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Conceived and designed the experiments: ET DJJ JWT. Performed the experiments: ET DJJ. Analyzed the data: ET. Wrote the paper: ET DJJ JWT.
A role for natural selection in reinforcing premating barriers is recognized, but selection for reinforcement of postmating barriers remains controversial. Organisms lacking evolvable premating barriers can theoretically reinforce postmating isolation, but only under restrictive conditions: parental investment in hybrid progeny must inhibit subsequent reproduction, and selected postmating barriers must restore parents' capacity to reproduce successfully. We show that reinforced postmating isolation markedly increases maternal fitness in the fungus Neurospora crassa, and we detect the evolutionary genetic signature of natural selection by quantitative trait locus (QTL) analysis of the reinforced barrier. Hybrid progeny of N. crassa and N. intermedia are highly inviable. Fertilization by local N. intermedia results in early abortion of hybrid fruitbodies, and we show that abortion is adaptive because only aborted maternal colonies remain fully receptive to future reproduction. In the first QTL analysis of postmating reinforcement in microbial eukaryotes, we identify 11 loci for abortive hybrid fruitbody development, including three major QTLs that together explain 30% of trait variance. One of the major QTLs and six QTLs of lesser effect are found on the mating-type determining chromosome of Neurospora. Several reinforcement QTLs are flanked by genetic markers showing either segregation distortion or non-random associations with alleles at other loci in a cross between N. crassa of different clades, suggesting that the loci also are associated with local effects on same-species reproduction. Statistical analysis of the allelic effects distribution for abortive hybrid fruitbody development indicates its evolution occurred under positive selection. Our results strongly support a role for natural selection in the evolution of reinforced postmating isolation in N. crassa.
Although Darwin believed that natural selection could not drive intersterility between species, it is now well established that there is a role for natural selection in the evolution of premating discrimination that reinforces barriers to hybridization. However, natural selection for postmating barriers, like hybrid inviability, is still controversial, because it can only occur when overall maternal fitness is increased by the inviability of hybrid offspring. Constraint on adaptive evolution of postmating barriers poses a problem when organisms without premating preferences must adapt to the presence of related species and ensure that reproduction occurs only between members of the same species. We studied the evolutionary genetics of a reinforced, postmating barrier between two species of mold, Neurospora crassa and N. intermedia. Although hybrids have low fitness, Neurospora females do not discriminate against different-species sex partners before mating. Instead, N. crassa has adapted to the presence of the N. intermedia in its range by selectively aborting hybrid fruitbodies. We show that abortion increases maternal fitness because N. crassa can mate again after hybridization only if fruitbodies abort. Abortion is controlled by 11 loci, whose genetic effects are consistent with an adaptive evolution model, confirming that abortion evolved via natural selection against hybridization.
The evolution of reproductive isolation between diverging lineages is a critical step in speciation. Most reproductive isolation barriers between taxa evolve as side effects of changes resulting from within-population processes, including, as Darwin recognized, natural selection [1]. Wallace reasoned that natural selection against maladaptive hybridization itself could drive the evolution of reproductive isolation barriers when taxa co-occur geographically (are sympatric) [2]. This mechanism, known as 'the Wallace effect', is also termed reinforcement, because preexisting reproductive isolation is 'reinforced' by natural selection for stronger barriers. Following an extensive correspondence with Wallace on this matter, Darwin remained skeptical [1]–[3]. Today, reinforcement of premating barriers by natural selection is widely accepted, but natural selection for reinforced postmating isolation still remains controversial [4]–[6].
Theoretically, reinforcement of postmating barriers can occur when 1) evolution of premating barriers is constrained, 2) there is substantial parental investment in the production and care of progeny, 3) individuals that are capable of mating more than once are unable to do so because the energetic costs of nurturing the unfit hybrids make subsequent reproduction less likely, and 4) reinforcement of the postmating barrier restores parents' capacity to successfully reproduce after hybridization [7]–[9].
After reinforcement, sympatric species or populations should show stronger barriers than those that are geographically separated (allopatric), because natural selection for stronger barriers only occurs when populations are overlapping. This biogeographic pattern has been observed for premating barriers in many animals, plants, and fungi where it has been investigated [6], [10], [11], but only a few instances of stronger postmating reinforcement in sympatry have been reported over the past 65 years [9], [12]–[14], including a microbial example involving abortion of hybrid fruitbodies (perithecia) in matings between sympatric populations of the haploid fungal species N. crassa and N. intermedia [15], [16].
The geographical ranges of N. crassa and N. intermedia are broadly overlapping, and individuals of both species can be collected from the same site [15], [17], [18]. Both species are largely outbreeding, and outbreeding is confirmed by population genetic analysis [19]–[22]. Hybrids of the two species can be obtained in laboratory crosses, but natural hybrids have not been encountered [15]. This absence may reflect the rarity of hybridization in nature, the low viability of hybrids, or both. Nevertheless, phylogenetic conflict between some gene trees and the species tree of the Neurospora genus indicates historical hybridization and introgression between N. crassa and N. intermedia [23].
In Neurospora, mating can occur only between individuals having different alleles at the mating-type locus (mat a or mat A). Under nutrient limited conditions, a haploid colony of Neurospora differentiates female reproductive structures (protoperithecia). Fertilization occurs when a specialized hypha (trichogyne) growing from a protoperithecium fuses with a cell from a colony of the opposite mating type. The attraction of trichogynes to fertilizing cells is mediated by mating-type specific pheromomones. After fertilization, nuclei from the fertilizing strain travel through the trichogyne to the protoperithecium, where karyogamy eventually occurs. A series of independent meiotic events give rise to the sexual progeny (ascospores), which develop within flask shaped fruitbodies (perithecia) on the maternal thallus. Upon maturity, the ascospores are forcibly ejected from the fruitbody.
In Neurospora, evolution of premating isolation is apparently constrained because the sequences of the mating-type–specific, peptide pheromones controlling attraction between trichogynes and fertilizing cells are conserved throughout the genus (as determined by BLAST analysis of the N. crassa, N. tetrasperma, and N. discreta genomes [24]) and even beyond [25]. Evolution of the extracellular, ligand-binding portions of mating-pheromone receptor proteins is also comparatively constrained [26]. In Neurospora, sex cells of mating-type–compatible partners usually fuse before incompatibilities are expressed, and incompatibility arises either prezygotically in the fusion cell and the subsequent dikaryotic cells that proliferate from it, or postzygotically during the meiosis that directly follows karyogamy and during the formation and development of the ascospores [17]. Since Neurospora progeny develop within fruitbodies composed entirely of maternal tissue, the maternal colony (mycelium) bears virtually the entire cost of reproduction. Because 98% of N. crassa×N. intermedia hybrid progeny are inviable [15], and because allocation of resources to developing fruitbodies on one part of the colony abolishes the fertility of uncrossed regions of the colony [27], hybridization is severely maladaptive.
The key questions are: 1) Does abortion of hybrid fruitbodies by N. crassa make subsequent reproduction possible for the maternal colony, thereby conferring a fitness advantage? and 2) Did this postmating barrier evolved by natural selection? First we show that hybrid fruitbody abortion is adaptive, because it preserves the fertility of maternal N. crassa. Then by quantitative trait locus (QTL) analysis of hybrid fruitbody abortion and statistical analysis of the allelic effects distribution for the detected loci, we show that the genetic architecture that we observe is consistent with evolution by positive natural selection.
We tested whether female fertility of colonies was preserved after abortion of sympatric hybrid fruitbodies in sequential mating experiments between the two Neurospora species. N. crassa colonies were grown in Petri plates on synthetic cross agar medium, which promotes sexual reproduction. One half of the receptive N. crassa colony was fertilized by either an allopatric or sympatric N. intermedia strain, and the effect of allopatric vs. sympatric fertilization on the reproductive success of subsequent conspecific fertilization on another portion of the maternal colony was assayed. Initial fertilizations by a conspecific strain or with water (pseudo-fertilizations) were also performed as controls. Each of the four initial-fertilization treatments (allopatric male, sympatric male, conspecific male, pseudo-fertilization) was replicated three times. Following Dettman, et al., reproductive success was scored on a seven-category scale incorporating fruitbody development and quality of ejected ascospores, if any [15]. The N. crassa and N. intermedia strains used are described in the Materials and Methods and listed in Table 1. In all sequential matings, both the maternal strain and conspecific fertilizing strains were N. crassa from the NcC clade endemic to India, hereafter referred to as NcC-India [15].
Fruitbody development on portions of the maternal colony fertilized by allopatric N. intermedia strains is normal and results in ascospore ejection, although a majority of the hybrid ascospores are unmelanized and inviable (reproductive success score (RSS)=4.33±0.14, Figure 1). Following allopatric hybridization, response to conspecific fertilization at the second time point is completely inhibited (RSS=0.00±0.00).
In contrast, fertilization by sympatric N. intermedia strains at the first time point yields only aborted fruitbodies (RSS=1.00±0.00), but the subsequent conspecific matings are fully fertile (RSS=6.00±0.00, Table 2). We performed semi-parametric regression using a proportional hazards survival model [28] to examine the effects of fertilization at the first time point (water control, conspecific control, or allopatric or sympatric heterospecific) on progression through the sexual cycle after fertilization at the second time point (measured as RSS). The first–time-point fertilization treatment had a significant effect (P<0.0028), whereas the nested effects of geographic origin and strain identity of the first–time-point male do not have significant effects (Table 3). We conclude that abortion of hybrid fruitbodies is selectively advantageous because abortion preserves maternal fertility of the colony after hybridization.
Previous research on the genetics of reinforcement focused on premating barriers in animals [29]–[31]. Here we investigate whether the genetic architecture of a reinforced, microbial, postmating barrier is consistent with evolution by directional natural selection [32], [33]. A 500-member, N. crassa mapping population was derived from an intraspecific, inter-clade cross between the NcC-India strain FGSC 8866, and a Louisiana, USA, strain, FGSC 8903, a member of the NcA clade, hereafter referred to as NcA-Louisiana. Neurospora are hermaphroditic so that the parental strains could be mated reciprocally. Based on the identity of the maternal parent we can infer that the mapping population contains both individuals with NcA-Louisiana cytoplasm (57%) and individuals with NcC-India cytoplasm (43%).
