.

Scientists this month unveiled evidence that an estimated 1 percent of all autism cases may stem from a structural change involving 25 to 30 genes on chromosome 16. On January 9, a team, led by researchers at Massachusetts General Hospital (MGH) and Children's Hospital Boston, announced that it had found copy number variations—deletions of duplications of segments of genetic code that alter the number of copies of a gene a person carries—in 12 of 1,400 autism sufferers it was studying. (A person normally receives two copies of each gene, one from each parent.) The researchers report in The New England Journal of Medicine (NEJM) that they replicated the finding in two other cohorts—one with 500 participants and another with 300 individuals diagnosed with autism. A week later, a Canadian research team announced in The American Journal of Human Genetics that it had uncovered genetic kinks in the same region in a sample of 927 people—427 of whom suffer from autism

The back-to-back findings come amid a stream of evidence pointing to genetic rearrangements as key culprits in autism. (Chromosome 16 is the second instance of a copy number variation to be fingered as a causative mutation of the condition. Scientists first reported a link between a surplus of genetic material on chromosome 15 and autism in 1994, a finding that has since been replicated and confirmed to be a copy number variation.

Less than a year ago the Autism Genome Project (AGP) Consortium, a collective of more than 120 scientists representing various institutions around the world, reported in Nature Genetics that it had found similar chromosomal variants in several autism patients. About the same time, scientists at Cold Spring Harbor Laboratory in Long Island, N.Y., focusing on families with one autistic child, reported that an estimated 10 to 30 percent of all reported cases of autism may be caused by new (or spontaneous) mutations in the number of copies of genes in children (that were not found in either parent).

Citing the growing body of evidence of links between copy number variations and diseases such as autism, an international science consortium announced yesterday that it plans to sequence the genomes of 1,000 people from around the world in an attempt to flush out genetic suspects. "The importance of these variants has become increasingly clear with surveys completed in the past 18 months that show these differences in genome structure may play a role in susceptibility to certain conditions, such as mental retardation and autism," the National Institutes of Health, one of the participant organizations, said in a statement.

Mark Daly, an assistant professor of medicine at Harvard University Medical School and MGH as well as co-author of the NEJM study, notes that "It is extremely unusual to see these spontaneous deletions and duplications in a region that's usually a copy number–stable region. This specific spontaneous mutation, which we found in a sufficient number of cases, announced itself as an autism risk factor."

"I'm really happy because we found the same result on [chromosome] 16 using a Canadian cohort," adds Stephen Scherer, director of The Center for Applied Genomics at The Hospital for Sick Children in Toronto, a co-author on the second study. "Validation of a complex disease is very exciting."

The search for autism-related genes has led to stunning evidence of the complexity of the disease, which is estimated to affect one in every 150 children born worldwide. Autism involves a spectrum of illnesses that all have similar symptoms, including Rett syndrome, which researchers have linked to a specific genetic mutation. The syndrome only strikes girls and is characterized by asocial behavior and cognitive deficits. But the exact causes of the vast majority of autism-related disorders remain a mystery: classic genetic studies, which tie the ailment to single nucleotide polymorphisms (SNPs—deletions, additions or substitutions of one unit in the genetic code), have returned a number of different markers with very few well-replicated candidates.

Some research teams over the past five years have used microarray or gene-chip technology to compare genomes and quickly scan them for variations of copy numbers on each chromosome. Daly says that microarray data provided his team with an early analysis of the vast data sets it is reviewing; he says scientists plan to follow up their initial findings with an SNP-association study. The Canadian researchers—some of whom were members of The AGP Consortium—specifically set out to examine copy number glitches.

Daly believes these structural events are likely behind only some cases of autism. "I think the middle ground is that some of the genetics underlying autism have their roots in these spontaneous deletions or duplications," he says. "At the same time, across our data set, we don't see regions like this. It's a part of the puzzle, but there's a lot more to it than just this type of event."

Michael Wigler, a geneticist at Cold Spring and the senior author of the 2007 Science paper, believes that successfully hunting down these lesions in the genome will prove crucial to unraveling autism. At the least, he says, they will point researchers in the right direction.

