text:
- Research
- Open Access
Pairwise and higher-order correlations among drug-resistance mutations in HIV-1 subtype B protease
- Omar Haq1,
- Ronald M Levy1, 2Email author,
- Alexandre V Morozov1, 3Email author and
- Michael Andrec1, 2
© Haq et al; licensee BioMed Central Ltd. 2009
- Published: 27 August 2009
The reaction of HIV protease to inhibitor therapy is characterized by the emergence of complex mutational patterns which confer drug resistance. The response of HIV protease to drugs often involves both primary mutations that directly inhibit the action of the drug, and a host of accessory resistance mutations that may occur far from the active site but may contribute to restoring the fitness or stability of the enzyme. Here we develop a probabilistic approach based on connected information that allows us to study residue, pair level and higher-order correlations within the same framework.
We apply our methodology to a database of approximately 13,000 sequences which have been annotated by the treatment history of the patients from which the samples were obtained. We show that including pair interactions is essential for agreement with the mutational data, since neglect of these interactions results in order-of-magnitude errors in the probabilities of the simultaneous occurence of many mutations. The magnitude of these pair correlations changes dramatically between sequences obtained from patients that were or were not exposed to drugs. Higher-order effects make a contribution of as much as 10% for residues taken three at a time, but increase to more than twice that for 10 to 15-residue groups. The sequence data is insufficient to determine the higher-order effects for larger groups. We find that higher-order interactions have a significant effect on the predicted frequencies of sequences with large numbers of mutations. While relatively rare, such sequences are more prevalent after multi-drug therapy. The relative importance of these higher-order interactions increases with the number of drugs the patient had been exposed to.
Correlations are critical for the understanding of mutation patterns in HIV protease. Pair interactions have substantial qualitative effects, while higher-order interactions are individually smaller but may have a collective effect. Together they lead to correlations which could have an important impact on the dynamics of the evolution of cross-resistance, by allowing the virus to pass through otherwise unlikely mutational states. These findings also indicate that pairwise and possibly higher-order effects should be included in the models of protein evolution, instead of assuming that all residues mutate independently of one another.
- Pair Correlation
- Connected Information
- Pair Level
- Maximum Entropy Distribution
- Accessory Position
The protease enzyme coded for by the pol gene of the Human Immunodeficiency Virus HIV-1 plays a critical role in the reproduction of the virus by cleaving the GAG precursor protein in a sequence-specific manner into its functional form, and as such, is a key target of several families of commonly used drugs used to control HIV infection [1]. Unfortunately, the virus has been able to evolve resistance to many of these drugs, in part due to the high mutation rates in the HIV genome [2]. The patterns of mutations in protease are complex, involving multiple key primary mutations that inhibit the action of drugs and a host of accessory mutations that can modulate the enzyme's stability or activity or otherwise enhance the fitness of the virus. It is now understood that these mutations do not occur independently of each other, but instead are correlated, resulting in complex patterns of co-evolving mutations [3–7].
Previous studies have mostly focused on correlations between mutations in the HIV protease gene at the pair level [3, 5–7]. However, recognition that the observed mutations may also be involved in higher-order interactions has led to a few studies in which correlated pairs of mutations are grouped using tools such as multidimensional scaling [3, 6], Bayesian networks [8], networks defined by patterns of conditional selection pressure [5], and clustering [9, 10]. The underlying assumption is that understanding higher-order interactions is important for a complete understanding of the evolution of resistance in HIV protease.
In this paper, we investigate correlations among HIV protease mutations at and beyond the pair level, and the impact of drug treatment on the nature of those correlations. We only consider the presence or absence of a non-synonymous mutation relative to a defined wild-type sequence, and not the precise base or amino acid substitution which has occurred. We develop a hierarchy of probabilistic log-linear models [11] that can in principle describe residue interactions of arbitrary order, and use those to analyze HIV protease sequence data obtained from patient cohorts with varying protease inhibitor (PI) treatment histories.
We use "connected information" [12] to quantify inter-residue interactions at the triplet and higher level. Unlike the Bayesian network approach [13], the information-theoretic methodology allows us to distinguish intrinsic three-body effects from the cases in which correlations between three random variables can be attributed mostly to pairwise interactions. The connected information viewpoint of higher order correlation has not been previously used in the analysis of mutational patterns in HIV protease, although it has been employed in a much more limited analysis of the V3 loop of the HIV envelope protein [14], and log-linear models have been used to study protein-protein interactions [15]. We find that pairwise interactions are necessary to achieve even qualitative agreement with the mutational data, while higher order interactions play an important role in predicting how frequently sequences with several mutations appear in the database. Simultaneous appearance of multiple mutations may play an important role in the phenomenon of multiple- or cross-resistance of the viral protease.
Increased mutation frequencies under drug exposure
It is of interest to ask which amino acid positions exhibit elevated mutation frequencies under drug treatment. Mutations at many of these positions are associated with decreased HIV-1 inhibitor susceptibility, and it is useful to classify mutations as belonging to "primary" vs "accessory" resistance classes. The terms "secondary" and "compensatory" have also been used as synonyms for "accessory". The specific criteria for such a classification are ad hoc in nature, but have generally been defined as follows.
Primary mutations are usually selected first in the presence of the drug and confer resistance, even when present as single point mutations [16, 17]. They can be structurally important, e.g. situated near the enzymatic active site, in which case their effect on inhibitor binding can be rationalized due to their physical proximity to the inhibitor [18]. In the case of protease, however, there are exceptions, as some mutations (such as positions 54, 76, 88 and 90) are situated far from the active site or have no direct contacts with the substrate, yet still reduce drug susceptibility [16]. The mechanism of action of these mutations is not clearly understood [19].
Accessory mutations confer resistance only when present with additional primary or accessory drug-resistance mutations and have little or no effect on inhibitor susceptibility on their own. Some of these mutations occur in the absence of drug treatment, but their frequency of occurrence is observed to increase in treated patients. Accessory mutations may "rescue" possible losses of activity or stability in the enzyme that may have been caused by a destabilizing primary resistance mutation, and therefore may have a compensatory function in restoring viral fitness [20].
We make use of a primary and accessory classification scheme based on the work of Shafer et al. [21–23, 17]. We define 17 primary drug resistance positions (residues 23, 24, 30, 32, 33, 46, 47, 48, 50, 53, 54, 73, 76, 82, 84, 88, 90) and 24 accessory drug resistance positions (residues 10, 11, 13, 20, 34, 35, 36, 43, 45, 55, 58, 60, 63, 71, 74, 75, 77, 79, 83, 85, 89, 91, 93, 95). The remaining positions are polymorphic mutations not associated with drug resistance or are conserved sites [24, 3, 25, 21, 26, 23].
Exhaustive analysis of residue pairs
Previous studies of pair correlations in protein families have indicated that coevolving pairs of residues tend to lie closer to each other in structure than random residue pairs [27–31]. In the case of the HIV protease, the distribution of ϕ values for drug-associated positions with distance shows a characteristic triangular shape, particularly for the PI2+ cohort (Figure 3B) [32]. In particular, distances associated with the most correlated residue pairs (30–88, 54–82, 32–47, and 37–77) are all within a few Ångstroms of each other. The triangular shape in Figure 3B is not surprising, given that most of the drug-associated positions are on the substrate cleft and thus tend to be relatively close to each other in space. As a result, 78 pairs (36% of the statistically significant pairs) are within 8 Å of each other. We also see no tendency for exposed residues to preferentially coevolve, in contrast to previous studies on mutation covariation in other protein families [27, 29, 15].
Of the 26 statistically significant negatively correlated pairs in the PI2+ cohort, 10 pairs involve either residues 30 or 88. Residue 30 is negatively correlated with positions 82, 10, 46, 90, 54, 73 and 84 (in decreasing order of the magnitude of the correlation), while position 88 is negatively correlated with positions 82, 73 and 54. It is interesting to note that position 63, which has a high mutation rate in both the PI0 and PI2+ cohorts, is negatively correlated with positions 80, 52, 5, 83, 64, and 61, of which only position 83 is associated with drug resistance [3]. In fact, of the 23 unique positions involved in the 26 negatively correlated pairs, most, but not all, are positions of drug resistance. It is possible that the 7 non-drug associated positions, 5, 15, 20, 52, 61, 64 and 80 play a role in the stability or function of the protein, even if they do not interfere with inhibitors [33–35]. Residue 80, in particular is negatively correlated with three residues, 63, 71 and 90, all of which play either primary or accessory roles in drug resistance.
