text:
- Research article
- Open Access
Genetic and epigenetic variations contributed by Alu retrotransposition
© de Andrade et al; licensee BioMed Central Ltd. 2011
- Received: 17 February 2011
- Accepted: 20 December 2011
- Published: 20 December 2011
De novo retrotransposition of Alu elements has been recognized as a major driver for insertion polymorphisms in human populations. In this study, we exploited Alu-anchored bisulfite PCR libraries to identify evolutionarily recent Alu element insertions, and to investigate their genetic and epigenetic variation.
A total of 327 putatively recent Alu insertions were identified, altogether represented by 1,762 sequence reads. Nearly all such de novo retrotransposition events (316/327) were novel. Forty-seven out of forty-nine randomly selected events, corresponding to nineteen genomic loci, were sequence-verified. Alu element insertions remained hemizygous in one or more individuals in sixteen of the nineteen genomic loci. The Alu elements were found to be enriched for young Alu families with characteristic sequence features, such as the presence of a longer poly(A) tail. In addition, we documented the occurrence of a duplication of the AT-rich target site in their immediate flanking sequences, a hallmark of retrotransposition. Furthermore, we found the sequence motif (TT/AAAA) that is recognized by the ORF2P protein encoded by LINE-1 in their 5'-flanking regions, consistent with the fact that Alu retrotransposition is facilitated by LINE-1 elements. While most of these Alu elements were heavily methylated, we identified an Alu localized 1.5 kb downstream of TOMM5 that exhibited a completely unmethylated left arm. Interestingly, we observed differential methylation of its immediate 5' and 3' flanking CpG dinucleotides, in concordance with the unmethylated and methylated statuses of its internal 5' and 3' sequences, respectively. Importantly, TOMM5's CpG island and the 3 Alu repeats and 1 MIR element localized upstream of this newly inserted Alu were also found to be unmethylated. Methylation analyses of two additional genomic loci revealed no methylation differences in CpG dinucleotides flanking the Alu insertion sites in the two homologous chromosomes, irrespective of the presence or absence of the insertion.
We anticipate that the combination of methodologies utilized in this study, which included repeat-anchored bisulfite PCR sequencing and the computational analysis pipeline herein reported, will prove invaluable for the generation of genetic and epigenetic variation maps.
- Methylation Status
- Bisulfite
- AluY Family
Repetitive elements constitute over 50% of the human genomic sequence [1]. The most prevalent repeats are the Alu family of SINEs, which comprise approximately 10% of the human genome. A typical Alu element is approximately 300 bp long and contains two almost identical arms separated by an A-rich sequence. The ancestor of the Alu monomer is the 7 SL RNA gene, which encodes the RNA component of the signal recognition particle (SRP) that is involved in the translocation of newly synthesized proteins [2, 3]. Similar to the 7 SL gene, Alu elements with intact promoters - namely A and B boxes - may be transcribed by RNA polymerase III [2, 4]. With the aid of the LINE-encoded retrotransposition machinery, Alu transcripts gain mobility and expand in genomes through a process involving reverse transcription and integration [5].
Alu retrotransposition has been an important molecular evolutionary force reshaping the primate genomes [6]. The expansion of the Alu elements in the primate genomes is dated at least 60 million years ago [7]. Based on their evolutionary history, Alu elements can be classified in three major subfamilies: AluJ, AluS, and AluY [8]. Among them, the youngest Alu elements - AluY and its variants AluYa-g - remain very active, and exhibit the highest rate of retrotransposition in the human genome [9–12]. While several recent studies have shown that LINE-1 elements contribute substantially to the structural variations observed in the human genome [13–15], the retrotransposition rate of Alu elements is ten times higher than that of LINE-1, with an estimated new insertion at every 21 births [16].
Decades of research have demonstrated that Alu elements play important roles in the genome and transcriptome [17–20]. Alu elements may contribute a large number of transcription factor binding sites [21], some of which may serve as enhancers involved in tissue development [22, 23]. In addition, some Alu elements may be expressed and Alu transcription affects nearby gene expression, distal gene expression, and global translation. For instance, the expression of an Alu in the promoter of an epsilon-globin gene was found to negatively regulate globin gene expression by transcriptional interference [24]. Recently, Alu RNA was found to be a modular transacting repressor of mRNA transcription [25]. Interestingly, such transcriptional suppression was found to be specific and limited to certain genes. Alu RNAs also affect translational initiation and were found to form stable, discrete complexes with the double-stranded RNA-activated kinase PKR, and to antagonize PKR activation [26]. Transcription derepression of otherwise active Alu elements, which so often reside within genes, may lead to the formation of double-strand RNA - if in antisense orientation - and ultimately to heterochromatinization and silencing of the gene [27].
