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Rates of spontaneous mutation have been estimated under optimal growth conditions for a variety of DNA-based microbes, including viruses, bacteria, and eukaryotes. When expressed as genomic mutation rates, most of the values were in the vicinity of 0.003–0.004 with a range of less than two-fold. Because the genome sizes varied by roughly 104-fold, the mutation rates per average base pair varied inversely by a similar factor. Even though the commonality of the observed genomic rates remains unexplained, it implies that mutation rates in unstressed microbes reach values that can be finely tuned by evolution. An insight originating in the 1920s and maturing in the 1960s proposed that the genomic mutation rate would reflect a balance between the deleterious effect of the average mutation and the cost of further reducing the mutation rate. If this view is correct, then increasing the deleterious impact of the average mutation should be countered by reducing the genomic mutation rate. It is a common observation that many neutral or nearly neutral mutations become strongly deleterious at higher temperatures, in which case they are called temperature-sensitive mutations. Recently, the kinds and rates of spontaneous mutations were described for two microbial thermophiles, a bacterium and an archaeon. Using an updated method to extrapolate from mutation-reporter genes to whole genomes reveals that the rate of base substitutions is substantially lower in these two thermophiles than in mesophiles. This result provides the first experimental support for the concept of an evolved balance between the total genomic impact of mutations and the cost of further reducing the basal mutation rate.
Author Summary
Spontaneous mutations are key drivers of evolution and disease. In microbes, most mutations are deleterious, some are neutral (without significant impact), and a few are advantageous. Because deleterious mutations reduce fitness, there should be constant selection for antimutator mutations that reduce rates of spontaneous mutation. However, such reductions are necessarily achieved at some cost. Therefore, a mutation rate should converge evolutionarily on a value that reflects this trade-off. For DNA microbes, the observed genomic mutation rate is remarkably (and mysteriously) invariant, in the neighborhood of 0.003–0.004, with a range of less than two-fold despite huge variation per average base pair in organisms with a wide diversity of life histories. Would an environmental condition that increased the average deleterious impact of a mutation be balanced by additional investments in antimutator mutations? It is widely observed that many mutations with mild impacts become strongly deleterious at higher temperatures, so mutation rates were measured in two thermophiles, a bacterium and an archaeon. Remarkably, both displayed average mutation rates reduced by about five-fold from the characteristic mesophilic value, most of the decrease reflecting a 10-fold reduction in the rate of base substitutions.
Citation: Drake JW (2009) Avoiding Dangerous Missense: Thermophiles Display Especially Low Mutation Rates. PLoS Genet 5(6): e1000520. https://doi.org/10.1371/journal.pgen.1000520
Editor: Josep Casadesús, Universidad de Sevilla, Spain
Received: March 18, 2009; Accepted: May 18, 2009; Published: June 19, 2009
This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose.
Funding: This research was supported by funds allocated to project number Z01 ES65016 of the Intramural Research Program of the NIH, National Institute of Environmental Health Sciences. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing interests: The author has declared that no competing interests exist.
It has become increasingly clear that the basal rate of spontaneous mutation per genome per replication is remarkably invariant in DNA microbes: using a classical correction factor for estimating the ratio of all base-pair substitutions (BPSs) to detected base-pair substitutions, genomic mutation rates (mutations per genome per replication) vary by less than twofold while genome sizes vary by ≈6,000-fold (Table 1). Thus, when mutation rates are expressed per average base pair, they also vary by a similarly large factor. Therefore, basal mutation rates characteristic of unstressed microbial populations can evolve to finely tuned values. The theory of mutation rates has its roots in Haldane's 1927 formulation of the impact of selection and mutation on fitness [1], followed by Sturtevant's 1937 conjecture that the deleterious character of most mutations would generate selective pressures that should lower mutation rates indefinitely [2]. In 1967, Kimura offered the hypothesis that there would be a "physiological cost" to each reduction in rate, leading to an equilibrium value when that cost outweighs the gain in fitness [3]. The surprise has been that the observed genomic rates are so narrowly distributed among DNA microbes despite a wide variety of life histories and genome sizes. An even deeper mystery, not to be addressed here, is why the particular microbial genomic rate of about 0.003–0.004 has been adopted by microbes of such diverse life histories and genome sizes.
If the Kimura conjecture is correct, then increasing the average deleterious impact of a spontaneous mutation (and thus converting many neutral or nearly-neutral mutations to deleterious mutations) would lower the rate of mutation, at least on an evolutionary time scale. The concept of an equilibrium basal mutation rate is difficult to test in a laboratory context because any imposed resetting of the equilibrium would probably require numbers of generations large even by microbial standards, and is difficult to test convincingly because only one or a few habitats could be explored. However, it has recently proven possible to test the concept by examining a natural evolutionary experiment, life at high temperatures. Those who gather mutants for fun or profit have often observed that the most common class of mutations is to temperature sensitivity, indicating that many missense mutations are well tolerated at the standard growth temperature but become much more deleterious, often to the point of lethality, at a temperature only 5°C–10°C higher. This widespread anecdotal observation implies that macromolecular stability becomes increasingly dependent on structural integrity as temperatures rise, a reasonable conjecture in keeping with the considerable constraints observed in the proteins of thermophilic microbes (e.g., [4]). It is therefore likely that the average missense mutation harms thermophiles more than mesophiles (the hypothesis of dangerous missense). This simple prediction was supported by the observation that missense mutations accumulated to a lesser extent (compared to synonymous mutations) in thermophiles than in mesophiles during the course of molecular evolution (dN/dS falling from 0.14 to 0.09), implying stronger purifying selection in thermophiles [5]. Here, direct measurements of the rate and character of spontaneous mutation are compared for mesophilic and thermophilic microbes.
Two Extrapolation Problems
The first phase of determining genomic mutation rates involves measuring a mutation frequency, converting the frequency to a rate, and taking precautions to exclude or take into account the impact of perturbations such as differential growth rates of mutants versus wild type and delayed expression of the mutant phenotype. In addition to measuring rates, it is crucial to identify the kinds of mutations that arise in order to exclude biases due to massive mutational hotspots or to bizarre classes of mutations. The typical result is a rate for a mutation-reporter gene, which is then extrapolated to the whole genome provided that the spectrum of mutations is fairly ordinary. However, there is a substantial problem here: while most indels are detected, most BPSs fail to produce a phenotypic change detectable in the laboratory. One must therefore estimate their full frequencies. (An exception is the still rare case that mutation detection is achieved with phenotype-blind genomic DNA sequencing.) Two methods have been applied. Both make the reasonable assumption that almost all indels and chain-termination (CT) BPSs are detected with high efficiency in protein-coding sequences. (Although exceptions occur, they are infrequent and tend to occur at the extreme downstream end of a gene.) The first method was based in part on the average relative frequencies of CT and non-CT BPSs in a handful of spectra and provided a correction factor for base substitutions of 4.726 [6]. This method was used for almost all of the entries in Table 1; however, the range of values averaging to 4.726 was large, reducing reliability. The second method is based exclusively on CT mutations. It involves examining the reporter sequence for all possible BPSs capable of generating CTs and then dividing the observed CT mutation frequency by that reduced target size and multiplying by 3 (to account for the three BPSs that can arise at any site) to obtain an average mutation rate per base pair. The CT method also has drawbacks. First, it cannot report A·T→G·C mutations, but these generally arise at approximately average BPS rates, suggesting a minimal problem. Second, CT mutations are typically a minority of all mutations, so that many spectra sport only a few CTs, reducing sampling accuracy.
The other major barrier to accurate extrapolation from a mutation-reporter gene to the whole genome becomes manifest when sequencing reveals a major hotspot. Mutation rates at particular sites vary greatly, but most mutational spectra display a range of site-specific numbers of mutations ranging from 1 to hotspots with from several percent to even a quarter of the whole collection. The impact of a hotspot containing a quarter of all the mutations is modest, but some genes contain single hotspots bearing the large majority of mutations; the classic example is the E. coli lacI gene, where ∼72% of all mutations are indels arising at a stretch of 13 BPSs consisting of 3.25 repeats of a tetramer [7]. However, such massive indel hotspots are infrequent among genes, and it is reasonable to post occasional genomic rates both including and removing them.
Thermophiles Versus Mesophiles