Additionally, the types of residues involved in pair correlations changes upon treatment, with accessory positions becoming more prominent: 10% of the correlated pairs in the PI0 cohort consist of a primary and an accessory position, and that this combination increases to 31% in the PI2+ cohort. Furthermore, 45% of the pairs in the PI0 cohort involve at least one accessory position, which increases to 61% in the PI2+ cohort. As expected, pairs of non-resistance-associated positions decrease from 52% to 35%. It is interesting to note that the same trend is not observed for primary positions: 57% of pairs in the PI0 cohort contain at least one primary position, and this is essentially unchanged in the PI2+ cohort (59%). Therefore, drug treatment causes correlated pairs involving primary and non-resistance associated positions to be replaced by pairs involving primary and accessory positions.
Exhaustive analysis of residue triples
We examined the degree of connected information in both the PI0 and PI2+ cohort sequences. The connected information was calculated for all = 88, 560 residue triples in the HIV protease as described in the Methods section below. In the PI0 cohort sequences, only 175 residue triples have statistically significant three-body interaction at the p = 0.001 level, while the PI2+ cohort sequences have 6,300 significant triples. Furthermore, the significant triples from the PI2+ cohort are enriched in drug resistance associated positions: 32% of them consist of only drug associated positions, compared to 12% of the full set of 88,560. In contrast, the significant triples from the PI0 cohort show no such enrichment: only 9% consist solely of drug resistance associated positions.
The 10 triples in HIV protease from the PI2+ cohort sequences with the largest (A, B, C) values (all three-body interactions are significant with p ≪ 10-6).
residue triple
I multi (A, B, C)
Another way in which the magnitude of the effect of the three-body interactions can be visualized is by considering conditional probabilities for the 46-54-90: P(54 m |46 m 90 m ) = 0.4186, P(54 m |46 m 900) = 0.5508, P(54 m |46090 m ) = 0.4359, and P(54 m |460900) = 0.0280. All of these probabilities differ significantly from the independent estimate of P(54 m ) = 0.1178. We also see clear evidence of "triplet correlation" in the data, in the sense that the probability of a mutation depends very strongly on the state of both of the other residues, e.g. residue 54 is much less likely to be mutated if both 46 and 90 are wild-type than if only one of them is wild-type.
Much of this probabilistic dependency, however, can be accounted for by pair interactions. The corresponding probabilities for the best-fit two-body model are (54 m |46 m 90 m ) = 0.6286, (54 m |46 m 900) = 0.2236, (54 m |46090 m ) = 0.2316, and (54 m |460900) = 0.0488. Even though this model contains no three-body interactions, a qualitative "triplet correlation" of the type seen above remains (i.e. (54 m |46 m 900) ≈ (54 m |46090 m ) > (54 m |460900)), indicating that non-trivial three-way probabilistic dependencies can arise purely from pair correlations [37]. Overall, three-body interactions do quantitatively modulate the probabilities, but only to a small degree, since is an order of magnitude smaller than I multi even for the triples in Table 1. In other words, the contribution of three-body interactions to any of the triplet distributions that describe the mutational patterns of protease taken three at a time is roughly 10% or less of the effect induced by the pairwise interactions acting on these positions. Nonetheless, there can be substantial effects on a "micro level", such as the more than twofold difference in P(54 m |46 m 900) seen here.
Increased higher-order interactions in larger residue groups
It should be noted that since the univariate marginals are preserved by all of the models, the mean total number of mutations is the same for all 4 curves in Figure 11A. Therefore, under- or overestimation of the total number of mutations in one part of the distribution must be compensated by over- or under-estimation (respectively) in another, implying that the curves must cross one other. To quantify this effect, we can compare the predicted probabilities for seeing 5 or more mutations under each model and comparing to the observed probability. Those probabilities are 0.0372, 0.1197, and 0.1030 for the independent, pair-term model, and pair+three-body model, respectively, compared to the observed probability of 0.1007. The deviations of the first two from the observed are highly statistically significant, while the latter has a p-value of ≈ 0.06. The qualitative distribution is well accounted for by the pair interaction model, with the three-body and higher interactions modulating the details of the shape of the distribution, such as the increase in frequency of 3–5 and 13–15 mutations, at the expense of a decrease in the frequency of 6–10 mutations.
To see if the synergistic effects seen for the 10-residue group become even stronger for 15 residues, we repeated this analysis for the 15-residue group 10-20-33-36-46-54-55-63-71-73-74-82-84-90–93 (Figure 10), which was chosen by selecting the residues with the largest change in mutation frequencies upon PI treatment. For the PI2+ cohort, we find that = S(ind) - S(pair) = 1.1810, the sum of the remaining connected information measures of third order and higher is S(pair) - S(obs) = 0.8249 and I multi = S(ind) - S(obs) = 2.0059. The contribution of three-body and higher-order interactions now make up 41% of the total correlation information. However, this result may somewhat overestimate the true amount of higher-order correlation. A fit with a three-body model (Equation 12) gives an estimate of of 0.2638, and the ratio represents a lower bound on the contribution from correlations beyond the pair level. A more complete account of the many issues involved in estimating the amount of higher order correlation and its precision and accuracy for finite data sets will be the subject of a future communication. The comparison of the predicted and observed distributions of total number of mutations bears out this result (Figure 11B), showing more pronounced differences between the observed (black) and pair-model (red) distributions. In addition to the overestimation of the upper tail similar to that seen for the 10-residue group, we now also see that the pair model cannot reproduce the bimodal shape seen in the observed data. Again, the same 15-residue group for the PI0 cohort shows considerably weaker overall correlation (Figure 10).
Discussion and conclusion
Treating HIV protease with drugs results in the appearance of complex mutational patterns: observed mutations are not limited to the active site and often occur in groups that involve two or more residues. Furthermore, some mutations occur even in the absence of drugs, presumably following neutral rather than adaptive evolution. To study correlations between different residue positions in HIV protease, we have developed a hierarchy of models that allows us to include inter-residue correlations of arbitrary order within a consistent framework. Using only HIV protease sequences as input, we find that pair interactions become common and quite strong after PI treatment. In fact, it is often impossible to achieve even qualitative agreement with the data without including the two-body terms (Figures 9, 11, and Additional File 2). This finding calls into question a common assumption employed in current probabilistic approaches to phylogeny [38] that most residues evolve independently.
We have developed an information-theoretic method to study interactions between mutations beyond the pairwise level. Our approach is based on the notion of the connected information (Equation 2) [12]. While there are a variety of quantitatively different measures of pair correlation [11, 39] that may differ in their sensitivity in various regimes, they all measure essentially the same qualitative feature of the observed data. On the other hand, no single summary statistic can capture all of the various characteristics of higher-order behavior, leading to multiple descriptions that provide complimentary information. Connected information is one intuitive statistic that provides insight into the degree of structure in the data beyond the pair correlation level.
Connected information provides information which is complementary to Bayesian network analyses based on factorizations of the joint probability. It can readily verified that = 0 if at least one of the random variables is independent of the other two, or if the joint distribution involves conditional independence (e.g. P(A, B, C) = P(A)P(B|A)P(C|A)). However, a joint distribution with triplet-level probabilistic dependencies (in the sense that two of the variables are independent of each other, but the third depends jointly on the state of the other two, e.g. P(A, B, C) = P(A)P(B)P(C|A, B), or if P(A, B, C) cannot be factorized into any simpler form) could still be consistent with no three-body connected information if the observed triplet distribution is the maximum entropy distribution relative to its marginals. Thus, even if a Bayesian network-style analysis shows that a given triple cannot be factorized into any simpler form, that "triplet correlation" could still be consistent with a very small or zero , indicating that the observed behavior is dominated by two-body interactions. In fact, it has been shown that very complex correlation patterns among random variables can arise from large numbers of weak pairwise interactions [37].
Other information-theoretic measures of "higher-order correlation" have also been proposed, including higher-order mutual information, which measures "frustration" or the degree of synergy vs redundancy among several random variables [40]. While this measure has been used in the analysis of HIV envelope protein sequence data [41], its interpretation is considerably less intuitive. Similarly, ad hoc methods for finding putative clusters of mutually correlated residues [3, 9, 8, 10] cannot reliably uncover sets that have intrinsic higher order interactions, as defined by large (data not shown).
Plotting ϕ values for pairs of residues as a function of the distance between them (Figure 3B) reveals that while some large pair correlations arise from direct contacts between residues (e.g. ϕ ≈ 0.8, d < 5 Å), there are also strong correlations (ϕ ≈ 0.5) between amino acids separated by 15 Å or more, making physical coupling between them very unlikely. To provide an example of the former, we consider mutations involving residues 30 and 88. The closest distance between heavy atoms of residues 30 and 88 is just 3.66 Å, making likely some sort of physical interaction between them. Mutations at residue 30 are strongly and uniquely associated with resistance to the protease inhibitor nelfinavir, and there exists a strong correlation between mutations at positions 30 and 88 [42] which may be due in part to a compensation of the loss of a surface negative charge from the D30N mutation being restored by N88D [9].