The N. crassa mapping strains were crossed maternally and paternally to N. intermedia strains from Tamil Nadu, India, which are sympatric to the NcC-India parent and allopatric to the NcA-Louisiana parent. NcC-India aborts fruitbodies after fertilization by N. intermedia from India, but NcA-Louisiana does not [15], [16]. The mapping strains were also crossed maternally and paternally to African N. intermedia strains, which are allopatric to both the NcC-India and NcA-Louisiana parents. Neither the NcA-Louisiana parent nor the NcC-India parent aborts fruitbodies after fertilization by the African N. intermedia strain. The four traits that we studied were fruitbody development in the four different types of matings. The four types of matings are defined by the parental role of the mapping strain (maternal or paternal) and the geographic relationship of the N. intermedia strain (Indian and therefore sympatric to the NcC-India parent of the mapping population, hereafter termed "sympatric"; or African, and therefore allopatric to both the NcC-India parent and the NcA-Louisiana parent, hereafter termed "allopatric"; see Table 4). Therefore each member of the mapping population was crossed twice to an Indian N. intermedia strain and twice to an African N. intermedia strain, once with the mapping strain in the maternal role and once with the mapping strain in the paternal role, for a total of four crosses per mapping strain. We examined fruitbody development in each cross, recording its fruitbody development score (FDS) 10 days after fertilization (see Materials and Methods).
The mapping strains were genotyped at 69 AFLP (Table 5 and Table 6) and 28 microsatellite (Table 7) markers as well as the mat locus. A genetic map containing seven linkage groups (LG) reflecting the seven chromosomes of Neurospora was estimated, with a total map length of 837.9 cM and an average intermarker distance of 9.2 cM (Figure 2).
Non-Mendelian segregation and non-random associations (NRA) among alleles at multiple loci can reflect genetic incompatibilities between the NcC and NcA genomes. Individuals in the mapping population inherited 53.6% of genotyped alleles from the NcA-Louisiana parent, and this is significantly higher than the 50% expected under Mendelian segregation (t Test, Pt=6.8721, DF=93<0.0001). The proportion of NcA-Louisiana alleles inherited varied across linkage groups (ANOVA, PF ratio=23.34, DF=93<0.0001), with linkage groups II, VI, and VII showing below 50% inheritance of NcA-Louisiana alleles (Figure 3). The skew towards NcA alleles was strongest on linkage group I, with 59.0% of marker alleles inherited from that parent. Of 22 markers showing significant segregation distortion (χ2, P<0.05), only one marker on LG IV (nc4L4) was significantly skewed in favor of the NcC-India allele, while the 21 other significantly distorted markers favored the NcA-Louisiana and were located on LG I (16 markers) and LG IV (5 markers).
Seven marker pairs representing four pairs of linkage groups exhibit significant non-random associations (Table 8). Positive non-random associations, reflecting an overrepresentation of parental haplotypes, are consistent with the existence of negative epistasis among clade specific alleles, as predicted by the Dobzhansky-Muller (D-M) model of the evolution of genetic incompatibilities first articulated by Bateson [34]. Per this model, incompatibilities between two loci, X and Y, must arise in in a two-step fashion, as follows: Consider that at the time that the NcA and NcC clades diverged, both populations contained ancestral (anc) alleles at every locus. The first fixation of a derived allele, e.g., Xanc→XNcA, will not cause incompatibility, because XNcA arises in an ancestral genetic background and must also be compatible with the ancestral alleles that comprise the NcC genome. However the next derived allele to be fixed by either population can be a source of incompatibility because the genetic backgrounds of the two clades are no longer identical. For example, if we consider fixation of a new Y allele in the NcC population, Yanc→YNcC, we see that YNcC evolves in the presence of Xanc, and may not be compatible with the previously derived allele XNcA in the NcA clade. Conversely, fixation of a new Y allele in the NcA population, Yanc→YNcA can also give rise to incompatibilities, since YNcA need not be compatible with the Xanc allele, which is still fixed in the NcC population."
Given partial intersterility between NcA-Louisiana and NcC-India and the segregation distortion that we observed, we predicted that observed significant non-random associations would be positive, indicating a deficit of recombinant haplotypes. However, only one significant marker pair (markers nc2L1 and tnf223 from LG II and IV, respectively) showed positive non-random associations (D'=0.19). Surprisingly, the loci on the three other significant linkage-group pairs (III and V, V and VI, and VI and VII) showed negative, non-random associations, with D' values ranging from −0.35 to −0.15, implying overrepresentation of recombinant haplotypes. Overrepresentation of recombinant haplotypes could be evidence that an optimal N. crassa genome would include some mixture of clade-specific alleles. Indeed, a population genomic study of N. crassa NcA from the Caribbean Basin and the south-eastern United States identified a haplotype in the Louisiana population consisting of a four-gene "migrant tract" originating from an unidentified, genetically diverged population or species, concluding that this tract was fixed in the Louisiana population via a selective sweep [35]. Alternatively, if alleles are not fixed in the clades, overrepresented recombinant haplotypes could be analogous to selectively advantageous, intrapopulation haplotypes.
We used composite interval mapping to identify QTLs for fruitbody development. The genetic basis of postmating reinforcement was revealed by mapping loci for maternal influence on sympatric fruitbody development (trait A, see Table 4). We identified 11 additive-effect QTLs for this trait (Figure 4a and Table 9; complete CIM scans for all traits are in Figure 5). Seven of the QTLs are located on LG I, including one of large effect, while LG II and V each contain a single broad QTL region of weak effect, and the left arm of LG VI harbors two other QTLs of large effect. The detected QTLs account for roughly 61% of trait variance, with the three loci of large effect accounting together for roughly 30% of trait variance. The inferred cytoplasm type (NcA-Louisiana or NcA-India) of the mapping strains did not significantly affect this trait.
For 10 of the 11 QTLs, the allele from the sympatric NcC-India parent has a negative effect on sympatric fruitbody development. Only the weak QTL on LG II has the opposite effect. The prevalence of negative alleles in the NcC-India background is consistent with evolution of abortion via directional natural selection. This inference can be statistically tested by the QTL sign test, which tests the null hypothesis that the observed genetic architecture was generated during neutral trait evolution, i.e., without selective advantage for negative alleles [32].
The QTL sign test assumes that all QTL effects are additive. We note that the accepted model of intrinsic, postzygotic isolation barriers involves negative among species-specific alleles in hybrids. However, our experiments were not designed to, nor can they, interrogate the genetics of hybrid dysfunction, but rather the genetics of evolutionary response to maladaptive hybridization. The genetic architecture of reinforcement may or may not involve epistatic effects. However, in contrast to the case of hybrid dysfunction, epistasis would involve within-genome, interaction effects among loci contributing to the reinforced barrier. To determine whether or not epistasis plays a role in the genetics of reinforced sympatric barriers in N. crassa, and to determine whether or not the genetic data conform to the assumptions of the QTL sign test, we performed a two-dimensional genome scan for interacting QTLs.
No significant interaction effect was detected. Moreover, the genome-wide maximum LOD score for any interaction effect was determined to be 16.7, well below the critical LOD score of 37.4 (for a Type I error of 0.05), which was estimated from 1000 permutations of the data. Because the two-way scan for genetic interactions among loci failed to find any significant or marginally significant interaction effects, the data are consistent with an additive genetic model and conform to the assumption of additivity required for the QTL sign test.
Given the observed number of positive and negative QTLs and the distribution of their effect magnitudes (gamma b=0.034, c=13.5), and conditioned on the parental difference in fruitbody development after sympatric fertilization (F.D.S=2), the null hypothesis of neutral trait evolution for abortion in NcC-India is rejected (QTL sign test, P=0.0099) (Figure 4b). This result implies that fruitbody abortion in sympatric NcC-India–maternal×N. intermedia-paternal crosses (trait A) evolved under positive natural selection via a reinforcement mechanism.
One of the major QTLs on LG VI is flanked by two microsatellite loci, nc6L15 and nc6L16, and can therefore be physically located to a 135,874 bp region of the N. crassa genome containing 24 ORFs (Table S1) [36].
We also investigated the genetics of fruitbody development in crosses not showing reinforced isolation (Traits B, C, and D, see Table 4). Note that NcC-Indian strains show enhanced isolation from Indian N. intermedia only when the NcC-Indian strain performs the maternal role [16]. We did not detect any QTLs for paternal influence on the development of sympatric fruitbodies (trait B), but in crosses to allopatric N. intermedia strains we detected two QTLs affecting maternal influence and three QTLs affecting paternal influence on fruitbody development (traits C and D, respectively; Figure 4a). All five of these loci are located on LG I. Four of the five allopatric QTLs co-localize with three of the sympatric QTLs on LG I, which could either indicate the presence of genes with pleiotropic effects or linked genes with trait-specific effects.
Linkage group I represents less than 24% of the genome, and it is striking that 75% of our QTLs map here. Interestingly, 73% of loci showing non-Mendelian segregation also map to linkage group I, so that every QTL on this linkage group is flanked by at least one marker showing segregation distortion. In all cases, the NcC alleles of the linkage group I QTLs have a negative effect on sympatric hybrid fruitbody development, and the NcC alleles of the linkage group I markers are underrepresented in the NcA×NcC N. crassa mapping population.
It is not known why one-fifth of genetic markers are distorted in favor of the NcA allele. It is possible that, because laboratory strains of N. crassa have historically been derived from the NcA clade, our crossing and progeny-isolation methods, which were developed for NcA-clade N. crassa, have inadvertently created a selective environment favoring NcA alleles at loci linked to distorted markers. It is also possible that distorter loci present in the NcA background are normally repressed through the action of NcA modifier loci, but become unrepressed and active in some recombinant NcA×NcC genotypes. Although segregation distortion can be caused by nuclear-cytoplasm incompatibilities, it is unlikely to be the cause in this case. The Neurospora mapping population comprises a mixture of individuals with NcA-Louisiana cytoplasm and NcC-India cytoplasm. Moreover, even in individuals with NcC-India cytoplasm, linkage group I markers are distorted in favor of the NcA allele, with an overall NcA allele frequency of 0.61 for markers on this linkage group.
Another hypothesis is that reinforcement alleles themselves can pleiotropically cause ascospore inviability in conspecific, inter-clade crosses. Laboratory crosses between members of the NcA and NcC clades are partially intersterile, usually resulting in <50% ascospore viability [15]. In Neurospora, all products of meiosis contribute to the ascospore cohort, so segregation distortion most likely results from inviability of hybrid ascospores carrying the disfavored NcC allele(s).
Pleiotropy for reinforcement and reproductive isolation between allopatric conspecifics has previously been observed in animals [37], [38]. Reduced conspecific fertility can present a challenge to the evolution of reinforced barriers, since the fitness advantage of avoiding hybridization must outweigh the cost of lower conspecific fertility. However, restricted migration between conspecific populations should reduce the incidence of interpopulation mating and reduce the fitness costs associated with pleiotropic effects on conspecific fertility. The NcC and NcA clades are geographically separated, so limited interclade migration would reduce the fitness cost to NcC of lower fertility with NcA and facilitate the spread of reinforcement alleles in the NcC clade. If the pleiotropy hypothesis is correct, the evolution of reinforcement QTL on linkage group I could be a partial explanation for the evolution of incomplete reproductive isolation between the NcC and NcA clades.
