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Minn. Sunday School Students Raise Money For Gay Marriage With Lemonade Stand
Last weekend, children from the First Universalist Church of Minneapolis set up a lemonade stand to support marriage equality during the church's fundraiser, the Golden Valley (Minn.) Patch reported.
Church officials called the event the "Lemonade Stand-ing on the Side of Love." Young Sunday school students charged thirsty customers a quarter for a glass of lemonade. All of the proceeds went to the Minnesota Unitarian Universalist Social Justice Alliance, an organization that is fighting against a proposed amendment to the state's constitution that would ban gay marriage.
"It was really neat -- a lot of the kids had made posters during their time in Religious Education class, and hung those up to direct people from the sanctuary to the social hall," Maren Ahlberg, a First Universalist member, told Patch.
Lauren Wyeth, the church's head of religious education, said several children were motivated to sell lemonade for the cause.
"A lot of the folks that are here at First Universalist want to raise their children in a way that honors the diversity of families and honors all kinds of love," Wyeth said.
In all, the group of children raised $150 for the organization.
Golden Valley Patch reported that the goal of the event was to "give voters personal examples of who stands to lose if the amendment passes." Wyeth also said that the event was to help the children feel confident with sharing stories about their parents, friend's parents or a LGBT relative.
"Those conversations come up on the playground," Wyeth said. "If other kids are repeating the anti-gay attitudes that they pick up from the broader culture, or their parents, or attack ads they'll usually have more confidence."
When the ultra-conservative website Free Republic posted the story, commenters reaction in blunt statements of hate typical of its far-right viewpoint that advocates against LGBT equality or even the legitimacy of same-sex attraction. Several commenters opined that this church is not truly "Christian."
"Yeah. Because this was a pro-homo thing, my guess is that yellow stuff wasn't really lemonade," a commenter wrote. "Church ought to be quotation marks. The unitarian universalist church is just a bunch of liberals gathering together to worship whatever feel like at the moment," another reader commented. "I think it was a LemonAids Stand," one user wrote.
Minnesota does not recognize same-sex marriage on a state level but domestic partnership ordinances have been legalized in 12 cities, including Minneapolis, Duluth and St. Paul. Additionally, the state has anti-discrimination and hate crime laws that protect on the basis of sexual orientation and gender identity. Same-sex couples are also allowed to adopt children.
The First Universalist Church of Minneapolis' website devotes an entire page to marriage equality and lists its goals on how to defeat the marriage amendment.


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I like him, he keeps his feet moving after first contact when blocking somebody also it looks like he has some pretty soft hands for a fb.
Check this guy out. He's the number 1 rated FB in the draft.
[ame="http://www.youtube.com/watch?v=0a4x6V31BMQ"]YouTube - Henry Hynoski #27 Pitt Fullback[/ame]
Great Googly Moogly
RyanD82 - Twitter
I like him, he keeps his feet moving after first contact when blocking somebody also it looks like he has some pretty soft hands for a fb.
Like him....
What round is he projected to go in? Might take a flyer on this guy, FB's aren't too coveted anymore, and He could probably do better than Tim Castille and Mike Cox.
Would be nice to draft one and keep him for awhile kinda like how T-Rich wash before we let him go to the Jets. (Dude is still kicking, slowly but he's kicking)
I can't imagine him being a highly projected pick. Just a guess and I have no stats to support this, but I would bet historically kickers and punters are probably more often drafted higher.
I wouldn't have a problem using a 5th or 6th on him if he is that good, but anything else I would rather take a flyer on hoping to land a diamond in the rough WR, O-lineman or even QB to replace Brodie. Oh don't forget taking a shot on a pass rushing LB or Nose tackle.
I like him! If he can lead block like that in the NFL for Jamal, Wow! Can we say we like pancakes!
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Id have to see more of this guy blocking before I would begin to say its something we should waste a draft pick on, those little FB passes and scrambles the guy wouldnt get 2 yards on in the NFL.
I would gladly spend a 4th round or later on him if we don't get a FB through free agency. We are going to be running a "meat and potatoes" offense and as much as i like castille and cox, the fb needs to be upgraded for our offense to improve. Ideally, we could grab him with one of our 5ths.
Great Googly Moogly
RyanD82 - Twitter

subset: commoncrawl


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PHOENIX, Nov. 14, 2012 /PRNewswire/ -- EastBridge Investment Group (OTCQB: EBIG) today announced that it has entered into an agreement to merge with Cellular Biomedicine Group Inc. (CBMG), a translational medicine company with serial preclinical and clinical research projects and multiple planned clinical trials in the Greater China market. The combined company will be headquartered in Palo Alto, California.
Under the terms of the merger agreement, CBMG shareholders will be issued shares of EastBridge constituting 70% of the combined company with the EastBridge shareholders retaining the other 30%. The parties anticipate that CBMG's current Chairman and CEO Steve Liu, will lead the combined company as Chairman and CEO; EastBridge's current Chairman and CEO Keith Wong will take up two new roles as Chief Business Development Officer, CBDO, and Board Director of the combined company; and Norm Klein, the current COO and CFO, will also take up two new roles as CFO and Board Director of the combined company. The transaction is expected to close early in the first quarter of 2013. Both companies have agreed to continue to operate as separate divisions which should allow them to continue to expand current business opportunities with each other's assistance. The combined company plans to apply for listing on the NASDAQ exchange as soon as practicable with the future Nasdaq ticker symbol "CBMG" reserved.
Led by a team of seasoned management personnel and scientists, CBMG has developed cellular medicine technologies featuring autologous and allogeneic platforms. CBMG's portfolio of intellectual property consists of in-house developed protocols for regenerative medicine therapies and in-licensed immunotherapy patents and know-how exclusively for Greater China. In collaboration with multiple Chinese AAA hospitals, CBMG maintains a pipeline of clinical trials based on US protocols, including treatments for knee osteoarthritis, liver cancer, skin cancer, and spinal muscular atrophy. CBMG operates two biologics GMP facilities in China, which are compliant with U.S. standards.
Keith Wong, Chairman and CEO of EastBridge, stated "We are happy to announce the merger with CBMG as there are many clear synergies between the two companies." Steve Liu, Chairman and CEO of CBMG stated, "We are very excited to be a part of China's $50 billion pharmaceutical market today. It is estimated that within five years this market will become the second largest healthcare market in the world. We look forward to working with EastBridge to augment CBMG's growth in these exciting times."
Conference call details:
When: Tuesday, November 20th, 2012
Time: 10:00 AM ET
Dial in: 1-605-475-6350
Passcode: 769117
About Cellular Biomedicine Group
Cellular Biomedicine Group Inc (CBMG) offers cell therapies for the treatment of cancer and degenerative diseases. Our developmental stem cell, progenitor cell, and immune cell products are the result of collaborative research and development between scientists and doctors from the U.S. and China. Our flagship cGMP facility, consisting of eight independent cell production lines, is designed, certified and managed according to U.S. standards. To learn more about CBMG, please visit: www.cellbiomedgroup.com
About EastBridge
EastBridge Investment Group focuses on high-growth companies in Asia and in the United States, offering IPOs, Joint Ventures and Merchant Banking services. The Company targets industries in electronics, real estate, auto, metal, energy, environmental, bioscience and food retail distribution. To learn more about EastBridge Investment Group go to our web site: www.EbigCorp.com. To receive EBIG's email alert, send a blank email to firstname.lastname@example.com.
Forward-Looking Statements
Statements in this press release relating to plans, strategies, economic performance and trends, projections of results of specific activities or investments, and other statements that are not descriptions of historical facts may be forward-looking statements within the meaning of the Private Securities Litigation Reform Act of 1995, Section 27A of the Securities Act of 1933 and Section 21E of the Securities Exchange Act of 1934. Forward-looking information is inherently subject to risks and uncertainties, and actual results could differ materially from those currently anticipated due to a number of factors, which include, but are not limited to, risk factors inherent in doing business. Forward-looking statements may be identified by terms such as "may," "will," "should," "could," "expects," "plans," "intends," "anticipates," "believes," "estimates," "predicts," "forecasts," "potential," or "continue," or similar terms or the negative of these terms. Although we believe that the expectations reflected in the forward-looking statements are reasonable, we cannot guarantee future results, levels of activity, performance or achievements. The Company has no obligation to update these forward-looking statements.
|SOURCE EastBridge Investment Group
Copyright©2012 PR Newswire.
All rights reserved