It is possible that chains of intermediate interactions result in long-range coupling between two coevolving yet physically distant residues [43]. However, because non-zero values of λ ij indicate a presence of direct interactions between residues i and j in our model [15], we can decompose such "energetically connected pathways" into contributions from separate pairs. In contrast to a previous study [15], we find that non-zero values of λ ij are only weakly correlated with distance (Additional File 3). This lack of correlation is not entirely surprising, since even direct interactions between a pair of residues need not have a purely physical origin. Indeed, if protein fitness is a non-linear function of its stability or enzymatic activity [44], two mutations can be correlated because they compensate each other by making independent and opposite contributions to the overall fitness, even if there is no direct or indirect physical interaction between them [45]. By the same argument, the three-body terms also result from a mixture of physical and epistatic (compensatory) origins.
We have shown that three-body and higher-order correlations have the largest effect on the probabilities of the simultaneous occurrence of multiple mutations in the HIV protease (Figure 11). Since both this and previous studies have found that the total number of mutated positions is correlated with treatment by multiple protease inhibitors (Figure 1) [3], the presence of higher-order interactions may influence how protease reacts to multiple drugs, and could have an important impact on the evolution of cross-resistance, for example, by providing the virus with an "escape hatch" of large numbers of mutations. Higher-order interactions could also impact the time evolution of mutations by allowing the virus to pass through otherwise unlikely mutational states. We have seen that the impact of higher-order interactions in 10 to 15 residue clusters is at least a factor of two larger than the largest values for residue triples (approximately 20% or more of the total entropy change). One of the outstanding questions raised by this work is whether the impact of higher-order interactions for HIV evolving under the pressure of multiple drugs continues to become stronger for larger residue groups (ultimately the set of all 41 drug-associated positions). Unfortunately, there is not enough sequence data to perform such an analysis. Short of obtaining additional data, it may also be possible to explore this question by constructing synthetic data sets using λ i and λ ij values consistent with an observed distribution at the level of residue triples (i.e. Figure 7C).
The sequence-based approach presented here is not limited to the HIV protease and its response to drug treatment, and should be equally useful in studies of the evolution of drug resistance in other systems. Moreover, it will be of interest to extend our techniques to other examples of short-term neutral and adaptive evolution, including controlled evolution in the lab accompanied by protein sequencing at different timepoints. Recent work has suggested that evolutionary pathways of proteins are relatively restricted and may be predictable in general [46], and specific methods for predicting the mutational dynamics of HIV protease have been proposed, based on Bayesian network models [47] or pairwise conditional selection pressure [5]. A better understanding of the nature of the probabilistic dependencies underlying the network models should lead to improved prediction strategies. However, our model cannot distinguish between physical and epistatic origins of the observed co-evolution. To do this, we need a different approach which would explicitly introduce protein fitness as a function of residue energies (including interactions across protein-protein and protein-ligand interface). These energies would be fit against the sequence data, resulting in a prediction that decomposes observed inter-residue correlations into the physical and epistatic parts. This approach is currently being pursued in our laboratories.
HIV sequence database
Aligned and annotated HIV-1 protease amino acid sequences were obtained using the web interface of the Stanford HIV Drug Resistance Database http://hivdb.stanford.edu/cgi-bin/PI_Form.cgi[16]. The sequences are all classified under the HIV-1 Main group, subtype B. For the purposes of this paper, we are considering only the number of protease inhibitors and not the specific combinations of protease inhibitors used in the treatment. There are 13,608 sequences in this curated set, of which 8,229 sequences are drug naive, 2,677 sequence are associated with PI monotherapy, and the remaining 2,702 sequences are associated with between 2 through 6 protease inhibitors.
Calculation of higher-order interactions
leading to a cubic equation in α [11].
To obtain the maximum entropy distributions for more than three binary random variables, nonlinear optimization is unavoidable. However, instead of directly maximizing the entropy subject to the marginal probability constraints, we maximize the likelihood subject to the constraints that the λ's vanish beyond a given order [48]. In most cases, the latter will be more computationally efficient since the number of λ variables grows polynomially with the number of variables, while the dimensionality of the null space defined by the marginal probability constraints (which is one-dimensional for three variables) increases exponentially with the number of variables.
where λ is the vector of parameters, the indices (i, j) and (i, j, k) run over all distinct combinations of {A, B, C,...} with i ≠ j and i ≠ j ≠ k, respectively, and I, J, and K are numerical values of the corresponding random variable (we use 0 for wild-type and 1 for mutant). For an n-variate distribution P(A, B, C,...) there are n(n - 1)/2 pair parameters λ ij and n(n - 1)(n - 2)/6 three-body parameters λ ijk (λ0 is a normalization constant). The independent model was determined by forming products of the observed univariate marginals. The magnitudes of the λ i and λ ij parameters in the two-body model are related to the mutation frequencies at site i and pair correlations between sites i and j, respectively. In fact, the magnitudes of λ ij in the context of a two-body model have been proposed as a measure of "direct information", i.e. the part of pair correlation resulting from direct coupling [15]. It should be noted that the relative magnitudes of λ ij is dependent on the choice of the numerical values assigned to the random variables I, J and K in Equations 11 and 12. In this work, we assign values of 0 and 1 for wild-type and mutant, respectively, for computational convenience. It has been argued, however, that a more appropriate choice of numerical values is one which is symmetric about zero, e.g. ± 1, which allows "gauge constraints" to be introduced [15]. While this choice will affect the the values of λ ij and their interpretation as "direct information", it will not change the best-fit two-body or three-body probabilities and consequently will have no impact on the values of and related measures of higher-order interactions.
where i is one of the 2 n states, N i is the number of times that state was observed, and P(i|λ) is the predicted probability for state i to be observed given the vector of parameters λ. Maximization was performed numerically using the "nlm" function of the R software package [50]. All entropy and connected information values are given in natural log units. Statistical significance of the three-body interactions was estimated using the likelihood ratio test under the null hypothesis that the data were generated by (A, B, C) by Monte Carlo sampling. For all of the residue triples in Table 1, the p-values for the observed likelihood ratio were too small to be estimated (p ≪ 10-6), indicating very strong statistical significance for the three-body interaction.
This work has been supported by a grant from the National Institutes of Health, GM-30580 (to RML) and by an Alfred P Sloan Research Fellowship (to AVM).
This article has been published as part of BMC Bioinformatics Volume 10 Supplement 8, 2009: Proceedings of the European Conference on Computational Biology (ECCB) 2008 Workshop: Annotation, interpretation and management of mutations. The full contents of the supplement are available online at http://www.biomedcentral.com/bmcbioinformatics/10?issue=S8.
Authors' Affiliations
- Swanstrom R, Erona J: Human immunodeficiency virus type-1 protease inhibitors: Thereputic successes and failures, suppression and resistance. Pharmacol Ther 2000, 86: 145–170. 10.1016/S0163-7258(00)00037-1View ArticlePubMedGoogle Scholar
- Rambaut A, Posada D, Crandall KA, Holmes EC: The causes and consequences of HIV evolution. Nat Rev Genet 2004, 5: 52–61. 10.1038/nrg1246View ArticlePubMedGoogle Scholar
- Wu TD, Schiffer CA, Gonzales MJ, Taylor J, Kantor R, Chou S, Israelski D, Zolopa AR, Fessel WJ, Shafer RW: Mutation patterns and structural correlates in human immunodeficiency virus type 1 protease following different protease inhibitor treatments. J Virol 2003, 77: 4836–4847. 10.1128/JVI.77.8.4836-4847.2003PubMed CentralView ArticlePubMedGoogle Scholar
- Hoffman NG, Schiffer CA, Swanstrom R: Covariation of amino acid positions in HIV-1 protease. Virology 2003, 314: 536–548. 10.1016/S0042-6822(03)00484-7View ArticlePubMedGoogle Scholar
- Chen L, Lee C: Distinguishing HIV-1 drug resistance, accessory, and viral fitness mutations using conditional selection pressure analysis of treated versus untreated patient samples. Biology Direct 2006, 1: 14. 10.1186/1745-6150-1-14PubMed CentralView ArticlePubMedGoogle Scholar
- Rhee SY, Liu TF, Holms SP, Shafer RW: HIV-1 subtype B protease and reverse transcriptase amino acid covariation. PLoS Comput Biol 2007, 3: e87. 10.1371/journal.pcbi.0030087PubMed CentralView ArticlePubMedGoogle Scholar
- Wang Q, Lee C: Distinguishing functional amino acid covariation from background linkage disequilibrium in HIV protease and reverse transcriptase. PLoS ONE 2007, 2: e814. 10.1371/journal.pone.0000814PubMed CentralView ArticlePubMedGoogle Scholar