"The general approach of looking at copy number variation as the cause for genetic disease has probably taken one of those exponential—it's probably hyper- —exponential leaps," he says. "So, in 2003 we published [a previous] Science paper, which showed that there is [a] relatively large amount of copy number variation among normal, healthy people."

Both of the new studies found that copy number events involving either duplication or deletion of the 25 to 30 chromosome-16 genes—several of which are known to play a role in the developing brain—appear to cause autism. "That region—or rather, a gene in that region—is apparently extremely sensitive to [copy-number] dosage," Daly says. "Too much or too little causes developmental differences."

In the case of a deletion of this DNA segment, the damaged gene likely will not produce enough protein. This can potentially cause myriad malfunctions, because proteins typically work in complexes. Hence, a deficiency of one can hobble the entire collaborative effort. By the same token, a surfeit of one protein caused by duplication would also cause malformed complexes.

There is another wrinkle in the Canadian data set, Scherer says: of the 427 autism sufferers assessed, 7 percent showed evidence of copy number variation. Within that group, 27 individuals (11 percent) had two or more of these spontaneous deletions or duplications (or one of each). One of them had a deletion on chromosome 22 that affected, among other genes, SHANK3, which has been implicated in mental retardation. This deletion was accompanied by a duplication of a genetic segment on chromosome 20.

Scherer calls the mutation on chromosome 20 a "modifier" that adds to the complexity of the phenotype. "There are some of these copy number changes that increase the risk of being autistic, but they may need to be inherited with other changes that culminate," in the disorder, he says. "It's going to vary based on your sex, your genetic background. and possibly the environment." He believes that a person without the chromosome-20 alteration is likely to suffer mental retardation and one who has it is more likely to develop autism.

That is in line with a unified genetic theory of autism proposed by Wigler, who performed a rigorous statistical analysis of a large data set cobbled together by the Autism Genetic Resource Exchange, a group of autism researchers who share data collected from families with autistic children. Wigler proposed the idea of modifying genes partly to account for the disparity in autism incidence between boys and girls. (Boys are four times more likely than girls to develop autism.)

So how significant is the latest finding about chromosome 16? "It's going to be the intense subject of functional studies, studies in model organisms, and genetic follow-up studies in human samples," Daly says. Clinicians monitoring children with the deletion or the duplication of material there, he explains, may eventually be able to find a matching set of specific symptoms that accompany those particular genetic events. "There are folks at the Children's Hospital," Daly notes, who "are already turning this into a critical screening tool."

For Scherer's part, he believes, from his preliminary observations of one family, that a deletion of DNA on the 16th chromosome may result in autism accompanied by mental retardation as well as disruptions in the aortic valve (one of the heart's four valves) that may cause seizures. "In fact," he says, "if you see…[this pair of symptoms], along with autism, it might predict the [chromosome-] 16 deletion."

If that bears out, then looking for the deletion ahead of time could be crucial in early intervention. "We tried to do a thorough study," Scherer says, "so that we could convince people to do a microarray analysis as part of their workup."

PR
Political Animals (Yes, Animals)

Just as there are myriad strategies open to the human political animal with White House ambitions, so there are a number of nonhuman animals that behave like textbook politicians. Researchers who study highly gregarious and relatively brainy species like rhesus monkeys, baboons, dolphins, sperm whales, elephants and wolves have lately uncovered evidence that the creatures engage in extraordinarily sophisticated forms of politicking, often across large and far-flung social networks.

Male dolphins, for example, organize themselves into at least three nested tiers of friends and accomplices, said Richard C. Connor of the University of Massachusetts at Dartmouth, rather like the way human societies are constructed of small kin groups allied into larger tribes allied into still larger nation-states. The dolphins maintain their alliances through elaborately synchronized twists, leaps and spins like Blue Angel pilots blazing their acrobatic fraternity on high.

Among elephants, it is the females who are the born politicians, cultivating robust and lifelong social ties with at least 100 other elephants, a task made easier by their power to communicate infrasonically across miles of savanna floor. Wolves, it seems, leaven their otherwise strongly hierarchical society with occasional displays of populist umbrage, and if a pack leader proves a too-snappish tyrant, subordinate wolves will collude to overthrow the top cur.