Notably, QTLs on other linkage groups (II, V and VI) are flanked by markers showing Mendelian segregation, so that for these QTLs there is no suggestion of pleiotropic negative effects on within-species, inter-clade reproduction. Moreover, the two reinforcement QTLs on linkage group VI lie in the vicinity of three markers (tnc088, nc6L13, nc6L2), which participate in non-random associations with loci on other linkage groups, such that recombinant, non-parental haplotypes are overrepresented in the mapping population. Therefore the patterns of marker inheritance near QTLs on these other linkage groups do not suggest any connection between reinforcement QTLs and isolation of conspecific allopatric N. crassa populations.
We note that linkage group I contains the mating-type locus of Neurospora, and that some studies have found that reproductive isolation loci are more prevalent on sex-determining chromosomes than on autosomes [39], [40]. It is true that in a closely related species, N. tetrasperma, recombination is suppressed over a large region of Linkage Group I in a process considered analogous to an early stage of sex-chromosome evolution [41]. However, no recombination block exists on linkage group I of N. crassa. Additionally, Neurospora species are hermaphroditic, so the mating-type locus determines mating compatibility, rather than sexual role. We therefore consider it unlikely that the same forces that cause reproductive isolation loci to preferentially accumulate on sex chromosomes can account for the prevalence of the observed QTLs on linkage group I.
Earlier genetic studies of reproductive isolation in Neurospora identified a QTL on linkage group I as the N. crassa member of a Dobzhansky-Muller incompatibility locus-pair responsible for a severe defect in hybrid perithecial development between allopatric N. crassa and N. intermedia when N. crassa acts as the male partner [42], [43]. These incompatibility loci were first identified in populations of N. crassa×N. intermedia hybrids evolved under divergent environmental conditions in a test of the hypothesis that ecological adaptation can incidentally drive reproductive isolation [42]. Subsequent mapping determined that the incompatibility was caused by interactions between an N. crassa locus (dma on linkage group I) and an N. intermedia locus (dfe on linkage group V) [43]. Considering that both the geographic relationship of the species and gender role of N. crassa differ between this study and ours, it is very interesting that the N. crassa dma locus maps to a region of linkage group I that potentially coincides with the locations of QTLs identified in our study. Direct comparison between mapping results is prevented by the absence of sequence anchored, microsatellite markers in this region of our map.
Sexual microbes are likely to have simple premating recognition mechanisms, but will nevertheless experience selective pressure to avoid maladaptive hybridization. When evolution of premating barriers is constrained, microbial reinforcement may be more likely to involve non-premating-recognition mechanisms, including differentiated substrate or host fidelities [10] or evolution of divergent mating kinetics [44]. Here we have shown that selective abortion of hybrid fruitbodies by N. crassa fertilized by sympatric N. intermedia had the potential to evolve by natural selection by demonstrating that maternal colonies that abort hybrid fruitbodies are subsequently able to mate normally with conspecifics and can have higher reproductive fitness. We then show that the genetic architecture of hybrid fruitbody abortion is consistent with evolution via directional natural selection. Plants and animals are known to sometimes selectively abort otherwise viable embryos, thereby restricting parental investment to offspring with higher potential fitness [45]. Our data show that microbes like Neurospora, which provide costly parental care and are capable of multiple matings, are capable of undergoing reinforcement selection for selective abortion of hybrid offspring. Further studies on the evolution and genetics of reproductive isolation in microbial eukaryotes will be needed to challenge this hypothesis.
The biology of Neurospora, the evolutionary relationships among species and clades, and the biogeography of reproductive isolation between N. crassa and N. intermedia have been described previously [15], [16]. Culturing, crossing, and isolation of ascospore progeny were performed as previously described [15], [17]. Table S1 lists the wild-collected Neurospora strains used in this study. The QTL mapping population created for this study has been deposited with the Fungal Genetics Stock Center, Kansas City, Missouri.
Sequential fertilization was performed according to the methods of Howe and Prakash [27], except that at the first mating time point (5 days after inoculation of the NcC-India maternal strain), the conidial suspension of one fertilizing strain (either an NcC-India strain as conspecific positive control; an allopatric N. intermedia (African (n=2) or Caribbean (n=2)); sympatric N. intermedia (Indian (n=2)); or water negative control) was applied to 50% of the plate, while at the second time point (10 days after maternal inoculation), the fertilizing suspension of NcC-India was applied to two 1 cm2 spots located 1.5 cm–2 cm from the edge of the first fertilization. Three replicates were performed for a total of 24 plates. Fertility was scored 20 days after maternal inoculation using a 0–6 reproductive success scale (RSS) [15]. The effects of cross type and geographic origin and strain identity of the first–time-point fertilizing males on reproductive success of the second–time-point crosses were analyzed using a semi-parametric, proportional hazards model, with nested effects, as implemented by JMP 5.0.1a.
We obtained genomic DNA for each member of the QTL mapping population following the protocol of Dettman et al. [15]. AFLP and microsatellite primer sequences are shown in Table 5, Table 6, and Table 7. The mat-a1 and mat-A1 loci were amplified by multiplex PCR with the following primers: Ba1-5, AAGAAGAAGGTCAACGGCTTCATG; Ba1-3, CCAGAGCCATGTTCTAGGAATCATT; Sa1-5, CGTCGATGGCAATCTTTTCTGGAA; and Sa1-3, ATTGGCATCGTAGTTGAGAAGCTT [46]. The mat-a1 and mat-A1 fragments were distinguished by agarose gel electrophoresis. Genomic DNA was prepared for amplification of AFLP loci with the Invitrogen AFLP Core Reagent Kit. Selective "E" primers were 5′-modified with either 6-FAM or HEX fluorescent dye, and products of selective amplification were electrophoretically separated on an ABI 3100 genetic analyzer, and data were collected and analyzed with the ABI software GeneScan and Genotyper (Applied Biosystems, Inc., Carlsbad, CA).
Microsatellite loci in targeted genomic regions (e.g., chromosome ends and QTL regions) were selected from a published list of SSR in N. crassa [47], and primers were designed with Primer3 on the web [48]. Forward primers were 5′-fluorescently labeled with NED, 6-FAM, or HEX dyes, and size data were collected as for AFLP markers, above.
Linkage analysis was performed with MAPMAKER/EXP 3.0 [49]. Loci were sorted into sets of linked markers iteratively at a linkage threshold of 3.0 LOD and 50 cM (Kosambi) and then 6.0 LOD and 30 cM using the "group" command. For each linkage group, the "order" command was used to identify the best marker order with a window of 7 markers and a log-likelihood exclusion threshold of 2.0, followed by attempted placement of excluded markers at log-likelihood 1.0. Marker order was confirmed using the "ripple" command to permute five neighboring loci at a time and flag any alternative orders within a log-likelihood of 2.0 of the best order. Markers that could not be placed confidently according to these criteria were discarded. Linkage groups were assigned to the seven chromosomes of N. crassa based on inclusion of multiple microsatellite markers targeted to that chromosome.
Mendelian segregation of markers was checked in R/qtl [50] with the "geno.table" command. Linkage disequilibrium in pairs of physically unlinked markers was tested in Genepop 3.4 [51], using option 2, which uses a Fisher exact test implementing a Markov chain to estimate an unbiased P-value. Experiment wide significance threshold (Type I error α=0.05) was determined by Bonferroni correction for the 21 non-identical linkage-group pairs.
We investigated the genetics of hybrid fruitbody development in four kinds of N. crassa×N. intermedia matings (Table 4). Crosses were performed on synthetic cross media in 16×100 mm tubes. At 10 days post-fertilization, fruitbody development scores (FDS) were recorded following a four-category scale: 0, no fruitbody development; 1, early abortion (small fruitbodies without apical pores (ostioles)); 2, late abortion (larger fruitbodies with ostioles, but lacking "beaks"); and 3, fully developed (large flask-shaped fruitbodies with ostioles and "beaks"). Some crosses resulted in a mixture of two consecutive fruitbody development stages and were recorded as half steps in the scale (e.g., crosses with early- and late-aborted fruitbodies were recorded as 1.5).
Quantitative trait loci (QTLs) were identified by composite interval mapping (CIM) using Windows QTL Cartographer, v2.5 [52]. CIM was performed under model 6 with 5 control markers and a window size of 20 cM using a 1 cM walk speed. At each step the likelihood ratio statistic (LR) testing the hypothesis that a QTL exists versus the null hypothesis that no QTL exists was determined. For each trait, a critical LR threshold reflecting a Type I error of 0.05 was estimated by permuting the data 1000 times. Significant QTLs were CIM maxima whose LR exceeded the critical threshold and whose 95% confidence intervals were discontinuous with those of other CIM maxima. Ninety-five percent support intervals were estimated as the area bounded by 1-LOD drops in the LR where LOD=log10(LR/2 ln 10).
The null hypothesis of neutral trait evolution for sympatric hybrid fruitbody abortion (the reinforcement trait) was tested by subjecting the genetic effects data to a QTL sign test [32], as implemented by the QTLsigntest software provided by H. A. Orr. Because the QTL sign test assumes an additive genetic model, we first scanned for epistatic loci using "scantwo" of R/qtl using the expectation-maximization, interval mapping algorithm and multipoint genotype probabilities calculated using the "calc.genoprob" command with a step size of 2.5 and an error probability of 0.01. For each chromosome position the likelihood ratio statistic comparing the full epistatic model to the two-locus additive model was determined For each trait, a critical likelihood ratio statistic threshold reflecting a Type I error of 0.05 was estimated by permuting the data set 1000 times.
QTLsigntest determines how likely the proportion of loci with positive vs. negative additive effects is under a neutral model of complex trait evolution, when conditioned on the magnitude of the trait difference in the parent strains, the number of detected QTLs, the threshold of detection, and distribution of the absolute value of additive effects, which are all empirically determined. QTLsigntest was parameterized as follows: Parental RSS difference=2; number of QTLs=11; detection threshold=0.25; effects distribution gamma (shape=13.5, scale=0.034).
Candidate genes for postmating reinforcement in N. crassa. The ORFs found between microsatellite markers nc6L15 and nc6L16 on linkage group VI, which flank a major female-fertility QTL affecting sympatric hybrid fruitbody development, are listed. The presence of these ORFs in publicly available reproductive EST libraries is noted. The Sexual, Perithecial, Sperithecial, and Westergaard EST libraries were constructed from cDNA harvested from mycelia undergoing sexual development. All data are from the Broad Institute Neurospora crassa database.
We thank Brianne H. Daniels, Kenneth Takeoka, Kwan Hong Teoh, and Eze Uzozike for valuable assistance with data collection.
The authors have declared that no competing interests exist.
This work was funded by NSF grant DEB-0516511. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
PubMed Central Canada is a service of the Canadian Institutes of Health Research (CIHR) working in partnership with the National Research Council's Canada Institute for Scientific and Technical Information in cooperation with the National Center for Biotechnology Information at the U.S. National Library of Medicine(NCBI/NLM). It includes content provided to the PubMed Central International archive by participating publishers.