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Read what I've highlighted in red!
New Brunswick town plagued for weeks by 'earthquake swarm' — and no one knows why
Joe O'Connor Apr 17, 2012
McAdam, New Brunswick, has been struck by over 35 minor tremours — called an "earthquake swarm" — in the past five weeks.
Springtime in McAdam, a tiny village in southwestern New Brunswick not far from the Maine border, is like springtime in most other parts of Canada. Locals chatter about the NHL playoffs, the garden they are planting, the grass that needs to be cut, the fish they can't wait to catch and the cottage they can't wait to get to, once the warm weather really settles in.
Lately, however, an interloper has elbowed its way into the community's daily dialogue. Pushing aside the playoffs. Pushing its way to the very top of the talking points.
"Everybody is talking about the earthquakes," says David Blair, a retired science teacher and lifetime McAdam resident from his home on Old Harvey Road, just east of downtown.
Courtesy of David Blair
Retired science teacher David Blair with a seismograph set up in his basement. "Everybody is talking about the earthquakes," he says.
"You'll be out and about and people will say, 'Did you feel the one last night, or did you feel the one this morning? Some people will say yes, others might say no. It really depends on what you are doing.'
"If you are quiet at home and there is not a lot of noise you are probably going to feel or hear it. But if you are banging around, or if the grandkids are banging around, you sometimes won't know if it's the kids — or if it was an earthquake — they are about the same magnitude, I guess."
Residents were initially rattled awake at 1:40 a.m. on March 10 by a 2.4-magnitude earthquake that was followed three minutes later by a 1.4-magnitude aftershock. People described hearing what sounded like an explosion. Pictures fell off walls. Window panes rattled. Floorboards creaked and groaned. Some houses even shook, while locals, initially, felt a surge of panic that eased, somewhat, by morning with the realization that a bomb had not gone off but a small earthquake had.
Three days later: two more earthquakes. And in the five weeks since there have been 35 additional shakes, a steady tide of minor tremours that is a popular topic of conversation among villagers and a seismic anomaly that scientists can't entirely explain.
"What is happening in McAdam is something called an earthquake swarm," says Stephen Halchuk, a seismologist with Natural Resources Canada. "It is a series of earthquakes, which is rare. But what is particularly unusual about what is happening in McAdam is that it is basically happening directly beneath the village — and at a depth of less than one kilometre.
"When these earthquake swarms occur they are typically in a remote area where nobody feels them."
Mr. Halchuk can't say how long the earthquakes will continue for, or if a big one, capable of causing serious property damage and endangering lives, could be lurking among all the little ones.
"We can never say never," he says. "I wouldn't rule out a very large event occurring but, typically, in situations like this, the magnitude of the earthquakes is modest and doesn't increase."
Researchers from the University of New Brunswick addressed community members at an atypical town hall meeting Monday night, answering questions and assuaging lingering fears. One working theory they have to explain the quakes is an early spring thaw. A rapid change in groundwater levels could, perhaps, be causing the underlying rocks to slip and stress, unleashing the multiple shocks.
'You sometimes won't know if it's the kids or if it was an earthquake'
Moodus, Connecticut, is another tiny town with an earthquake problem, one dating back hundreds of years and which, today, is the basis of the local high school's team name: The Noises. The noises in McAdam, meanwhile, have been described by the locals as a loud "bang," a "boom," "like dynamite being blown up," and "a loud thud, like somebody falling out of bed."
Natural Resources and UNB have installed three seismographs in the village to monitor the situation and to flesh out their working hypotheses. One of the machines is in David Blair's basement.
"They are not paying me for it," he says, laughing. "It's just volunteer. The government doesn't have any money except to pay its MPs and MLAs."
Connie Klein is the village clerk. She doesn't really like talking to the media, or the academics, because she doesn't really know what to say. But, when she does talk, she explains how the quakes, to her, have become another noise — in her otherwise tranquil village — a sharp "bang" as though someone were upstairs in her house and dropped an encyclopedia on the floor while she was downstairs washing the dishes.
"You might jump for a second," she says.
But then you get on with your day.
And Tuesday, in McAdam, was one for the ages. The sun was blasting hot and talk of the unusual April weather — and not whether the Earth was about to split open and swallow the place whole — was on everybody's lips.
"It has been really nice here," Ms. Klein says. "In fact, it's actually too hot."


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Radioactive waste leaking
AT LEAST six underground tanks containing radioactive waste are leaking at a site that was used to make Cold War-era bombs in the US state of Washington.
Authorities in the state are urging more federal help to clean up the site, which first produced fuel for nuclear bombs in World War II and closed 25 years ago
Washington's governor, Jay Inslee, said the extent of the leaks at the Hanford Site, more than eight kilometres from the Columbia River, was disturbing.
''There is no immediate or near-term health risk associated with these newly discovered leaks …'' he said, after meeting the US energy secretary, Steven Chu.
''But nonetheless this is disturbing news for all Washingtonians.''
Details of exactly what kind of waste was involved were not immediately available. The Hanford nuclear site, 300 kilometres south-east of Seattle, was used to produce plutonium for the bomb that brought an end to World War II.
Millions of litres of leftover waste is contained in 177 tanks at the site.
Agence France-Presse
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The Institute communicates its scientific and educational information in a number of ways. One major outlet is peer-reviewed publications and reports. Cary Institute staff regularly publish in the best-rated journals in their respective fields.
Export 1 results:Sort by: Author Title [ Type] Year
Filters: Author is Lewin, N. [Clear All Filters]
Journal Article
The roles of small rodents in creating patchy environments", Pol. Ecol. Stud., vol. 20, p. 265-276, 1994., "