All informative microbial mutation rates obtained before 2000 were for mesophilic species, but rates and spectra are now available for two genes in each of two very different thermophiles, the crenarchaeon Sulfolobus acidocaldarius [6] and the bacterium Thermus thermophilus [8], both growing at close to 75°C. In the first study, with S. acidocaldarius, BPSs were a smaller fraction of the spectrum than in mesophiles, and this observation prompted the hypothesis of dangerous missense. Note, however, that if greater fractions of missense mutations are phenotypically detectable in thermophiles than in mesophiles, then the historical method of correcting for undetected BPSs becomes inappropriate when based on mesophiles. It is therefore advisable to resort exclusively to the CT method for estimating total BPS rates, which is the central result for this report.
Table 2 lists genomic mutation rates estimated using the CT method (or its lacZα equivalent), sometimes based on the same sources as for Table 1 but excluding some reports whose sequencing information was inadequate for the CT method. The nine entries at the top are for mesophiles and reveal no significant departures from the values in Table 1, providing empirical confidence in the robustness of the CT method. The two entries at the bottom are for thermophiles, whose numbers of CTs are small. (The data for the two mutation-reporter genes are combined in each organism because of the small number of CTs.) The thermophile BPS rates are substantially lower, by about 10-fold, than their mesophile counterparts. When major indel hotspots are included, indel rates are less than twofold lower in thermophiles, while total genomic rates are about fivefold lower. (When the indel hotspots are removed from the analysis, the indel rate decrease is three-fold and the total genomic rate decrease is seven-fold.) Although these ratios are somewhat uncertain because of the small numbers of CTs for five of the seven mesophiles and both thermophiles, the mean difference is large enough to support the inference that BPS rates are lower in thermophiles. The mesophile and thermophile values were compared using randomization t-tests [9], a nonparametric test that requires no assumptions about normality or equal variances of the mutation rates. The resulting one-sided p values are 0.018 for both the total mutation rate and its BPS component, and 0.27 for the indel values that include the hotspots.
The Central Result
Genomic mutation rates have long been suspected to evolve as a balance between the deleterious impact of the average mutation and the cost of further reducing the mutation rate. A test of this conjecture on the evolutionary scale could consist of estimating mutation rates in organisms whose environment increases the impact of the average mutation. Because many base substitutions do greater harm at higher temperatures, thermophiles were suitable candidate organisms. For both a bacterium and an archaeon, the thermophiles display sharply reduced rates of base pair substitutions compared to the typical mesophile.
The lower mutation rates in thermophiles are likely to reflect their higher optimal growth temperatures. There is no obvious hint of a particular aspect of life history other than temperature that sets the two thermophiles apart from the mesophiles. The %(G+C) values for the ten organisms in Tables 1 and/or 2, listed monotonically with the two thermophile values in bold, are 35–36–37–38–41–50–50–51–68–69, providing no hint of a role for this variable, as also noted in the earlier molecular-evolution study [5]. Thus, the Kimura conjecture, that the equilibrium mutation rate reflects a balance between the impact of the average mutation compared to the cost of keeping mutations in check, is supported in a natural experiment.
The hypothesis of dangerous missense predicts that BPS rates will be reduced in thermophiles but does not speak directly to indel rates. However, indel rates are also reduced, although less strongly than are BPS rates and with a p value of 0.27 for these data. One candidate explanation for this difference is that the reduction in BPS rates is achieved by the accumulation of modifiers selected to target BPS mutagenesis but at most incidentally targeting indel mutagenesis. Because single-base additions and deletions tend to be the large majority of indels in mesophiles (35 single-base indels/38 total indels in phage λ, 20/23 in phage T4, 45/45 in Herpes simplex virus, 604/641 in E. coli, 88/97 in S. cerevisiae, and 24/32 in S. pombe) and are similarly frequent in thermophiles (84/95 in S. acidocaldarius and 46/54 in T. thermophilus), these small indels must be the main targets of antimutagenic modifiers acting on indels generally. Both single-base indels and BPSs result from errors of insertion followed by failures of proofreading and DNA mismatch repair in well studied model organisms such as E. coli and S. cerevisiae, but little is known about the sources of spontaneous mutations in S. acidocaldarius and T. thermophilus.
New Fishing Holes
Are there likely to be other outliers with informative deviations from the mutational pattern that is consistently displayed among the mesophilic microbes examined to date with respect to either the mutation rate or the BPS:indel ratio?
Mutations to cold sensitivity are rarely reported and are anecdotally described as difficult to discover. If they are indeed rare, perhaps fewer missense mutations produce mutant phenotypes in psychrophiles than in mesophiles. One evolutionary consequence might then be a relaxation to a higher spontaneous rate of BPS mutation, perhaps with little effect on the rate of indel mutation.
Because of incomplete buffering against the impacts of their environments, halophiles and acidophiles experience relative high internal concentrations of Na+ and H+, respectively, compared to other microbes. These ionic environments might be unusually stressful to mutants carrying missense mutations, resulting in adjustments to their mutational patterns in the same direction as seen for thermophiles. Although without significance because of sampling constraints, Table 2 attributes a five-fold lower BPS mutation rate to the acidophile S. acidocaldarius than to the non-acidophile T. thermophilus. Unfortunately, an attempt to characterize mutation in the halophilic archaeon Haloferax volcanii failed, probably because this mesophile is highly polyploid [10].
The lactic acid bacterium Oenococcus oeni, used in wine making to convert malic acid to lactic acid, lacks the usual bacterial DNA mismatch repair (MMR) system and has a high mutation rate as judged by mutations conferring resistance to rifampin and erythromycin, as does Oenococcus kitaharae [11]. These results suggest a powerful genus-wide mutator condition, which would normally be highly deleterious. The question then arises whether the lack of MMR is so strongly adaptive in these species as to outweigh the sharply decreased fitness of the mutator condition, or whether the species have been unable to re-acquire the MMR genes by horizontal transfer.
Whereas the above two species lack MMR function and display mutator phenotypes, the crenarchaeons as a whole, including S. acidocaldarius, lack all known bacterial MMR genes, but S. acidocaldarius, at least, displays an antimutator phenotype compared with mesophiles. How can this be? In Escherichia coli, the mutation rate per average base pair ≈8×10−10 (Tables 1 and 2). Based on the strengths of mutator mutations, replication infidelity can be estimated as the product of three components during DNA replication: insertion errors ≈0.9×10−5, proofreading failures ≈1.7×10−2, and MMR failures ≈5×10−3 [12],[13]. In bacteriophage T4, which does not employ a general MMR system, the mutation rate per average base pair ≈2×10−8 (Tables 1 and 2). Based on the strengths of mutator mutations, replication fidelity can be estimated as the product of two components during DNA replication: insertion errors ≈1×10−5 and proofreading failures ≈2×10−3 [13]. Thus, T4 makes up for the lack of MMR by a proofreading potency about an order of magnitude greater than that operating in E. coli. The mutation rate per base pair for S. acidocaldarius ≈3×10−10, which might be achieved by a product of factors applied to the T4 insertion and proofreading accuracies that together produce a 70-fold improvement. Alternatively, S. acidocaldarius may possess an MMR system so distinct from the standard mutHLS model as to have escaped recognition by genomic scans. Note also that both thermophiles have genomes about twofold smaller than the E. coli genome.
General Procedures
We begin in possession of values for the following:
G = the genome size in bases or base pairs.
T = the number of bases or base pairs in the target (the mutation-reporter sequence).
μT = the measured mutation rate at T, corrected where necessary for mutants expressing the characteristic phenotype but revealed by sequencing to lack mutations in the reporter gene, but not corrected for mutants with two or more mutations (which are infrequent and sometimes absent). In many cases, μT = f/ln(μTN) where f = the measured mutation frequency for the given target, N = the final population size, and the median μT over several cultures is used [14], a method that is robust compared to the classical fluctuation test provided the average number of mutational events per culture is ≥30 [15].
M = number of sequenced mutants = B+I, where B = number of BPS mutants and I = number of indel mutants, the latter also including complex mutants (a minority, if present at all) regardless of their components.
For the "historical" method, we correct for undetected BPSs by multiplying the number of detected BPSs by 4.726 [6]. Then the average mutation rate per base or base pair μb = [μT corrected upwards by (I+4.726B)/M]/T = (I+4.726B)( μT/MT). The genomic mutation rate μg = Gμb.
For the "CT" method, the indel genomic mutation rate μg(I) is calculated as above ignoring the BPS component, B becomes BCT = number of mutations to a chain-terminating codon (TAG, TGA, or TAA), and P = number of possible mutational pathways to a CT mutation within T (there being three mutational BPS pathways per base or base pair). Then the BPS genomic mutation rate μg(B) = μT (3BCT/MP)G. The total genomic rate μg = μg(I)+μg(B).
Phage M13.