One of the key mechanisms controlling Alu expression is DNA methylation. The human genome has approximately 28 million CpG dinucleotides, 7 million of which are found within Alu elements [1]. In most somatic tissues, the CpG dinucleotides within the Alu sequence are heavily methylated to suppress Alu expression [28, 29]. It has been demonstrated that the A and B boxes (5-16 bp, and 75-84 bp from the 5' terminus, respectively) are critical cis-elements for Alu expression. In particular, methylation of the B box is thought to inhibit protein binding and hence block Alu transcription [30]. Albeit not sufficient, demethylation and consequently transcription of Alu elements is required for occurrence of de novo retrotransposition [28]. Methylated CpGs can undergo deamination and thereby lead to mutations that render them unable for retrotransposition [8, 9].
Although much effort has been made to identify structural variations resulting from Alu integration, much less is known with regard to the epigenetic status of newly inserted elements and of their flanking genomic sequences. Here we report the utilization of an Alu-anchored bisulfite PCR strategy to generate methylation maps for thousands of Alu elements in human cerebellum and in ependymomas [31, 32]. In this approach, most of the targeted Alu elements are members of the active AluY subfamilies. In this study, we analyzed the aforementioned datasets to identify newly integrated Alu elements, to investigate sequence characteristics and commonalities of their integration sites, to uncover their methylation statuses, and to determine whether the methylation patterns of the sequences surrounding their integration sites would be altered in the alleles harboring the insertion in individuals hemizygous for the Alu retrotransposon.
Identification of recent Alu insertions
The method developed by Xie and colleagues was initially designed to generate a methylation map for a subset of young Alu elements [32]. The strategy applied a primer targeting CpG-rich Alu repeats to simultaneously amplify thousands of Alu elements and their 5' flanking sequences. Unequivocal mapping of these repeats was therefore achieved through their - most often unique - 5' flanking sequences. Eight Alu libraries were derived with this strategy, six from ependymomas and two from normal brain tissues [31, 32].
327 clusters comprised in 1,762 sequence reads.
Sample ID*
289,816 (140,865**)
sequenceRead_Alu insertion
genomeRegion_Alu insertion
498 (327**)
Ratio (genomeRegion_AluInsertion/
To investigate whether these putatively new Alu insertions had been identified in previous studies, we extracted 1,763 and 795 known polymorphic Alu elements from dbSNP (The Single Nucleotide Polymorphism database, NCBI) [33] and dbRIP (Database of Retrotransposon Insertion Polymorphisms) [34], respectively. This analysis revealed that 316 of the 327 clusters were novel, i.e. they corresponded to yet undocumented de novo retrotransposition events. The putative integration sites of 140 of such clusters (42.8%) were found to localize to intronic regions, except for one, which mapped to the 3'-UTR of TOMM40, a gene that codes for the translocase of the mitochondrial outer membrane (TOM) complex. We further analyzed these genes with NCBI's DAVID functional annotation tool to examine whether any specific gene category was more likely to harbor these Alu insertions. One hundred thirty-two genes were found annotated in the NCBI database. Compared to all genes annotated in the human genome, no significant enrichment was identified for this set of 132 genes in terms of biological process, cellular localization or molecular function (Additional File 2, Table S2).
Verification of recent Alu insertions
To validate the evolutionarily recent Alu de novo retrotransposition events identified in this study, we randomly selected twenty-one genomic loci encompassing such putative new Alu insertions. For each genomic locus, we designed primers based on the upstream and downstream sequences surrounding the predicted integration sites. With these primers, the PCR products were expected to be ~120 bp (without Alu insertion) or ~420 bp (with Alu insertion). Due to the diploidy of the human genome, three kinds of PCR results were expected: (1) hemizygous Alu insertion: PCR products of two different sizes were expected, one fragment with the Alu insertion and another without it (spanning ~420 bp and ~120 bp, respectively); (2) homozygous Alu insertion: only one PCR product was expected, this fragment containing an Alu element (spanning ~420 bp); (3) nulizygous Alu insertion: no Alu insertion was present in either homologous chromosome, hence just one small PCR product (spanning ~120 bp) was expected.
Genomic features and sequence characteristics of Alu elements and their flanking sequences
It has been shown that polymorphic Alu elements and their flanking sequences may share some distinct sequence features [34, 35]. The Alu transcripts derived from the ones with conserved structure would interact productively with SRP9/15 host proteins and gain the ability to retrotranspose [12]. The AluY subfamily and its variants Yc1, Yc2, Ya5, Ya5a2, Ya8, Yd8, Yb8, and Yb9, are the ones considered to be very active due to the conservation of its structure. To conclude the analysis, we classified the Alu elements identified in this study according to its family of origin. We found that the new insertions identified in this study belong to the relatively recent family of AluY elements or to the subfamilies AluYa5, AluYb8, AluYb9, and AluYg6. It has also been shown that the occurrence of a longer poly(A) tail might facilitate Alu retrotransposition [35]. Our analysis revealed that all twenty-two new Alu insertions that were sequence verified in this study have an A-tail that ranged from 11 to 45 nucleotides, with an average length of 29 bp.