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Dear Colleague,
The Society for Molecular Biology & Evolution is now accepting proposals
for symposium topics for the 2014 Annual Meeting, taking place in San Juan
Puerto Rico, June 8th - 12th 2014.
Continue Reading →
FeaturedMeeting Announcements
Hi folks,
As you probably know, Twitter uses Hashtags to group tweets together and if you look for a particular hashtag you can see all/many tweets that have used this hashtag.
Facebook has now introduced these as well. Therefore, we are suggesting some hashtags that you can use on twitter and facebook so that we can add to the discussion and instantaneous analysis. Also, if somebody cannot get to the meeting or if somebody cannot make a particular talk, then these hashtags can be useful in gathering the information together for the meeting.
Dear SMBE Members:
SMBE is accepting proposals to host its International Meeting in 2015. The meeting is usually held in June or July and attracts ~1500 scientists from throughout the world. The Society will accept proposals from any location, but the successful location will need to have a lecture venue of sufficient size to host plenary lectures for all attendees and will also need to be near housing, preferably with a wide variety of options.
In the past 25 years, stretching from the dawn of personal computing to the social media age, the "always free" molecular evolutionary genetics analysis (MEGA) tool has been downloaded 1.6 million times worldwide.
Vincent L. Cannataro and Jeffrey P. Townsend
AbstractSelective sweep is a phenomenon of reduced variation at presumably neutrally evolving sites (hitchhikers) in the genome that is caused by the spread of a selected allele at a linked focal site, and is widely used to test for action of positive selection. Nonetheless, selective sweep may also provide an unprecedented opportunity for studying nonequilibrium properties of the neutral variation itself. We have demonstrated this possibility in relation to ancient selective sweep for modern human-specific changes and ongoing selective sweep for local population-specific changes.
AbstractThe Molecular Evolutionary Genetics Analysis (Mega) software implements many analytical methods and tools for phylogenomics and phylomedicine. Here, we report a transformation of Mega to enable cross-platform use on Microsoft Windows and Linux operating systems. Mega X does not require virtualization or emulation software and provides a uniform user experience across platforms. Mega X has additionally been upgraded to use multiple computing cores for many molecular evolutionary analyses. Mega X is available in two interfaces (graphical and command line) and can be downloaded from www.megasoftware.net free of charge.
AbstractIn this perspective, we evaluate the explanatory power of the neutral theory of molecular evolution, 50 years after its introduction by Kimura. We argue that the neutral theory was supported by unreliable theoretical and empirical evidence from the beginning, and that in light of modern, genome-scale data, we can firmly reject its universality. The ubiquity of adaptive variation both within and between species means that a more comprehensive theory of molecular evolution must be sought.
AbstractKimura's neutral theory provides the whole theoretical basis of the behavior of mutations in a Wright–Fisher population. We here discuss how it can be applied to a cancer cell population, in which there is an increasing interest in genetic variation within a tumor. We explain a couple of fundamental differences between cancer cell populations and asexual organismal populations. Once these differences are taken into account, a number of powerful theoretical tools developed for a Wright–Fisher population could be readily contribute to our deeper understanding of the evolutionary dynamics of cancer cell population.
AbstractHIV is one of the fastest evolving organisms known. It evolves about 1 million times faster than its host, humans. Because HIV establishes chronic infections, with continuous evolution, its divergence within a single infected human surpasses the divergence of the entire humanoid history. Yet, it is still the same virus, infecting the same cell types and using the same replication machinery year after year. Hence, one would think that most mutations that HIV accumulates are neutral. But the picture is more complicated than that. HIV evolution is also a clear example of strong positive selection, that is, mutants have a survival advantage. How do these facts come together?
AbstractImportance of chance, finiteness, and history in evolution is pointed out with special reference to the neutral theory.
AbstractThe evolution of viral pathogens is shaped by strong selective forces that are exerted during jumps to new hosts, confrontations with host immune responses and antiviral drugs, and numerous other processes. However, while undeniably strong and frequent, adaptive evolution is largely confined to small parts of information-packed viral genomes, and the majority of observed variation is effectively neutral. The predictions and implications of the neutral theory have proven immensely useful in this context, with applications spanning understanding within-host population structure, tracing the origins and spread of viral pathogens, predicting evolutionary dynamics, and modeling the emergence of drug resistance. We highlight the multiple ways in which the neutral theory has had an impact, which has been accelerated in the age of high-throughput, high-resolution genomics.
AbstractResearch in population genetics and evolutionary biology has always provided a computational backbone for life sciences as a whole. Today evolutionary and population biology reasoning are essential for interpretation of large complex datasets that are characteristic of all domains of today's life sciences ranging from cancer biology to microbial ecology. This situation makes algorithms and software tools developed by our community more important than ever before. This means that we, developers of software tool for molecular evolutionary analyses, now have a shared responsibility to make these tools accessible using modern technological developments as well as provide adequate documentation and training.
AbstractGenetic differences between species and within populations are two sides of the same coin under the neutral theory of molecular evolution. This theory posits that a vast majority of evolutionary substitutions, which appear as differences between species, are (nearly) neutral, that is, these substitutions are permitted without a significantly adverse impact on a species' survival. We refer to them as evolutionarily permissible (ePerm) variation. Evolutionary permissibility of any possible variant can be inferred from multispecies sequence alignments by applying sophisticated statistical methods to the evolutionary tree of species. Here, we explore the evolutionary permissibility of amino acid variants associated with genetic diseases and those observed in personal exomes. Consistent with the predictions of the neutral theory, disease associated amino acid variants are rarely ePerm, much more biochemically radical, and found predominantly at more conserved positions than their non-disease counterparts. Only 10% of amino acid mutations are ePerm, but these variants rise to become two-thirds of all substitutions in the human lineage (a 6-fold enrichment). In contrast, only a minority of the variants in a personal exome are ePerm, a seemingly counterintuitive pattern that results from a combination of mutational and evolutionary processes that are, in fact, broadly consistent with the neutral theory. Evolutionarily forbidden variants outnumber detrimental variants in individual exomes and may play an underappreciated role in protecting against disease. We discuss these observations and conclude that the long-term evolutionary history of species can illuminate functional biomedical properties of variation present in personal exomes.
AbstractAmong the multitude of papers published yearly in scientific journals, precious few publications may be worth looking back in half a century to appreciate the significance of the discoveries that would later become common knowledge and get a chance to shape a field or several adjacent fields. Here, Kimura's fundamental concept of neutral mutation-random drift, which was published 50 years ago, is re-examined in light of its pervasive influence on comparative genomics and, more specifically, on the contribution of transposable elements to eukaryotic genome evolution.
AbstractI detail four major open problems in microbial population genetics with direct implications to the study of molecular evolution: the lack of neutral polymorphism, the modeling of promiscuous genetic exchanges, the genetics of ill-defined populations, and the difficulty of untangling selection and demography in the light of these issues. Together with the historical focus on the study of single nucleotide polymorphism and widespread non-random sampling, these problems limit our understanding of the genetic variation in bacterial populations and their adaptive effects. I argue that we need novel theoretical approaches accounting for pervasive selection and strong genetic linkage to better understand microbial evolution.
AbstractKimura's neutral theory argued that positive selection was not responsible for an appreciable fraction of molecular substitutions. Correspondingly, quantitative analysis reveals that the vast majority of substitutions in cancer genomes are not detectably under selection. Insights from the somatic evolution of cancer reveal that beneficial substitutions in cancer constitute a small but important fraction of the molecular variants. The molecular evolution of cancer community will benefit by incorporating the neutral theory of molecular evolution into their understanding and analysis of cancer evolution—and accepting the use of tractable, predictive models, even when there is some evidence that they are not perfect.
AbstractKimura's neutral theory of molecular evolution has been essential to virtually every advance in evolutionary genetics, and by extension, is foundational to the field of conservation genetics. Conservation genetics utilizes the key concepts of neutral theory to identify species and populations at risk of losing evolutionary potential by detecting patterns of inbreeding depression and low effective population size. In turn, this information can inform the management of organisms and their habitat providing hope for the long-term preservation of both. We expand upon Avise's "inventorial" and "functional" categories of conservation genetics by proposing a third category that is linked to the coalescent and that we refer to as "process-driven." It is here that connections between Kimura's theory and conservation genetics are strongest. Process-driven conservation genetics can be especially applied to large genomic data sets to identify patterns of historical risk, such as population bottlenecks, and accordingly, yield informed intuitions for future outcomes. By examining inventorial, functional, and process-driven conservation genetics in sequence, we assess the progression from theory, to data collection and analysis, and ultimately, to the production of hypotheses that can inform conservation policies.
AbstractDAMBE is a comprehensive software package for genomic and phylogenetic data analysis on Windows, Linux, and Macintosh computers. New functions include imputing missing distances and phylogeny simultaneously (paving the way to build large phage and transposon trees), new bootstrapping/jackknifing methods for PhyPA (phylogenetics from pairwise alignments), and an improved function for fast and accurate estimation of the shape parameter of the gamma distribution for fitting rate heterogeneity over sites. Previous method corrects multiple hits for each site independently. DAMBE's new method uses all sites simultaneously for correction. DAMBE, featuring a user-friendly graphic interface, is freely available from http://dambe.bio.uottawa.ca (last accessed, April 17, 2018).
AbstractFish mitochondrial genome (mitogenome) data form a fundamental basis for revealing vertebrate evolution and hydrosphere ecology. Here, we report recent functional updates of MitoFish, which is a database of fish mitogenomes with a precise annotation pipeline MitoAnnotator. Most importantly, we describe implementation of MiFish pipeline for metabarcoding analysis of fish mitochondrial environmental DNA, which is a fast-emerging and powerful technology in fish studies. MitoFish, MitoAnnotator, and MiFish pipeline constitute a key platform for studies of fish evolution, ecology, and conservation, and are freely available at http://mitofish.aori.u-tokyo.ac.jp/ (last accessed April 7th, 2018).
AbstractHere, we present a synthetic view on how Kimura's Neutral theory has helped us gaining insight on the different evolutionary forces that shape human evolution. We put this perspective in the frame of recent emerging challenges: the use of whole genome data for reconstructing population histories, natural selection on complex polygenic traits, and integrating cultural processes in human evolution.
AbstractPrimates have traditionally been regarded as vision-oriented animals with low olfactory ability, though this "microsmatic primates" view has been challenged recently. To clarify when and how degeneration of the olfactory system occurred and to specify the relevant factors during primate evolution, we here examined the olfactory receptor (OR) genes from 24 phylogenetically and ecologically diverse primate species. The results revealed that strepsirrhines with curved noses had functional OR gene repertoires that were nearly twice as large as those for haplorhines with simple noses. Neither activity pattern (nocturnal/diurnal) nor color vision system showed significant correlation with the number of functional OR genes while phylogeny and nose structure (haplorhine/strepsirrhine) are statistically controlled, but extent of folivory did. We traced the evolutionary fates of individual OR genes by identifying orthologous gene groups, demonstrating that the rates of OR gene losses were accelerated at the ancestral branch of haplorhines, which coincided with the acquisition of acute vision. The highest rate of OR gene loss was observed at the ancestral branch of leaf-eating colobines; this reduction is possibly linked with the dietary transition from frugivory to folivory because odor information is essential for fruit foraging but less so for leaf foraging. Intriguingly, we found accelerations of OR gene losses in an external branch to every hominoid species examined. These findings suggest that the current OR gene repertoire in each species has been shaped by a complex interplay of phylogeny, anatomy, and habitat; therefore, multiple factors may contribute to the olfactory degeneration in primates.
AbstractAlthough the neutral theory of molecular evolution was proposed to explain DNA and protein sequence evolution, in principle it could also explain phenotypic evolution. Nevertheless, overall, phenotypes should be less likely than genotypes to evolve neutrally. I propose that, when phenotypic traits are stratified according to a hierarchy of biological organization, the fraction of evolutionary changes in phenotype that are adaptive rises with the phenotypic level considered. Consistently, molecular traits are frequently found to evolve neutrally whereas a large, random set of organismal traits were recently reported to vary largely adaptively. Many more studies of unbiased samples of phenotypic traits are needed to test the general validity of this hypothesis.
AbstractIn its initial formulation by Motoo Kimura, the neutral theory was concerned solely with the level of variability maintained by random genetic drift of selectively neutral mutations, and the rate of molecular evolution caused by the fixation of such mutations. The original theory considered events at a single genetic locus in isolation from the rest of the genome. It did not take long, however, for theoreticians to wonder whether selection at one or more loci might influence neutral variability at linked sites. Once DNA sequence variability could be studied, and especially when resequencing of whole genomes became possible, it became clear that patterns of neutral variability in genomes are affected by selection at linked sites, and that these patterns could advance our understanding of natural selection, and can be used to detect the action of selection in genomic regions, including selection much weaker than could be detected by direct measurements of the relative fitnesses of different genotypes. We outline the different types of processes that have been studied, in approximate order of their historical development.
AbstractSmall insertions and deletions (INDELs; ≤50 bp) are the most common type of variability after single nucleotide polymorphism (SNP). However, compared with SNPs, we know little about the distribution of fitness effects (DFE) of new INDEL mutations and how prevalent adaptive INDEL substitutions are. Studying INDELs has been difficult partly because identifying ancestral states at these sites is error-prone and misidentification can lead to severely biased estimates of the strength of selection. To solve these problems, we develop new maximum likelihood methods, which use polymorphism data to simultaneously estimate the DFE, the mutation rate, and the misidentification rate. These methods are applicable to both INDELs and SNPs. Simulations show that they can provide highly accurate results. We applied the methods to an INDEL polymorphism data set in Drosophila melanogaster. We found that the DFE for polymorphic INDELs in protein-coding regions is bimodal, with the variants being either nearly neutral or strongly deleterious. Based on the DFE, we estimated that 71.5–83.7% of the INDEL substitutions that took place along the D. melanogaster lineage were fixed by positive selection, which is comparable with the prevalence of adaptive substitutions at nonsynonymous sites. The new methods have been implemented in the software package anavar.
AbstractMammalian diversification has coincided with a rapid proliferation of various types of noncoding RNAs, including members of both snRNAs and snoRNAs. The significance of this expansion however remains obscure. While some ncRNA copy-number expansions have been linked to functionally tractable effects, such events may equally likely be neutral, perhaps as a result of random retrotransposition. Hindering progress in our understanding of such observations is the difficulty in establishing function for the diverse features that have been identified in our own genome. Projects such as ENCODE and FANTOM have revealed a hidden world of genomic expression patterns, as well as a host of other potential indicators of biological function. However, such projects have been criticized, particularly from practitioners in the field of molecular evolution, where many suspect these data provide limited insight into biological function. The molecular evolution community has largely taken a skeptical view, thus it is important to establish tests of function. We use a range of data, including data drawn from ENCODE and FANTOM, to examine the case for function for the recent copy number expansion in mammals of six evolutionarily ancient RNA families involved in splicing and rRNA maturation. We use several criteria to assess evidence for function: conservation of sequence and structure, genomic synteny, evidence for transposition, and evidence for species-specific expression. Applying these criteria, we find that only a minority of loci show strong evidence for function and that, for the majority, we cannot reject the null hypothesis of no function.
AbstractThe genetic basis of parallel evolution of similar species is of great interest in evolutionary biology. In the adaptive radiation of Lake Victoria cichlid fishes, sister species with either blue or red-back male nuptial coloration have evolved repeatedly, often associated with shallower and deeper water, respectively. One such case is blue and red-backed Pundamilia species, for which we recently showed that a young species pair may have evolved through "hybrid parallel speciation". Coalescent simulations suggested that the older species P. pundamilia (blue) and P. nyererei (red-back) admixed in the Mwanza Gulf and that new "nyererei-like" and "pundamilia-like" species evolved from the admixed population. Here, we use genome scans to study the genomic architecture of differentiation, and assess the influence of hybridization on the evolution of the younger species pair. For each of the two species pairs, we find over 300 genomic regions, widespread across the genome, which are highly differentiated. A subset of the most strongly differentiated regions of the older pair are also differentiated in the younger pair. These shared differentiated regions often show parallel allele frequency differences, consistent with the hypothesis that admixture-derived alleles were targeted by divergent selection in the hybrid population. However, two-thirds of the genomic regions that are highly differentiated between the younger species are not highly differentiated between the older species, suggesting independent evolutionary responses to selection pressures. Our analyses reveal how divergent selection on admixture-derived genetic variation can facilitate new speciation events.
AbstractIdentifying the genomic basis underlying local adaptation is paramount to evolutionary biology, and bears many applications in the fields of conservation biology, crop, and animal breeding, as well as personalized medicine. Although many approaches have been developed to detect signatures of positive selection within single populations and population pairs, the increasing wealth of high-throughput sequencing data requires improved methods capable of handling multiple, and ideally large number of, populations in a single analysis. In this study, we introduce LSD (levels of exclusively shared differences), a fast and flexible framework to perform genome-wide selection scans, along the internal and external branches of a given population tree. We use forward simulations to demonstrate that LSD can identify branches targeted by positive selection with remarkable sensitivity and specificity. We illustrate a range of potential applications by analyzing data from the 1000 Genomes Project and uncover a list of adaptive candidates accompanying the expansion of anatomically modern humans out of Africa and their spread to Europe.
AbstractDetecting selection on codon usage (CU) is a difficult task, since CU can be shaped by both the mutational process and selective constraints operating at the DNA, RNA, and protein levels. Yang and Nielsen (2008) developed a test (which we call CUYN) for detecting selection on CU using two competing mutation-selection models of codon substitution. The null model assumes that CU is determined by the mutation bias alone, whereas the alternative model assumes that both mutation bias and/or selection act on CU. In applications on mammalian-scale alignments, the CUYN test detects selection on CU for numerous genes. This is surprising, given the small effective population size of mammals, and prompted us to use simulations to evaluate the robustness of the test to model violations. Simulations using a modest level of CpG hypermutability completely mislead the test, with 100% false positives. Surprisingly, a high level of false positives (56.1%) resulted simply from using the HKY mutation-level parameterization within the CUYN test on simulations conducted with a GTR mutation-level parameterization. Finally, by using a crude optimization procedure on a parameter controlling the CpG hypermutability rate, we find that this mutational property could explain a very large part of the observed mammalian CU. Altogether, our work emphasizes the need to evaluate the potential impact of model violations on statistical tests in the field of molecular phylogenetic analysis. The source code of the simulator and the mammalian genes used are available as a GitHub repository (https://github.com/Simonll/LikelihoodFreePhylogenetics.git).
AbstractWhen a substitution model is fitted to an alignment using maximum likelihood, its parameters are adjusted to account for as much site-pattern variation as possible. A parameter might therefore absorb a substantial quantity of the total variance in an alignment (or more formally, bring about a substantial reduction in the deviance of the fitted model) even if the process it represents played no role in the generation of the data. When this occurs, we say that the parameter estimate carries phenomenological load (PL). Large PL in a parameter estimate is a concern because it not only invalidates its mechanistic interpretation (if it has one) but also increases the likelihood that it will be found to be statistically significant. The problem of PL was not identified in the past because most off-the-shelf substitution models make simplifying assumptions that preclude the generation of realistic levels of variation. In this study, we use the more realistic mutation-selection framework as the basis of a generating model formulated to produce data that mimic an alignment of mammalian mitochondrial DNA. We show that a parameter estimate can carry PL when 1) the substitution model is underspecified and 2) the parameter represents a process that is confounded with other processes represented in the data-generating model. We then provide a method that can be used to identify signal for the process that a given parameter represents despite the existence of PL.