- Deforche K, Silander T, Camacho R, Grossman Z, Soares MA, Van Laethem K, Kantor R, Moreau Y, Vandamme A-M: Analysis of the HIV-1 pol sequences using Bayesian Networks: implications for drug resistance. Bioinformatics 2006, 22: 2975–2979. 10.1093/bioinformatics/btl508View ArticlePubMedGoogle Scholar
- Svicher V, Ceccherini-Silberstein F, Erba F, Santoro M, Gori C, Bellocchi MC, Gianella S, Trotta MP, Monforte A, Antinori A, Perno CF: Novel human immunodeficiency virus type 1 protease mutations potentially involved in resistance to protease inhibitors. Antimicrob Agents Chemother 2005, 49: 2015–2025. 10.1128/AAC.49.5.2015-2025.2005PubMed CentralView ArticlePubMedGoogle Scholar
- Liu Y, Eyal E, Bahar I: Analysis of correlated mutations in HIV-1 protease using spectral clustering. Bioinformatics 2008, 24: 1243–1250. 10.1093/bioinformatics/btn110PubMed CentralView ArticlePubMedGoogle Scholar
- Bishop YM, Fienberg SE, Holland PW: Discrete Multivariate Analysis: Theory and Practice. The MIT Press; 1975.Google Scholar
- Schneidman E, Still S, Berry MJ II, Bialek W: Network information and connected correlations. Phys Rev Lett 2003, 91: 238701. 10.1103/PhysRevLett.91.238701View ArticlePubMedGoogle Scholar
- Pearl J: Probabilistic Reasoning in Intelligent Systems: Networks of Plausible Inference. Morgan Kaufmann Publishers; 1988.Google Scholar
- Ahn C, Koch GG, Paynter L, Preisser JS, Seillier-Moiseiwitsch F: Log-linear models for mutations in the HIV genome. J Stat Plan Inf 2007, 137: 3227–3239. 10.1016/j.jspi.2007.03.007View ArticleGoogle Scholar
- Weigt M, White RA, Szurmant H, Hoch JA, Hwa T: Identification of direct residue contacts in protein-protein interaction by message passing. Proc Natl Acad Sci USA 2009, 106: 67–72. 10.1073/pnas.0805923106PubMed CentralView ArticlePubMedGoogle Scholar
- Kantor R, Betts BJ, Ravela J, Rhee SY, Gonzales MJ, Shafer RW: Human immunodeficiency virus reverse transcriptase and protease sequence database. Nucleic Acids Res 2003, 31: 298–303. 10.1093/nar/gkg100PubMed CentralView ArticlePubMedGoogle Scholar
- Johnson VA, Brun-Vézinet F, Clotet B, Günthard HF, Kuritzkes DR, Pillay D, Schapiro JM, Richman DD: Update of the drug resistance mutations in HIV-1: December 2008. Topics in HIV Medicine 2008, 16: 138–145.PubMedGoogle Scholar
- Clemente JC, Hemrajani R, Blum LE, Goodenow MM, Dunn BM: Secondary mutations M36I and A71V in the human immunodeficiency virus type 1 protease can provide an advantage of the emergence of the primary mutation D30N. Biochemistry 2003, 42: 15029–15035. 10.1021/bi035701yView ArticlePubMedGoogle Scholar
- Muzammil S, Ross P, Freire E: A major role for a set of non-active site mutations in the development of HIV-1 protease drug resistance. Biochemistry 2003, 42: 631–638. 10.1021/bi027019uView ArticlePubMedGoogle Scholar
- Shafer RW, Dupnik K, Winters MA, Eshleman SH: A guide to HIV-1 reverse transcriptase and protease sequencing for drug resistance studies. In Human retroviruses and AIDS, theoretical biology and biophysics. Los Alamos, NM: Los Alamos National Laboratories; 2001:1–51.Google Scholar
- Rhee SY, Taylor J, Wadhera G, Ben-Hur A, Brutlag DL, Shafer RW: Genotypic predictors of human immunodeficiency virus type 1 drug resistance. Proc Natl Acad Sci USA 2006, 103: 17355–17360. 10.1073/pnas.0607274103PubMed CentralView ArticlePubMedGoogle Scholar
- Shafer RW, Schapiro JM: HIV-1 drug resistance mutations: An updated framework for the second decade of HAART. AIDS Rev 2008, 10: 67–84.PubMed CentralPubMedGoogle Scholar
- Shafer RW, Rhee SY, Pillay D, Miller V, Sandstrom P, Schapiro JM, Kuritzkes DR, Bennett D: HIV-1 protease and reverse transcriptase mutations for drug resistance surveillance. AIDS 2007, 21: 215–223. 10.1097/QAD.0b013e328011e691PubMed CentralView ArticlePubMedGoogle Scholar
- Hertogs K, Bloor S, Kemp SD, Van de Eynde C, Alcorn TM, Pauwels R, Van Houtte M, Staszewski S, Miller V, Larder BA: Phenotypic and genotypic analysis of clinical HIV-1 isolates reveals extensive protease inhibitor cross-resistance: A survey of over 6000 samples. AIDS 2000, 14: 1203–1210. 10.1097/00002030-200006160-00018View ArticlePubMedGoogle Scholar
- Ohtaka H, Schon A, Freire E: Multidrug resistance to HIV-1 protease inhibition requires cooperative coupling between distal mutations. Biochemistry 2003, 42: 13659–13666. 10.1021/bi0350405View ArticlePubMedGoogle Scholar
- Johnson VAA, Brun-Vézinet F, Clotet B, Günthard HFF, Kuritzkes DRR, Pillay D, Schapiro JMM, Richman DDD: Update of the drug resistance mutations in HIV-1 2007. Topics in HIV Medicine 2007, 15: 119–125.PubMedGoogle Scholar
- Pollock DD, Taylor WR, Goldman N: Coevolving proteing residues: Maximum likelihood indentification and relationship to structure. Journal of Molecular Biology 1999, 287: 187–198. 10.1006/jmbi.1998.2601View ArticlePubMedGoogle Scholar
- Tiller ERM, Lui TWH: Using muliple interdependency to separate functional from phylogenetic correlations in protein alignments. Bioinformatics 2003, 19: 750–755. 10.1093/bioinformatics/btg072View ArticleGoogle Scholar
- Gloor GB, Martin LC, Wahl LM, Dunn SM: Mutual information in protein multiple sequence alignments reveals two classes of coevolving positions. Biochemistry 2005, 44: 7156–7165. 10.1021/bi050293eView ArticlePubMedGoogle Scholar
- Flares MA, Travers SAA: A novel method for detecting intramolecular co-evolution: Adding a further dimension to selective constraints analyses. Genetics 2006, 173: 9–23. 10.1534/genetics.105.053249View ArticleGoogle Scholar
- Perez-Jimenez R, Godoy-Ruiz R, Parody-Morreale A, Ibarra-Molero B, Sanchez-Ruiz JM: A simple tool to explore the distance distribution of correlated mutations in proteins. Biophys Chem 2006, 119: 240–246. 10.1016/j.bpc.2005.09.010View ArticlePubMedGoogle Scholar
- Fodor AA, Aldrich RW: Influence of conservation on calculations of amino acid covariance in multiple sequence alignments. Proteins 2004, 56: 211–221. 10.1002/prot.20098View ArticlePubMedGoogle Scholar
- Nijhuis M, Schuurman R, de Jong D, Erickson J, Gustchina E, Albert J, Schipper P, Gulnik S, Boucher CAB: Increased fitness of drug resistant HIV-1 protease as a result of acquisition of compensatory mutations during suboptimal therapy. AIDS 1999, 13: 2349–2359. 10.1097/00002030-199912030-00006View ArticlePubMedGoogle Scholar
- Perno CF, Cozzi-Lepri A, Balotta C, Forbici F, Violin M, Bertoli A, Facchi G, Pezzotti P, Cadeo G, Tositti G, Pasquinucci S, Pauluzzi S, Scalzini A, Salassa B, Vincenti A, Phillips A, Dianzani F, Appice A, Angarano G, Monno L, Ippolito G, Moroni M, Monforte A, Italian Cohort Naive Antiretroviral (I.CO.N.A.) Study Groupa: Secondary mutations in the protease region of human immunodeficiency virus and virologic failure in drug-naive patients treated with protease inhibitor-based therapy. The Journal of Infectious Diseases 2001, 184: 983–991. 10.1086/323604View ArticlePubMedGoogle Scholar
- Piana S, Carloni P, Rothlisberger U: Drug resistance in HIV-1 protease: Flexibility-assisted mechanism of compensatory mutations. Protein Science 2002, 11: 2393–2402. 10.1110/ps.0206702PubMed CentralView ArticlePubMedGoogle Scholar
- Sugiura W, Matsuda Z, Yokomaku Y, Hertogs K, Larder B, Oishi T, Okano A, Shiino T, Tatsumi M, Matsuda M, et al.: Interference between D30N and L90M in selection and development of protease inhibitor-resistant human immunodeficiency virus type 1. Antimicrob Agents Chemother 2002, 46(3):708–715. 10.1128/AAC.46.3.708-715.2002PubMed CentralView ArticlePubMedGoogle Scholar
- Schneidman E, Berry MJ II, Segev R, Bialek W: Weak pairwise correlations imply strongly correlated network states in a neural population. Nature 2006, 440: 1007–1012. 10.1038/nature04701PubMed CentralView ArticlePubMedGoogle Scholar
- Durbin R, Eddy S, Krogh A, Mitchison G: Biological Sequence Analysis. Cambridge University Press; 1998.View ArticleGoogle Scholar
- Gower JC: Measures of Similarity, Dissimilarity, Distance, Encyclopedia of Statistical Sciences. 1985, 5: 397–405.Google Scholar
- Matsuda H: Physical nature of higher-order mutual information: Intrinsic correlations and frustration. Phys Rev E 2000, 62: 3096–3102. 10.1103/PhysRevE.62.3096View ArticleGoogle Scholar
- Gilbert PB, Novitsky V, Essex M: Covariability of selected amino acid positions for HIV type 1 subtypes C and B. AIDS Res. Human Retrovir 2005, 21: 1016–1030. 10.1089/aid.2005.21.1016View ArticleGoogle Scholar
- Patick AK, Duran M, Cao Y, Shugarts D, Keller MR, Mazabel E, Knowles M, Chapman S, Kuritzkes DR, Markowitz M: Genotypic and phenotypic characterization of human immunodeficiency virus type 1 variants isolated from patients treated with the protease inhibitor nelfinavir. Antimicrob Agents Chemother 1998, 42: 2637–2644.PubMed CentralPubMedGoogle Scholar
- Steve W: Lockless and Rama Ranganathan. Evolutionarily conserved pathways of energetic connectivity in protein families. Science 1999, 286: 295–299. 10.1126/science.286.5438.295View ArticleGoogle Scholar
- Mustonen V, Kinney J Jr, Callan CG, Lässig M: Energy-dependent fitness: A quantitative model for the evolution of yeast transcription factor binding sites. Proc Natl Acad Sci USA 2008, 105: 12376–12381. 10.1073/pnas.0805909105PubMed CentralView ArticlePubMedGoogle Scholar
- DePristo MA, Weinreich DM, Hartl DL: Missense meanderings in sequence space: A biophysical view of protein evolution. Nat Rev Genet 2005, 6: 678–687. 10.1038/nrg1672View ArticlePubMedGoogle Scholar
- Weinreich DM, Delaney NF, DePristo MA, Hartl DL: Darwinian evolution can follow only very few mutational paths to fitter proteins. Science 2006, 312: 111–114. 10.1126/science.1123539View ArticlePubMedGoogle Scholar
- Deforche K, Camacho R, Van Laethem K, Lemey P, Rambaut A, Moreau Y, Vandamme A-M: Estimation of an in vivo fitness landscape experienced by HIV-1 under drug selective pressure useful for prediction of drug resistance evolution during treatment. Bioinformatics 2008, 24: 34–41. 10.1093/bioinformatics/btm540View ArticlePubMedGoogle Scholar
- Good IJ: Maximum entropy for hypothesis formulation, especially for multidimensional contingency tables. Ann Math Stat 1963, 34: 911–934. 10.1214/aoms/1177704014View ArticleGoogle Scholar
- Press WH, Teukolsky SA, Vetterling WT, Flannery BP: Numerical Recipes in C: The Art of Scientific Computing. Cambridge University Press, Cambridge; 1992.Google Scholar
- R Development Core Team: R: A Language and Environment for Statistical Computing. R Foundation for Statistical Computing, Vienna, Austria; 2005.Google Scholar
This article is published under license to BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