Sequence features of newly inserted Alu repeats.
Chromosome coordinate
Alu subfamily
Putative cleavage site
Poly-A sequence
In addition to the Alu sequence itself, the genomic sequence adjacent to the recent Alu insertions encompass at least two typical sequences. As a hallmark of a recent retrotransposition event, the sequences immediately flanking the Alu elements corresponded to short direct repeats, ranging from 4-17 nucleotides. The insertion mechanism generates direct target site duplications (TSDs) flanking the newly inserted element. These TSDs have variable length and are highly suggestive of LINE mediated endonucleolytic cleavage [12, 38]. Such short direct repeats, also called AT-rich target site duplications, were present in 19 of the sequence-verified genomic loci (Table 2).
Methylation status of recent Alu elements
All sequences generated in our previous studies, encompassing Alu elements and their 5' flanking sequences, were derived from bisulfite converted genomic DNA [31, 32]. Due to the high frequency of C-to-T transitions in CpG dinucleotides of Alu repeats caused by deamination of the methylated cytosines, in the absence of a reference genomic sequence, one cannot determine the methylation status of a novel Alu insertion by this method. Hence, to examine the methylation pattern of the newly integrated Alu elements, we aligned the sequences generated in this study for nineteen of such Alu elements with their bisulfite converted sequences from our previous studies [31, 32] (Additional File 3, Figure S1). Our results showed that the recently inserted Alu elements are heavily methylated, with an average methylation level of 90.7%; this is similar to the average methylation level observed for evolutionarily young non-polymorphic Alu elements [31, 32]. We further examined the methylation status of the two important promoter regions inside the Alu elements, the A and B boxes. Alu elements have a bipartite structure, which is similar to that of tRNA elements. It has been shown that the A box is responsible for determining the strength of the Pol III promoter and the B box is important in enabling transcription [30, 39]. Also, deletion of the B box sequence completely abolished transcription of the elements, while deletion of the A box reduced the efficiency of transcription [40]. In almost all cases, these promoter sequences were methylated (Additional File 3, Figure S1). This result suggests that transcription of most newly inserted Alu elements is suppressed by DNA methylation.
Recent studies demonstrated that major structural variants in the human genome are derived from retrotransposons, Alu elements in particular [16, 41]. Due to the extensive sequence homology that exists among young Alu repeats, the identification of such structural variants remains a challenging task. To date, a total of 2,558 polymorphic Alu retrotransposons have been reported to occur in human populations, 1,763 of which have been deposited in dbSNP and 795 in dbRIP. In this study we implemented a computational pipeline to identify recent Alu insertions, and examined the methylation status of the newly inserted Alu retrotransposons and their flanking sequences. At the time we developed this strategy the Genome Sequencer FLX System was the most suitable alternative available, given the greater length of the sequence reads that it generates, and the fact that sequences would encompass an Alu repeat and would be derived from bisulfite-converted genomic DNA. Altogether, the longer reads generated with the FLX System greatly facilitated their mapping back to the reference genome sequence. Notwithstanding this advantage, however, we anticipate that our approach may be adapted to take advantage of competing next generation sequencing platforms that have a higher throughput and that can now generate sufficiently long sequence reads.
Using this strategy a total of 327 putative Alu elements were identified. We found that 42.8% of their insertion sites fell within intronic regions, while one integration site mapped to the 3'-UTR of the TOMM40 gene. TOMM40 is a component of the preprotein translocase complex of the outer mitochondrial membrane, which consists of at least 7 different proteins (TOMM5, TOMM6, TOMM7, TOMM20, TOMM22, TOMM40, and TOMM70). These results are consistent with previous studies indicating that Alu retrotransposons tend to be inserted within intragenic regions [1, 42].
Out of the twenty-one insertion events that were randomly selected for validation analysis, nineteen were successfully verified. A limitation of the Alu-anchored bisulfite PCR approach that needs to be acknowledged is the fact that only 5' flanking sequences are obtained. The right arm of the Alu retrotransposons and their 3' flanking sequences are not represented in the sequence reads that are generated. Hence, in order to design primers for the validation experiments, we used the reference sequence of the human genome as source of putative 3'-flanking sequences for the Alu insertions. Accordingly, it is possible that the two cases that could not be verified may have been caused by the utilization of an incorrect 3' flanking sequence for primer design. Notwithstanding this limitation, the lowest estimated accuracy for the analysis pipeline that we have implemented in this study for the identification of de novo Alu retrotransposition events would be of 90.5% (19/21).