AbstractThe olfactory receptor (OR) gene families, which govern mammalian olfaction, have undergone extensive expansion and contraction through duplication and pseudogenization. Previous studies have shown that broadly defined environmental adaptations (e.g., terrestrial vs. aquatic) are correlated with the number of functional and non-functional OR genes retained. However, to date, no study has examined species-specific gene duplications in multiple phylogenetically divergent mammals to elucidate OR evolution and adaptation. Here, we identify the OR gene families driving adaptation to different ecological niches by mapping the fate of species-specific gene duplications in the OR repertoire of 94 diverse mammalian taxa, using molecular phylogenomic methods. We analyze >70,000 OR gene sequences mined from whole genomes, generated from novel amplicon sequencing data, and collated with data from previous studies, comprising one of the largest OR studies to date. For the first time, we demonstrate statistically significant patterns of OR species-specific gene duplications associated with the presence of a functioning vomeronasal organ. With respect to dietary niche, we uncover a novel link between a large number of duplications in OR family 5/8/9 and herbivory. Our results also highlight differences between social and solitary niches, indicating that a greater OR repertoire expansion may be associated with a solitary lifestyle. This study demonstrates the utility of species-specific duplications in elucidating gene family evolution, revealing how the OR repertoire has undergone expansion and contraction with respect to a number of ecological adaptations in mammals.
AbstractWith advances in transcript profiling, the presence of transcriptional activities in intergenic regions has been well established. However, whether intergenic expression reflects transcriptional noise or activity of novel genes remains unclear. We identified intergenic transcribed regions (ITRs) in 15 diverse flowering plant species and found that the amount of intergenic expression correlates with genome size, a pattern that could be expected if intergenic expression is largely nonfunctional. To further assess the functionality of ITRs, we first built machine learning models using Arabidopsis thaliana as a model that accurately distinguish functional sequences (benchmark protein-coding and RNA genes) and likely nonfunctional ones (pseudogenes and unexpressed intergenic regions) by integrating 93 biochemical, evolutionary, and sequence-structure features. Next, by applying the models genome-wide, we found that 4,427 ITRs (38%) and 796 annotated ncRNAs (44%) had features significantly similar to benchmark protein-coding or RNA genes and thus were likely parts of functional genes. Approximately 60% of ITRs and ncRNAs were more similar to nonfunctional sequences and were likely transcriptional noise. The predictive framework established here provides not only a comprehensive look at how functional, genic sequences are distinct from likely nonfunctional ones, but also a new way to differentiate novel genes from genomic regions with noisy transcriptional activities.
Abstractβ-Catenin acts as a transcriptional coactivator in the Wnt/β-catenin signaling pathway and a cytoplasmic effector in cadherin-based cell adhesion. These functions are ancient within animals, but the earliest steps in β-catenin evolution remain unresolved due to limited data from key lineages—sponges, ctenophores, and placozoans. Previous studies in sponges have characterized β-catenin expression dynamics and used GSK3B antagonists to ectopically activate the Wnt/β-catenin pathway; both approaches rely upon untested assumptions about the conservation of β-catenin function and regulation in sponges. Here, we test these assumptions using an antibody raised against β-catenin from the sponge Ephydatia muelleri. We find that cadherin-complex genes coprecipitate with endogenous Em β-catenin from cell lysates, but that Wnt pathway components do not. However, through immunostaining we detect both cell boundary and nuclear populations, and we find evidence that Em β-catenin is a conserved substrate of GSK3B. Collectively, these data support conserved roles for Em β-catenin in both cell adhesion and Wnt signaling. Additionally, we find evidence for an Em β-catenin population associated with the distal ends of F-actin stress fibers in apparent cell–substrate adhesion structures that resemble focal adhesions. This finding suggests a fundamental difference in the adhesion properties of sponge tissues relative to other animals, in which the adhesion functions of β-catenin are typically restricted to cell–cell adhesions.
AbstractThe evolution of new biochemical activities frequently involves complex dependencies between mutations and rapid evolutionary radiation. Mutation co-occurrence and covariation have previously been used to identify compensating mutations that are the result of physical contacts and preserve protein function and fold. Here, we model pairwise functional dependencies and higher order interactions that enable evolution of new protein functions. We use a network model to find complex dependencies between mutations resulting from evolutionary trade-offs and pleiotropic effects. We present a method to construct these networks and to identify functionally interacting mutations in both extant and reconstructed ancestral sequences (Network Analysis of Protein Adaptation). The time ordering of mutations can be incorporated into the networks through phylogenetic reconstruction. We apply NAPA to three distantly homologous β-lactamase protein clusters (TEM, CTX-M-3, and OXA-51), each of which has experienced recent evolutionary radiation under substantially different selective pressures. By analyzing the network properties of each protein cluster, we identify key adaptive mutations, positive pairwise interactions, different adaptive solutions to the same selective pressure, and complex evolutionary trajectories likely to increase protein fitness. We also present evidence that incorporating information from phylogenetic reconstruction and ancestral sequence inference can reduce the number of spurious links in the network, whereas preserving overall network community structure. The analysis does not require structural or biochemical data. In contrast to function-preserving mutation dependencies, which are frequently from structural contacts, gain-of-function mutation dependencies are most commonly between residues distal in protein structure.
AbstractThe visual systems of snakes are heavily modified relative to other squamates, a condition often thought to reflect their fossorial origins. Further modifications are seen in caenophidian snakes, where evolutionary transitions between rod and cone photoreceptors, termed photoreceptor transmutations, have occurred in many lineages. Little previous work, however, has focused on the molecular evolutionary underpinnings of these morphological changes. To address this, we sequenced seven snake eye transcriptomes and utilized new whole-genome and targeted capture sequencing data. We used these data to analyze gene loss and shifts in selection pressures in phototransduction genes that may be associated with snake evolutionary origins and photoreceptor transmutation. We identified the surprising loss of rhodopsin kinase (GRK1), despite a low degree of gene loss overall and a lack of relaxed selection early during snake evolution. These results provide some of the first evolutionary genomic corroboration for a dim-light ancestor that lacks strong fossorial adaptations. Our results also indicate that snakes with photoreceptor transmutation experienced significantly different selection pressures from other reptiles. Significant positive selection was found primarily in cone-specific genes, but not rod-specific genes, contrary to our expectations. These results reveal potential molecular adaptations associated with photoreceptor transmutation and also highlight unappreciated functional differences between rod- and cone-specific phototransduction proteins. This intriguing example of snake visual system evolution illustrates how the underlying molecular components of a complex system can be reshaped in response to changing selection pressures.
In the murky blackwaters of the peat swamp forests of Southeast Asia lives the world's smallest fish, the dwarf minnow of the genus Paedocypris. This extreme environment, characterized by low oxygen and high acidity, is home to several miniaturized fish species. Paedocypris adults are a mere 8–12 mm in length and resemble zebrafish larvae, a phenomenon known as progenesis. In the case of Paedocypris, this developmental truncation is extreme, with over 40 bones found in zebrafish adults that never develop at all in Paedocypris.
AbstractWe report here the complete genome sequences of seven Vibrio anguillarum strains isolated from multiple geographic locations, thus increasing the total number of genomes of finished quality to 11. The genomes were de novo assembled from long-sequence PacBio reads. Including draft genomes, a total of 44 V. anguillarum genomes are currently available in the genome databases. They represent an important resource in the study of, for example, genetic variations and for identifying virulence determinants. In this article, we present the genomes and basic genome comparisons of the 11 complete genomes, including a BRIG analysis, and pan genome calculation. We also describe some structural features of superintegrons on chromosome 2 s, and associated insertion sequence (IS) elements, including 18 new ISs (ISVa3 − ISVa20), both of importance in the complement of V. anguillarum genomes.
Pádraic Corcoran, Toni I. Gossmann, Henry J. Barton, The Great Tit HapMap Consortium, Jon Slate, and Kai Zeng
Wesley C. Warren, Lukas Kuderna, Alana Alexander, Julian Catchen, José G. Pérez-Silva, Carlos López-Otín, Víctor Quesada, Patrick Minx, Chad Tomlinson, Michael J. Montague, Fabiana H.G. Farias, Ronald B. Walter, Tomas Marques-Bonet, Travis Glenn, Troy J. Kieran, Sandra S. Wise, John Pierce Wise Jr, Robert M. Waterhouse, John Pierce Wise Sr
Abstract"Candidatus Fokinia solitaria" is an obligate intracellular endosymbiont of a unicellular eukaryote, a ciliate of the genus Paramecium. Here, we present the genome sequence of this bacterium and subsequent analysis. Phylogenomic analysis confirmed the previously reported positioning of the symbiont within the "Candidatus Midichloriaceae" family (order Rickettsiales), as well as its high sequence divergence from other members of the family, indicative of fast sequence evolution. Consistently with this high evolutionary rate, a comparative genomic analysis revealed that the genome of this symbiont is the smallest of the Rickettsiales to date. The reduced genome does not present flagellar genes, nor the pathway for the biosynthesis of lipopolysaccharides (present in all the other so far sequenced members of the family "Candidatus Midichloriaceae") or genes for the Krebs cycle (present, although not always complete, in Rickettsiales). These results indicate an evolutionary trend toward a stronger dependence on the host, in comparison with other members of the family. Two alternative scenarios are compatible with our results; "Candidatus Fokinia solitaria" could be either a recently evolved, vertically transmitted mutualist, or a parasite with a high host-specificity.
AbstractTaste perception is crucial in monitoring food intake and, hence, is thought to play a significant role in human evolution. To gain insights into possible adaptive signatures in genes encoding bitter, sweet, and umami taste receptors, we surveyed the available sequence variation data from the 1000 Genomes Project Phase 3 for TAS1R (TAS1R1-3) and TAS2R (TAS2R16 and TAS2R38) families. Our study demonstrated that genes from these two families have experienced contrasting evolutionary histories: While TAS1R1 and TAS1R3 showed worldwide evidence of positive selection, probably correlated with improved umami and sweet perception, the patterns of variation displayed by TAS2R16 and TAS2R38 were more consistent with scenarios of balancing selection that possibly conferred a heterozygous advantage associated with better capacity to perceive a wide range of bitter compounds. In TAS2R16, such adaptive events appear to have occurred restrictively in mainland Africa, whereas the strongest evidence in TAS2R38 was detected in Europe. Despite plausible associations between taste perception and the TAS1R and TAS2R selective signatures, we cannot discount other biological mechanisms as driving the evolutionary trajectories of those TAS1R and TAS2R members, especially given recent findings of taste receptors behaving as the products of pleiotropic genes involved in many functions outside the gustatory system.
AbstractDuring the demographic history of the Pan clade, there has been gene-flow between species, likely >200,000 years ago. Bonobo haplotypes in three subspecies of chimpanzee have been identified to be segregating in modern-day chimpanzee populations, suggesting that these haplotypes, with increased differentiation, may be a target of natural selection. Here, we investigate signatures of adaptive introgression within the bonobo-like haplotypes in chimpanzees using site frequency spectrum-based tests. We find evidence for subspecies-specific adaptations in introgressed regions involved with male reproduction in central chimpanzees, the immune system in eastern chimpanzees, female reproduction and the nervous system in Nigeria-Cameroon chimpanzees. Furthermore, our results indicate signatures of balancing selection in some of the putatively introgressed regions. This might be the product of long-term balancing selection resulting in a similar genomic signature as introgression, or possibly balancing selection acting on alleles reintroduced through gene flow.
AbstractBiases in mutation rate can influence molecular evolution, yielding rates of evolution that vary widely in different parts of the genome and even among neighboring nucleotides. Here, we explore one possible mechanism of influence on sequence-specific mutation rates, the electron–hole, which can localize and potentially trigger a replication mismatch. A hole is a mobile site of positive charge created during one-electron oxidation by, for example, radiation, contact with a mutagenic agent, or oxidative stress. Its quantum wavelike properties cause it to localize at various sites with probabilities that vary widely, by orders of magnitude, and depend strongly on the local sequence. We find significant correlations between hole probabilities and mutation rates within base triplets, observed in published mutation accumulation experiments on four species of bacteria. We have also computed hole probability spectra for hypervariable segment I of the human mtDNA control region, which contains several mutational hotspots, and for heptanucleotides in noncoding regions of the human genome, whose polymorphism levels have recently been reported. We observe significant correlations between hole probabilities, and context-specific mutation and substitution rates. The correlation with hole probability cannot be explained entirely by CpG methylation in the heptanucleotide data. Peaks in hole probability tend to coincide with mutational hotspots, even in mtDNA where CpG methylation is rare. Our results suggest that hole-enhanced mutational mechanisms, such as oxidation-stabilized tautomerization and base deamination, contribute to molecular evolution.
AbstractDPANN archaea are primarily known based on genomes from metagenomes and single cells. We reconstructed a complete population genome for Candidatus "Forterrea," a Diapherotrite with a predicted symbiotic lifestyle probably centered around nucleotide metabolism and RuBisCO. Genome-wide analysis of sequence variation provided insights into the processes that shape its population structure in the deep subsurface. The genome contains many transposons, yet reconstruction of a complete genome from a short-read insert data set was possible because most occurred only in some individuals. Accuracy of the final reconstruction could be verified because the genome displays the pattern of cumulative GC skew known for some archaea but more typically associated with bacteria. Sequence variation is highly localized, and most pronounced around transposons and relatively close to the origin of replication. Patterns of variation are best explained by homologous recombination, a process previously not described for DPANN archaea.
AbstractChloroplast genomes have undergone tremendous alterations through the evolutionary history of the green algae (Chloroplastida). This study focuses on the evolution of chloroplast genomes in the siphonous green algae (order Bryopsidales). We present five new chloroplast genomes, which along with existing sequences, yield a data set representing all but one families of the order. Using comparative phylogenetic methods, we investigated the evolutionary dynamics of genomic features in the order. Our results show extensive variation in chloroplast genome architecture and intron content. Variation in genome size is accounted for by the amount of intergenic space and freestanding open reading frames that do not show significant homology to standard plastid genes. We show the diversity of these nonstandard genes based on their conserved protein domains, which are often associated with mobile functions (reverse transcriptase/intron maturase, integrases, phage- or plasmid-DNA primases, transposases, integrases, ligases). Investigation of the introns showed proliferation of group II introns in the early evolution of the order and their subsequent loss in the core Halimedineae, possibly through RT-mediated intron loss.
AbstractThe world's smallest fishes belong to the genus Paedocypris. These miniature fishes are endemic to an extreme habitat: the peat swamp forests in Southeast Asia, characterized by highly acidic blackwater. This threatened habitat is home to a large array of fishes, including a number of miniaturized but also developmentally truncated species. Especially the genus Paedocypris is characterized by profound, organism-wide developmental truncation, resulting in sexually mature individuals of <8 mm in length with a larval phenotype. Here, we report on evolutionary simplification in the genomes of two species of the dwarf minnow genus Paedocypris using whole-genome sequencing. The two species feature unprecedented Hox gene loss and genome reduction in association with their massive developmental truncation. We also show how other genes involved in the development of musculature, nervous system, and skeleton have been lost in Paedocypris, mirroring its highly progenetic phenotype. Further, our analyses suggest two mechanisms responsible for the genome streamlining in Paedocypris in relation to other Cypriniformes: severe intron shortening and reduced repeat content. As the first report on the genomic sequence of a vertebrate species with organism-wide developmental truncation, the results of our work enhance our understanding of genome evolution and how genotypes are translated to phenotypes. In addition, as a naturally simplified system closely related to zebrafish, Paedocypris provides novel insights into vertebrate development.
AbstractDinoflagellates are a group of unicellular protists with immense ecological and evolutionary significance and cell biological diversity. Of the photosynthetic dinoflagellates, the majority possess a plastid containing the pigment peridinin, whereas some lineages have replaced this plastid by serial endosymbiosis with plastids of distinct evolutionary affiliations, including a fucoxanthin pigment-containing plastid of haptophyte origin. Previous studies have described the presence of widespread substitutional RNA editing in peridinin and fucoxanthin plastid genes. Because reports of this process have been limited to manual assessment of individual lineages, global trends concerning this RNA editing and its effect on the biological function of the plastid are largely unknown. Using novel bioinformatic methods, we examine the dynamics and evolution of RNA editing over a large multispecies data set of dinoflagellates, including novel sequence data from the peridinin dinoflagellate Pyrocystis lunula and the fucoxanthin dinoflagellate Karenia mikimotoi. We demonstrate that while most individual RNA editing events in dinoflagellate plastids are restricted to single species, global patterns, and functional consequences of editing are broadly conserved. We find that editing is biased toward specific codon positions and regions of genes, and generally corrects otherwise deleterious changes in the genome prior to translation, though this effect is more prevalent in peridinin than fucoxanthin lineages. Our results support a model for promiscuous editing application subsequently shaped by purifying selection, and suggest the presence of an underlying editing mechanism transferred from the peridinin-containing ancestor into fucoxanthin plastids postendosymbiosis, with remarkably conserved functional consequences in the new lineage.
Abstractβ-lactamases are enzymes which are commonly produced by bacteria and which degrade the β-lactam ring of β-lactam antibiotics, namely penicillins, cephalosporins, carbapenems, and monobactams, and inactivate these antibiotics. We performed a rational and comprehensive investigation of β-lactamases in different biological databases. In this study, we constructed hidden Markov model profiles as well as the ancestral sequence of four classes of β-lactamases (A, B, C, and D), which were used to identify potential β-lactamases from environmental metagenomic (1206), human microbiome metagenomic (6417), human microbiome reference genome (1310), and NCBI's nonredundant databases (44101). Our analysis revealed the existence of putative β-lactamases in the metagenomic databases, which appeared to be similar to the four different molecular classes (A–D). This is the first report on the large-scale phylogenetic diversity of new members of β-lactamases, and our results revealed that metagenomic database dark-matter contains β-lactamase-like antibiotic resistance genes.
AbstractSex determination systems are exceptionally diverse and have undergone multiple and independent evolutionary transitions among species, particularly reptiles. However, the mechanisms underlying these transitions have not been established. Here, we tested for differences in sex-linked markers in the only known reptile that is polymorphic for sex determination system, the spotted snow skink, Niveoscincus ocellatus, to quantify the genomic differences that have accompanied this transition. In a highland population, sex is determined genetically, whereas in a lowland population, offspring sex ratio is influenced by temperature. We found a similar number of sex-linked loci in each population, including shared loci, with genotypes consistent with male heterogamety (XY). However, population-specific linkage disequilibrium suggests greater differentiation of sex chromosomes in the highland population. Our results suggest that transitions between sex determination systems can be facilitated by subtle genetic differences.
AbstractLevels and patterns of genetic diversity can provide insights into a population's history. In species with sex chromosomes, differences between genomic regions with unique inheritance patterns can be used to distinguish between different sets of possible demographic and selective events. This review introduces the differences in population history for sex chromosomes and autosomes, provides the expectations for genetic diversity across the genome under different evolutionary scenarios, and gives an introductory description for how deviations in these expectations are calculated and can be interpreted. Predominantly, diversity on the sex chromosomes has been used to explore and address three research areas: 1) Mating patterns and sex-biased variance in reproductive success, 2) signatures of selection, and 3) evidence for modes of speciation and introgression. After introducing the theory, this review catalogs recent studies of genetic diversity on the sex chromosomes across species within the major research areas that sex chromosomes are typically applied to, arguing that there are broad similarities not only between male-heterogametic (XX/XY) and female-heterogametic (ZZ/ZW) sex determination systems but also any mating system with reduced recombination in a sex-determining region. Further, general patterns of reduced diversity in nonrecombining regions are shared across plants and animals. There are unique patterns across populations with vastly different patterns of mating and speciation, but these do not tend to cluster by taxa or sex determination system.
AbstractA gene which carries a bona fide loss-of-function mutation effectively becomes a functionless pseudogene, free from selective constraint. However, there is a number of molecular mechanisms that may lead to at least a partial preservation of the function of genes carrying even drastic alleles. We performed a direct measurement of the strength of negative selection acting on nonsense alleles of protein-coding genes in the Zambian population of Drosophila melanogaster. Within those exons that carry nonsense mutations, negative selection, assayed by the ratio of missense over synonymous nucleotide diversity levels, appears to be absent, consistent with total loss of function. In other exons of nonsense alleles, negative selection was deeply relaxed but likely not completely absent, and the per site number of missense alleles declined significantly with the distance from the premature stop codon. This pattern may be due to alternative splicing which preserves function of some isoforms of nonsense alleles of genes.