text:
Often we want to love people based on how perfect they are, how worthy they are. But this kind of love doesn't last very long. Who can live up to it? The Bible's letter to the Colossians (verse 3:14) looks at it a different way: we choose to love first (an active choice, not only an emotion), and this makes the "bond of perfection," one that can unite imperfect people.
This new print is available in a variety of sizes, starting at only $15 on my Society 6 print shop. Check it out!
It will also be available in an upcoming collection of postcard sized prints. More info on that here.

subset: commoncrawl


text:
China Jade Mongolian Grill
7308 International Dr. Send to Phone
407-363-9797 | Make Restaurant Reservations | Menu
OpenLunch & Dinner daily
- Parking available
- Dress code: Casual
* Click here for rating key
China Jade Mongolian Grill Restaurant Review: Hungry tourists can fill up at this I-Drive Chinese buffet that also has a Mongolian barbecue. The concept is simple: grab an empty bowl and fill it with your choice of raw meats, vegetables and sauces before handing it to the chef who stir-fries it on the grill. There are several meats, including beef, chicken and pork, sliced super thin. The vegetable selection varies, but usually includes broccoli, carrots, onions, snow peas and cabbage. The generous buffet has plenty of old standbys including beef and broccoli, fried rice, sweet-and-sour chicken, egg rolls and sushi. Your meal includes a salad bar, the buffet, the Mongolian barbecue, desserts and a scoop-it-yourself ice cream bar. The dinner buffet often includes several seafood items, including sautéed shrimp and crab legs. This location is near Universal Studios and Belz Factory Outlet World.
GAYOT's Annual Restaurant Awards honor America's best chefs, as well as promising newcomers and a top restaurateur.


text: Some of you may know that I've recently accepted a promotion at my job. The promotion has required me to move from Illinois to Oklahoma. My wife and kids won't be joining me for a few months, so I'm on my own down here. Some of the extra time on my hands has been spent writing music, playing guitar, singing, and recording. I don't have much of a studio, or much experience with recording, mixing, or producing. Ubuntu Studio helps, but there's a bit of a learning curve.Anyway, I wrote this song today for my wife. Feel free to check out the couple other songs on the channel and the couple of non-music videos there. One of the songs has too much gain on the vocal tracks so I plan to do it over sometime.


text:
The quest by the Federal Government to attract investment into the Energy Sector and boost electricity supply in the country may have received additional boost as consortium of American Investors have indicated interest to inject about 100 Mega Watts of electricity in Abuja and Kano State.
The value of the renewable energy investment is N35billion about ($212 million) and is expected to come on stream by the third quarter of 2015. The consortiums of American companies who have indicated interest are Global Business Resources USA, Global Resources Network USA, Flatbush Solar USA and Charbourne & Parke, LLP USA.
The Group which was led by Mr. James C. Nicholas, executive partner of Global Business Resources USA disclosed this yesterday when they paid a courtesy visit on the executive secretary of the Nigerian Investment Promotion Commission (NIPC), Mrs. Saratu Umar, in her office. The group is in Nigeria to explore opportunities in renewable energy development under the Umbrella of Power Africa Initiative of President Obama.
The group has already begun discussion with Abuja Distribution Company and would be visiting Kano to discuss on where to site the project and other logistics. They are also desirous to embark on rural electrification project across the country with a view to supplying electricity to rural dwellers and provide relevant tools or device that will assist in monitoring and securing of electricity infrastructure in the country.
Responding, Umar commended them for their interest to invest in Nigeria and help develop the power sector which is very critical to the economic development of the country.
She assured of the Commission's readiness to provide them with all necessary assistance and support for the smooth take-off of the project in Abuja and Kano, stressing that NIPC as an agency of the federal government has the mandate to encourage, promote, coordinate and monitor all investments in the Nigerian economy.
Source: Leadership


text:
Welcome to Agribiz Corporation
Manufacturer & Suppler of Plastic Mulch Film Laying Machine
Established in 2010 Agribiz Corporation is a well known and trusted name in hi-tech mechanised agriculture equipments. Known for introducing latest agri-equipments Agribiz corporation was the first company to introduce plastic mulch and Drip pipes laying machine and fertiliser spreaders in Indian markets.Read more about us
Our Works
Leading Manufacturer of latest mechanized agriculture equipment in India. Agribiz has been designing farming equipment for raised bed farming.
Founder's Message
Our founder have a vision of increasing prosperity by adopting mechanized farming by using latest technological innovation by using latest agriculture equipment.


text:
rane xp2016 hum and new psu...
i am getting a hum through my speakers when i have my xp2016 plugged into the mp2016, when i power down the xp2016 the hum goes away.
also, the external power supply for the units, the one i am using on the mp is about 2 metres from the 230v end to the rj45 end, but the one i am using on the xp is about 300mm from the 230v end to the rj45 end, is is possible to purchase longer power supplies?
|All times are GMT -4. The time now is 06:25 AM.
Powered by vBulletin Version 3.0.0
Copyright ©2000 - 2019, Jelsoft Enterprises Ltd.
©2006 Wave Music


text:
It's another Unscripted Sketches post with another awesome sketch! And we have a piggyback challenge if you want to add it - include numbers on your creation. Here's the sketch and my card:
I flipped the sketch sideways and used paper that already had numbers on it - hey, that's not cheating! The only stamping is the ink pen and splotches and the little "Hi" - a simple notecard that can be used in so many ways! All products from Papertrey Ink.
Come join us this week! We announce a top three every week and it could be you!
Favorite Things: January 2019
1 day ago


text:
Hasbo Playskool Busy Ball-Tivity
The Hasbo Playskool Busy Ball-Tivity Centre is is one of the hottest gifts you can give your child this year.
When I checked this playskool toy out I wanted to get one for my son immediately. The first thing I usually do is go to Walmart to find out how much they are. But this time I did a little more research and was so stunned about the results that I needed to create a page for.
I put a price comparison at the bottom of this page. First, let me show you why I was so thrilled about this product.
Product Features
Treat baby to some poppin', droppin' funtime with this air-powered activity set.
Cute jungle animals and a ladybug entice little fingers to grasp and explore
As the air starts to flow, lively music plays and animal sounds seem to bring this activity set to "life"
It's a jungle out there – and that's what your baby will love so much about this air-powered play center
Activity set comes with monkey and palm tree snap-in piece, 3 balls, 3 stacking pieces and instructions
Hasbo Playskool Busy Ball-Tivity Price Comparison
Ok, I guess the shopping fever has really gotten into me this year but it paid off. Check out the prices below.
Walmart: $29.96
Target: 23.09
Playskool: $32.99
Amazon: $9.99 plus free shipping if you buy $25.00 worth of items
Just click on the pictures to see enlarged versions


text:
+263 242 748621 - 7+263 772 151 657 / 8
Our VIP Flush Toilet Units are luxury self contained mobile toilet units which are ideal for any event be it a wedding / corporate cocktail party / conferences / sporting event / fairs etc. The VIP Flush Units consist of two toilets, each with it's own hand basin, LED lighting and all accessories required for the unit. The units are 100% self contained, they don't need to be connected to water mains or electricity which makes them idea for any site.
Event Mobility offers refrigerated units which are ideal for onsite catering services and bar services. The units run off 15amp power and are able to be cooled down to 1 degree Celsius. The units have internal stainless steel shelving which assist in optimising space inside the trailer.
Rooney's Hire Service144 Seke Road, Graniteside, Harare, Zimbabwe.Tel: +263 242 748621 - 7Cel: +263 772 151 657 / 8
Copyright © 2019, Rooney's - The Hiring Specialists
Designed and developed by 3 Degrees Design


text:
ATK Natural & Hairy presents: Beyonce has a big red dildo and she loves plunging it into her wet hairy hole. She does a sexy strip tease, then spreads her furry box open. Finally sliding her big red in. Roman K. photographed this young lovely for you to enjoy. show more
Date: October 16th, 2014; Site: ATK Natural & Hairy; Network: ATK; Pictures in set: 214; Dimension: 2000x3000
Tags: hairy, brunette, caucasian, curvy, hairy ass, hairy pits, hairy pussy, long hair, masturbation, painted fingernails, panties, small tits, spreading pussy, toys, vibrator.
|Dirty Love
|Retro Sex Symbols
|Hairy Women
|Hairy Classic
|Hairy Beauty
|Mature Pix
|Pussy Fur
Visit ATK for other quality adult sites besides ATK Natural & Hairy.