The sequence features (TSD, TT/AAAA cleavage sequence, and A-rich Alu tail) that are typically observed in newly inserted Alu elements constitute hallmarks of retrotransposition [5, 10, 36, 37]. Indeed, further analysis of the aforementioned nineteen PCR-cloned Alu elements and flanking sequences revealed the presence of both the TSD and TT/AAAA sites. Alu A-tails seem to be an important factor to enable Alu element retrotransposition [4, 35]. Roy-Engel et al. reported that the average A-tail length of active Alu elements is 26 [35]. Consistent to their finding, the Alu A-tail sizes of the Alu elements described in this study ranged from 11-45 with an average of 29 bp.
Most cancer genomes are characterized by localized hypermethylation as well as by global hypomethylation [43, 44]. This hypomethylation process may enable transcription and de novo retrotransposition of Alu elements which, in turn, may lead to genome instability [45]. Our previous study demonstrated that the methylation level of Alu elements decreased in ependymomas, and most significantly in recurrent tumors [31]. To examine whether some Alu insertions represented somatic events limited to recurrent ependymomas, which could have occurred in consequence of the loss of DNA methylation, we generated and compared PCR products from ten genomic loci in primary and in recurrent tumors derived from one individual. The same results were obtained in all ten genomic loci. In addition, five of the ten Alu insertions were also found in other individuals. These results suggested that such validated Alu insertions most likely represent germ-line rather than somatic events.
In this study, in addition to identifying structural variants in the genome of 6 individuals, we investigated epigenetic variations that might result from de novo retrotransponsition events. The Alu elements identified in this study were heavily methylated, as it was previously shown by high-throughput bisulfite sequencing and herein validated by cloning and sequencing analyses. The analysis of methylation throughout the mapped Alus and among the 19 loci verified revealed that there were no methylation differences among tissues (Additional File 4, Table S3). This result indicates that at least by the time these DNA samples were obtained most of the newly inserted Alu elements were already transcriptionally repressed. This finding is further supported by the fact that the promoters of the Alu elements, i.e. their A and B boxes, were found to be methylated. However, there was one exception. We found an Alu insertion that was partially unmethylated (chr9:37594172-37594310). Interestingly, the insertion of this Alu element occurred 1,576 bp downstream from a CpG island and 1,674 bp downstream from the transcription start site of TOMM5, a gene encoding the translocase of the outer mitochondrial membrane 5. With a completely unmethylated promoter (both the A box and the B box were unmethylated), it is conceivable that this Alu element may have remained transcriptionally active and hence have served as source for additional retrotransposition events. Another interesting finding was that a CpG island that is upstream of the element - i.e., that of TOMM5 - may be influencing the methylation pattern of this Alu repeat. Indeed, the methylation status of the CpG island was similar to that of the 5' end sequences of this Alu repeat, i.e. both were unmethylated. It would be interesting to explore the functional impact of this particular Alu on the nearby TOMM5 gene. Additionally, 3 Alu repeats and 1 MIR element that are localized between the newly inserted Alu repeat and the CpG island were found to exhibit very low methylation levels. Such striking pattern of DNA methylation may indeed be an indication of the influence exerted by the adjacent CpG island. It is also possible that other epigenetic factors might be affecting the methylation statuses of these Alu elements, such as nucleosome positioning. Two previous studies have reported the influence of nucleosome positioning, within and around Alu element, in Alu activity [46, 47]. Accordingly, it is noteworthy that an AluJo that is localized downstream of this newly inserted Alu exhibits a similar pattern of DNA methylation, i.e. its 5' half is unmethylated while its 3' half is methylated. In our previous study [32], we found that genomic localization has a profound impact on Alu methylation status. In this study, the identification of both methylated and unmethylated Alu elements provided additional support to there being a positional effect on Alu methylation. Last, but not least, it is noteworthy that two of the novel Alu insertions herein reported map within or near genes encoding members of the preprotein translocase complex of the outer mitochondrial membrane, namely TOMM40 and TOMM5, respectively. It is conceivable that given their housekeeping function and ubiquitous expression pattern, hence commonly open chromatin status, these genes may be more vulnerable to uptake de novo retrotransposition events.
To explore the epigenetic impact of Alu insertion on adjacent genomic sequences, we examined the methylation statuses for two loci harboring hemizygous insertions, and - in one case - obtained the methylation patterns of CpG dinucleotides flanking the Alu insertion sites. Both alleles - irrespective of the presence of an inserted element - were found to be heavily methylated, and no significant epigenetic variation was observed in association with the presence of the additional Alu element.