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Are you kidding?? This is the solution to Obesity Nation Wide! Use your own fat to power all your mobile devices.. i'm down!
What calories? Pizza is a vegetable!
There's nothing like a good explication to completely ruin what was already a bad joke! Thank you!
Ruining a bad joke is like ruining a turd. It can't be done. A joke that incredibly stupid must be taken seriously, and its spewer stomped on. Not only does pizza have vegetables, but vegetables have calories (how can anyone at
Anyone know where I can find a good nerd site that keeps out the normtards?
First, let's skip the term "nutrients", which marketing and new-age blather have rendered meaningless for public discussion. This wouldn't consume vitamins and minerals. It would consume blood sugar or lipids, both of which average Americans have in great excess.
I've been calling for this (blood-sugar power for implanted devices) for decades. It's easy to point out how obvious this solution is, because I'm not in a field that gives me any insight into the actual technical problems.
If I had to power my computing and communication tools from my own body's stores of energy, I'd get to eat a lot more, and I'd probably still lose the spare tire.
It's easy to point out how obvious this solution is, because I'm not in a field that gives me any insight into the actual technical problems.
I love this statement. I will use this in the future without reference to you in meatspace, sadly.
If we can only weaponize it, Americans would be all over it.
"Paint a target with the LipidLaser 2000 and our global network of satellites will obliterate it from orbit with microwaves! Lose weight AND your enemies!"
Are you kidding me? If you could convert unnecessary body mass into useful energy, the average American would become self-sufficient. And probably invincible.
The Matrix 4
You finally discover that really it was humans that lived above ground and developed the heat feeding system. People of Zion are really the robots.
It's already been tested in the lab [slashdot.org], or did everyone forget the story from earlier this year? Well, not quite ATP, but a fuel cell utilizing glucose and oxygen at blood concentrations [bbc.co.uk]. It worked quite well in a rat in 2010.
Just so I can say, "My bugs have bugs"
Batteries are more efficient in energy/mass ratio than feeding is. Which means, the insect either has to bring food with it, which is inefficient, or have to find some and feed on spot, in which case it can't be remote controlled.
Until someone builds an evolution chamber and a bunch of spine crawlers show up.
I've seen this before on some of the colonies.
They do have crappy silk that doesn't involve killing the worms...
The vegan (as opposed to vegetarian) "philosophy" prohibits using or consuming anything animal derived, even if the animal itself isn't killed. That's why vegans won't drink milk or eat honey. So even silk that didn't involve killing the worms still came from worms, and is therefore still a vegan taboo.
Organic compost of plant matter? Just because, in the U.S.A., agriculture has a hard-on for pollution, chemical fertilizers, herbicides and pesticides, does not mean that no alternative exists.
Yeah, they want humane treatment for insects too
I wonder how many bugs, worms, rodents, amphibians and assorted other animals are killed when some farmer runs a combine or plough over a field during the course of a year
Car analogy time:
Yeah, they want humane treatment for humans too
I wonder how many people die on the roads each year
Hah, idiots. They think they can be for driving but against murder. It makes no sense!
Some forms of vegan don't mind using anything the animal naturally casts off. Or they could be a level five vegan and not eat anything that casts a shadow.
Yeah, they want humane treatment for insects too, i have a vegan friend that wont even eat honey or use silk
Really? Wow. I deal with that by not hanging out with fucking lunatics, myself.
You too can be the "fly on the wall" watching and listening through a tiny video camera and mic attached to the fly.
Can it power my TV?
Perhaps not the TV, but imagine a self powering bug zapper.
In the future, all the pretty girls will have flies circling around them. (remote controlled by
It might actually turn into sign of desirability. More flies == more fly (pun intended).
Probably a sign of cognitive decline.
So our first steps into bridging the biological/electronic divide involve what is essentially an artificial parasite? Somehow, I fear it will not be long before some medical researcher who has read too much Douglas Hofstadter thinks of taking it to the next level of abstraction by monitoring intestinal health using parasitic instruments attached to tapeworms.
"parasitic instruments attached to tapeworms"
Well, now that you mention it, tapeworms are a lot better at controlling our immune systems than we are...
I for one welcome our new cyborg helminth overlords!
There's the core of an SF-horror movie for ya.
In English, every word can be verbed. Would that it were so in our programming languages.