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Gateway Classic Cars is a premier classic and exotic car sales company. We are in a massive growth state with multiple showrooms nationwide. If you have a vehicle for sale, please give us a call at 866-430-2345.
For sale in our Detroit showroom is this clean low mile 2004 Harley Davidson Sportster XL883. Dressed in deep burgundy metallic with black trim... More
2002 Kawasaki Vulcan 800 Trike for sale in our Indianapolis Showroom. This Kawasaki Vulcan 800 is powered by an 800cc engine. Options on this... More
For sale is a 2007 Honda VTX1300C Motorcycle with just 1288 miles! This is a two owner all original bike that was well taken... More

subset: commoncrawl


text:
Analyst Research
|Report Title
Provider: Pechala's Reports
Marcus & Millichap Inc: Business description, financial summary, 3yr and interim financials, key statistics/ratios and historical ratio analysis.
Provider: Reuters Investment Profile
Provider: Thomson Reuters Stock Report
Validea Guru Analysis Report for MMI. Analysis using Validea's interpretation of the published quantitative strategies of well-known Wall Street experts including Peter Lynch, Warren Buffett, Ben Graham and Ken Fisher, among others.
Provider: Validea
NYSE and AMEX quotes delayed by at least 20 minutes. NASDAQ delayed by at least 15 minutes. For a complete list of exchanges and delays, please click here.
Marcus & Millichap Inc Prices Initial Public Offering
Marcus & Millichap Inc announced that its initial public offering of 6,000,000 shares of its common stock at a price to the public of $12.00 per share. The shares will begin trading on October 31, 2013, on the New York Stock Exchange under the ticker symbol "MMI." Of the shares in the offering, 2,726,587 shares are being offered by Marcus & Millichap, Inc. and 3,273,413 shares are being offered by selling stockholders. In addition, the Company has granted the underwriters a 30-day option to purchase up to an additional 900,000 shares of common stock on the same terms and conditions. Total net proceeds to the Company from the offering, after deducting the underwriting discount and estimated offering expenses, will be approximately $34.6 million. The Company intends to use the net proceeds from the offering for general corporate purposes, including capital expenditures and working capital to expand its services and potential acquisitions of real estate businesses or companies. The Company will not receive any proceeds from the sale of shares by the selling stockholders. Upon the completion of the initial public offering, Marcus & Millichap Real Estate Investment Services, Inc. will be a wholly owned subsidiary of Marcus & Millichap, Inc. Upon the completion of the initial public offering, Marcus & Millichap Real Estate Investment Services, Inc. will be a wholly owned subsidiary of Marcus & Millichap, Inc.
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We had a brilliant night in this gorgeous funky little cocktail party that has a brilliant Karaoke set up and delicious cocktails. Will definitely be back! Great for a girlie...
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We had a brilliant night in this gorgeous funky little cocktail party that has a brilliant Karaoke set up and delicious cocktails. Will definitely be back! Great for a girlie night out
I like chic bars with nice décor, great lighting, fancy drinks, and nicely dressed clientele. The other bars we had visited were very dressed down and casual and we were looking for something nicer. Roskos is gorgeous! very chic and lovely interior. It was exactly what we looking for, for an after dinner outing
Anyway we found the bar through...
Been for nights out in Lagos three years now, and this place is without doubt the best bar in the town! Great cocktails, cracking dj's and a great party atmosphere. Loved it!!
A small hip bar in the centre of lagos cocktails 2 for 1 from 9 to 11 that's 5 euro for two! Helpful and friendly staff, great sounds and nice crowd. Don't miss this one......
Wonderful staff and amazing cocktails make Roskos a must visit! Thank you for great drinks, conversation and fun!
Best cocktails we had in lagos and more upmarket than the surrounding back packer bars. Can reccomend the daiquiris in particular
Good cocktails and inexpensive, good ambience and fun, women feel safed, good music and space. Very clean comparing the most of the bars at Lagos. FUN! I recommend!
Thank you, we like people to feel at home...Christine.
Came here twice during our recent stay, both times we thoroughly enjoyed.
The first time we drank excessively and went through the cocktail menu quite possibly, from what I can remember I recommend the espresso martini as that one stands out.
The second time we went on a Wednesday night, 3€ mojitos all night! Very nice indeed for the price!...
Thank You for your kind words, this is Robin's wife, I'm so glad you enjoyed your holiday and we hope you come back to Lagos and pop in and see us. Christine.
Okay, I need to be clear, we are not the target market for this bar, it was our first time in Lagos and we didn't have a clue where to eat, we were lucky to come across this Bar, we arrived at 7.0 they had just opened and were busy preparing for the evening ahead, we had a couple of...
Thank you for taking the time to do a review and and all the kind words. Hopefully we will see you again very soon. Christine.
Love love love this place! Went during my holiday and cocktails were only 4 euro! Amazing cocktails presented and could definatly taste the alcohol!
Chilled out atmosphere and music that you can dance to or have a chat through. Friendly staff and lighting and seating very classy and chic. Would definately recommend this place if you are near lagos!
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|By PR Newswire
Article Rating:
|November 14, 2012 03:44 PM EST
MONTREAL, Nov. 14, 2012 /PRNewswire/ -- Wajam (www.wajam.com), a leader in social search technology, announced today its selection by Deloitte's Technology Fast 50 as the Jury's Favorite "Coup de Coeur" Award recipient. Launched 15 years ago, the Technology Fast 50™ program celebrates the world-class achievements and tremendous evolution of the Canadian technology sector. The jury was impressed with Wajam's leadership, business model, ethics and vision. Companies recognized by the jury must possess the management experience and superior technology that mark a successful early-stage company poised for continued growth and displaying the potential to make the Fast 50 in the near future.
"We are so honored to be recognized by Deloitte for our team's innovation and contribution to the Canadian technology sector," said Wajam CEO Martin-Luc Archambault. "The Wajam team appreciates all of the awards we have received this year, which validate the need and relevance social search brings to internet navigation. We hope to some day be a future Technology Fast 50 company, and will continue to recruit talent to expand our leadership in innovation."
Last week, Wajam was selected to the CIX Top 20 elite list. This week, the company was recognized as a finalist in the Red Herring Global 100, and just a two weeks ago, Wajam CEO Martin-Luc Archambault was named an Ernst & Young Emerging Entrepreneur Award recipient for 2012 in Quebec. Wajam was also selected as a Red Herring Americas Top 100 Winner and recipient of the C2MTL Emerging Entrepreneurs Award this year.
About Wajam – Great Minds Search Alike – www.wajam.com
Wajam (www.wajam.com) is a social search engine that gives you access to the knowledge of your friends when you need it. The Wajam Social Search Everywhere Plugin makes searching the web more personal by adding results from friends to standard searches on Google, Bing or Yahoo! It elevates search relevance by featuring your friends' likes and reviews for products, places, links, photos and videos within your regular search habits, even in places like Amazon, Yelp, Trip Advisor, YouTube and Ebay. The Wajam Search site gives users the ability to search their network of trusted friends and followers across multiple social networks all in one place.
Wajam is an International Stevies Award Silver Winner for Most Innovative Company in the U.S. and Canada as well as Best Social App; a Red Herring Americas Top 100 Winner and recipient of the C2MTL Emerging Entrepreneurs Award. Wajam received the highest security certifications from TRUSTe, McAfee and Norton, and is committed to protecting privacy and keeping user info safe. Wajam launched in private beta in January 2011 and was founded by Martin-Luc Archambault, who was previously CEO of Zango Canada and named Finalist in Ernst & Young Entrepreneur of the Year Award. Wajam is privately held and funded by Bolidea. Visit us on Facebook at http://www.facebook.com/wajam or Twitter at https://twitter.com/#!/wajam. Please refer also to our award-winning Press Page or visit our Blog.
Contact: John Sperrazzo, Eye-To-Eye Communications, Inc. 858.361.4495, [email protected]
SOURCE Wajam
Internet of Things (IoT) will be a hybrid ecosystem of diverse devices and sensors collaborating with operational and enterprise systems to create the next big application. In their session at @ThingsExpo, Bramh Gupta, founder and CEO of robomq.io, and Fred Yatzeck, principal architect leading product development at robomq.io, discussed how choosing the right middleware and integration strategy from the get-go will enable IoT solution developers to adapt and grow with the industry, while at the same time reduce Time to Market (TTM) by using plug and play capabilities offered by a robust IoT ...
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17th Cloud Expo, taking place Nov 3-5, 2015, at the Santa Clara Convention Center in Santa Clara, CA, will feature technical sessions from a rock star conference faculty and the leading industry players in the world. Cloud computing is now being embraced by a majority of enterprises of all sizes. Yesterday's debate about public vs. private has transformed into the reality of hybrid cloud: a recent survey shows that 74% of enterprises have a hybrid cloud strategy. Meanwhile, 94% of enterprises are using some form of XaaS – software, platform, and infrastructure as a service.
Jul. 4, 2015 02:00 PM EDT Reads: 1,053
SYS-CON Events announced today that Secure Infrastructure & Services will exhibit at SYS-CON's 17th International Cloud Expo®, which will take place on November 3–5, 2015, at the Santa Clara Convention Center in Santa Clara, CA. Secure Infrastructure & Services (SIAS) is a managed services provider of cloud computing solutions for the IBM Power Systems market. The company helps mid-market firms built on IBM hardware platforms to deploy new levels of reliable and cost-effective computing and high availability solutions, leveraging the cloud and the benefits of Infrastructure-as-a-Service (IaaS...