In this work we have identified a few novel Alu insertions sites. We used DNA samples from normal and from tumor tissues, but the data obtained did not show any tissue preference for these insertions. More studies are highly desired to further scrutinize the functional aspects of structural variants in the human genome, including epigenetic variations that might arise in consequence of a de novo retrotransposition event.
High-throughput bisulfite sequencing datasets for Alu elements
The high-throughput bisulfite sequencing data were derived from Alu-anchored bisulfite PCR libraries derived from tissues samples, including a normal cerebellum, a normal 4th ventricle lining, two primary non-aggressive, two primary aggressive and two recurrent ependymomas [31, 32]. Briefly, genomic DNA is first digested with AluI restriction enzyme, ligated to adaptors and then subjected to bisulfite treatment. Bisulfite treated DNA is amplified with adaptor and Alu-specific primers, the latter targeting a large pool of CpG-rich Alu elements. Thus, each PCR product contains the 5'end of an Alu element and its (most often) unique flanking genomic sequence, which makes it possible for each sequence to be unambiguously mapped to the reference human genome. Primary non-aggressive ependymomas are defined as primary tumors from patient free of disease progression for more than 4 years and primary aggressive ones are defined as primary tumors from patients with recurrent disease within 3 years or deceased of disease.
Computational pipeline for the identification of recent Alu insertions
To identify putatively recent Alu insertions, sequence reads rejected in previous studies were selected. Briefly, after removal of primer and adaptor sequences, sequences greater than 40 bp were aligned to the in silico bisulfite converted reference genome using multiple cycles of MegaBLAST. The word size used in Megablast was set to 100 for the first cycle, it was decreased by 20 for every cycle thereafter until the last, for which the minimum length of best perfect match was set to 40. In addition, the identity percentage cutoff for a significant alignment was set to be 100 for the last cycle and 95 for all other cycles of Megablast [32]. The sequence reads that mapped to genomic loci within 10 bp from an Alu element were considered as a putative recent Alu insertion.
PCR, cloning, and sequencing
For PCR primer design, the original (not bisulfite converted) DNA sequences flanking the predicted Alu insertion sites were extracted from the UCSC reference human genome, based on their genomic coordinates [48]. PCR primers were designed in the region surrounding the Alu insertion sites. PCR reactions were performed using HotStartTaqR Plus Master Mix from QIAGEN. Each reaction was prepared as follows: 12.5 μL of HotStartTaqR mix, 30 ng of DNA, 14 μM of each primer, and enough water for 25 μL. The PCR reactions were performed on a MJ Research machine (model PTC 225). Reactions were subjected to an initial activation step of 95°C for 15 min, then by a denaturation step of 94°C for 1 min, followed by 40 cycles of 1 min at 94°C, 30 s at optimal annealing temperature, and 40 s at 72°C, followed by a final extension step of 10 min at 72°C. PCR product annealing temperatures (Tm) and primers used on each reaction are listed on Additional File 5, Table S4. After reactions were completed the amplified fragments were separated using 1.5% agarose gel electrophoresis that was stained with ethidium bromide and visualized using UV fluorescence system. Running was carried out until a good separation of bands was obtained. After separation in the 1.5% agarose gel the bands were excised off the gel and purified using a gel purification kit from QIAGEN, QIAquickR PCR Purification Kit. The purified PCR products were cloned using the TOPO TA CloningR System from Invitrogen. Sequencing reactions for individual colonies were conducted at the Sequencing Core Facility of the Children's Memorial Research Center of Northwestern University's Feinberg School of Medicine.
Bisulfite PCR
Bisulfite conversion of genomic DNA was performed with EZ DNA Methylation Gold kit (Zymo Research Corporation, Irvine, CA) following the manufacturer's instructions. 300 ng of genomic DNA was treated and eluted with 10 μL of elution solution. After this step, DNA from the chr10:72275361-72275449 genomic locus was amplified using the pair of primers: 5'- GGA TTA AGT TTT TTT TTT GTT T -3' and 5'- CTA CAA AAA AAA ATA ACT CAT A -3'; the chr2:48129974-48130105 genomic locus was amplified using the pair of primers: 5'- CCT TAC CAT TTA AAA ATA AAA AAT CAA -3' and 5'- GTT TAA GAT TTA AAG GAA TGA GTT AG -3'. PCR reactions were prepared using the same reagents and conditions described above. The PCR program used was: activation step of 95°C for 15 min, then by a denaturation step of 94°C for 1 min, followed by 40 cycles of 1 min at 94°C, 30 s at optimal annealing temperature (49°C for locus chr10:72605338-72605440 and 42°C for locus chr2:48276482-48276601), and 40 s at 72°C, followed by a final extension step of 10 min at 72°C. PCR amplified fragments were separated in 1.5% agarose and excised from it as described above.