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*NOTE - Please use the File Attachments options to upload your entry*
This one is a play on words. Kan is "cap" and well, this does have a sort of cap/hood and Kamoku is "shy".
So obviously, this weird character uses its little hood to hide from the outer world (indicating that it isn't a good character).
I guess some of you will think this is a tortoise or a jellyfish, which was my impression when I looked at it. I didn't intend it to look like that one bit, but I don't feel like changing it knowing the contest ends today and also because it took me five hours to do it all.
So anyway, Paint is not my fave art program, but it is the only one I have. I am sure it won't win although my lrgest desirte is to get one of those American V2s so all I can do is cross my fingers and hope BanDai takes a shine on Kanokutchi.
EDIT: Thetre is a problem.It is all correct, the format is PNG (graficos de red portattiles is PNG in Spanish , portable net graphics) and the size is 26 KB but I get an error.
This means take the followijg link and I shall change its format to JPG which can be viewed here, but the original picture is on the link (JPG makes colours go blurry)
Photobucket link
Original picture in there. Modified format VV
JPEG JPG will not work.
Sorry, there is an uploading problem. I am afraid Admin, that you will have to take my link.
Sorry if this causes any bbothers.
Started by spiffy , Nov 13 2005 10:31 AM


text:
Laila Ali: How I Got My Bikini Body Back
Don Flood
After delivering her second child, daughter Sydney, in 2011, Laila Ali wasn't stressed about getting her pre-baby body back.
"The second time around I knew what to expect so I was a little easier on myself," says Ali, who also has son CJ, 4, with her husband, former NFL star Curtis Conway. "Once I saw that you just have to eat right and exercise that's what I did and it worked."
It worked really well: In this week's issue of PEOPLE, Ali, 35, shows off her toned physique for her first ever bikini shoot.
"I've never done a bikini shoot before," she admits.
"I'm kind of shy, I'm not one of those people whose comfortable walking around with my butt out. But I had a baby, I got my weight down and I'm always talking about fitness and health. It's time to show people what I'm working with."
"I'm naturally a big girl," Ali explains. "I have to work to be fit. I love running. I run three times a week and I do strength training and weights. And sometimes I throw a little spinning in there."
Apart from her time in the gym, Ali watches what she eats by sticking to lean proteins and vegetables most of the time.
"It's what you do 90 percent of the time that matters," she tells PEOPLE. "I'm not really into junk food, but I like to have a cheat day no matter what, usually on the weekend. I might have pizza, frozen yogurt or a brownie."
Though the discipline matters to Ali — it's also a much-needed dose of personal time for the mom of two.
"I know how to work hard and push myself," she says. "It's my time for myself. I need to get it in."
For more on how Ali stays fit and healthy, pick up the latest issue of PEOPLE — featuring our Bodies special — on newsstands now.
— Raha Lewis