G = 6407. This system is unique among popular mutation reporters. It consists of an E. coli lacZα transgene embedded in the single-stranded DNA of the M13 genome and carrying both an upstream regulatory region and the beginning of the lacZ gene. Because thousands of mutants have been sequenced, it has become apparent which mutations are detectable when present singly and which are not [14],[16]. The target sizes for base substitutions (TB = 245) and for single-base indels (T±1 = 177) are thus well defined, and we further assume that the infrequent larger indels are fully detectable (TL = 239). The measured mutation frequency f was 5.86×10−4 [17], M = 117, B = 67, I±1 = 11 and IL = 39. Assuming that virtually all replication occurs by a rolling circle mechanism, the mutation rate is calculated as for RNA viruses, μ = f/2c where c is the number of consecutive cycles of infection [18]. The following protocol was used to grow the stock ([17] and T. A. Kunkel, personal communication). The contents of one plaque (≥1013 pfu) were added to l L of medium containing E. coli cells diluted from an overnight culture to about 107 cells/ml, so that the multiplicity of infection was about 103. The input of infected cells from the plaque was ≤108, so that the input concentration of infected cells = 108/103/ml = 105/ml, that is, no more than 105/107 = 0.01 of all cells. c≈2.5 in the plaque +1 in the liquid culture = 3.5. Then μb = (f/2c)Σ(proportion of mutations of type i = Ni/117)(1/Ti) = (5.86×10−4/7)[(3×67/117)(1/245)+(11/117)(1/177)+(39/117)(1/239)] = 7.48×10−7. μg(B) = (f/2c)(3×proportion of BPSs)(G/TB) = (5.86×10−4/7)(3×67/117)(6407/245) = 0.00376, μg(I+L) = (f/2c)[(proportion of ±1 indels)(1/T±1)+(proportion of larger indels)(1/TL)](G) = (5.86×10−4/7)[(11/117)(1/177)+(39/117)(1/239)](6407) = 0.00103, and μg = μg(B)+μg(I+L) = 0.00479.
Phage λ.
G = 4.850×104. For the cII gene, T = 294, f = 5.36×10−5, and, for 93 mutants, B = 55 (BCT = 8 and P = 35) and I = 38 ([19] and J. Wagner, personal communication). Then μT = f/ln(μT N) = 6.10×10−6. Using the historical method, μb = (6.10×10−6)(38+55×4.726)(1/93)(1/294) = 6.64×10−8 and μg = 0.00304. μg(I) = (6.10×10−6)(38/93)(4.85×104/294) = 0.00041. Using the CT method, μg(B) = (6.10×10−6)(8/93)(3×4.85×104/35) = 0.00218 and μg = μg(I)+μg(B) = 0.00259.
Herpes simplex virus type 1.
G = 1.523×105. For the tk gene, T = 1131, f = 6×10−5, N = (0.3–20)×108 and, for 67 mutants, B = 22 (BCT = 5 and P = 90) and I = 45 [20]. Replication proceeds by a mixture of exponential and linear replication, for which we use μT = f/ln(μTN) and μT = f/2c (with c = 2), respectively. The corresponding values are μT = 8.40×10−6 and 10×10−6, giving a mean of 9.20×10−6. Using the historical method, μb = (9.20×10−6)(45+22×4.726)(1/67)(1/1131) = 1.81×10−8 and μg = 0.00275. μg(I) = (9.20×10−6)(45/67)(1.52×105/1131) = 0.00083. Using the CT method, μg(B) = (9.20×10−6)(5/67)(3×1.52×105/90) = 0.00348 and μg = μg(I)+μg(B) = 0.00432.
Phage T2.
See [15].
Phage T4.
G = 1.689×105. For the rI gene, T = 294, μT = 2.82×10−6 and, for 66 mutants, B = 34 (BCT = 6 and P = 43) and I = 32 [21]. Using the historical method, μb = (2.82×10−6)(32+34×4.726)(1/66)(1/294) = 2.80×10−8 and μg = 0.00473. μg(I) = (2.82×10−6)(32/66)(1.689×105/294) = 0.00079. Using the CT method, μg(B) = (2.82×10−6)(6/66)(3×1.69×105/43) = 0.00302 and μg = μg(I)+μg(B) = 0.00381.
Thermus thermophilus.
G = 2.127×106. For the pyrEF genes, T = 1326, μT = 3.21×10−7 and, for 73 mutants, B = 19 (BCT = 2 and P = 103) and I = 54 (or 18 without the hotspot) [8]. The historical method is inappropriate for thermophiles (see text). μg(I) = (3.21×10−7)(54/73)(2.127×106/1326) = 0.000381. Using the CT method, μg(B) = (3.21×10−7)(2/73)(3×2.127×106/103) = 0.000545. μg = μg(I)+μg(B) = 0.000926 (or 0.000672 without the indel hotspot).
Sulfolobus acidocaldarius.
G = 2.226×106. For the pyrEF genes, T = 1240, μT = 3.37×10−7 and, for 108 mutants, B = 13 (BCT = 1 and P = 184) and I = 95 (or 46 without the hotspot) [6]. The historical method is inappropriate for thermophiles (see text). μg(I) = (3.37×10−7)(95/108)(2.226×106/1240) = 0.000532. Using the CT method, μg(B) = (3.37×10−7)(1/108)(3×2.226×106/184) = 0.000113. μg = μg(I)+μg(B) = 0.000645 (or 0.000371 without the indel hotspot).
Escherichia coli.
G = 4.639×106. For the lacI gene, μT = 6.043×10−7 (excluding 10 IS insertions) [15]. T = 1083 and, for 721 mutants, B = 80 (BCT = 24 and P = 110) and I = 641 (or 116 without the indel hotspot) [7]. Using the historical method, μb = (6.043×10−7)(641+80×4.726)(1/721)(1/1083) = 7.89×10−10 and μg = 0.00366. μg(I) = (6.043×10−7)(641/721)(4.639×106/1083) = 0.00230. Using the CT method, μg(B) = (6.043×10−7)(24/721)(3×4.639×106/110) = 0.00255 and μg = μg(I)+μg(B) = 0.00485 (or 0.002961 without the indel hotspot).
Saccharomyces cerevisiae.
G = 1.246×107 and calculations are as above.
For the URA3 gene, T = 804, P = 123, and four sets of values are available. For the first [22], μT = 2.77×10−8 and, for 106 mutants, B = 89 (BCT = 39) and I = 17; for the historical method, μb = 1.42×10−10 and μg = 0.00177; μg(I) = 0.00007; for the CT method, μg(B) = 0.00310; and μg = 0.00317. For the second [23], μT = 6.25×10−8 and, for 20 mutants, B = 15 (BCT = 4) and I = 5; for the historical method, μb = 2.95×10−10 and μg = 0.00368; μg(I) = 0.00024; for the CT method, μg(B) = 0.00380; and μg = 0.00404. For the third [23], μT = 3.50×10−8 and, for 106 mutants, B = 89 (BCT = 39) and I = 17; for the historical method, μb = 1.56×10−10 and μg = 0.00195; μg(I) = 0.00017; for the CT method, μg(B) = 0.00022; and μg = 0.00038. For the fourth [24], μT = 4.75×10−8 and, for 106 mutants, B = 89 (BCT = 39) and I = 17; for the historical method, μb = 2.37×10−10 and μg = 0.00295; μg(I) = 0.00014; for the CT method, μg(B) = 0.00446; and μg = 0.00460. The respective averages are, for the historical method, μb = 2.08×10−10 and μg = 0.00259; μg(I) = 0.00015; and, for the CT method, μg(B) = 0.00289 and μg = 0.00305.
For the CAN1 gene, T = 1773, P = 226, and three sets of values are available. For the first [25], μT = 2.77×10−7 and, for 20 mutants, B = 11 (BCT = 1) and I = 9; for the historical method, μb = 5.18×10−10 and μg = 0.00645; μg(I) = 0.00095; for the CT method, μg(B) = 0.00249; and μg = 0.00344. For the second [26], μT = 3.01×10−7 and, for 23 mutants, B = 17 (BCT = 5) and I = 6; for the historical method, μb = 4.13×10−10 and μg = 0.00514; μg(I) = 0.00036; for the CT method, μg(B) = 0.00701; and μg = 0.00737. For the third [24], μT = 1.52×10−7 and, for 227 mutants, B = 150 (BCT = 70) and I = 77 (including 13 complex mutations); for the historical method, μb = 2.97×10−10 and μg = 0.00370; μg(I) = 0.00036; for the CT method, μg(B) = 0.00775; and μg = 0.00811. The respective averages are, for the historical method, μb = 4.09×10−10 and μg = 0.00510; μg(I) = 0.00056; and, for the CT method, μg(B) = 0.00575 and μg = 0.00631.
The averages of the four URA3 plus three CAN1 values (sum÷7) are: for the historical method, μb = 2.94×10−10 and μg = 0.00366; μg(I) = 0.00033; and, for the CT method, μg(B) = 0.00412 and μg = 0.00444.
Schizosaccharomyces pombe.
G = 1.252×107, values are from [27] and S. Davey (personal communication) and calculations are as above. For the ura4 gene, T = 795, μT = 4.56×10−8 and, for 39 mutants, B = 22 (BCT = 5, P = 116) and I = 17; for the historical method, μb = 1.78×10−10 and μg = 0.00223; μg(I) = 0.00031; for the CT method, μg(B) = 0.00189; and μg = 0.00221. For the ura5 gene, T = 648, μT = 8.44×10−8 and, for 49 mutants, B = 34 (BCT = 5, P = 96) and I = 15; for the historical method, μb = 4.67×10−10 and μg = 0.00585; μg(I) = 0.00050; for the CT method, μg(B) = 0.00337; and μg = 0.00387. The average values for the two genes are: for the historical method, μb = 3.23×10−10 and μg = 0.00404; μg(I) = 0.00041; for the CT method, μg(B) = 0.00263; and μg = 0.00304.
Neurospora crassa:.
G = 3.804×107. Using the old mutation data (see [15]), for ad-3AB, μb = 4.11×10−11 and μg = 0.00172. For mtr, μb = 9.15×10−11 and μg = 0.00383. The average values are μb = 6.63×10−11 and μg = 0.00278.
I thank Grace Kissling for the statistical analysis, and Marilyn Diaz, Dennis Grogan, Libertad García Villada, Tom Kunkel, Susan Rosenberg, and Roel Schaaper for their helpful suggestions regarding intermediate versions of this article.
Author Contributions
Conceived and designed the experiments: JWD. Analyzed the data: JWD. Wrote the paper: JWD.
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- 11. Marcobal AM, Sela DA, Wolf YI, Markarova KS, Mills DS (2007) Role of hypermutability in the evolution of the genus Oenococcus. J Bacteriol 190: 564–570.
- 12. Schaaper RM (1993) Base selection, proofreading, and mismatch repair during DNA replication in Escherichia coli. J Biol Chem 268: 23762–23765.
- 13. Bebenek A, Dressman HK, Carver GT, Ng S, Petrov V, et al. (2001) Interacting fidelity defects in the replicative DNA polymerase of bacteriophage RB69. J Biol Chem 276: 10387–10397.
- 14. Drake JW (1991) A constant rate of spontaneous mutation in DNA-based microbes. Proc Natl Acad Sci U S A 88: 7160–7164.
- 15. Foster PL (2006) Methods for determining spontaneous mutation rates. Methods Enzymol 409: 195–214.
- 16. Bebenek K, Kunkel TA (1995) Analyzing fidelity of DNA polymerases. Methods Enzymol 262: 217–232.
- 17. Kunkel TA (1985) The mutational specificity of DNA polymerase-β during in vitro DNA synthesis. J Biol Chem 260: 5787–5796.
- 18. Drake JW, Holland JJ (1999) Mutation rates among lytic RNA viruses. Proc Natl Acad Sci U S A 96: 13910–13913.
- 19. Wagner J, Nohmi T (2000) Escherichia coli DNA polymerase IV mutator activity: genetic requirements and mutational specificity. J Bacteriol 182: 4587–4595.
- 20. Drake JW, Hwang CBC (2005) On the mutation rate of herpes simplex virus type 1. Genetics 170: 969–970.
- 21. Schultz GE Jr, Drake JW (2008) Templated mutagenesis in bacteriophage T4 involving imperfect direct or indirect sequence repeats. Genetics 178: 661–673.
- 22. Lee GS-F, Savage EA, Ritzel RG, von Borstel RC (1988) The base-alteration spectrum of spontaneous and ultraviolet radiation-induced forward mutations in the URA3 locus of Saccharomyces cerevisiae. Mol Gen Genet 214: 396–404.
- 23. Pursell ZF, Isoz I, Lundström E-B, Johansson E, Kunkel TA (2007) Yeast DNA polymerase ε participates in leading-strand DNA replication. Science 317: 127–130.
- 24. Lang GI, Murray AW (2008) Estimating the per-base-pair mutation rate in the yeast, Saccharomyces cerevisiae. Genetics 178: 67–82.
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- 26. Venkatesan RN, Hsu JJ, Lawrence NA, Preston BD, Loeb LA (2006) Mutator phenotypes caused by substitution at a conserved motif A in eukaryotic DNA polymerase δ. J Biol Chem 281: 4486–4494.
- 27. Fraser JLA, Neill E, Davey S (2003) Fission yeast Uve1 and Apn2 function in distinct oxidative damage repair pathways in vivo. DNA Repair 2: 1253–1267.