To amplify the 5'end of the CpG island and the sequence flanking the 3' end of the Alu element (AI19 chr9:37594172-37594310) we used nested PCR. The primers used for the CpG island were: external primers (TOMM5-eFor 5'- AAG TTG GGA GAA TTA GGA TGA TT -3' and TOMM5-eRev 5'- CTA ATT TTT ATA TAA CAA ATA TTA TTA AAA ACA AC -3'), internal primers (TOMM5-iFor 5'- GTA TTT TTA GAG TTA AGG GGT GT -3' and TOMM5-iRev 5'- CAC TTC AAA TCA ACT AAA TCA AAA C -3'). The primers used for the 3'Alu flanking sequence were: external primers (ch9Down-eFor 5'- TTT GTA GTG ATG TTG AAA GTA GTA AGA- 3' and ch9Down-eRev 5'- AAT ATA TAC CTT CCC TTT CCA ACT -3'), internal primers (ch9Down-iFor 5'- TTT ATT TTA GAT TGA GTT TTG TTT TGT -3' and ch9Down-iRev 5'- CTT AAA CCC AAA AAT ATA AAA TTA CAA TAC -3'). The PCR program used was the same mentioned above, the TM temperatures were 51°C for external primers and 50°C for internal primers.
Alu classification
As it was mentioned above, the initial data used for this study were derived from Alu-anchored bisulfite PCR libraries. These libraries were constructed using primers specially designed to target the most recent and active Alu elements, which are the ones that belong to AluY family. To classify the Alu insertions verified in this study within the AluY family, we aligned the sequences generated in this study with those contained in the UCSC Genome browser. From this alignment, the element with the highest score was used to classify a newly identified Alu insertion into the AluY family.
This study was supported by the Everett/O'Connor Charitable Trust; Dr. Ralph & Marian C. Falk Medical Research Trust; Gus Foundation; The Maeve McNicholas Memorial Foundation and Medical Research Institute Council. We are most grateful to Selva Musa for her outstanding technical support and invaluable contributions to this work.
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- Xie H, Wang M, Bonaldo Mde F, Rajaram V, Stellpflug W, Smith C, Arndt K, Goldman S, Tomita T, Soares MB: Epigenomic analysis of Alu repeats in human ependymomas. Proc Natl Acad Sci USA. 2010, 107 (15): 6952-6957. 10.1073/pnas.0913836107.View ArticlePubMedPubMed CentralGoogle Scholar
- Xie H, Wang M, Bonaldo Mde F, Smith C, Rajaram V, Goldman S, Tomita T, Soares MB: High-throughput sequence-based epigenomic analysis of Alu repeats in human cerebellum. Nucleic Acids Res. 2009, 37 (13): 4331-4340. 10.1093/nar/gkp393.View ArticlePubMedPubMed CentralGoogle Scholar
- Sherry ST, Ward MH, Kholodov M, Baker J, Phan L, Smigielski EM, Sirotkin K: dbSNP: the NCBI database of genetic variation. Nucleic Acids Res. 2001, 29 (1): 308-311. 10.1093/nar/29.1.308.View ArticlePubMedPubMed CentralGoogle Scholar
- Wang J, Song L, Grover D, Azrak S, Batzer MA, Liang P: dbRIP: a highly integrated database of retrotransposon insertion polymorphisms in humans. Hum Mutat. 2006, 27 (4): 323-329. 10.1002/humu.20307.View ArticlePubMedPubMed CentralGoogle Scholar
- Roy-Engel AM, Salem AH, Oyeniran OO, Deininger L, Hedges DJ, Kilroy GE, Batzer MA, Deininger PL: Active Alu element "A-tails": size does matter. Genome Res. 2002, 12 (9): 1333-1344. 10.1101/gr.384802.View ArticlePubMedPubMed CentralGoogle Scholar
- Feng Q, Moran JV, Kazazian HH, Boeke JD: Human L1 retrotransposon encodes a conserved endonuclease required for retrotransposition. Cell. 1996, 87 (5): 905-916. 10.1016/S0092-8674(00)81997-2.View ArticlePubMedGoogle Scholar
- El-Sawy M, Deininger P: Tandem insertions of Alu elements. Cytogenet Genome Res. 2005, 108 (1-3): 58-62. 10.1159/000080802.View ArticlePubMedGoogle Scholar
- Hagan CR, Sheffield RF, Rudin CM: Human Alu element retrotransposition induced by genotoxic stress. Nat Genet. 2003, 35 (3): 219-220. 10.1038/ng1259.View ArticlePubMedGoogle Scholar
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- Lupski JR: Retrotransposition and structural variation in the human genome. Cell. 2010, 141 (7): 1110-1112. 10.1016/j.cell.2010.06.014.View ArticlePubMedGoogle Scholar
- Grover D, Mukerji M, Bhatnagar P, Kannan K, Brahmachari SK: Alu repeat analysis in the complete human genome: trends and variations with respect to genomic composition. Bioinformatics. 2004, 20 (6): 813-817. 10.1093/bioinformatics/bth005.View ArticlePubMedGoogle Scholar
- Deng G, Nguyen A, Tanaka H, Matsuzaki K, Bell I, Mehta KR, Terdiman JP, Waldman FM, Kakar S, Gum J, et al: Regional hypermethylation and global hypomethylation are associated with altered chromatin conformation and histone acetylation in colorectal cancer. Int J Cancer. 2006, 118 (12): 2999-3005. 10.1002/ijc.21740.View ArticlePubMedGoogle Scholar