Jul. 4, 2015 01:00 PM EDT Reads: 1,263
The 17th International Cloud Expo has announced that its Call for Papers is open. 17th International Cloud Expo, to be held November 3-5, 2015, at the Santa Clara Convention Center in Santa Clara, CA, brings together Cloud Computing, APM, APIs, Microservices, Security, Big Data, Internet of Things, DevOps and WebRTC to one location. With cloud computing driving a higher percentage of enterprise IT budgets every year, it becomes increasingly important to plant your flag in this fast-expanding business opportunity. Submit your speaking proposal today!
Jul. 4, 2015 12:30 PM EDT Reads: 1,127
"We have a tagline - "Power in the API Economy." What that means is everything that is built in applications and connected applications is done through APIs," explained Roberto Medrano, Executive Vice President at Akana, in this SYS-CON.tv interview at 16th Cloud Expo, held June 9-11, 2015, at the Javits Center in New York City.
Jul. 4, 2015 12:30 PM EDT Reads: 1,360
The 5th International DevOps Summit, co-located with 17th International Cloud Expo – being held November 3-5, 2015, at the Santa Clara Convention Center in Santa Clara, CA – announces that its Call for Papers is open. Born out of proven success in agile development, cloud computing, and process automation, DevOps is a macro trend you cannot afford to miss. From showcase success stories from early adopters and web-scale businesses, DevOps is expanding to organizations of all sizes, including the world's largest enterprises – and delivering real results. Among the proven benefits, DevOps is corr...
Jul. 4, 2015 12:15 PM EDT Reads: 1,101
The basic integration architecture, as defined by ESBs, hasn't changed for more than a decade. Most cloud integration providers still rely on an ESB architecture and their proprietary connectors. As a result, enterprise integration projects suffer from constraints of availability and reliability of these connectors that are not re-usable across other integration vendors. However, the rapid adoption of APIs and almost ubiquitous availability of APIs amongst most SaaS and Cloud applications are rapidly redefining traditional integration approaches and their reliance on proprietary connectors. ...
Jul. 4, 2015 12:00 PM EDT Reads: 1,050
The Internet of Things is not only adding billions of sensors and billions of terabytes to the Internet. It is also forcing a fundamental change in the way we envision Information Technology. For the first time, more data is being created by devices at the edge of the Internet rather than from centralized systems. What does this mean for today's IT professional? In this Power Panel at @ThingsExpo, moderated by Conference Chair Roger Strukhoff, panelists addressed this very serious issue of profound change in the industry.
Jul. 4, 2015 10:45 AM EDT Reads: 1,499
Internet of Things is moving from being a hype to a reality. Experts estimate that internet connected cars will grow to 152 million, while over 100 million internet connected wireless light bulbs and lamps will be operational by 2020. These and many other intriguing statistics highlight the importance of Internet powered devices and how market penetration is going to multiply many times over in the next few years.
Jul. 4, 2015 09:00 AM EDT Reads: 2,338
Today air travel is a minefield of delays, hassles and customer disappointment. Airlines struggle to revitalize the experience. GE and M2Mi will demonstrate practical examples of how IoT solutions are helping airlines bring back personalization, reduce trip time and improve reliability. In their session at @ThingsExpo, Shyam Varan Nath, Principal Architect with GE, and Dr. Sarah Cooper, M2Mi's VP Business Development and Engineering, will explore the IoT cloud-based platform technologies driving this change including privacy controls, data transparency and integration of real time context wi...
Jul. 4, 2015 09:00 AM EDT Reads: 1,417
WebRTC converts the entire network into a ubiquitous communications cloud thereby connecting anytime, anywhere through any point. In his session at WebRTC Summit,, Mark Castleman, EIR at Bell Labs and Head of Future X Labs, will discuss how the transformational nature of communications is achieved through the democratizing force of WebRTC. WebRTC is doing for voice what HTML did for web content.
Jul. 4, 2015 08:45 AM EDT Reads: 1,072
To many people, IoT is a buzzword whose value is not understood. Many people think IoT is all about wearables and home automation. In his session at @ThingsExpo, Mike Kavis, Vice President & Principal Cloud Architect at Cloud Technology Partners, discussed some incredible game-changing use cases and how they are transforming industries like agriculture, manufacturing, health care, and smart cities. He will discuss cool technologies like smart dust, robotics, smart labels, and much more. Prepare to be blown away with a glimpse of the future.
Jul. 4, 2015 08:30 AM EDT Reads: 1,215
Explosive growth in connected devices. Enormous amounts of data for collection and analysis. Critical use of data for split-second decision making and actionable information. All three are factors in making the Internet of Things a reality. Yet, any one factor would have an IT organization pondering its infrastructure strategy. How should your organization enhance its IT framework to enable an Internet of Things implementation? In his session at @ThingsExpo, James Kirkland, Red Hat's Chief Architect for the Internet of Things and Intelligent Systems, described how to revolutionize your archit...
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It is one thing to build single industrial IoT applications, but what will it take to build the Smart Cities and truly society-changing applications of the future? The technology won't be the problem, it will be the number of parties that need to work together and be aligned in their motivation to succeed. In his session at @ThingsExpo, Jason Mondanaro, Director, Product Management at Metanga, discussed how you can plan to cooperate, partner, and form lasting all-star teams to change the world and it starts with business models and monetization strategies.
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SYS-CON Events announced today that BMC will exhibit at SYS-CON's 16th International Cloud Expo®, which will take place on June 9-11, 2015, at the Javits Center in New York City, NY. BMC delivers software solutions that help IT transform digital enterprises for the ultimate competitive business advantage. BMC has worked with thousands of leading companies to create and deliver powerful IT management services. From mainframe to cloud to mobile, BMC pairs high-speed digital innovation with robust IT industrialization – allowing customers to provide amazing user experiences with optimized IT per...
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There will be 150 billion connected devices by 2020. New digital businesses have already disrupted value chains across every industry. APIs are at the center of the digital business. You need to understand what assets you have that can be exposed digitally, what their digital value chain is, and how to create an effective business model around that value chain to compete in this economy. No enterprise can be complacent and not engage in the digital economy. Learn how to be the disruptor and not the disruptee.
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The Internet of Things is not only adding billions of sensors and billions of terabytes to the Internet. It is also forcing a fundamental change in the way we envision Information Technology. For the first time, more data is being created by devices at the edge of the Internet rather than from centralized systems. What does this mean for today's IT professional? In this Power Panel at @ThingsExpo, moderated by Conference Chair Roger Strukhoff, panelists will addresses this very serious issue of profound change in the industry.
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Business as usual for IT is evolving into a "Make or Buy" decision on a service-by-service conversation with input from the LOBs. How does your organization move forward with cloud? In his general session at 16th Cloud Expo, Paul Maravei, Regional Sales Manager, Hybrid Cloud and Managed Services at Cisco, discusses how Cisco and its partners offer a market-leading portfolio and ecosystem of cloud infrastructure and application services that allow you to uniquely and securely combine cloud business applications and services across multiple cloud delivery models.
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Buzzword alert: Microservices and IoT at a DevOps conference? What could possibly go wrong? In this Power Panel at DevOps Summit, moderated by Jason Bloomberg, the leading expert on architecting agility for the enterprise and president of Intellyx, panelists peeled away the buzz and discuss the important architectural principles behind implementing IoT solutions for the enterprise. As remote IoT devices and sensors become increasingly intelligent, they become part of our distributed cloud environment, and we must architect and code accordingly. At the very least, you'll have no problem fillin...
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text: Aviation SAS secures survival deal Add to ... Reuters Published Tuesday, Nov. 20, 2012 6:31AM EST Last updated Tuesday, Nov. 20, 2012 6:33AM EST Scandinavian airline SAS wrapped up a deal with the last of eight unions on a plan to reduce costs, clearing the way to finalise a $515-million (U.S.) dollar rescue deal. Hayley Platt reports. Watch More Video: Greeks defy Europe with overwhelming referendum 'No' Greek negotiator says bailout offer was easy to refuse Facebook unveils subtle changes to its logo Whole Foods CEOs apologize for overcharging Countdown on to Greek vote Vancouver housing heats up, but it's not too hot to handle Market Call Tonight: Top Picks from Middlefield Group's Andy Nasr Canadian bank stocks underperform in first half of year Is China a bubble about to burst? Potash Corp. wants face time with K+S Blaming 'foreign puppets' for China's stock markets rout Plan now to avoid a financially-uncertain retirement Another Sears Canada CEO out: A turnaround plan is long overdue BP settles U.S. oil spill claims for $18.7 billion K+S rejects $8.6B takeover proposal by Potash Corp. U.S. adds 223,000 jobs in June Creditors to Greece: No more talks until after referendum Strike begins at nine Loblaw-owned stores Athens is 'like winter,' say local business owners Bull vs. Bear: Should you give your kids an allowance? Comments AA More Related to this Story Economy Video: U.S. economy most dynamic in the world: Harper Labour Video: Court order preserves Hostess for now