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subset: commoncrawl


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The annual Bloomin' Tulips Festival is Waratah-Wynyard's annual celebration of everything that is colourful, beautiful and charming about not only the tulips that adorn Table Cape, but also our municipality.
The event - the major event within the three-week calendar of events will be the Bloomin' Tulips Festival - a one day event to be held on Saturday 12 October 2019.
The Bloomin' Tulips Foreshore Market is an integral part of the festival and is held on the first and third Sunday of each month from 8.30am until 2pm.
For more information on any of these events contact the Waratah-Wynyard Council on 6443 8320.
How we got here - In 1991 the Wynyard Tourism and Promotion Association ran the first Tulip Festival in Wynyard, offering a weekend of entertainment, stalls and attractions to promote the area and attract visitors to our town and the North West Coast.From the outset, Wynyard was labeled by news reporters as "Tulip Town" and this has become somewhat of a continuing connection.
By 1997 the Wynyard Tulip Festival had formed a committee in its own right and had taken on an incorporated status, which in 2005 a Special Committee of the Waratah-Wynyard Council. The Festival is now coordinated by a Tulip Festival Project Officer who is employed on a part-time basis by the Waratah-Wynyard Council.
Bloomin' Tulips! is a festival of sense stimulating events celebrating the flush of spring colour in the small coastal town of Wynyard on the north west coast of Tasmania. The event is themed Colours of Wynyard to celebrate the spring flowering of the tulips at Van Diemen Quality Bulb Farm on Table Cape.
During Bloomin' Tulips you will marvel at the wonderland created by tulips in full bloom along our scenic coast. The Bloomin' Tulips Festival itself will offer you the chance to enjoy local art, craft, and music to suit all tastes; experience some magic at our at the spectacular fireworks over the Inglis River on Saturday evening; sample the culinary delights of our fresh island produce or just become a local during your brief time with us!
Volunteers form an integral part of the Festival's success and a huge thank you must go to the Wynyard Fire Brigade, Wynyard Lions Club, and all the people that roll up their sleeves every year.
For more information on how to become a volunteer click here.
The major event within the three week calendar of events will be the Bloomin' Tulips Festival - a one day event to be held on SATURDAY 12 OCTOBER 2019.
The Bloomin' Tulips Colour Run is also now part of the festival and will be held on Friday 11 October 2019. Colour Run Entry Forms Link
The Bloomin' Tulips Foreshore Market is an integral part of the festival and is held on the first and third Sunday of each month from 8.30am until 2pm.
The program for the 2019 Bloomin' Tulips Festival is available via the following link.
Keep an eye on this website as well as our Facebook page for more information on this year's program.


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Archive for November, 2008
Apparently Buying Five Billion Dollars Worth Of Arms From Israel Since 2002 Made India A Terror Target!
Posted by Job on November 29, 2008
Posted in Christianity | Tagged: Chabad House, hostage, India, Islam, Israel, Mumbai, Muslim, terrorism, war on terror | 15 Comments »
Thanksgiving For The Sovereignty of God
Posted by Job on November 28, 2008
Posted in Christianity | Leave a Comment »
Birthright Taglit Screening Out Messianic Jews
Posted by Job on November 28, 2008
Posted in Christianity | Tagged: birthright, Israel, Judaism, Messianic Judaism, taglit | Leave a Comment »
The Meaning Of The Song Of Solomon For Christians
Posted by Job on November 26, 2008
Want to know the meaning of the Song of Solomon for Christians? Well here is part 1 of a teaching from Messianic Perspectives, a Messianic Jewish Ministry, that teaches what this book means for Christians!
Posted in Bible, Christianity, Jesus Christ | Tagged: jesus in the old testament, Judaism, Messianic Judaism, Old Testament, shepherd, shulamite, shulamite girl, Song of Solomon, typology | 1 Comment »
Horrid Hindu Atrocities Against Christians Challenge India's Pluralism
Posted by Job on November 24, 2008
Posted in Jesus Christ | Tagged: Christian Persecution, extremism, Hinduism, India, Orissa, terrorism | Leave a Comment »
God Helps Those Who Cannot Help Themselves
Posted by Job on November 24, 2008
Posted in Bible, Christianity, Jesus Christ | Tagged: evangelism, gospel, salvation, video | Leave a Comment »
Recession May Force Some Christian Colleges To Close
Posted by Job on November 24, 2008
Posted in Christianity | Tagged: Christian colleges, education, recession | 2 Comments »
Hindu Extremists Paying To Kill Christians In Orissa India
Posted by Job on November 24, 2008
Please read: Being paid to kill Christians
Posted in Christianity | Tagged: Christian Persecution, Hinduism, India, Orissa | Leave a Comment »
Endtimes Alert: Indonesian AIDS patients face microchip monitoring
Posted by Job on November 24, 2008
Indonesian AIDS patients face microchip monitoring
By NINIEK KARMINI
JAKARTA, Indonesia (AP) — Lawmakers in Indonesia's remote province of Papua have thrown their support behind a controversial bill requiring some HIV/AIDS patients to be implanted with microchips — part of extreme efforts to monitor the disease.
Local health workers and AIDS activists called the plan "abhorrent."
"People with AIDS aren't animals; we have to respect their rights," said Tahi Ganyang Butarbutar, a prominent Papuan activist.
But legislator John Manangsang said by implanting small computer chips beneath the skin of "sexually aggressive" patients, authorities would be in a better position to identify, track and ultimately punish those who deliberately infect others with up to six months in jail or a $5,000 fine.
The technical and practical details still need to be hammered out, but if the proposed legislation gets a majority vote as expected, it will be enacted next month, he and others said.
Indonesia is the world's fourth most populous country and has one of Asia's fastest growing HIV rates, with up to 290,000 infections out of 235 million people, fueled mainly by intravenous drug users and prostitution.
But Papua, the country's easternmost and poorest province with a population of about 2 million, has been hardest hit. Its case rate of almost 61 per 100,000 is 15 times the national average, according to internationally funded research, which blames lack of knowledge about sexually transmitted diseases.
"The health situation is extraordinary, so we have to take extraordinary action," said another lawmaker, Weynand Watari, who envisions radio frequency identification tags like those used to track everything from cattle to luggage.
A committee would be created to decide who should be fitted with chips and to monitor patients' behavior, but it remains unclear who would be on it and how they would carry out their work, lawmakers said Monday.
Since the plan was initially proposed, the government has narrowed its scope, saying the chips would only be implanted in those who are "sexually aggressive," but it has not said how it would determine who fits that group. It also was not clear how many people it might include.
Nancy Fee, the UNAIDS country coordinator, said the global body was not aware of any laws or initiatives elsewhere involving HIV/AIDS patients and microchips.
Though she has yet to see a copy of the bill, she said she had "grave concerns" about the effect it would have on human rights and public health.
"No one should be subject to unlawful or unnecessary interference of privacy," Fee said, adding that while other countries have been known to be oppressive in trying to tackle AIDS, such policies don't work.
They make people afraid and push the problem further underground, she said.
Tahi Ganyang, the Papuan activist, said the best way to tackle the epidemic was through increased spending on sexual education and condom use.
Associated Press Writer Irwan Firdaus contributed to this report.
Posted in Christianity | Tagged: AIDS, endtimes, eschatology, Indonesia, prophecy | 1 Comment »
Examples of Scientific Accuracy In The Bible
Posted by Job on November 23, 2008
Posted in Bible, Christianity | Tagged: Apologetics | Leave a Comment »
THE GOSPEL WORTHY OF ALL ACCEPTATION by Andrew Fuller
Posted by Job on November 23, 2008
The Gospel Worthy of All Acceptation
Part I
By Andrew Fuller, 1786
The Gospel Worthy of All Acceptation
Part II
By Andrew Fuller, 1786
The Gospel Worthy of All Acceptation
Part III
By Andrew Fuller, 1786
The Gospel Worthy of All Acceptation
Concluding Reflections
By Andrew Fuller, 1786
The Gospel Worthy of All Acceptation
By Andrew Fuller, 1786
Posted in Christianity | Leave a Comment »
What Genesis 1 Tells Christians About Evangelism
Posted by Job on November 19, 2008
Please direct your attention to the account of creation in Genesis 1. After you have done so, please notice what it says about God's creations of living things: plants, animals and humans. Five times, in Genesis 1:11, Genesis 1:12, Genesis 1:21, Genesis 1:24, Genesis 1:25 and it is mentioned that after God created living things, He gave living things the ability to reproduce after their kind (using the King James Version manner of speaking). That means that every living thing that God made has the ability to bear fruit, to produce offspring that is similar to its parents, to replicate itself. This applies not only to the plant and animal kingdoms or to human beings in natural terms. There is a spiritual principle here too. After all, who can deny that the church is a living thing? The church is the Body of Christ. Jesus Christ is alive, not dead. Upon His resurrection, the angels at His tomb bore witness of this. They stated "why do you seek the living among the dead?" Jesus Christ before He went to the cross told this to the Saduccees who challenged Him about the resurrection: that God and the Kingdom of heaven have rule and dominion over the living, not the dead. Those in the Body of Christ will never truly die, but will have life everlasting. Meanwhile, those outside the Body of Christ will suffer not only a physical natural death, but the second death in the lake of fire. Just as the Body of Christ is most certainly alive, then its members, individual Christians, are alive too. Not only are we alive, but we are new creatures! So yes, Genesis 1:11, Genesis 1:12, Genesis 1:21, Genesis 1:24, Genesis 1:25 speaks to us. As living beings that are new creations of God in Our Lord Jesus Christ, we will also reproduce after our kind. That is, we will heed the command of our Lord and Savior Jesus Christ and bear good fruit by fulfilling the Great Commission, testifying of the Lord Jesus Christ and His great salvation to any and all that will hear, and produce disciples that will enter the Body of Christ just as we did. After all, how did you enter the Body of Christ. Perhaps your parents took you to church and you heard a sermon. Perhaps a family member or a coworker shared the gospel with you. Maybe you heard the gospel on radio or television. Or perhaps it was a stranger who took a minute of your time that changed you for an eternity, and maybe he or she even gave you a tract. No matter the method, these people were fulfilling the mandate – not option but mandate – that Jesus Christ gave His true followers to spread the gospel to any and all. And this mandate was not some new thing, but rather a restatement and application to the church what was already given in a place like Genesis 1:26-28, which told all living creation including humankind to multiply, reproduce and fill the earth. To the church in particular, Genesis 1:26-28 is tied to Matthew 28:19-20, the Great Commission! As living things, as living men, we are able to fulfill this mandate through evangelism. Even those of us who are not able to bear natural children due to God's not willing us to bear them for one reason or another will produce much fruit for the Holy and righteous God in evangelism, and in this way fulfill the prophesy given in Isaiah 54:1. (By the way, amillennial, dominion, and official theologists please take note of Matthew 28:17-18, which states that the dominion of the earth, all things, has been given to CHRIST JESUS. So the great commission does not refer to politics, economics and culture, only evangelism.) So this is yet another way that the new covenant is a greater spiritual fulfillment of the lesser natural covenants that preceded it, and how the old covenant prophets bore witness that just such a thing would happen. So Christian, set aside all of your worldly distractions that Satan and his workers have erected before you to hinder you from accomplishing good works that glorify the Father in the Name of His Son Jesus Christ and with the empowering of the Holy Spirit, and concentrate on the task at hand, which is spreading the gospel of Jesus Christ among our family, neighbors, cities, nations, and to the ends of the earth. Heed the command of your Savior and your Lord Jesus Christ. Go and reproduce living things after your kind which God the Father will then conform into the image of His Son.
Posted in Christianity, false religion, religious right | Tagged: creation, Genesis 1 | 3 Comments »
A Christian Haiku: Jesus Please Help Me
Posted by Job on November 15, 2008
Submitted by a ministry supporter.
Jesus please help me.
I feel as if I am running
out of time
Spreading your message
has made me visible to the
I have gone too far now to ask your
Put me back to where I was
before I got your sign
A prophet you have called
to go anywhere
Only with the call,
the demonic forces were made
Follow me as I follow Christ is the message I am to ssppread
Even if it means your life
Nothing about it is dread
Posted in Christianity | Tagged: haiku, poetry, prayer | 2 Comments »