- Roman-Gomez J, Jimenez-Velasco A, Agirre X, Castillejo JA, Navarro G, Garate L, Jose-Eneriz ES, Cordeu L, Barrios M, Prosper F, et al: Promoter hypermethylation and global hypomethylation are independent epigenetic events in lymphoid leukemogenesis with opposing effects on clinical outcome. Leukemia. 2006, 20 (8): 1445-1448. 10.1038/sj.leu.2404257.View ArticlePubMedGoogle Scholar
- Daskalos A, Nikolaidis G, Xinarianos G, Savvari P, Cassidy A, Zakopoulou R, Kotsinas A, Gorgoulis V, Field JK, Liloglou T: Hypomethylation of retrotransposable elements correlates with genomic instability in non-small cell lung cancer. Int J Cancer. 2009, 124 (1): 81-87. 10.1002/ijc.23849.View ArticlePubMedGoogle Scholar
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- Tanaka Y, Yamashita R, Suzuki Y, Nakai K: Effects of Alu elements on global nucleosome positioning in the human genome. BMC Genomics. 11: 309.Google Scholar
- Kuhn RM, Karolchik D, Zweig AS, Trumbower H, Thomas DJ, Thakkapallayil A, Sugnet CW, Stanke M, Smith KE, Siepel A: The UCSC genome browser database: update 2007. Nucleic Acids Res. 2007, D668-673. 35 DatabaseGoogle 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.


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Praesent augue arcu, ornare mattis a libero morbi tristique senectus fames ac turpis egestas...
At UniqueClick, we offer a wide range of web based solutions that become a part of your Business.
Symbol, Monogram, Mascot logo, Abstract logo, The emblem
Business card, Letterhead, Envelope, Hoarding, Brochure
Wireframe, Custom website layout design, Mobile apps ui/ux design
PSD to Responsive HTML. HTML5, CSS3, Bootstrap
Custom WP website development, Theme installation
Custom E-Commerce website development, Theme installation
Web Banner ad design, Animated GIF banner design
Facebook, Twitter, Google+, YouTube cover design. Facebook ad
Looking for a reliable web design company! UniqueClick is a trusted name to accomplish the custom website design requirements from 9+ Years'.
More than 100 clients from 25+ Countries like India, Australia, Brazil, UK, UAE, USA, Singapore, Spain, are using our web design, internet marketing, psd to html, social media, corporate branding, web hosting and responsive websites services.
Hundreds of awesome projects delivered in more than 25 countries earned us the respect we proudly cherish today.
Hurry, this special price is for limited period and our service is 100% guaranteed, no risk.
I would highly recommend UniqueClick to anyone that requires exceptional work to be completed. It's been a very enjoyable process for a large and complicated project.
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Uniqueclick was doing his job in a very professional manner. Highly recommend him. Thanks!
A quick and seamless project. Great work and design. Followed brief perfectly. Lovely to deal with.
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Most of the time, a meeting isn't necessary and the work is completed with only discussions over email. We upload our work to private websites that only clients can access. We then work with you to discuss the site and review changes. The site isn't made available to the public until it looks and works exactly as you wish.
The time it takes to build a site depends on the complexity of the site. If you have a deadline, we will work hard to meet it. The most common cause of delay is the content (text/images) from the client.
Please provide all text, photos and graphics in electronic form. For example, the output from a digital camera, Microsoft Word or a digital scanner. If this is not possible, we can help get your information ready for the website.
If you are not satisfied with the initial design, we will work with you further and come up with another design. Our aim is to design a website that you are completely happy with.
The cost of a website varies depends on its complexity. We are happy to discuss your requirements and provide a quote at any time.
We give as much latitude as we possibly can. After you've identified the general format and color schemes of your preferred design, we'll complete up to 2 rounds of major revisions at no charge. All minor revisions are provided at no charge.
No. We discuss your website requirements and price accordingly. Once you have placed your order with us, we will adhere to our quoted price.