text:
The design collaboration Design Marketo, currently curating their first corner shop at the Barbican Art Gallery in London, hosted a design dinner in the Barbican's Garden room. Based on the theme Spring and Sprouts, this one-off meal gave diners the chance to see and experience their new selection of design products, crafted and commissioned for the evening, along with a few edible design elements.
To begin, guests were greeted at tables set with Refurbished Cutlery and Glass by use.dev.org. Unwanted, mismatched cutlery and glassware are coated with a layer of thin rubber to create a matching set.
Amandine Allessandra created in Rainbow Veritas, a tablecloth printed with floral patterns that are revealed when something is spilled on it. We had fun trying...
Peter Marigold's Swede Lights used vegetables as a base for votive candles and bulbs held high by upcycled forks. Other vegetables were carved for the occasion to hold candles. Handmade Glass Knuckle jugs by Nicola Zocca were filled with fresh, chilled water.
Some of the food was designed for the evening as well. Notable were the Bang ! bread volcanos by Olivia Decaris. These experimental treats are the result of a careful calibration of different types of bread dough, resulting in a thick, crusty volcano with soft, flavoured bread lava that expands from the crater during baking.
To start, a sprout and shoot salad served with spring greens and grapefruit was followed by a stoplight of three different vegetable soups. Smoked haddock and spinach in broth made up the main course, followed by a pannacotta and rhubarb dessert. This part of the menu was created by Quentin Fitch of the Barbican's Searcys restaurant.
To finish the meal, tea and coffee were served with the In Between saucers by Hina Thibaud. Double sided, one is a smaller saucer to hold espresso cups and the other is large enough to hold a mug of tea. Often created from shortbread, this evening the saucers were moulded in sweet sesame brittle, and guests were happy to nibble these little pedestals into oblivion.
Design Marketo is a platform to help young designers reach a wider audience. Their Food Marketo installation, cooking courses and pop-up shop created in conjunction with Apartamento magazine were a hit in Milan in 2010. We are looking forward to the next course in the Marketo menu...


text:
5 Minutes of What?
5-Minute Batman Stories and 5-Minute Marvel Stories are new hardcover books from HarperCollins that each feature twelve stories that each take about five minutes to read. But trying to find out more about these volumes is like pulling teeth!
At first I thought these hardcovers might be featuring reprints of stories that appeared originally in comic books but then I read in their descriptions that the books have full-page illustrations and spot illustrations throughout. This means that they're more like Big Little Books than comic books.
Still I'd like to know what writers and artists were involved, but that information is not offered. Anonymity is overrated and I know that if my writing or artwork were featured in a book I would not want it being kept a secret.


text:
2019-01-12 (10 Days Old)Total Views: 39 | Last 7 Days: 5
This week at the Tel Aviv airport: An Israeli-Arab teacher who believes education is the path to coexistence, and a British teacher who lets her students' creativity run wild
BBC: How has Liberia's George Weah performed as a president?
WashingtonPost: Saints owner says team was 'unfairly deprived' of chance at Super Bowl
CNN: Model claiming to have election meddling intel on trial in Russia
CBS: Dr. Martin Luther King Jr., in his own words
CBS: Hospital pricing lists may be misleading patients with inflated costs
BBC: Indonesia reviews release of radical cleric Abu Bakar Ba'asyir
BBC: Year of the Pig? Peppa takes China by storm
BBC: Davos 2019: 'I'm the boss, he's the spouse'
ABC: PHOTOS: A look back at the life of Martin Luther King Jr.
CNN: Lawmaker: Giuliani sounds like a crazy, old uncle
CNN: After BuzzFeed article, Trump legal team reached out to Mueller's office, Giuliani says
CNN: International tourism hits 2020 forecast two years early
BBC: The teen Primark model with vitiligo
BBC: Bedford Prison: Inmate killed cell rats during inspection
BBC: The secret to a good deal, according to the professionals
BBC: Being awake during brain surgery: 'You are hovering in between worlds'
ABC: Giuliani walks back statements of Trump's involvement in Moscow project
ABC: 2 doctors for incapacitated patient who gave birth leave
FOX: 'Dog the Bounty Hunter' star Beth Chapman undergoing chemotherapy for throat cancer, lawyer says
CNN: Exclusive: Uber memo describes issues inside company
CBS: New video changes conversation around incident between teen and Native American man
CBC: Galchenyuk unsure of Montreal reception in return with Coyotes
CBC: Potent power play fuels Flames' rise to top of West


text:
So now that i am 5 months pregnant with baby number 2, i have been sleeping long hours. I go in with my daughter M, to put her down at her bedtime, around 9ish and then dont come out at all till Morning!!
So today hubby dear tries to wake me up at around 7:30 in the morning.
Hubby: Your highness, if you are done with your beauty sleep, are you going to wake up now? I need to be at work by 9 and i have to drop you off before that.
Me: uggggg...i am so sleepy...
Hubby: What, you have slept for close to 10 hours now!!
Me: Well, I am sleeping for two....remember that!
I am highly amused at my own witticism and laugh.
Hubby hauls me out of bed. (Gently ofcourse...i am 5 months pregnant after all!!)


text:
I am always looking for new meals that are quick and easy to cook during the week. It's nice to be able to come home and do very little to prepare a meal and have the end result be something delicious. When I saw this pasta in the October issue of "Everyday with Rachael Ray" I knew I had to try it; it looked great and very low fuss.
This recipe hit the mark! It was sooo good, very light but filling. I loved the juicy sweetness from the tomatoes, the tartness of the dill and the ricotta topped everything off and mixed nicely with all the flavors. This is a great meal for the weeknight; just serve with some nice, crusty bread and even a salad and you're all set. This would also make a great side dish to accompany some type of chicken or pork, no matter what you serve this with you will not be disappointed.
Ribbon Pasta with Tomato, Dill and Ricotta
Salt and Pepper
1 Pound egg tagliatelle, fettuccine, or pappardelle pasta
3 Tablespoons extra virgin olive oil
2 Cloves garlic, crushed
2 Pints cherry tomatoes, halved
1/4 Cup finely chopped fresh dill
4 Cups loosely packed baby arugala leaves
3/4 Cup fresh ricotta cheese
Bring a large pot of water to a boil. Once water comes to a boil, add salt and pasta and cook until al dente (follow directions on pasta box for al dente pasta). Drain pasta and return to pot, reserving 3/4 cup of pasta water.
While the pasta is cooking, heat olive oil in a large skillet and add garlic and cook over medium-high heat for about 2 minutes, stirring often. After 2 minutes discard the garlic and add the tomatoes and season with salt and pepper, cook for 4 minutes and then add dill and reserved pasta water.
Add the tomatoes and arugala to the pasta and season with salt and pepper. Serve the pasta in shallow dishes topped with ricotta.
Recipes Source: "Everyday with Rachael Ray" October 2010
Saturday, November 13, 2010
Ribbon Pasta with Tomato, Dill and Ricotta
Posted by Lynsey
Labels: Healthy Eating, Main Dishes, Pasta, Quick and Easy, Tomatoes
Subscribe to: Post Comments (Atom)


text:
As many of you know, I've been working on a short story entitled Kisses for Tati (formerly known as The Drums) for a couple years now. I don't know why it's taken so long for me to finish this particular piece, especially when I'm able to see the story so very clearly in my mind.
In any case, I feel I should share something with you guys, and since I've made some progress on Kisses for Tati, I've decided that's what I'm gonna share. For those who don't know, Kisses for Tati is about Tati (natch), a socially awkward young woman who meets the man of her dreams but whose hopes of happiness are threatened by her domineering older sister.
Check out the excerpt below: Continue reading


text:
Fenna Berry is a self-taught American multimedia artist born in 1995. She lives with schizophrenia, but uses her art and creativity as an outlet for therapy.
Most of her works are done using surrealist techniques such as automatism or they are created with a highly personal concept in mind.
Browse the gallery below to see some of her creative expressions, and to learn more – visit her website by clicking here.


text:
- Kevin-V's Activity
Please update your blog, Kevin! We miss hearing from you. And I hate using twitter. lol
Kevin, I've given up with tweeting GS's ignorant Twitter account, but may I point out that ever since the website redesign, left clicking links on Firefox doesn't open them. Have to open in new tab!
Good luck on your new adventure....don't be a stranger
Best of luck to you man and thanks for all the great work you've done here. You are avery talentend person and you'll be missed here.
Why the fuck did you leave?. I was buying the games because of your reviews but now my life is in the ruins.
He gets in one final '10' on his way out... glad he could have a good last game to review...
Best of Lucks to you, Kevin. Even though many of your reviews got critics but I was in the same page with you when I tried the games that you reviewed. Much oblige ...
Just found out your leaving. Not cool at all. I wish you every success in your future. Thanks for all the great reviews(even the ones I disagreed with :)).
Sad to see you leave, you are the best reviewer in GameSpot. Good luck in your future proyects!
Going to sound cheesy here but Kevin were one of the main reasons I started using this site. Your reviews have been a great influence and I'v played alot of new games because of them. Thanks and gl.
What are you leaving to do Kevin? I love RPG's and valued every single review you carried out for this site. Best and most trusted reviews on Gamespot in my opinion. Thank you!!!
Goodbye , Kevin! Best RPG reviewer ever. I forgive you about you overrating for Dragon Age 2, because your Killzone 2 and WItcher 3 reviews are some of the best critic work ever. Good luck, dude!
Goodbye, Kevin! Thanks for all the great reviews, and your insight on games. You will be missed. Your writing (along with Greg Kasavin) set the standard Gamespot has been known for. Good luck.
Hey Kevin. I feel sorry for you man. For a review you have to play awful games like Godzilla. Thanks for the review, atlas us gamers are spared the horror.
When are you going to post the Godzilla PS4 review?
Fucked up review after fucked up review.
I leik chickannnnn
Your the reason I came back to Gamespot after Gerstmann-gate. I think you are an exquisite writer and are my favorite reviewer working today. Keep your chin up!
Just posting here to let you know how disappointed I am with the numbers out there on certain reviews, I tend to curl up in my bed and put my thumb in my mouth while lying in the foster position cryin
Thanks for the honest Batman review; seems some other media outlets are a bit /swayed/