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Hey!! What's the story?
What stories/books/poems/novels have you recently completed or had published?
I'll go first:
^This is a trailer for my new book!
Views: 169
Scary trailer - I didn't even know a book could have a trailer. I will look out for your book.
Thanks! Book trailers aren't super common, but with the internet age we see them a little more these days. On rare occasions I have seen book trailers on TV. But they're usually just like: "Anne Rice's new book has been called horrific and sexy. In stores this October." Not too much imagery.
This isn't new, but I really liked your trailer so here's one of mine for a humorous short story:
The inmates are revolting, and it's not going to be pretty in this tension-filled escapade that will surprise you. Edge-of-your-seat action with a bit of humor thrown in, and an ending that can't be missed.
One of my short horror stories "Thrill of the Chase" is included in the "Once Bitten" anthology edited by Steve Lewis due to be published by Knight Watch Press on April 11th 2015.
This is the arachnophobe's nightmare of a book cover! Great artwork from Stephen Cooney.


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Jenna Lyon's Home (Creative Director of JCrew)
I am currently in bed and don't intend to leave my house after an ambitious day yesterday. My "Path" went something like this: Palo Alto to Potrero, a relaxing massage and spa day with the girls at Mizu, lunch at Source (this place is SO good for vegan/vegetarian food), finally got to see Cirque Du Solei's Totem by the ballpark (the acts are insane! Definitely recommend to go see this show!) Rushed back to Liz's for champagne and got dressed for the Sara Bareilles concert at Bill Graham--she is one of my favorite artists of all time. Such a great personality and good voice! After that we headed down a few blocks to eat at the Boxing Room since it was open till 1am. I haven't had a back to back day like that in so long, I'm beat.


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MAGISTRATES have granted extra time for detectives to quiz five men in connection with a near #2 million robbery.
The men, aged 21 to 31 and all from Coventry, West Midlands, are being questioned in relation to the raid at the NatWest cash compound in Tamworth, Staffordshire, on October 18.
They were arrested in dawn raids on Tuesday by Staffordshire Police on suspicion of conspiracy to rob and police now have until 4.30am tomorrow to question them.
The robbery involved a skip lorry being driven through security gates and smashing into the side of the complex.
It was driven by masked men wearing high-visibility jackets.
A #50,000 reward has been offered for information leading to the conviction of the offenders who struck at the compound on the city's Amington industrial estate at around 10pm.


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Registered charity number 1140291
Hands On London, a registered charity with the Charity Commission, was founded in 2010. We are the local affiliate of the global umbrella organisation, Hands on Network (HON), which is headquartered and registered in the United States. Hands on London (HOL) strives to be the first point of contact for individuals in London who want to be involved in community service.
Hands On London offers a variety of events to participate in and raise valuable donations to support the charity. Check out the events below for more information.
Tel: 020 7424 5520
How to advertise | Webmaster | Site Map | Contact Us | Privacy & Cookie Policy | All Charities Strategic Partners | Fundraising Advice | Work For Us | Fitness Advice | Kit Advice
Copyright Skyline Promotions Ltd 2019 All Rights Reserved