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Originally written on Fox Sports North | Last updated 5/31/13
MINNEAPOLIS All offseason, the Twins preached the importance of bolstering a starting pitching rotation that was among the worst in baseball in 2012. Through two months of the 2013 season, Minnesota is still near the bottom of many starting pitching categories. Prior to Thursday, Twins starters had a combined 5.64 ERA, tied with Houston for the worst in baseball. By comparison, Minnesota's rotation had a collective 5.40 ERA by the end of last season. Only Colorado's 5.80 ERA was worse. Opponents batted .329 and slugged a remarkable .515 against Twins starters in the first 50 games prior to Thursday, both of which are easily the highest of any starting staff. The Astros are next at .304 and .502, respectively. The Chicago White Sox boast the best opponent batting average in the American League Central at a paltry .229, second-best in baseball. Minnesota signed two veteran free agents and traded for another this past winter, but only one of the three has been reliable to date. Tha...
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Select Local Merchants
Victory Christian SchoolRead MoreSinger-songwriter Pat Green takes his winning mesh of rocking country music back on the road, fortifying his army of fans while spreading Texas pride across the land on his current tour. From massive hits such as ?Wave on Wave? through his surplus of honky-tonk heartbreakers and keg-juggling anthems, Pat rides a brakeless gravy train fueled by a strange talent called ?talent? and the storytelling skills of a straight-shooting Aesop. The Rally on the Ravine concert celebrates a football rivalry between the Ouachita Baptist Tigers and the Henderson State Reddies, who will be clashing on the gridiron across Highway 67 before the show (football tickets are not included with this Groupon).1244 Maul RdCamden, AR US
White House CafeRead MoreLocated in Camden, White House Cafe is a well-known restaurant that delivers appetizing coffee and good steak. This restaurant delivers an unforgettable dining experience set in a relaxed ambience. It's a culinary destination for visitors that want a delicious meal. There's no specific recommended attire, so feel free to dress casually and comfortably. There really is something for everyone, with gluten-free options, as well as low-fat and vegetarian items on the robust menu. Plus, if you're looking for the perfect spot for a get-together between family or friends, it's been reviewed as a great local option for both big groups and families with kids. In addition to its convenient take-out menu, the restaurant even provides catering for events around town. Specializing in both lunch and dinner, White House Cafe definitely stands out from the rest. Visitors to the restaurant have access to a private lot nearby or can park on the street.323 S Adams AveCamden, AR US
Postmasters GrillRead MoreA top-rated steakhouse in Camden, Postmasters Grill offers a fun atmosphere with a fresh menu that features seafood. It's a local favorite for those seeking outstanding food. There's not really a recommended attire, so feel free to dress comfortably. It's been described as a solid option for a romantic occasion and large groups, in addition to doing business and special occasions. In addition to its convenient take-out offerings, the restaurant even provides catering for events around town. Or, if you just want to stop by for a beverage, the restaurant has a good selection at its bar. A good spot for dinner, Postmasters Grill definitely stands out from the rest. Visitors to the restaurant have access to a private lot nearby or can park on the street. Would you rather ride there? Bicycle parking is also provided.133 W Washington StCamden, AR US
Sand BarRead MoreThe premier destination for fabulous pub food and tasty seafood, Sand Bar in Camden on S Adams Ave is one of the area's top-rated restaurants. It's a popular restaurant for customers interested in good food. It's often reported that the background noise can get a bit loud here. There really is something for everyone, with gluten-free options, as well as low-fat and vegetarian items on the robust menu. Plus, it's been tabbed by the locals as a really good option for large groups. If you just feel like stopping by for a beverage, the restaurant has helpful bar staff (and a good selection behind the bar). One of the tastier franchises in the area, a trip to this Sand Bar is definitely worthwhile.245 S Adams AveCamden, AR US
Sonic Drive-InRead MoreStop by Sonic Drive-In after work for a quick burger and fries. Low-fat and gluten-free options are featured on the menu. Surround yourself with the wonderful weather at your next night out at Sonic Drive-In. Not a popular place for dress-up dining, most Sonic Drive-In patrons come in casual attire. If you're in a hurry, place an order for pickup instead. Through their catering service, Sonic Drive-In can also set out a delicious spread for your next party. null You won't break the bank at Sonic Drive-In, with a meal typically hovering below the $15 mark. Sonic Drive-In serves breakfast, lunch, and dinner, so stop by whenever is most convenient for you.670 Cash Rd SWCamden, AR US
A Cut AboveRead MoreA perfect tan can change your look at the drop of a hat. Visit A Cut Above in Fordyce for an exciting transformation! Look brighter, feel happier with this salon's full tanning salon. They have you covered entirely, with their services including tanning beds, spray tan, and airbrush tan. Refresh and rediscover the natural beauty of your hair with a customized hair treatment. If alternative treatments are more your style, this salon has a range of options perfect for you, including acne treatment. For the most natural looking color, A Cut Above takes the cake in top-of-the-line tanning.1203 Moro StFordyce, AR US