text:
San Francisco Campus Medical Specialties Doctoral Programs | Medical & Naturopathic Schools
A wide range of Medical & Naturopathic Schools & universities offer campus Medical Specialties Doctoral programs. Whether you are interested in a Doctor of Chiropractic, a Doctorate in Acupuncture and Oriental Medicine or an allopathic medical program, studying in person may place you in an enriching environment where you receive hands-on learning. You don't have to be the student in the front row, but classroom learning tends may be interactive, make use of demonstration, and give you the opportunity to take part in live lectures. Search for doctorate degrees in medical specialties at a local college, study out of state or abroad depending on your unique needs.
Campus Medical Specialties Doctoral Programs: Basics
Campus Medical specialties doctoral programs may be offered as research (Ph.D) and clinical (First-professional) doctorate degrees. First professional degrees represent a category of qualifications in a professional subject matter that require students to have previously earned an undergraduate degree or completed specific coursework in before enrolling.[i] Holders of first-professional degrees are considered to have earned entry-level education and may take graduate study subsequently. While some of these degrees are called "Doctor", unlike the Doctor of Philosophy, candidates do not have the same research and dissertation requirements. However, there may be a clinical component, residency, externship or supervised practicum. That said, other medical specialties doctorate degree programs may be open to individuals who have completed a masters degree or who are licensed and experienced professionals.
DID YOU KNOW? In 2014, 70 Professional Doctorate degrees in Acupuncture and Oriental Medicine were awarded[ii]
Why Earn a Doctorate Degree in a Medical Specialty on Campus?
Campus Medical specialties doctoral programs are
likely to be distinguished by unique features depending on where you attend and what you study.
There is no substitute for your own homework to determine what is important for you, after all, this is somewhere you may be spending 4 years! A few reasons that may compel you to choose a residential program might include:
- You may find a school with a research facility that impresses you;
- You may wish to be mentored by faculty members at a specific school;
- You may find a school where the faculty has been published and you may be able to take seminars or lectures with these professors;
- Enrolling in an on-campus program gives you access to libraries, student services, and the ability to participate in college activities;
- You may prefer the social component, the opportunity to network professionally, or just meet new people.
- You may be able to complete your clinical experience on campus;
- You may have more options than you would for online programs because of the practical requirements of your program.
Explore Campus Medical Specialties Doctoral Programs
Search our site for Campus medical specialties doctoral programs such as:
- Alternative Medicine on campus doctorate degree programs (e.g. Doctor of Naturopathy (N.D.), Doctor of Acupuncture & Oriental Medicine (DAOM)
- Chiropractic on campus doctorate degree programs (e.g. Doctor of Chiropractic (D.C. or D.C.M.)
- Dental Medicine on campus doctorate degree programs (e.g. Doctor of Dental Science (D.D.S.) or Doctor of Dental Medicine (D.M.D.)
- Medicine & Premed on campus doctorate degree programs (e.g. Doctor of Medicine (M.D.), Doctor of Osteopathic Medicine/Osteopathy (D.O.)
- Optometry on campus doctorate degree programs (e.g. Doctor of Optometry (O.D.)
- Physician Assistant on campus doctorate degree programs
Medical & Naturopathic Schools: Professional Education on Campus
Working towards a professional doctorate degree on campus may be a rewarding way to experience classroom work with supervised clinical experience. Some of the sponsored listings you might find on GradSchools.com might include the following alternative medical professional doctorate degree programs:
Professional Education for Naturopathic Physicians
Licensed naturopathic physicians attend 4-year graduate-level naturopathic medical school. N.D.s study many of the same basic sciences as an M.D., with the addition of holistic approaches to disease prevention, optimal wellness and healing. In terms of accreditation, the Association of Accredited Naturopathic Medical Colleges (AANMC), established in 2001, offers several accredited naturopathic medical programs on different campus locations within North America.[iii] Naturopathic practice might include[iv]:
- Clinical and laboratory diagnostic testing
- Nutritional Medicine
- Botanical Medicine
- Public Health Measures
- Hygiene
- Counseling
- Minor Surgery
- Homeopathy
- Acupuncture
- Prescription medication
- Intravenous and Injection Therapy
- Naturopathic obstetrics
Potential N.D. Coursework
Per the American Association of Naturopathic Physicians (AANP), students working toward a Doctor of Naturopathy might learn some of the following potential subjects[v]:
- Standard Medical Curriculum
- Disease prevention
- Clinical nutrition
- Homeopathic medicine
- Botanical medicine
- Psychology
- Counseling
Licensing Requirements
At present, 17 states, the District of Columbia, and the United States territories of Puerto Rico and the United States Virgin Islands have licensing or regulation laws for naturopathic doctors. In these areas, naturopathic doctors are required to graduate from an accredited four-year residential naturopathic medical school and pass an extensive postdoctoral board examination (NPLEX) to receive a license[vi].
Professional Education for Acupuncturists
Students who aspire to practice Acupuncture in the United States sometimes need to have first completed 2 years of study at the baccalaureate level although some schools require students to have earned their Bachelors degree. Acupuncture schools may be accredited by the Accreditation Commission for Acupuncture and Oriental Medicine (ACAOM)[vii] which is recognized by the U.S. Department of Education as the authority in this field.
Potential DAOM Coursework
Students in Doctor of Acupuncture programs might study[viii]:
- Oriental medical theory
- Pulse diagnosis
- Chinese medical anatomy
- How to use acupuncture needles
- Oriental herbal studies
- Pain management
- Professional ethics
Licensing Requirements
Some states require acupuncturists to obtain a license to practice and may either have their own licensing examinations (California) or require prospective candidates to pass the National Certification Commission for Acupuncture and Oriental Medicine (NCCAOM) exams or complete their certification program.[ix] Certification is usually contingent of graduating from an accredited ACAOM program and completing a clean needle technique course offered by the Council of Colleges of Acupuncture and Oriental Medicine (CCOAM)[x], and pass certification exams in some of the course topics listed above.
Professional Education for Chiropractors
Chiropractic colleges with campus Doctor of Chiropractic degree programs are typically 4-years. Students often have a Bachelors degree or at least 3 years of undergraduate school prior to enrolling.[xi] Chiropractic programs are accredited by the Council of Chiropractic Education[xii].
Potential D.C. Coursework
Students in a Doctor of Chiropractic degree program may take courses in:
- Anatomy
- Physiology
- Biology
- Spinal assessment
- Spinal adjustment techniques
- Diagnosis
Licensing Requirements
All states and the District of Columbia require Chiropractors to be licensed, and some states require chiropractors to have earned a Bachelors degree. All jurisdictions require students to pass the National Board of Chiropractic Examiners exam.[xiii]
Find a Campus Medical Specialties Doctoral Program
Search for a Campus Medical Specialties Doctoral program by selecting a program category (subject). You might then look up medical, naturopathic or chiropractic schools by location (city, state, or country). Bear in mind the dialogue on state licensure and do your due diligence.
Take the Next Step
Looking for a school where you might graduate in 4-years with a professional doctorate could be an exciting venture whether you study at a local college campus or relocate for your studies. Browse our selection of sponsored programs then take that next step and contact prospective schools for more information.
Sources: [i] https://www.ed.gov/international/usnei/edlite-index.html |[ii] datausa.io/profile/cip/513301/ |[iii] https://www.naturopathic.org/education |[iv] naturopathic.org/content.asp?contentid=59 |[v] https://www.naturopathic.org/education |[vi] naturopathic.org/content.asp?contentid=57[vii] https://acaom.org/ [viii]https://nces.ed.gov/ipeds/cipcode/cipdetail.aspx?y=55&cipid=87679 |ix] https://www.nccaom.org/ |[x] https://www.ccaom.org/ |[xi] bls.gov/ooh/healthcare/chiropractors.htm |[xii] https://www.cce-usa.org/[xiii] bls.gov/ooh/healthcare/chiropractors.htm |
Sponsored Listings
- Oakland, CAOakland, CA
Samuel Merritt University
Doctor of Podiatric Medicine
The California School of Podiatric Medicine is one of nine accredited podiatric medical schools in the nation and the only one in the San Francisco Bay Area.
- Hayward, CAHayward, CA
Life Chiropractic College West
Doctor of Chiropractic Degree Program
Offering a wide variety of chiropractic techniques, state-of-the-art technology and one-on-one attention, graduates voted us #1 in Alumni Satisfaction...
- Vallejo, CAVallejo, CA
Touro University College Of Osteopathic Medicine
Touro University, California is located in the San Francisco Bay Area. As a branch campus of Touro College, NY, TU, California is fully accredited by Middle States and offers doctoral programs in osteopathic medicine and pharmacy; Masters in Physician A...
- San Francisco, CASan Francisco, CA
University of The Pacific
Dental DDS
The Arthur A. Dugoni School of Dentistry is the only dental school in the United States where you can complete a four-year academic curriculum in just three calendar years...
- Stanford, CAStanford, CA
Stanford University
Medical Doctor Program
At Stanford we believe that the best training for a career in medicine - whether in clinical practice or research - encourages...
- San Jose, CASan Jose, CA
Palmer College of Chiropractic
Chiropractic - West Campus
Palmer's Doctor of Chiropractic degree program offers you a rigorous academic education and hands-on learning in both...
- San Francisco, CASan Francisco, CA
University of California - Berkeley
Vision Science
The Vision Science Graduate Program prepares students for careers in teaching and research in vision science, optometry, ophthalmology, psychology, bioengineering, biophysics, and other disciplines. Areas of specialization in the program include visual...
- San Francisco, CA
- San Francisco, CASan Francisco, CA
California Institute of Integral Studies
Doctorate of Acupuncture and Chinese Medicine (DACM)
The Doctor of Acupuncture and Chinese Medicine (DACM) is designed for individuals who wish to enter the acupuncture and Chinese medical field at the doctoral level and do not have prior training in acupuncture and Chinese medicine...
- San Francisco, CA