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Christine Gregoire Reflects on Her Accomplishments as Governor of Washington
"Chinese companies will find it easy do business in Washington with the dedicated support of local and state government."
~Christine Gregoire, outgoing governor of the state of Washington
Christine Gregoire served as Washington State's 22nd governor from 2005 through 2012. Born Christine O'Grady, she was raised in Auburn, Washington by her mother, who worked as a short-order cook to support the family. She rode horses, picked blueberries and learned the value of hard work and a good education.
After graduating from Auburn High School, she entered the University of Washington. She graduated with a teaching certificate and Bachelor of Arts degree in speech and sociology. She met her husband, Mike Gregoire, in September 1971 when they both worked for the Washington Department of Social and Health Services. She received her law degree in 1977 from Gonzaga University.
Chris and Mike Gregoire have two daughters, Courtney and Michelle, whose achievements are both equally distinguished. Courtney is married to Scott Lindsay, and is an attorney who has served in the administrations of two US presidents and currently works at Microsoft. Michelle currently attends law school.
Governor Gregoire describes herself as striving to deliver real results to the people of Washington. She combines effective leadership with a tireless work ethic. As she leaves office, she takes time to reflect on her many accomplishments. In this article, she talks about Washington's trade relationship with China.
Q: Governor Gregoire, please share some highlights of what you've done to build trade between Washington and China during your term in office.
A: Washington has a distinguished history of trade and cultural exchange with China and I am proud to have had the opportunity to maintain and enrich those ties. During my term in office I've worked on a number of projects and initiatives that have not only helped to strengthen our existing relationships in China but also to build new ones.
Chief among those is the Washington Export Initiative, launched in 2010 to increase the number of Washington state companies exporting by 30 percent and help 5,000 Washington businesses achieve $600 million in new export sales.
As a prime US gateway to Asia, opportunities for Washington companies in China and the entire region are a focus. During a 2010 trade mission, we met with many Washington companies that have success stories in China, including some in clean energy and information communication technology. We participated in a number of meetings and signed MOU agreements to further education, research, trade and investment, particularly in agriculture, biomedical and life sciences.
We participated in a Washington Day at the World Expo in Shanghai and showcased our tourism opportunities. More than $10 million in sales is attributed to that 2010 trade mission. That number is expected to grow as business opportunities seeded on that trip come to fruition.
I had the opportunity to return again for the National Governor's Association last year, where I led the dialogue between US governors and the leaders from China's provinces.
While these individual missions help promote trade between Washington and China, our efforts are also supported by the ongoing work of our Departments of Commerce and Agriculture, and the countless efforts of other organizations and friends of the State who have made Washington's valued relationship with China what it is today.
Q: Where do you predict these efforts will lead?
A: Washington companies such as Starbucks, Microsoft, Boeing and Almond Roca are already household names across China. We believe that our efforts will continue to expand this list.
Companies like PopCap, a creator of games for mobile devices, have had tremendous success in the Chinese market and their story began with one of our earlier trade missions. Companies like Impinj, an RFID manufacturer, used Washington State Department of Commerce services for their distributor search. That assistance led to rapid sales growth and helped them establish of a growing sales force in the market.
On the 2010 mission, Demand Energy, a global provider of distributed energy systems, met and followed up with Chinese partners to plan for installation of their systems in China. These are just a few examples of how our efforts have led to successes in China and we have many more to come.
Q: What is the China trade potential for technology products and services?
A: Current policies in China stress innovation to help lead development. Washington companies can benefit from this demand if they adopt a prudent approach. Our companies are innovation leaders, especially those involved in information technology, clean energy technology, and life sciences.
Washington companies like Enprecis, a provider of feedback analysis tools in the automotive industry, have used Federal STEP (State Trade and Export Promotion) grant funding to take advantage of our China Accelerator program for a low-risk entry to the Chinese market. Over the past year, they've been able to secure deals and pilot programs with major Chinese automotive companies and are now planning to further expand their current operations.
There are many success stories like these, but the key is developing an effective China strategy to deal with concerns such as intellectual property rights and local protectionism. Our state government works closely with federal officials and with our companies to assist in this process.
Q: What advice might you give to those in China about conducting business successfully with Washington State?
A: I would encourage Chinese business leaders to look to our state as one of the leading centers for entrepreneurship and innovation in the US. We also have had a number of Chinese companies, ranging from Mindray and adSage to iSoftStone, that have chosen to invest in Washington and take advantage of our talented workforce and forward-looking businesses environment.
We also have a thriving overseas Chinese community and can provide numerous connections for our friends in China who want to establish a presence or conduct business in our state.
Chinese companies will find it easy do business here with the dedicated support of local and state government. If Chinese companies or entrepreneurs are looking to do business in the US, the State's Department of Commerce can provide them with the tools they need to succeed. Washington also has offices in China to facilitate trade and exchange between Chinese and Washington State companies, and the people in those offices can provide advice about doing business successfully here.
For More Information
Washington State Governor's Official Website at http://www.governor.wa.gov
State of Washington Department of Commerce http://www.commerce.wa.gov/Pages/default.aspx


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The next one involves finally getting on with my daughter's room redo. It is turning out great and I will be sure to share tidbits with you until the room is completed...well, as much as rooms get completed ;)
So here's some tidbits and I'll be sharing the final projects soon!
So how about you guys...what projects have you been wrapping up?


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"Training is needed in order to love properly; and to be able to give happiness and joy, you must practice DEEP LOOKING directed toward the other person you love. Because if you do not understand this person, you cannot love properly. Understanding is the essence of love. If you cannot understand, you cannot love. That is the message of the Buddha."
~ Thich Nhat Hanh


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After 3 babies and quite a few years of sleep deprivation I have done a few things since becoming a mummy that I'm not proud of, and I feel it is time to come clean. Please don't judge me, I'm not a bad person really.
- I have used my children to get out of things I have been invited to.
- I sometimes watch my partner sleeping peacefully while I have been up all night with the baby and debate whether punching him in the face is a good idea.
- Some days I have cried more than the baby.
- I have bribed my children with biscuits, sweets, chocolate, in return for their good behaviour. Mummy 0 - Children 1.
- Some days I don't get dressed until it is almost dinner time, and on those days I mean have a quick shower and put on new pjs.
- When a mummy friend tells me her baby is sleeping through the night, I mentally punch her in the face.
- Running out of coffee reduces me to tears.
- I have often told the children that mummy has a headache because I dont want to hear the recorder being played for the umpteenth time.
- I have skipped pages in a bedtime story just so that the children could go to bed abit earlier.
- Have picked sleep over nookie, Terrible, Terrible wife I hear you say, it's not that I don't love my partner, but parenting is exhausting work at times!
- Running out of wine reduces me to tears.
- I have taken the batteries out of a noisy toy and have claimed it was broken.
- I would love some Me Time, I have forgotten the meaning of this term. At the end of the day my needs are simple, having a shower, going to the toilet or finishing that hot coffee *ahem* Wine *ahem* in peace.
- Everyone tells you to enjoy the time you have with your kids while they are still young, but sometimes I am just so tired and worn out, enjoying them just feels like more work!
- Most days I feel like I have no idea what I'm doing, everyone thinks I have it all together, good wife, good mother, but I really don't. I'm ready to stop pretending to be perfect now.
I may moan about my children, they drive me mad most days, they keep me up all night, but I bloody love my children, I love being a stay at home mum and I wouldn't change that for the world.


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MedPharm Ltd has announced the expansion of its US Center of Excellence in Durham, North Carolina in response to increasing service demand for its topical and transdermal formulation development and performance testing services.
An investment of $4M in facility expansion and equipment will more than triple the existing footprint of the current US facility to 25,000 sq/feet. In parallel, the company has substantially increased its LC/MS capacity and the automation of sample handling.
The expansion creates additional laboratory space for MedPharm to de-risk its customers' development programs by carrying out expanded topical dosage formulation development services and innovative performance testing on topical and transdermal pharmaceutical products. Recently published FDA and EMEA guidance's on demonstrating generic equivalence for topical medicines, as well as the growing interest in developing new topical drugs, is fueling this demand.
"This expansion aligns with MedPharm's strategy to keep increasing the sophistication and relevance of our proprietary ex vivo human skin models and further increase our responsiveness to clients in formulation development, especially in ocular delivery," commented Eugene Ciolfi, MedPharm's President & CEO. He added, "The expansion complements the recent expansion we have completed in our facility at our headquarters in Guildford, UK and reflects the positive demand for our services globally."
As part of the investment, MedPharm has installed the latest Waters LC-MS/MS to manage the increasing demand for bio-analysis. This new model is equipped with the latest UPLC for rapid separation and method development and offers mass spectrometry detection at picogram levels.
"The FDA has set stringent validation criteria for any performance test if they are being used to substantiate generic equivalence", said Professor Marc Brown MedPharm's co-founder and Chief Scientific Officer. "Customers are recognising that MedPharm has the depth of experience to achieve positive outcomes from these complex experiments and has the strategic knowledge to support successful regulatory submissions of their topical and transdermal products."
For more information on MedPharm and its services, please visit: www.medpharm.com
About MedPharm
MedPharm Ltd. is a leading, global provider of contract topical and transdermal product design and formulation development services. MedPharm are experts at reducing risk and accelerating development times for generic and proprietary pharmaceutical customers through the use of proprietary, industry-leading performance testing models. Well-established as a global leader in dermatology, nail, mucosal membrane, and transdermal product development, MedPharm also offers innovative solutions for ophthalmic and airway preparations. These solutions are recognised for their scientific rigor by regulators and investors. MedPharm has fully established R&D centres in the US and UK and GMP clinical manufacturing at its global headquarters facility in Guildford, UK. For more information visit our website www.medpharm.com


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William J. Simione, Jr.
FHHC, Founding Member
The Simione team provides superior guidance, helping our clients achieve their business goals. Our full-time staff of committed industry veterans includes home care and hospice executives, registered nurses, accountants, information system specialists, certified valuation analysts, and experts in sales and marketing, compliance and Medicare reimbursement. With decades of experience with major government agencies and contractors that support healthcare initiatives, our nationally known industry leaders providing frequent presentations to state and national groups. The Simione team also serves on the committees and task forces that continue to shape the industry.


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WASHINGTON, D.C. April 28, 2010 – Michael J Feeney of the New York Daily News is this year's Emerging Journalist, selected by the National Association of Black
Journalists announced at its spring Board of Directors meeting. Feeney
will join other top honorees at the association's Salute to Excellence
Gala, July 31 in San Diego, during NABJ's 35th Annual Convention and
Career Fair, the largest gathering of minority journalists in the
"I'm honored and thrilled to receive such a prestigious award," said
Feeney. "I've been to these (NABJ) conferences since 2002, and I know
what these awards mean. This means a great deal to me."
Feeney joined The New York Daily News in 2009. His work on night
shift spans from crime to celebrity news. Prior to working with The New
York Daily News, Feeney was a breaking news reporter for The Record (of
Bergen County, N.J.). During his two-and-a-half years at the paper he
covered municipal government and police. He also crossed over into the
new- age of journalism and worked as a Web and multimedia journalist
for NorthJersey.com. Feeney started his career at the Associated Press
in Baltimore and Detroit. He attended the Knight Multimedia Journalism
workshop at the University of California, Berkeley.
"Michael represents where journalism is and where it's going. He has
covered multiple beats in print and lives in a multimedia world. His
fresh energy and eagerness to learn are the perfect ingredients for an
emerging journalist," said NABJ President Kathy Y. Times.
Feeney is a graduate of Delaware State University. He also runs his
own pop culture blog. Feeney credits the help of NABJ with much of the
success in his career and the valuable mentors and friendships he has
gained over the years.
"I remember Michael as a young student from Delaware State and a
NABJ newbie, he was hungry and wanted to succeed," said NABJ Treasurer
Gregory Lee Jr. "He is the success story that we hope for in each of
our students. He has obviously carried with him everything that he
learned early on from his NABJ family and capitalized on it."
The NABJ Board of Directors isn't the only group recognizing Feeney
for his work. His peers describe him as "a tremendously hard worker who
is an aggressive journalist."
The Salute to Excellence Awards Gala recognizes journalism that best
covered the black experience or addressed issues affecting the
worldwide black community during 2009. Feeney will be joined by other
top honorees, CNN's Soledad O'Brien for Journalist of the Year, and
NABJ Founder Paul Delaney for Lifetime Achievement.
NABJ's 35th Annual Convention and Career Fair will take place July
28-August 1 in San Diego, Calif. For additional information, ticket
sales, registration, please visit us at www.nabj.org, or contact Ryan Williams at rwilliams@nabj. org or (301) 405-0248.
Remember Me
9/12/2019Brian Jones Departs FOX Business Network
9/1/2019NABJ Advocacy: Addressing Diversity and Sensitivity Issues in the Media
8/29/2019NABJ President Dorothy Tucker Addresses KOCO-TV Anchor Comparing Black Co-Anchor to Gorilla
10/15/2019NABJ Chapter Grant Deadline- Oct. 15, 2019
12/31/2019NABJ Graduation Stoles Order
NABJ Journal
Summer 2018
Scripps Journalism
Career Program