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1-10 of 16 properties for sale found
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3 bedroom semi-detached house for sale Storers Quay, Isle of dogs, London, E14 3BZ
New to the market is this stunning home which has been extensively redesigned to a high specification by its owners. Rarely available in this popular location the humble exterior leads through to a superb and unexpected open plan reception kitchen area encased in glass walls and ceiling offering ... More details ›
£699,950 Under Offer
3 bedroom terraced house for sale Hesperus Crescent, Isle of dogs, London, E14 3AB
Quietly located in a pleasant residential cul-de-sac, this charming three bedroomed house is arranged over three floors, boasts stylish interior, bright accommodation and off-street parking. In fantastic order and recently refurbished the property briefly comprises of a separate reception, t... More details ›
£635,000 Under Offer
3 bedroom house for sale Lockesfield Place, Isle of Dogs, London, E14 3AJ
We are proud to present this delightful 3 bedroom house situated to the rear of the popular Lockesfield Place development. Entering the ground floor you are welcomed by the hallway offering access to the reception, kitchen and Wc. The reception opens out to the beautifully landscaped and maintain... More details ›
£506,000 Offers in Excess of
Offers in Excess of £506,000
2 bedroom flat for sale Chart House, Burrells Wharf Square, Isle of Dogs, London, E14 3TW
Holland Properties is delighted to offer this two bedroom apartment set on the 5th floor with underground parking, two balconies and partial river view. The property consists of two double bedrooms, large reception, en-suite bathroom, shower room and fitted kitchen. Burrells Wharf offers 24 hour ... More details ›
3 bedroom house for sale Prioress Crescent, Ingress Park, Greenhithe, DA9 9GE
Holland Properties are delighted to offer this three bedroom house situated on the popular and sought after Ingress Park development. The property's position is sure to impress being situated along side Ingress Abbey, a stunning building itself, particularly at night with additional views to ... More details ›
2 bedroom maisonette for sale Glengarnock Avenue, Isle of Dogs, London, E14 3BP
Holland Properties are delighted to offer to the market this spacious 2 double bedroom ex-local property boasting open plan kitchen, wooden flooring and having been decorated to a high standard. More details ›
£340,000 Under Offer
1 bedroom flat for sale Blake Building, Hornsey, London, N8 7QF
Located on the second floor of the Blake Building within the popular New River Village development is this spacious one bedroom apartment. Comprising of reception with open plan modern kitchen, large bedroom with fitted wardrobes and stylish bathroom. Plus, large balcony accessed via reception ... More details ›
£335,000 Sold STC
Offers in Excess of £335,000
2 bedroom apartment for sale Cadbury Way, London, SE16 3UZ
Holland Properties are pleased to offer for sale this split level ex local authority purpose built maisonette. The accommodation is ideally situated over the ground and first floors offering two bedrooms, open plan kitchen/dining area to the first floor and own front garden. More details ›
£335,000 Sold STC
1 bedroom flat for sale Blake Building, New River Avenue, London, N8 7QF
A lovely one bedroom apartment situated on the third floor of 'Blake Apartments' within the popular New River Village development. The apartment has been fitted to a very high specification and offers a private balcony, an on-site concierge, gym and restaurant / bar. This apartment is sub... More details ›
£300,000 Offers in Excess of
Offers in Excess of £300,000
2 bedroom flat for sale Castleton House, Pier Street, London, E14 3HU
A recently re-modernized two bedroom second floor apartment with accommodation that comprises; reception room with south facing balcony, separate kitchen, bathroom and separate cloakroom. This property is offered to the market chain free. More details ›


text:
Sweet Freezer Pickle Chips
5 Feb 2011
|2 1⁄2 Pound (5 Medium Sized)
|White onion
|1 Medium
|2 Tablespoon
|Ice cubes
|8 Cup (128 tbs)
|4 Cup (64 tbs)
|Cider vinegar
|2 Cup (32 tbs)
Insert slicing disc.
Slice cucumbers; transfer to a large mixing bowl.
Slice onion.
Add onion and salt to cucumbers; stir to mix.
Cover with ice cubes and refrigerate for 2 to 3 hours.
Drain water from bowl and discard any un-melted ice cubes, but do not rinse cucumber mixture.
Pack cucumber mixture in freezer containers or jars, filling each to within 1 1/2 inches of rim.
In a 2 to 3-quart pan, combine sugar and vinegar.
Bring to a boil over high heat, stirring until sugar is dissolved.
Pour just enough hot syrup over cucumber mixture to cover.
Put lids in place and freeze pickles for at least 1 week.
Before serving, thaw in refrigerator for about 8 hours.
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Sweet Freezer Pickle Chips Recipe


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Library Blog
Featured Authors
John Adams: On the Best-Read Founding Father and the Woman Behind Him
October 28, 2010
What would you do if you had to build a nation? Start Googling? Download an app? When John Adams strode onto the world stage by joining the Continental Congress as a representative from Massachusetts, he had no iPhone or MacBook Air. Contemporary European books (often in their original languages) and Greek and Latin classics were his RSS and HTML.
Library Life
Book Club Profile: Mysterious Undertakings at Waldo
October 28, 2010
Welcome to the first of our KCPL Book Group profiles. Mysterious Undertakings at the Waldo Branch gladly agreed to accommodate a drop-in visitor who took copious notes and photographs. But then again, Waldo welcomes all newcomers, and on this particular night, there was more than one newbie.
Library Life
Video: It's Only Civil to Remember Missouri
October 27, 2010
Earlier this month, Kansas City Public Library Executive Director Crosby Kemper III was officially sworn in to the Missouri Civil War Sesquicentennial Commission. In addition to having a tongue-twisting name, the MCWSC aims to commemorate Missouri's historically significant yet often overlooked role in the Civil War, and to recognize how that role reverberates today. The Commission was established in April 2010 by Missouri Governor Jay Nixon.
Library Life
And the Top Chef Is...
October 26, 2010
Our Food for Fines Week (October 18 through 24) has come to a close. And while we're still running the numbers to see how much food Library patrons donated to our community partner Harvesters and how much we forgave in overdue fines, we'd like to recognize one person with exceptionally good taste.
Featured Authors
Garry Trudeau's Kansas City Connection
October 22, 2010
The world may have never known Doonesbury if it weren't for Jim Andrews and John McMeel. The founders of Andrews McMeel Universal (then called Universal Press Syndicate) were headquartered in a rented house in Leawood when they discovered a young cartoonist named Garry Trudeau.


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The paper 'Energy Babble: Mixing Environmentally-Oriented Internet Content to Engage Energy Community Groups' received an Honourable Mention Award at CHI 2015 – ACM Conference on Human Factors In Computer Systems. The paper was co-authored with Bill Gaver, Mike Michael, Tobie Kerridge, Andy Boucher, Liliana Ovalle and Matthew Plummer-Fernandez.
Honourable Mention Award
With Jennifer Gabrys, Michael Guggenheim, Noortje Marres, Dan Neyland and Evelyn Ruppert I'm involved in launching and editing a new journal that brings together contributions that span science and technology studies with art and design.
Call for Contributions for the Launch of
'Demonstrations: Journal for Experiments in the Social Studies of Technology'
Demonstrations is a new open access online platform for publications at the intersection of science and technology studies, art and design, to be launched in the autumn of 2015. For our inaugural issue, we invite contributions exploring the theme of 'demonstration' by intellectual and creative means, in the form of research articles, review essays as well as more reflective contributions.
About the journal
Demonstrations is an interdisciplinary platform that aims to publish peer-reviewed contributions that combine creative approaches with science and technology studies. It provides this new and sprawling area an appropriate platform for publication, experiment and reflection. We hope to publish both more conventional articles on methodological, theoretical and empirical issues, but also and primarily items that combine and play with methods, media, materials and theories in novel ways. As an online platform, Demonstrations has a number of advantages for such publications over conventional journals, most obviously the fact that it is easy to include high resolution audio-visual material with decreasing space limitations. We then hope that Demonstrations becomes the main hub for a number of emerging traditions across the social studies of science, technology, art and design which are difficult to classify and which combine ethnography and art and design as well as digital and environmental methods.
Demonstrations is conceived as a platform for varieties of contributions, rather than as a traditional journal. We plan to offer a set of different possible templates for content and we will publish contributions not in issues, but continuously as part of annual volumes. Some of these will be assembled in thematic collections but on a dynamic basis, as part of or after publication. This will allow us to publish items in a more timely fashion. Each contribution will have a stable DOI, which makes it equivalent to a journal article.
Launching Demonstrations
For the platform launch, we invite contributions that examine, review and provoke intellectual and creative contexts for Demonstrations. Since its very inception, social studies of science and technology (STS) have taken a special interest in the public staging of knowledge and innovation, and over the last decades this work has been extended into spaces of creative and curatorial practice by leading figures in design, art and STS. For the inaugural issue of Demonstrations, we invite contributions to showcase these new directions in creative and critical research on and deployment of demonstrations across STS, art and design.
We invite three types of contributions:
a) Research contributions that showcase the critical and creative analysis of demonstrations as well as the deployment of demonstrations as devices of social and cultural research and intervention.
b) Review contributions that present and review the state of the art in demonstrations across the fields of STS, art and design.
c) Reflections on the theme of demonstration in relation to STS literatures and performative traditions in art and design.
Key Deadlines
Submission of contributions: May 15, 2015
Review of contributions: July 15, 2015
Final submission: September 15, 2015
Submission Format
Provided it follows the Demonstrations formatting guidelines, a submission can contain all kinds of media, including text, images and video. Detailed formatting guidelines will appear on the journal website soon. If you plan to submit a visually complex contribution, please get in contact with the editors (firstname.lastname@example.com). The submissions guidelines can be found at http://bit.ly/1CpwlUZ.
Journal info
Demonstrations is housed in the Centre for the Study of Invention and Social Process (CSISP) in the Sociology Department, Goldsmiths and hosted by Goldsmiths Library.
Editorial Collective: Jennifer Gabrys, Michael Guggenheim, Noortje Marres, Dan Neyland, Evelyn Ruppert, Alex Wilkie (Goldsmiths)
Editorial Assistant: Laurie Waller
Designer: Danah Abdulla
Editorial Board: Carl DiSalvo (Atlanta), Matt Fuller (Goldsmiths), Lucy Kimbell (London), Bernd Kräftner (Vienna), Javier Lezaun (Oxford), Daniel Lopez (Barcelona), Amade M'Charek (Amsterdam), Katja Mayer (Vienna), Anna Munster (Sydney), Michelle Murphy (Toronto), Tahani Nadim (Berlin), María Puig de la Bellacasa (Leicester), Israel Rodriguez Giralt (Barcelona), Tomas Sanchez Criado (Barcelona), Alex Taylor (Oxford), Nina Wakeford (Goldsmiths), Gisa Weszkalnys (London).
I've been interviewed as part of an AHRC commissioned project 'ProtoPublics'. As described on the site, during the interview I talk "about 'putting pressure on the social', the potentials in the interface between computational technology and politics and how he uses Actor Network Theory (ANT) in his work". ProtoPublics also features interviews with Rob Imrie, Rachel Aldred, Tim Schwanen, Noortje Marres, Pelle Ehn and Cat Rossi.
Engagement, Implications & Relevance of Design Research
On the 11th of March doctoral students in design from Brighton University and Goldsmiths convened at the Victoria & Albert Museum to discuss the topics of engagement, implications and relevance of doctoral research in design. Here's the blurb from the handout:
"During this session at the V&A we will discuss the topics of 'engagement', 'implications' and 'relevance' of doctoral research. We will consider the epistemic value of PhD research in design, whether practice-based, written or some combination of the two. We will explore the kinds of knowledge produced as part of your research, who or what this knowledge addresses, whom or what is affected or implicated by the outcomes of your research as well as the relevance of your research to the discipline of design as well as other disciplines, stakeholders, implicated parties etc.
It is worth noting that we are using the terms engagement, implications and relevance as indicators of the practical concerns of design researchers as well as theoretical concepts that can be used to sensitise us to the epistemic politics of our research. The notion of engagement has, for example, has been taken up by scholars on the Public Understanding of Science (within the field of Science and Technology Studies) to denote how the researched (e.g. publics) can contribute to and are shaped in knowledge production (Wynne, 2006; DiSalvo, 2009; Michael, 2012). In terms of implications, Paul Dourish (2006) provides a useful provocation in which the role of ethnography in design can be better understood as providing an analytic sensibility as well as a theoretical orientation to understanding social practices and technology in order to inform IT design. Arguably, there is little debate about the 'relevance' of design research and its contribution and accountability to policy objectives. Although 'users' play a conspicuous and normative role In design discourse (as primarily essentialist entities) it is, perhaps, to the social sciences (e.g. Rappert, 1999; Burawoy, 2005) that we can look to provoke debates around the users of our own knowledge practices as well as a more reflexive discussion about the epistemic politics and utility of design."
The seminar was organised by Guy Julier and myself.
Energy Babble: Mixing Environmentally-Oriented Internet Content to Engage Community Groups
The paper 'Energy Babble: Mixing Environmentally-Oriented Internet Content to Engage Community Groups' has been accepted for the CHI 2015 conference, authored by Bill, Mike, Tobie, myself, Andy, Liliana and Matthew.