text:
Husband-and-wife cooking team Debbie Gold and Michael Smith-renowned for innovative menus featuring the seasons' finest offerings-harvest tomatoes, squash and potatoes at a Mennonite farm and gather eggs at Campo Lindo Farms. Recipes: Poached Egg Soup with Squash Blossoms & Tomatoes; Chicken Breast with Sour Cherries & Asparagus Salad.
Visit the Website: www.chefsafield.com
Episode #204 / Length: 26 minutes


text:
Data Card Office
The Data Card office has requested the following:
Would like to have a permanent workstation at the Cartersville Charger Card Office set up with IDMS and Blackboard for making IDs -Would like to have a printer/fax/copier at the Cartersville location as well -need more network drops in Charger Card office on main campus -Need VTS and IP converter (have both on hand) hooked up in library at North Metro but officials there are not keen on this effort for some reason is what I'm told -Need more copy/activity/vending readers on hand in case one goes out (but this comes out of Charger Card Office budget so probably not an issue for you) -Would like to upgrade copy readers on copiers at Heritage Hall, S. Poly, N. Metro, and Rome campus (in front of bookstore and activity reader inside bookstore)-again, this is probably not an issue for I.T. although it is technology-related -would like to add vending readers to all coke machines & vending machines on all campuses (another C. Card office issue) -We need additional memory for our server and we need to perform a firmware upgrade (free if we do it).
Most of this is stuff that would need to come out of the Charger Card office budget but just letting you know as most of it is "technology-related".
Status: 2/12/09
These items would come from the Auxiliary Budget.


text:
Events Listing
The RockBades course presents an alternate model for IASTM work- what we define as "Instrument Assisted Neuro Sensory Modulation" or IANSM. This draws from current thinking of the way in which fascia behaves and the influence that manual therapy has on these tissues.
Our IANSM model does not rely on traditional, at times aggressive treatment strategies using instruments, and shows how similar, if not better, results can be achieved with more gentle methods. This enhances patient safety and minimises the potential for adverse treatment effects.
Locations in TAS, NSW
CPD Excellence
Orthopaedic & Neurological Syndromes of the CERVICAL SPINE AND TMJ
Locations in TAS, NSW
Elixr Education Presents the Selective Functional Movement Assessment Courses
SFMA Level 1 & 2 – Sydney, May 2018
Elixr Education is excited to once again present the Selective Functional Movement Assessment (SFMA) Level 1 course in August, and for only the second time in Sydney, SFMA Level 2, both facilitated by the highly-respected Greg Dea.
The course teaches the SFMA movement based diagnostic system and provides healthcare professionals with an efficient and systematic tool to reach a comprehensive movement diagnosis.
It is a clinical assessment for people who experience pain. It finds the cause of pain - not just the source - by logically breaking down dysfunctional movement patterns in a structured, repeatable assessment.
Locations in TAS, NSW
Multimorbidity in Primary Care: An Interdisciplinary Challenge
Webinar dates
Clinical Assessment of Cervical Nerve Root Syndromes
Webinar dates
Getting Ahead of the Curve- The Modern Principles Of Scoliosis Management
Introduction to ClinicalKey
Webinar dates
RockTape 1/2 day course - Kinesiology & Sports Taping Seminar
Our half day introduction (level 1) courses are attractive to busy practitioners, and those that are keen to just learn the basic principles of kinesiology taping. This course provides the basics of kinesiology taping: regional taping for pain, taping for swelling and recovery and taping for posture. In addition this course includes an introduction to the concepts of "Fascial Chain Taping": utilising the benefits of kinesiology taping to tape the muscles associated with a particular activity. For example, linking the muscles of the appropriate kinetic chain, ie lower back, hamstring and calf muscles that make up the posterior chain.
Locations in TAS, NSW
Essentials of Effective Scoliosis Management
Scoliosis is a complex condition.
There is no one size fits all approach to its treatment.
Locations in TAS, NSW
Introduction to Eating Disorders, for Osteopaths
Webinar dates
Pain Management in Practice Workshops-Empower Rehab.
Do you have clients overly focused on their pain?
Is your client not following your advice?
Would you like your client to achieve ambitious goals?
Master your skills in managing clients with complex and chronic pain conditions in this 2 day workshop.
This 2 day workshop is designed to challenge and extend your skills to manage clients with persistent pain by providing osteopaths with a broader range of effective and practical treatment options.
Locations in TAS, NSW
Ergonomic Risk Assessment Training for Allied Health Professionals
This course is designed for those practitioners in both private practice and for those who would like to conduct ergonomic risk assessments within the OHS/WHS setting.


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As fun as Wolfenstein: The New Order looks, the id Software series we're most keen to hear more of is Doom. The vagueness continues. Publisher Bethesda today announced that The New Order will launch on May 20, and pre-ordering will get you access to a beta for "Doom." Which Doom? Doom 4? And what's the beta? Oh, don't worry your pretty little head with all that.
The Wolfenstein site refers merely to "Doom," as if we'll understand that. Yes, we're well aware of what Doom is, but what is this? Bethesda is slightly more verbose on Twitter, at least, saying it's beta access to "the next Doom game." Whatever that may be. One assumes the beta will multiplayer in some form, but that's purely speculation.
The New Order's coming to PC, PlayStation 4, Xbox One, Xbox 360, and PS3. It's developed by MachineGames, a studio formed by Starbreeze founder Magnus Hogdahl and several other Starbreeze folks.
"Running and gunning and killing Nazis are the essence of the franchise, and MachineGames so far has done an admirable job of paying tribute to the id games of yore," we wrote after seeing it in May, though we were a little more sceptical of its attempts at a story. Catch a glimpse of that story stuff in this new trailer, released today to celebrate all the news:


text: Ok so my LO is just short of his first bday and we have been successfully nursing this whole year. My goal was to nurse for a year since I didn't want to give him formula. Since I'm a SAHM he never got too many bottles since it was just easier to give him the breast. Up until now I have absolutely loved nursing him. It's honestly been the most amazing bonding experience with him. I never thought I would say this but I am sooo ready to wean him off the breast. He has become so much more aggressive and he bites me and honestly I don't enjoy it at all anymore it's terrible. I've tried pumping and giving him bottles but he isn't satisfied. Not to mention he can't see me pump cause he cries. I can't go into the other room and pump and leave him alone. Are there any ways to gently wean him off the breast? I will give him breast milk until he turns one and maybe after that. I don't mind pumping but I really just feel like its time to move on. Help!