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Creating Healthy, Happy, Sanitised Homes and Offices!
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We use the latest products that allow Carpets, Upholstery and Rugs to be cleaned as never before-"Removing Deep Down Soiling, Grime and Various Soft Stains.
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Wednesday, November 6, 2013
Biju Janata Dal seeks President\'s intervention over delay in cyclone assistance
Bhubaneswar: The ruling Biju Janata Dal (BJD) in Odisha yesterday sought the intervention of President Pranab Mukherjee over delay in getting financial assistance from the central government for the Cyclone Phailin-ravaged state.
A delegation of the party, comprising several ministers and legislators, met state Governor SC Jamir at Raj Bhavan in state capital Bhubaneswar, and submitted a memorandum to the president through him.
\"While the state government, on its part, has done its best to protect the life and properties of the people from this natural calamity, the central government has not provided any assistance, though duty-bound in our federal system,\" the party stated in the memorandum.
Soon after Cyclone Phailin struck the state on October 12, Chief Minister Naveen Patnaik requested the prime minister to sanction at least Rs. 1,000 crore advance, out of the National Disaster Response Fund (NDRF). Not a rupee has yet been sanctioned, the party alleged.
The United Progressive Alliance (UPA) government is forgetting that Odisha is a state within the union of India. The government of India is constitutionally duty-bound to come to the rescue of the people, the BJD delegation said.
Unless the central assistance is immediately sanctioned, the state will not be able to take up reconstruction and restoration of the damage caused by the cyclone, and subsequent floods, the party said.
In its memorandum, the party requested the president to intervene and direct the central government to immediately sanction the required amount.
The severe tropical Cyclone Phailin in the Bay of Bengal, which made landfall on October 12 night near Gopalpur in state\'s Ganjam district, killed at least 50 and left behind a trail of destruction in several districts of Odisha, bringing heavy rain and causing floods.
News From: http://www.7StarNews.com
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Updates to Future Projects are listed in Caboose News. Click on link to the left. Call or email to place your reservations!
Looking for models to sell on Consignment!! (Nothing under $250 retail value.) Consign fee is 25% of selling price. Call or Email today!
Phone calls Monday - Friday 9 am to 4 pm Eastern time. All Visits are by appointment only. Hours are subject to change without notice.
The Caboose no longer accepts American Express.
We do not take credit card information through our web site. Nor do we store it in our database. Credit card payments must be called in.
The Caboose, LLC
5 Mohawk Drive
Wolcott, CT 06716
United States
Phone: 203-879-9797
Fax: 203-879-4878
Monday - Friday
8am - 5pm EST


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Page 1: Introduction
Everyone knows about Cadbury and what it represents. The Cadbury Masterbrand is the flagship for a variety of well known distinctive products including Flake, Dairy Milk, Crunchie and Roses. Each of these products has its own place in the public imagination because we have grown up with them and they have associations for us with events in our personal histories.
Since 1990, Cadbury has developed and implemented an expanding programme of presence marketing as an effective way to promote the Cadbury Masterbrand in the UK. The strategy has been to select high profile sites in theme parks, shopping malls and airports to communicate Cadbury values and increase the availability of products.
This case study examines the creation of a new and exciting brand - Café Cadbury - which shows how detailed thought has been applied to making this a successful venture which extends the Cadbury reputation by providing a high profile presence and by giving consumers even more reasons to choose Cadbury.
The pilot Café Cadbury was opened on 8th October 2000 in an attractive three-storey 18th century Georgian building in the centre of Bath, a busy tourist destination and World Heritage City. The location was carefully chosen to position the new brand as a premium experience for the discerning customer. Café Cadbury is an exciting all day premium café and gift experience also offering a take-out service on the ground floor.


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Mystery, BABYLON the Great...
Almost four thousand years ago, a couple hundred years beyond the flood of
Noah, there was an attempt by people to unify their world into a "one world"
government, as well as religion. This first, great, unification of mankind
marked the synchronization of their political and religious thought into one
conglomerate - one ideology. This conglomerate became the foundation for
the establishment of the famous Tower of Babel, also known as Babylon.
The founders of this "one world" system were a couple of characters in the
Bible: Cush and Nimrod. This tower became mankind's symbol of vanity - a
declaration of independence from any authority of God. Cush and Nimrod
also began to push for control over all the proceedings. Utilizing the religious
and political thoughts they helped establish, a system began; a system which
would continue to pass on from one government to the next.
Despite attempts by Godly people to destroy this control and influence, the
system would continue on, often in the shadows of everyday life. It even, in
rare occasions, had to go undercover - the reason was that, at one time, the
entire system of Babylon was almost destroyed (as we'll see).
The physical tower, even the city of Babylon, would crumble over time,
however. Their system of thought and control would continue, slowly
manifesting into what the Bible calls the Mystery form of Babylon. This
Mystery form would be assimilated into a variety of religious, political and
cultural beliefs many of us know and follow.
Throughout time, up to today, there were people in positions of authority
and influence who wanted a unification of the world in thought and deed. If
"history repeats itself," as it often does, then understanding what went on in
our history is truly the key to diagnosing what might be going on in our
world as we speak. The lifestyles of Noah's day truly appears to be
re-surfacing again, possibly through the descendants of the very people who
once practiced it, long ago!


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A Free Multidisciplinary Arts Event located at the Stream Trail of Redwood Regional Park, Oakland, California
Art in Nature is a full day interdisciplinary, interactive, site-specific, multi-sensory, inter-cultural, and inter-generational immersion into art, nature, and the nature of art.
Stroll along the mile-long Stream Trail, experience art in nature, and delve into the nature of art throughout 12 theme areas featuring music, dance, sculpture, painting, martial arts, poetry, body painting, circus arts, theater, visual arts, storytelling, arts & crafts, kids activities and more.
Experience an immersion into art, nature and the beauty of creating. Take a walk, enjoy nature, explore your own creativity and see a variety performers and artists in action!


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Article by Judith Schrut, email Judith
As well as those eagerly awaited royal nuptials, Spring 2018 marks the 72nd anniversary of Prime Minister Winston Churchill's 1946 'special relationship' speech, in which he publicly bravo-ed the close US-UK cultural and historical ties. Although that relationship has been celebrated, tested, challenged and changed over the decades since, there is no doubting the many strong and enduring American Air Museum, Duxford, copyright IWM British-American connections.
Whether you have an academic or more personal interest in World War II US-UK links, the American Air Museum near Cambridge is a highly recommended must-see. With striking, award-winning architecture by Norman Foster, the Museum houses the largest collection of American aircraft outside the USA and is a fitting tribute to the 30,000 Americans who died after flying from UK air bases during the war.
You can also pay a visit to the nearby Cambridge American Cemetery and Memorial, where thousands of American military personnel are buried or honoured. This includes over 5,000 names engraved in stone on the Walls of the Missing. And it's always worth a detour to Cambridge itself, where we recommend a pint at The Eagle, the city's oldest pub and steeped in historic charm. Be sure to check out the pub's unique RAF Bar, where American and British fighter pilots socialised before setting off on their dangerous missions. Although many never did return, their names and moving messages are etched forever into the ceiling with Zippo lighters, candles and lipstick.
There are lots of other memorials to noted Americans dotted around the UK, including Pocahontas, Tom Paine and Whistler. You'll also find statues of FDR and Ike in Grosvenor Square, Abraham Lincoln in Parliament Square, George Washington outside the National Gallery, and Martin Luther King as one of the 20th century Christian martyrs carved above the doorway of Westminster Abbey. Virginia Quay Settlers Monument in Blackwall, East London, remembers the 105 "adventurers"– men, women and children - who sailed from here in 1606 in three small ships and went on to found Jamestown, the first English colony in North America.
Further information:
www.abmc.gov/cemeteries-memorials/ europe/cambridge-americancemetery