Wherever animals must pool their talents and numbers into cohesive social groups, scientists said, the better to protect against predators, defend or enlarge choice real estate or acquire mates, the stage will be set for the appearance of political skills — the ability to please and placate, manipulate and intimidate, trade favors and scratch backs or, better yet, pluck those backs free of botflies and ticks.

Over time, the demands of a social animal’s social life may come to swamp all other selective pressures in the environment, possibly serving as the dominant spur for the evolution of ever-bigger vote-tracking brains. And though we humans may vaguely disapprove of our political impulses and harbor “Fountainhead” fantasies of pulling free in full glory from the nattering tribe, in fact for us and other highly social species there is no turning back. A lone wolf is a weak wolf, a failure, with no chance it will thrive.

Dario Maestripieri, a primatologist at the University of Chicago, has observed a similar dilemma in humans and the rhesus monkeys he studies.

“The paradox of a highly social species like rhesus monkeys and humans is that our complex sociality is the reason for our success, but it’s also the source of our greatest troubles,” he said. “Throughout human history, you see that the worst problems for people almost always come from other people, and it’s the same for the monkeys. You can put them anywhere, but their main problem is always going to be other rhesus monkeys.”

As Dr. Maestripieri sees it, rhesus monkeys embody the concept “Machiavellian” (and he accordingly named his recent popular book about the macaques “Macachiavellian Intelligence”).

“Individuals don’t fight for food, space or resources,” Dr. Maestripieri explained. “They fight for power.” With power and status, he added, “they’ll have control over everything else.”

Rhesus monkeys, midsize omnivores with ruddy brown fur, long bearded faces and disturbingly humanlike ears, are found throughout Asia, including in many cities, where they, like everybody else, enjoy harassing the tourists. The monkeys typically live in groups of 30 or so, a majority of them genetically related females and their dependent offspring.

A female monkey’s status is usually determined by her mother’s status. Male adults, as the ones who enter the group from the outside, must establish their social positions from scratch, bite, baring of canines and, most importantly, rallying their bases.

“Fighting is never something that occurs between two individuals,” Dr. Maestripieri said. “Others get involved all the time, and your chances of success depend on how many allies you have, how wide is your network of support.”

Monkeys cultivate relationships by sitting close to their friends, grooming them at every possible opportunity and going to their aid — at least, when the photo op is right. “Rhesus males are quintessential opportunists,” Dr. Maestripieri said. “They pretend they’re helping others, but they only help adults, not infants. They only help those who are higher in rank than they are, not lower. They intervene in fights where they know they’re going to win anyway and where the risk of being injured is small.”

In sum, he said, “they try to gain maximal benefits at minimal cost, and that’s a strategy that seems to work” in advancing status.

Not all male primates pursue power by appealing to the gents. Among olive baboons, for example, a young male adult who has left his natal home and seeks to be elected into a new baboon group begins by making friendly overtures toward a resident female who is not in estrous at the moment and hence not being contested by other males of the troop.

“If the male is successful in forming a friendship with a female, that gives him an opening with her relatives and allows him to work his way into the whole female network,” said Barbara Smuts, a biologist at the University of Michigan. “In olive baboons, friendships with females can be much more important than political alliances with other males.”

Because males are often the so-called dispersing sex, while females stay behind in the support network of their female kin, females form the political backbone among many social mammals; the longer-lived the species, the denser and more richly articulated that backbone is likely to be.

With life spans rivaling ours, elephants are proving to possess some of the most elaborate social networks yet observed, and their memories for far-flung friends and relations are well in line with the species’ reputation. Elephant society is organized as a matriarchy, said George Wittemyer, an elephant expert at the University of California, Berkeley, with a given core group of maybe 10 elephants led by the eldest resident female. That core group is together virtually all the time, traveling over considerable distances, stopping to dig water holes, looking for fresh foliage to uproot and devour.

“They’re constantly making decisions, debating among themselves, over food, water and security,” Dr. Wittemyer said. “You can see it in the field. You can hear them vocally disagree.” Typically, the matriarch has the final say, and the others abide by her decision. If a faction disagrees strongly enough and wants to try a different approach, “the group will split up and meet back again later,” said Dr. Wittemyer.

Age has its privileges, he said, and the older females, even if they are not the biggest, will often get the best spots to sleep and the best food to eat. But it also has its responsibilities, and a matriarch is often the one to lead the charge in the face of conflicts with other elephants or predatory threats, sometimes to lethal effect.