text:
Website Worth and Statistics for Domain: astaro.de
Last update: Sep 16, 2015
Find the best content from astaro.de right here. Direct fast access to astaro.de. Our network of dedicated servers connects via a fast connection to any website. We do not host any files on our server. Visit astaro.de. This website contains information about astaro.de and www.astaro.de. The data come from various sources. astaro.de is not affiliated with us in any way. We offer statistics, hosting information, server information, google pagerank information. Google pagerank of this site can be found below.
There are 7,279,637 sites with a better three-month global Alexa traffic rank than Astaro.de, and the site is located in Germany. It is in the "Anbieter" category.
|Google Page Speed Rank
|Where are this site's visitors located?
|Alexa Rank?
|Google Pagerank ?
|Google trends
Bounce Raten/a
Daily Pageviews per Visitorn/a
Daily Time on Siten/a
|Top searches
|% of traffic from searches
|IP Address
|Unique visits/day
|Income per day
|$ 2
|Income per month
|$ 60
|Income per year
|$ 730
|Site value
|$ 1,198
Copy/Paste this code to your forum
Copy/Paste this code to your website


text:
Goodyear Tires
Known for their signature blimp and Nascar racing heritage, Goodyear tires currently boasts sales of nearly $20 billion globally. The company that started out with a little factory in Akron, Ohio making bicycle tires, is now the world's largest tire manufacturer. Goodyear tires are on more new vehicles in North America than any other brand. The brand is also the tire of choice for NASCAR, NASCAR Truck and dozens of other professional and amateur race series. This racing heritage, tire building expertise, and consistent product quality keeps drivers, racers and the original equipment vehicle brands coming back to Goodyear time and time again.
Goodyear has an impressive track record of excellent customer satisfaction for the tires that come original equipment on new vehicles, as well as for replacement tires. Goodyear classifies the dozens of tire styles for passenger cars, SUVs, Crossovers, trucks and commercial vans according to the following categories:
Excellent Value All-Season Tires
Goodyear's all-season tires help provide stable handling and even tread-wear in wet, icy and dry conditions. The Assurance TripleTred All-Season is their best all-season tire for confident traction in changing weather conditions. Based on Goodyear internal test results, the Assurance TripleTred All-Season gets, on average, 21% better wet traction than the leading competitor. For a smooth ride, the Assurance ComforTred Touring is their best option. Compared to the leading competitor, the Assurance ComfortTred Touring offers 20% more cushion and outperforms the leading competitor in ride comfort. These lines come in a huge number of sizes for passenger cars, SUVs and Crossovers, so we are confident there is a Goodyear Assurance all-season tire to meet your needs
Great Winter Tires
If you live in an area that has cold and snowy winters, then you know how important good tires are when the weather turns bad. Goodyear has a full range of studdable and studless winter tires. All Goodyear winter tires, like the Ultragrip Ice WRT, feature rubber that stays pliable at low temperatures, as well as lots of biting edges in the tread for more traction on ice and snow covered roads..
Sport Performance Tires
Goodyear high-performance tires help provide handling, maneuverability, and enhanced wet traction. There are treads that are pure summer tires, and treads that are all-season. The best summer tire is the Eagle F1 Asymmetric 2. The Eagle F1 Asymmetric 2 is the ultimate in wet and dry grip. In fact, according to Goodyear tests, it stops 6% shorter on wet asphalt than a leading competitor. tires. There is also an all-season version of the Eagle F1 Asymmetric, so if you run one set of tires all year round, and live in a climate with occasional snow and ice, or frequent cold weather, the Eagle F1 Asymmetric All-season is a smart choice for your sport or high performance luxury car.
Wrangler All-Terrain and M/T Tires
No matter how often, or how far, you venture off-road, there is a tough Goodyear Wrangler for you. Goodyear's best extreme off-road tire is the Wrangler MT/R with Kevlar. It features DuPont Kevlar in the steel belts of the crown of the tire to help resist punctures when powering over rocks, mud, and through sand. The most versatile Wrangler tire is the Wrangler All-Terrain Adventure With Kevlar. It has the Kevlar reinforcement for strength, plus superior traction through rain and snow for both everyday highway driving and going off-road at a moment's notice. For commercial applications, we recommend the Wrangler Duratrac. This is the workhorse Goodyear Tire. It has a bold aggressive tread design which is great for work truck towing and hauling.
Super Fuel-efficient Tires
Thanks to a casing construction that reduces the amount of heat generated, and low rolling resistance of the tread rubber, these tires can actually help save you gas, without any sacrifice in wet or dry traction. The Assurance Fuel Max and the Assurance CS Fuel Max can help you save up to 2,600 mi/4,000 Km worth of gas over the life of 4 tires (based on Goodyear's internal testing).
So no matter what type of vehicle you drive, or your particular driving needs, Tires Easy has the best prices on Goodyear Tires, always in stock and ready for fast shipping.


text:
"I'm a decent striker, but ended the season with quite a poor goals tally. How can I improve my finishing over the summer?"
Sam Cormack, via email
More after the break
Ole Gunnar Solskjaer says:
"Finishing is so many things. I used to watch strikers all the time. The best training session is simply to watch them, then go out and copy them.
To be a good goalscorer and a finisher you need to know what to do first, and then you go and train to become as good as you can be.
I used to always work on finishing techniques – hitting the ball correctly and getting a good strike on the ball.
One of the best exercises is just hitting the ball against a wall, getting it back, hitting it against the wall again, getting it back, striking it on the half-volley and on the volley and making sure you get your head and your knee over the ball.
I used to go out by myself all the time and practise in the afternoons when everyone else had gone back home. I'd throw the ball in the air, take a touch to the side and finish.
As a striker you have to be a bit selfish and you have to be ready to take the responsibility, so maybe the best thing to do is practise by yourself so that you're mentally ready."
For more football tips see:
Andy Cole: Finishing a one-on-one
Jermain Defoe's six-step guide to striking
Michu: Break through the defence
Demba Ba: Hit the target every time
Rickie Lambert: How to hit the back of the net


text:
UN call to tackle landfill Africa
12 dead in toxic waste scandal
A United Nations Environment Programme (UNEP) meeting this week will discuss the growing threat from toxic electronic waste being dumped on developing countries.
Last week, UNEP reported that criminal dumping of in Abidjan, Cote D'Ivoire, had caused the deaths of at least 12 people and forced 100,000 to seek medical attention. $30m of international financial assistance would be needed to clean up one of the world's poorest countries, it said. More on the incident here.
Opening the 8th Conference of the Parties to the Basel Convention in Kenya today, UNEP executive director Achim Steiner said Western countries were effectively using Africa as a landfill for useless electronics. Up to 75 per cent of the used TVs, CPUs, and mobile phones which end up in African ports like Lagos in Nigeria were defunct and represent no more than "long-distance dumping", he added.
Steiner said: "It also serves as a reminder that even the best laws are only as strong as the enforcement mechanisms and willingness of governments to act."
He called for better enforcement of the Basel Convention; a recent survey found 48 per cent of European waste shipments were illegal. UNEP's emergency fund should be increased in light of the Cote D'Ivoire crisis and more manufacturer-led recycling programmes were essential, Steiner added.
The Basel Convention came into force in 1992 and was designed to force nations to deal with their own toxic waste rather than pass the buck to poorer regions.
The EU's WEEE directive should finally come into force in 2007. It puts responsibilty for recycling and safe disposal on producers of electronic equipment. ®


text:
hi~ a while ago, Megan of Health Enterprises sent me a CVS 6-in-1 Manicure Tool.
the tool contains 'a dual-sided cuticle pusher and cleaning stick, nail brush, nail clippers and a dual-sided nail buffer and nail file' and is sold in CVS stores.
here's how the manicure tool looks from above and from the side.
i've had the chance to use the cuticle pusher and i like it. its very smooth and works really well~ ooh, also, the handle is made of silicon so its really comfy.
a big thanks to Megan of Health Enterprises for the tool. be sure to check out the Health Enterprises facebook.
have a good one!,


text:
Re: Imperial Conquest
"Hold that thought." Mattias said. Two stormtroopers walked over to the booth they were sitting at. "You there, you are under arrest for asso-" "Wow, hold on there. These guy are working with me, Mattias Felth, I was the one leading the moisture farm raid. They're assisting me in a covert op." "Yes, I remember you. The captain said something about a buisness partner. You, considering you didn't shoot back in the hangar, I'll let you go. For now. Good day, Mr. Felth." The two stormtroopers turned around and left. "Buisness partner?" Uriel asked. "Yea, that jawa." Mattias pointed to Yurt at the bar. <Yee!> Yurt shouted as he spun on his stool drinking a fifth jawa beer. "Really? Uriel asked. "Afraid so." Mattias replied. "What favor do I owe you Devon? I wanted to kill you."


text:
Copyright © 2014
Arbor Group is the premier film and video production company located in St. Louis, Missouri. As a director based production company, we are known for our skills in direction of real people as well as professional talent, and for being the top lifestyle directors in the Midwest. Our team has been responsible for award winning videography for television, commercial and corporate production for more than 23 years and has been a leader in the Midwest in producing and directing high definition video. Add to that fact, that we are St. Louis' premier tabletop directors and comedy directors and you know the Arbor Group can deliver excellence no matter what your requirements.
phone: 314 • 367 • 1111 email: firstname.lastname@example.com or email@example.com