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this is a discussion within the Saints Community Forum; NFL's Saints may drill in Jackson By Laura Hipp email@example.com Fans of the New Orleans Saints may not need to drive too far to see their team practice. The Mississippi Veterans Memorial Stadium Commission will consider a contract with the ...
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Join Date: Apr 2005
Location: new orleans
Posts: 584
NFL's Saints may drill in Jackson
NFL's Saints may drill in Jackson
By Laura Hipp
Fans of the New Orleans Saints may not need to drive too far to see their team practice.
The Mississippi Veterans Memorial Stadium Commission will consider a contract with the NFL team by Wednesday, said Watt Whatley, executive director. Officials hope the event leads to a return of exhibition games in the future.
The NFL franchise may play a scrimmage Aug. 6 in Jackson.
"I think it will be awesome just from the standpoint of getting New Orleans in the area," Whatley said. "They want to try to get more involvement out of this area."
For months, the commission has worked to bring the team to the capital city. Bringing All-Pro running back Deuce McAllister to play in front of thousands of fellow Mississippians sweetens the deal.
"It'd be great for Mississippi and Jackson," House Appropriations Chairman Johnny Stringer, D-Montrose, said.
Saints officials and McAllister could not be reached for comment.
Stadium officials and the governor's staff have spoken with Stringer and Senate Appropriations Chairman Jack Gordon, D-Okolona, about securing $150,000 for new lockers and training tables.
The equipment is needed before Jackson State University's football season starts this fall. Though no funds have been found, officials are certain lockers will be in place.
"We'd do it for Jackson State and this game," Stringer said.
An estimated 20,000 to 25,000 could attend the scrimmage.
"Any pro football team, I'd go to," said Garland Bridges, 19, of Brandon. He travels a few times in the fall to see the Saints play.
State officials hope a successful scrimmage will bring an exhibition game to Jackson, which last had one in the mid-1990s.
"We really need to show the Saints we'd be prepared to host an exhibition game," said Charlie Williams, chief of staff for Gov. Haley Barbour.
The Saints would arrive in the Jackson area on Aug. 5, and players and coaches would host clinics for area school children, Williams said.
Tickets would be in the "$10 range," and there would be a deal for families, he said. Those details have not been approved.
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text:
Command line operations
Most Java programmers use an Integrated Development Environment (IDE) such as IntelliJ, NetBeans, or Eclipse. Modern IDEs are usually pleasant to use, and go a long way in simplifying things for a typical programmer.
However, being familiar with Java's basic command line tools is still very useful. It not only gives you a better understanding of what's happening, it's essential in situations where you need to use the command line tools directly (scripts, tools, and so on).
The most commonly used tools are:
- javac - the java compiler
- java - launching programs on the Java Runtime Environment
- javadoc - generating documentation for your project
Here are some examples of running command line tools. It assumes a project with the following directory structure:
PROJECT_HOME -> lib (jar files) -> src (java code) -> hirondelle (top-level package; no .java files) -> ante (.java files) -> deluvian (.java files)
Compiling With javac
PROJECT_HOME>javac -cp lib\* src\hirondelle\ante\*.java src\hirondelle\ante\deluvian\*.javaThis compiles in place, and creates .class files beside .java files. If you want to place generated class files elsewhere, use the -d option to put them into an existing directory:
PROJECT_HOME>javac -cp lib\* -d build src\hirondelle\ante\*.java src\hirondelle\ante\deluvian\*.javaAlso, notice that all jars in a directory can be succinctly referenced using 'lib\*'. However, referencing the source files that you need to compile is a different story: all of your packages need to be listed one by one. That is, you can't simply specify a single root directory for your source files. This is a rather serious defect of javac.
If your jars are in various directories, then the classpath is a list delimited by semi-colons:
-cp lib\*;C:\abc\one.jar;C:\xyz\two.jar
Running With javaAny class having a main method may be run as a Java program. The main method must have the form:
public static void main(String... aArgs){...}An application is run by referencing a fully-qualified class name, but without the .class suffix. Assuming that the classes have been compiled in place, our example application can be run using:
PROJECT_HOME>java -cp lib\*;src hirondelle.ante.LauncherPassing 3 arguments to the program:
PROJECT_HOME>java -cp lib\*;src hirondelle.ante.Launcher arg1 arg2 "Arg Three"Setting a System property value with -D:
PROJECT_HOME>java -Dblah=whatever -cp lib\*;src hirondelle.ante.LauncherSet the initial and maximum size of the object allocation pool to 5 Meg and 100 Meg, respectively:
PROJECT_HOME>java -cp lib\*;src -Xms5m -Xmx100m hirondelle.ante.LauncherRun a jar file whose manifest specifies a Main-Class attribute:
C:\@build\dist>java -jar example-app-1.1.jarThe javaw command is the same as the java command, with the only difference being that javaw has no associated console window.
Using javadoc
For files under the src directory, the following command will javadoc all packages starting with 'hirondelle.ante', and output the result to C:\@build\javadoc:
javadoc -sourcepath src -subpackages hirondelle.ante -classpath lib\* -d C:\@build\javadocThe javadoc tool has many options. You may find these especially useful:
- -linksource: links your javadoc to a static snapshot of the underlying source code (very nice).
- -windowtitle, -header and -footer: often useful for showing the name and version of your application.
- -link: link to the JDK's javadoc as well
- -noqualifier: remove tedious package prefixes from JDK classes
- define the minimum class scope to be processed, using one of: -private, -package, -protected, -public.
javadoc -sourcepath src -subpackages hirondelle.ante -package -classpath lib\* -d C:\@build\javadoc -linksource -link http://docs.oracle.com/javase/7/docs/api/ -noqualifier java.*:javax.* -windowtitle "My App 1.0" -header "My App 1.0" -footer "My App"Javadoc can also be controlled using the following items, placed beside your source code:
- package-info.java - Each package can include a package-info.java file. It contains package-level documentation. Despite its extension, a package-info.java file is not a Java class file.
- doc-files subdirectories - each package can include a 'doc-files' subdirectory, containing items that can be linked from javadoc (images, html files, and so on) using ordinary hypertext links.
- overview.html - a single file describing general aspects of your project. (This file doesn't necessarily reside in your source tree, but it makes sense to place it there, in analogy with the package-info.java files.)
Would you use this technique?


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Unicorn & Seahorse Liberty Print Bag based on the Estelle pa...
Added Jun 29, 2011
Cornwall, United...
I found the Estelle bag pattern by fashion1 a fantastic pattern to use and easy to adapt. I made a bag for my mum using some Liberty fabric I had stashed away, and used the wrong side of some dyed velvet for the lining.
I changed the handles to suit the strange way my mum carries her handbags. :D On the front of the bag, I sewed on a few charms of my nan's and used some gold fabric paint to highlight the horns and eyes of the unicorns/seahorses on the print.
I would really recommend this pattern and many thanks to fashion1 for making such a novice-friendly and useful bag with such clear instructions.
Blogged here – the foxling Estelle Bag
Material Notes
Unicorn & Seahorse Liberty Print Cotton twill ( about a metre)
The wrong side (dyed poly) of some dyed velvet for the lining.
- For
- Women
- Garment Type
- Accessories
- Estelle Bag by Fashion1
3 Comments Sign in to add a post
Oct 16, 2011, 03.41 PMby Kate Coley
I love the way you've added the charms to this bag. Really makes it something special and given me lots of ideas. Thanks for posting.
Jul 13, 2011, 03.06 AMby hidngat39
love all the trinkets-really personalizes it! great job!
Jun 29, 2011, 03.43 PMby cupcakedreammachine
What a beautiful fabric!
Add to Favorites no one added it


text:
A group trying to recall Republican Gov. Kenny Guinn was having trouble collecting enough signatures to qualify for the Nevada ballot, an organizer said Wednesday.
"I don't know if we'll make it," Christopher Hansen, vice chairman of the Recall Guinn Committee, said of a Nov. 25 deadline.
The Nevada recall effort faces a tougher challenge than in California, he said, and accused state officials of blocking volunteers from collecting signatures in state and county government buildings.
Officials denied the obstruction charge, and said the recall effort was simply unpopular.
"They're making excuses for a lack of organization and a lack of ability to gather signatures," Guinn spokesman Greg Bortolin said. "Whether you agree or disagree with the governor, I would think the overwhelming majority of voters in our state just believe the recall is flat wrong."
Hansen said the Nevada recall petition, begun Aug. 27, faces a stiffer challenge than in neighboring California where voters ousted Gov. Gray Davis on Tuesday and elected actor Arnold Schwarzenegger.
"It's very much more difficult here," he said.
Nevada recall proponents must collect signatures equal to 25 percent of the number of ballots cast in the last governor's race compared with 12 percent in California.
Hansen disputed reports that his group of conservative and anti-tax advocates only had 12,000 signatures - or fewer than one-tenth the number needed. But he said he could not provide an accurate number. He said names from northern Nevada had not been tallied, and added that he thought he had at least 250 signatures in his car.
The Nevada group has to collect 128,109 signatures to force a recall of Guinn, who was re-elected to his second term in 2002 with 68 percent of the vote.
Guinn released a statement Wednesday congratulating Schwarzenegger and citing challenges faced by governors across the nation.
"Every elected official has been called upon to make difficult decisions which are not always popular," Guinn said.
Hansen said the Nevada recall drive was hamstrung by Nevada Secretary of State Dean Heller's rulings that two state Department of Motor Vehicles offices and the Clark County Government Center in Las Vegas did not have to allow signature-gathering inside buildings.
Steve George, spokesman for Heller, said state law lets officials direct signature-gatherers to a reasonable place inside or outside a government building.
County spokesman Erik Pappa said officials offered to let Hansen collect signatures outside the main door of the Clark County building, but Hansen refused. On Wednesday, officials let Hansen and his wife set up a table inside the building.
"He just wanted to challenge our policy to garner publicity for his recall effort," Pappa said.
The Nevada recall was launched after Guinn pushed for the largest tax increase in state history, an $836 million package passed after two contentious special legislative sessions and a legal battle that went to the state Supreme Court.


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