Hal Whitehead of Dalhousie University and his colleagues have found surprising parallels between the elephant and another mammoth mammal, the sperm whale, possessor of the largest brain, in absolute terms, that the world has ever known. As with elephants, sperm whale society is sexually segregated, the females clustering in oceanic neighborhoods 40 degrees north or south of the Equator, and the males preferring waters around the poles.

As with elephants, the core social unit is a clan of some 10 or 12 females and their offspring. Sperm whales also are highly vocal. They communicate with one another using a Morse code-like pattern of clicks. Each clan, Dr. Whitehead said, has a distinctive click dialect that the members use to identify one another and that adults pass to the young. In other words, he said, “It looks like they have a form of culture.”

Nobody knows what the whales may have to click and clack about, but it could be a form of voting — time to stop here and synchronously dive down in search of deep water squid, now time to resurface, move on, dive again. Clans also seem to caucus on which males they like and will mate with more or less as a group and which ones they will collectively spurn. By all appearances, female sperm whales are terrible size queens. Over the generations, they have consistently voted in favor of enhanced male mass. Their dream candidate nowadays is some fellow named Moby, and he’s three times their size.

Can Web-based worlds teach us about the real one?

Researchers are eager to study millions of online gamers that live in 'virtual' communities such Second Life, hoping to gain insight into our 'real world' behaviors.

With last week's stock market sputter and re­­newed warnings of a recession, policymakers and presidential candidates are hawking countless plans to jump-start the economy. These proposals are often complex, sometimes controversial, and almost always conjectural.

If only there were a way to take them for a test drive.

Robert Bloomfield is tinkering with such a plan. As an accounting professor at Cornell University, in Ithaca, N.Y., he researches virtual worlds – a nascent, but growing, field in the social sciences.

His studies in economic policy lead him into digital realms where the laws of gravity don't apply. But what about the laws of supply and demand?

Immersive online worlds such as Second Life and World of Warcraft attract populations that outnumber Sweden's. And now, scientists are following players down the rabbit hole in hopes of learning more about the real world.

By tapping into the behavior of an estimated 73 million online gamers, Mr. Bloomfield and others hope to study the effects of public policy with an ease and specificity that only computers can deliver. The tools are not yet perfected. But the potential is too strong to ignore, says Bloomfield.

"I got into this because I was talking to the Financial Accounting Standards Board," which develops standards for publicly traded companies in the US, he says. "They have a lot of questions about the effect of legislation, and it's very difficult for them to see before the fact what the policy effects will be."

So, inspired by the popularity of online worlds, several economists imagined creating their own immersive environments. They would design two identical worlds with the same virtual currency. And, sticking to the scientific method, the worlds would differ in some subtle variable.

Then, attract players. Let them loose. Sit back and watch.

The Standards Board loved the idea. But there was a hitch.

"What's missing? Lots and lots of money," Bloomfield says with a laugh. While games like World of Warcraft have multimillion-dollar budgets and teams of programmers, most researchers rely on grants and grad students.

For now, experiments have been repurposed to take place within the commercially successful worlds. And even though this setting is not ideal, several big reports have emerged.

Educators and epidemiologists have published studies on how players react to pandemics in World of Warcraft and the social game Whyville, which markets to young teens. IBM found that team captains in fast-paced fantasy games develop strong leadership skills – talents that the company says are applicable and highly prized in the corporate world.

Bloomfield is combing through data from a virtual stock exchange within Second Life, where avatars buy and sell shares in digital companies, earning in-game currency that is tied to real-life dollars. He's studying how unregulated markets behave. (Early analysis shows that small investors don't fare very well compared with the CEOs of the companies in which they invest – especially companies with heavy concentration of power in one person. The more distributed the control of a company, the better the returns for investors.)

Many skeptics, however, say that results found online don't mean anything in the real world. While virtual worlds are more realistic and immersive than Pong, they are still video games. Motivations and incentives are purposefully skewed to make the experience fun.

The second consideration is the test subjects themselves, says Danah Boyd, a social-technology expert and doctoral student at the University of California, Berkeley. One thing that attracted many researchers was the sheer size of the population. Major national polls consider 1,000 respondents to be sufficient, but Second Life offers 11.7 million avatars that can be scanned for data.