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This week in tech… The past couple of weeks we have been giving our guides a familiarisation on sidemount tech diving and twinset configurations. The guys like to play on something different from time to time and it also makes them appreciate that sometimes it takes a little bit longer to get ready.
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This week in Tec .. it has been a bit quieter after the holidays which of course means that we offered a Tech try dive in the pool to the resort guests! It being a bit chilly only our hardy Australian guests, Tarn and Cara from Melbourne gave it a go, and I can think of no better indication that they liked the configuration other than the first words out of Tarn's mouth, "How much is one of these?"
In Feb 2013, Two Fish Divers contracted with Blue Label Diving, a premier PADI/TDI facility for TEC Courses and diving, to run all TEC Diving services at Two Fish Divers locations, under the name Blue Label Diving Indonesia (BLDI).
One of the most important service that BLDI provides is Sidemount Diving – providing courses and diving for recreational use to 40m as well as services for more deeper diving to 100m-plus. When you visit us, you may be interested in our FREE weekly Intro to Tec that we hold at our resorts, or the 2-day/3dive Sidemount Course for only Eu140 (supplement on top of normal accomm/dive package) – and during Jan-Mar 2014 we are running our TEC Promo that gives you 20% discount on all our TEC courses!!
Read More
The last weeks have all gone under one keyword – planning – despite there being resorts and dive centers on Bunaken and Lembeh the process of getting helium and oxygen in all desired combinations is still quite slow. That in turns means we need to look quite far ahead with our planning (specially for divers wanting to dive deeper on trimix) to have stock of all gases for all dives and all situations.
After returning from the wonderful Singapore and ADEX show it was time to get down and dirty, and over the last week we have done lots of sidemount diving in Bunaken.


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Short web commercial I directed, shot, and edited for Brewsees, a company that makes bottle-opening sunglasses. The commercial was shot as part of AdLab, Boston University's student-run ad agency.
Shot on a 7D, edited on Adobe Premiere.
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Celebrities Who Have Voted Republican
Credit: Getty Images
Republican Celebrities: Clint Eastwood
5 of 10
Traditionally Clint has voted Republican. After backlash from his infamous Super Bowl ad this year, Eastwood made a statement saying it was not an endorsement to Obama after being slammed by Republicans. "l am certainly not politically affiliated with Mr Obama," he told Fox. "It was meant to be a message about just about job growth and the spirit of America."


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->''"Mean Street has gotta have a figure of authority. And I got the perfect figure for it."''
--> --'''[[{{Pete}} Big Bad Pete]]''', ''VideoGame/EpicMickey''
->''"Your leaders are just... taller than everyone else?"''
--> --'''Dib upon learning of the Irken command structure''', ''WesternAnimation/InvaderZim''
->Heavily built and massive, there was a suggestion of uncouth physical inertia in the figure, but above this unwieldy frame there was perched a head so masterful in its brow, so alert in its steel-gray, deep-set eyes, so firm in its lips, and so subtle in its play of expression, that after the first glance one forgot the gross body and remembered only the dominant mind.
-->--Introduction to [[Literature/SherlockHolmes Mycroft Holmes]], "The Bruce-Partington Plans"


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5th Annual Atlanta Hip Hop Day Festival
- Music
- 91 Peachtree St. Atlanta, GA 30303, US
AFP-Georgia: A Patriots Christmas
Americans for Prosperity - Georgia
Friday, December 11, 2009 at 7:00 PM (EST)
Join Americans for Prosperity and tea party friends statewide for the event of the holiday benefiting the work of Americans for Prosperity Georgia and Operation HomeFront in serving Georgia's military families. It's A Patriot's Christmas Featuring acoustic Christmas performances by Lauren St. Jane and others with the Atlanta hometown Premiere of TEA PARTY: The Documentary Film!
Date Friday, December 11 at 7PM
Location Georgia World Congress Center - Room C101 (Northside Drive Entrance)
Parking Gold ($10) or Yellow ($8) LotsTickets
General Admission $16.00 (plus 6% sales tax)
$19.00 at the door
Please print off your receipt after submitting order and bring it with you to claim tickets at event along with Photo ID of person whose name appears on the above card. Email address is opt in agreement to receive FREE weekly AFP GA newsletter. All sales final.
About The Film "Tea Party: The Documentary Film" follows the struggles of five grassroots individuals and their transformation from home town activists and rally organizers to national activists in the 9/12 March on Washington. In the process, the film reveals what is at the heart of this nationwide surge of civic engagement - a return to and respect for a limited government, personal responsibility and fiscal restraint.
Tea Party Movie Channel - Click HERE to watch reaction from the Right and the Left
Lauren St. Jane is known around Atlanta as a personality on 104.7 The Fish and a member of the On Time Traffic Team at Intelligent Talk 920am WGKA. She is also an aspiring and inspiring musician performing in local venues with a beautiful acoustic sound that blends folksy with bluegrass country influences.
Americans for Prosperity - Georgia (AFP-GA) is committed to educating citizens about economic policy and mobilizing those citizens as advocates in the public policy process. AFP is an organization of grassroots leaders who engage citizens in the name of limited government and free markets on the local, state, and federal levels. AFP grassroots activists advocate for public policies that champion the principles of entrepreneurship and fiscal and regulatory restraint. To that end, AFP supports:
Georgia Chapter of AFP
2675 Mall of GA Blvd, Bldg 100
Buford, GA 30519, Suite 101
Phone: (770) 315-4304
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In order to purchase these tickets in installments, you'll need an Eventbrite account. Log in or sign up for a free account to continue.


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HTC Files To Ban U.S. iPhone, iPad, iPod Sales
Policy Counsel
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NEW YORK (Reuters) – HTC said on Wednesday it filed a patent infringement case against Apple and asked the U.S. International Trade Commission to ban U.S. sales of iPhones, iPads and iPods.


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Castle regular Chris Bevins has become the first British climber to solo The Nose on El Capitan (Yosemite, California, USA) in under 24 hours. Read the full story on UKClimbing in which Chris graciously acknowledged the role The Castle played in his preparations. Awesome effort, Chris!
Hooray for us!
The Castle won the Sustainable Building and Sustainable Procurement Awards at the 2011/2012 Sustainable City Awards.
More sustainability...
Saturday 11th July - 12pm to 7pm
Including pizzas, burgers, cakes, cocktails, mini comp and free dyno workshops
**Update: Live music from Antarma & the family tree from 1.30pm!
Take a tour of the Castle Climbing Centre and discover what we're all
Castle Climbing Center - A Day in the Life from Tim Ewins on Vimeo.


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