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DANIELSVILLE – Billy McGilvary, 79, died Tuesday, May 18, 2010 at his residence.
Born in Barbour County, Ala., on January 26, 1931, he was the son of the late William Huey McGilvary and Ethel Agnes Land McGilvary. He was retired from the United States Army where he served tours of duty in the Korean Conflict and the Vietnam War. Mr. McGilvary was a member of the Bluestone Baptist Church.
Survivors include his wife, Bobbie Scroggins McGilvary, Danielsville; daughters, Joan Cross, Angie Brown, both of Danielsville and Kathy Widner, Phenix City, Ala.; brother, Fred McGilvary, Clayton, Ala.; sister, Dorothy Baker, Clayton, Ala.; seven grandchildren and 15 great-grandchildren.
Funeral services will be held at 2 p.m., Thursday, May 20, at the Pruitt Funeral Home Chapel with the Revs. George Lovley, Talmadge Ethridge and Ray Jenkins officiating. Graveside services will follow Friday, May 21, at 2 p.m. at the Lakeview Memory Gardens in Phenix City, Ala., with full military honors.
The body is at the funeral home where the family will receive friends Thursday, May 20, from 12 p.m. until the time of service. The family is at the home.
In lieu of flowers, memorial contributions may be made to the American Lung Association, 1301 Pennsylvania Ave., NW, Washington, D.C. 20004.
Pruitt Funeral Home, Royston, is in charge of arrangements.
Billy McGilvary
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The Democratic Society Congress (DTK) is a platform of Kurdish associations and movements that has proclaimed democratic self-government. The 850 delegates of the DTK have first met on july 14, 2011 in Amed, the capital of North Kurdistan, while operations, arrests and attacks were going on in all of the Kurdish region and the so-called "KCK" operations had already started with the arrestation of thousands of politicians, activists, party members, civil society association members.
The Democratic Autonomy model was developed as a status so to sensitize the government towards democracy and find a solution to the Kurdish problem and as a response to the fact that the kurds were not recognised any social status and didn't not want to live without a status anymore. The DTK called on the international community to recognize that right:
"In the light of international conventions on human, respect for the territorial integrity of a common land and the prospect of a democratic nation, we, the Kurdish people, pledge our Democratic Autonomy, as well as national commitment to unity of the peoples of Turkey" says the final declaration. The congress has implemented a plan and program for short, medium and long terms, articulated around 8 fields: Political, Legal, Self-Defense, Social, Economical, Cultural, Ecological and Diplomatical.
Indicating that democratic self-government is the natural system of all communities, the movement stresses that this autonomy is not intended to destroy a state and build another.
For the DTK, autonomy is not a democratic state system, but a system in which the Kurdish people could govern themselves.
"We are announcing our power and will to govern ourselves. Democratic autonomy is not for the Kurdish people alone, but is rather a model for a solution that will allow all of Turkey's peoples to freely express their beliefs and culture."
In this context, it was stated in the final document that:
"Democratic Autonomy, which is a unity in diversity, never means to build up a power against the power and the state. It means an organization of the society out of the state against being socialized and nationalized, in other words it means less state and more society. In this sense, democratic autonomy is in a process of a continuous development and construction just like a living organism. With a method of bilateral law, it designs and secures the state which will block its free development and legal structure".
To express the situation once again in a very clear and open way: '
'We have made the declaration of our social democratic solution model in the face of the statalist mentality and structures that impose a deadlock on all issues, on the Kurdish problem in particular. We project the autonomy for the Kurdish people and all peoples".
This decision of the Kurdish people and their the most legitimate and comprehensive enactment has initiated a new process and invited everyone to show its true colors in this process.
In addition, works will be continued to build and institutionalize the democratic autonomy regardless of the approaches and responses to this declaration of intention. "The struggle will continue to include this solution in the new constitution to be prepared."


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Bangalore – NOVEMBER 23, 2016 – Kenscio was recognized as one among 93 businesses to receive '2016 HOT In Bangalore' Award conferred by Owler. Owler recognizes the top trending companies in cities around the world. They filtered through more than 15 million companies and picked 4,500 award winners across 600 cities worldwide. The selection was done from Owler's community in Bangalore that features 5,100+ registered Bangalore companies.
Recipients were chosen based on several different metrics, including number of followers on Owler, insights collected from our community, social media followers, and blog posts over the past year.
"We've sorted through database of millions of contributions from our community and landed on the top trending companies from around the world," said Jim Fowler, CEO at Owler. "Being Hot In 2016 is an accomplishment to be proud of."
To see Kenscio's company profile on Owler, go to https://www.owler.com/iaApp/1576630/kenscio-company-profile
To see the official release from Owler, go to https://www.owler.com/reports/kenscio/kenscio-wins-owler-s-hot-in-bangalore-award/1479794760284
To read about the other winners, go to http://blog.owler.com/hot-in-bangalore/
About Owler: Owler is the crowdsourced competitive intelligence platform that business professionals use to outsmart their competition, gain competitive insights, and uncover the latest industry news and alerts. Owler is powered by an active community of 800K business professionals that contribute unique business insights such as competitors, private company revenue, and CEO ratings.
Launched in 2014, and funded by Norwest Venture Partners and Trinity Ventures, Owler is headquartered in San Mateo, CA with offices in Coimbatore, India. To know more, visit www.owler.com


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International Funds - Investing overseas without leaving home
To offset the risk of the Australian economy running flat, you can get international exposure quite easily through managed funds. Here's a couple worth a further look.
Platinum Capital(PMC)
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Platinum has a low exposure to Asia and a strategy of picking undervalued stocks with upside potential. It too invests with a long-term outlook in mind, but short-term volatility can cause underperformance when compared to the benchmark index.
Like BT Global, the buy-back program will be beneficial for investors by narrowing the discount to NTA. If the shares trade at an average weighted discount of more than 15% to NTA on the last business day of each of the 12 calendar...
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Platinum Capital rights issueQ.
PLATINUM CAPITAL LTD (PMC) Received the Rights Issue Offer Documents today at $1.55. Market is $1.595 and I'm not sure how much the offer will "water down" the issued capital. In other words, is it a deal? Can we have your thoughts...Read full Q&A +


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This post's title is a giveaway, isn't it? By reading the title alone you know what this is all about, but I'm telling you just the same. With the start of a new month, I decided to start something new on this blog as well. While many of you may have seen similar posts from other blogs, I give you my own version of it. Everyone, welcome to Life in Squares, a sneak peek of what's happening in this mundane life of mine through, well, everyone's favorite social media app, that is Instagram. If you haven't yet, you can follow me along here and let's be friends.
Life in Squares: Vol. 1
Tuesday, February 2, 2016
Posted by Karla ♥ at 15:42
Labels: Life, life in squares
Life is full of surprises. Why not allow it to surprise you?
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text: KIRKUS REVIEW Leonard returns to the Florida coastline for his weakest novel since Touch (1987)--a bumpily humorous but unfocused seriofarce about a probation officer and the eccentric judge she gets entangled with. As one of Leonard's very few heroines in 29 novels, spunky Kathy Baker of the Florida Dept. of Corrections blows a whiff of fresh air into the Leonard canon--as does the outspoken, aging hanging-judge Bob Gibbs--but not enough to put the spring into a slack plot that begins when skirt-chasing Gibbs takes a fancy to Kathy as she shows up in his courtroom with probation-violator Dale Crowe Junior. Gibbs throws the book at Dale, then asks former psychology-major Kathy out on a date under the guise of her talking to his wife, a former showgirl who seems to be possessed by the spirit of a 12-year-old 18th-century slave girl. Before Kathy can visit Gibbs, however, a hugh alligator appears on his property and sends his wife scurrying for northern climes. And that's just fine by Gibbs, who turns out to have imported the gator to get rid of his loony wife. But when Gibbs double-deals Dickey Campau, who brought the gator, Campau drives out to the judge's home and shoots up the house. Which is just as well, because the shots scare off Elvin Crowe, Dale's mean and flaky uncle, who's been hired by another of the judge's irate courtroom-victims, a crack-addicted M.D., to kill the judge. Caught up in the investigation into the gator-attack and shooting, Kathy matches up professionally and romantically with cool cop Gary Hammond--until a jarring note of raw violence takes out Gary and sets Kathy up for an anticlimactic confrontation with Elvin and the M.D. Nicely realized characters, the usual smart Leonard dialogue, a few moments of brisk high/low humor--but the meandering plot lacks drive, Gibbs rolls around like a loose wheel, and the whole affair seems more like a pale Carl Hiaasen imitation than true-blue Leonard: It's all a big disappointment after Leonard's crackling last, Get Shorty (1990). Pub Date: Aug. 1st, 1991ISBN: 0-385-30142-1Page count: 304ppPublisher: DelacorteReview Posted Online: May 20th, 2010Kirkus Reviews Issue: May 1st, 1991


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I've frequently discussed the unseen victims of tariffs imposed on imports into the United States. Often these new taxes on US consumers are imposed in the name of "fair trade." This new Cato Institute video sums up the problems surrounding those tariffs in a great new video that's so clear and simple that even a Congressman can understand it. (Well, maybe....)
Cato's Dan Ikenson discusses the video and some of Cato's research on US trade remedies laws here. Yesterday, he warned us of a particularly egregious decision over wood flooring from China, and unfortunately, his fears were confirmed today. (Higher prices at Lowes and Home Depot are sure to follow. Whoopee.)


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How to Pick a Password
I am a firm believer in using strong, hard to crack passwords. No sir, nobody's ever going to guess my passwords! The only way someone's going to get access to my bank account is to beat it out of me.
Wow, I probably shouldn't have said that.
You can comment on this comic on Facebook.
As always, thanks for using my Amazon Affiliate links (US, UK, Canada).


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Fans of the original and connoisseurs of isometric sci-fi base builder management sims rejoice! We have just released Space Colony: Steam Edition, our remaster of the original game that adds new missions, Steam Workshop support and several other cool Steam features. Developed and self-published by the folks here at Firefly, the new Steam Edition is now available at a special discounted 'launch week' price.
The new HD patch for Space Colony is available to download from various websites, hit Read More for a list of links to download the patch.
All Media
[External]Space Colony HD Official Site
System Requirements
[External]Space Colony [Take 2]
[External]Space Colony Fans
Contact Us
For a quick answer you can tweet your question to @fireflyworlds anytime from 10am-6pm GMT, Monday to Friday.