But think about the people behind the avatars, Ms. Boyd says. They are young – on average 26 to 28 years old. They are early adopters. "They are not a random sample of Americans," she says. "When I'm looking at teenagers, I don't speak about senior citizens.... And when you're talking about Second Life, you're not talking about the population at large."

Proponents of online research counter with figures that the audience for today's "massively multiplayer" games mirror the general public far more closely than most other video games. And in hopes of quelling the skeptics, several studies are attempting to see if reality shines through in the online worlds of pixels and pixies.

"If something that we know is true doesn't work in one of these virtual worlds, then we know that there's a problem," says Edward Castronova, a telecommunications professor at Indiana University in Bloomington. He's working to conclusively document principles such as supply and demand in digital worlds.

Other studies look into which human social quirks still turn up when character movement is controlled by mouse clicks.

Before graduating with a PhD from Stanford University last year, Nick Yee found that concepts of personal space from real life have seeped into Second Life.

"There is a well-known rule in the physical world that both personal distance and eye gaze are indicators of intimacy," Mr. Yee says. "So, when you're in an elevator, because you're already so close to people around you, it would be incredibly uncomfortable to look them in the eye unless you were very intimate with them. And so, in an elevator, everyone just tries to look at the blinking numbers."

Yee found the same phenomenon in Second Life. Within a distance of about 12 (virtual) meters, avatars who don't know one another generally look away.

Nothing is conclusive so far, concedes Professor Castronova. But he's certain that with time and funding, major research will emerge from virtual worlds.

"We're building petri dishes for social science," he says. "And if we're able to calibrate this machine correctly, I don't have any doubt that the results will be huge." 

New study links caffeine to miscarriages

Many obstetricians already advise women to limit caffeine, though the subject has long been contentious, with conflicting studies, fuzzy data and various recommendations given over the years.

The new study, being published Monday in the Journal of Obstetrics and Gynecology, finds that pregnant women who consume 200 milligrams or more of caffeine a day - the amount in 10 ounces of coffee or 25 ounces of tea - may double their risk of miscarriage. Ten ounces is equivalent to about 300 milliliters.

Pregnant women should try to give up caffeine for at least the first 3 or 4 months, said the lead author of the study, Dr. De-Kun Li, a reproductive and perinatal epidemiologist at the Kaiser Permanente Division of Research in Oakland, California.

"If, for whatever reason, they really can't do it, think of cutting to one cup or switching to decaf," Li said. "Stopping caffeine really doesn't have any downside."Click here to find out more!

Professional groups like the American College of Obstetricians and Gynecologists and the American Society for Reproductive Medicine have not taken official positions on caffeine, representatives said.

On Friday, the March of Dimes Web site said most experts agreed that the amount of caffeine found in 8 to 16 ounces of coffee a day was safe. It noted that some studies had linked higher amounts to miscarriage and low birth weight, but stated: "However, there is no solid proof that caffeine causes these problems. Until more is known, women should limit their caffeine intake during pregnancy."

Now, having reviewed the new study, the March of Dimes plans to change its message, to advise women who are pregnant or trying to conceive to limit their daily caffeine intake to 200 milligrams or less, said Janis Biermann, its senior vice president of education and health promotion.

Li's study included 1,063 pregnant women who were interviewed once about their caffeine intake. At the time of the interview, their median length of pregnancy was 71 days. But 102 had already miscarried - not surprising, because most miscarriages occur very early in pregnancy. Later, 70 more women miscarried, for an overall miscarriage rate of 16 percent for the group, a typical rate.

Of 264 women who said they used no caffeine, 12.5 percent had miscarriages. But the miscarriage rate was 24.5 percent in the 164 women who consumed 200 milligrams or more per day. The increased risk was associated with caffeine itself and not with other known risk factors like the mother's age or smoking habits, the researchers said.

Li said the study had answered an important question that previous research had left unresolved. Women who have morning sickness are less likely to miscarry than those who do not, possibly because the same hormonal changes that cause nausea and vomiting contribute to a healthy pregnancy. But some researchers said morning sickness could lead to misleading results in caffeine studies.

These researchers argued that because they feel ill, some women may consume less caffeine. That tendency may make it appear that they are less likely to miscarry because they avoid caffeine, when the real reason is actually that they started out with healthier pregnancies.

Li said he and his colleagues had carefully analyzed the data and determined that the risk from caffeine was real and could not be explained away by different rates of morning sickness.

Dr. Carolyn Westhoff, a professor of obstetrics and gynecology, and epidemiology, at Columbia University Medical Center in New York, had reservations about the study, noting that miscarriage is difficult to study or explain.

She said most miscarriages resulted from chromosomal abnormalities, and there was no evidence that caffeine could cause those problems.

FDA to Parents: Do Not Give Tots Cough and Cold Meds

The Food and Drug Administration warns that over-the-counter medications can produce potentially life-threatening side effects
 
 
JUST SAY NO TO COLD DRUGS: FDA warns parents again never to give over-the-counter cold medications to kids under two years of age.

The Food and Drug Administration this week issued a stern health advisory once again warning parents not to give babies under two years of age over-the-counter (OTC) cold and cough medicine because of potentially "serious and life-threatening side effects." This includes decongestants, expectorants, antihistamines and antitussives (cough suppressants) that you can pick up at pharmacies and supermarkets, including Wyeth's Robitussin, Novartis, AG's Triaminic and Johnson & Johnson's Tylenol Plus Cold.

"The FDA strongly recommends to parents and caregivers that OTC cough and cold medicines not be used for children younger than two," Charles Ganley, director of the FDA's Office of Nonprescription Products said. "These medicines, which treat the symptoms and not the underlying condition, have not been shown to be safe or effective in children under two."

The announcement comes on the heels of a high-profile meeting last October at which experts warned of dangerous side effects in children and recommended they not be given to those younger than six years old. Besides fatalities, adverse effects reported include convulsions, rapid heart rates, and reduced levels of consciousness.

The FDA noted in its public advisory that it is "aware of reports of serious side effects" in children between two and 11 years old, but is still reviewing information about risks in that age group. The agency in August warned parents not to give over-the-counter cough and cold remedies to tykes under two years old, but issued the current alert because it was worried parents did not get or heed the message.

After last year's huddle, many drug companies voluntarily pulled 14 cough and cold products targeted at toddlers from store shelves. The FDA in the past has not required pharmaceutical manufacturers to prove the elixirs, which have been sold for decades, work in children, allowing dosing to be gleaned from adult data. But pediatricians have been  increasingly concerned about the safety of these products in their young patients.

"Children metabolize and react to medications differently than adults, often in unanticipated ways,'' the American Academy of Pediatrics said in a statement released after the FDA alert. "Studies have shown cough and cold products are ineffective in treating symptoms of children under six years old and may pose serious risks.''

Physicians have suggested using old-fashioned remedies to help ease cold symptoms, including drinking lots of fluids (among them, Mom's favorite chicken soup), humidifiers and rubbing babies' backs in a steamed-up bathroom.

The FDA is expected to rule by the spring on whether the drugs should also be nixed for kids between the ages of two and 11. In the meantime, it cautions parents who use them in that age group to, among other things, carefully follow dosing directions, only use measuring spoons or cups that come with or are specifically designed to be used with the drugs, and remember that the drugs, at best, temporarily mask symptoms of but neither cure nor shorten the duration of colds or coughs.

Calendar
04 2024/05 06
S M T W T F S
1 2 3 4
5 6 7 8 9 10 11
12 13 14 15 16 17 18
19 20 21 22 23 24 25
26 27 28 29 30 31
Timepiece
タグホイヤー フォーミュラー1 ドリームキャンペーン
Blog Plus
SEO / RSS
Podcast
by PODCAST-BP
New TB
Bar Code
Data Retrieval
Oldest Articles
(09/30)
(09/30)
(09/30)
(09/30)
(09/30)
Photo Index
Reference
Latina




RSS Reader
無料RSSブログパーツ
Misc.
◆BBS


◆Chat


◆Micro TV


Maps



顔文字教室




Copyright © Info All Rights Reserved.
Powered by NinjaBlog
Graphics by 写真素材Kun * Material by Gingham * Template by Kaie
忍者ブログ [PR]