For small projects, 50% of the estimated fee must be provided prior to beginning work, with the remaining payment due upon completion of the project. For larger projects, an initial payment representing one-third of the estimated total fee is required. The second payment is required upon the halfway point, with the final third due upon completion of the project.
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We would be more than happy to answer any and all questions you might have. Give us a call, shoot us an email, or drop in for some wonderfully brewed coffee
Email : email@example.com

subset: commoncrawl


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Estate Planning is a Matter of Life and Death… Make Sure You Get out of the Weeds
It is natural that you'd rather plan a fun family vacation than your funeral. I hear this frequently. But while your family will enjoy the memory of family vacations for years to come, they'll also remember if you leave them without the protection of an estate plan. Studies show that as more than half of Americans die with no will or estate plan in place. That involves many families who must cope with not just grieving the loss of a loved one, but the financial and legal burden of untangling an estate.
Kiplinger's recent article, "How Wills and Trusts Work, and Where to Start," suggests that you think of estate planning in two basic categories: the death side of legal documents and the living side of legal documents.
First, if you don't have a will, you will die "intestate." The state's laws will determine who gets your property. The second and far better option is drafting a valid will with the help of an estate planning attorney like DANIEL T. FLEISCHER. Don't try to create a valid will on your own. The details are too important. A will must go through probate. While some people may contend that probating a will isn't a big deal in their state, it may be a big deal depending on your circumstances. I agree, probating a will is not a big deal in Florida, however you do want to minimize what assets are subject to probate.
What about trusts? The purpose of a trust is to avoid probate and make it easier on the remaining spouse and heirs to settle your affairs. A revocable living trust can be an excellent choice. Part of this includes having the names removed from the assets and retitled into the name of the trust. The grantor (creator) of the trust is still in control as the trustee. The trust "owns" the assets, but you still make all the decisions for assets in the trust.
A trust isn't the solution in all situations. A will can be perfect for families who don't own a home or any real property assets. Banking accounts or investment accounts have to be distributed. You usually can make those P.O.D. (pay on death) or T.O.D. (transfer on death) accounts to simplify things.
The "living documents" are valid while you are living and they die when you do. These living documents create a list of people in order of those you want to make decisions on your behalf if you are incapacitated. The documents protect your family from having to go to court to be appointed your legal guardian, which can be expensive and time-consuming. The basic living documents are:
- Durable power of attorney for finance;
- Medical power of attorney with a HIPAA waiver;
- Living will;
- Guardianship document for adults, which appoints a guardian of your person and a guardian of your estate;
- Declaration-of-intent document stating that you intended to put all your titled and non-titled assets into your trust; and
- Certificate of trust stating the trustees and successor trustees.
Most people don't want to leave their families with an expensive, stressful and time-consuming mess. However, procrastinating because they don't expect to die soon (even in their 80s!) or they'd rather not deal with their mortality, can lead to an added layer of hardship for your family. Set up an appointment with DANIEL T. FLEISCHER today!!!! Your family will be grateful that you did.
Do you live in Miami-Dade, Broward, or Palm Beach counties in Florida? Laws are constantly changing-- has your estate plan been reviewed in the last 2-3 years? Call me (954-888-1747) right away for peace of mind. I can help! Please visit www.411probate.com for valuable information.
- My practice is exclusively estate planning and probate,
- I have prepared numerous estate plans in 16 years of practice,
- I have administered estates and trusts through Probate all over Florida,
- I am a Certified Financial Planner Professional™, and
- I am here for YOU today and there for your FAMILY tomorrow.
Reference: Kiplinger (May 2017) "How Wills and Trusts Work, and Where to Start"


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Over Sh1.6bn might have been lost in NYS scandal, probe team tells Uhuru
Thursday February 18 2016
The amount of money lost in the National Youth Service scandal could be as much as Sh1.6 billion, according to a new report seen by the Nation.
The report reveals that further investigations into the NYS scandal had shown that hundreds of millions of shillings were paid to 15 additional companies beyond the Sh791 million already reported.
The report, the outcome of a meeting chaired by President Uhuru Kenyatta at State House in Nairobi on Monday, shows that the Ethics and Anti-Corruption Commission (EACC) was investigating payments amounting to Sh869 million to the additional companies.
This brings the total of the amounts suspected to have been paid out irregularly to Sh1.66 billion.
The report was compiled by a team comprising representatives from the Office of the President, the Office of the Director of Public Prosecutions, the Directorate of Criminal Investigations (DCI), the Kenya Revenue Authority and the Asset Recovery Agency.
The report also says that the Central Bank of Kenya had fined Family Bank Sh1 million for its role in handling the NYS money.
In the face of claims by one of the business people facing charges arising from the scandal that Mr Joseph Mugwanja, the head of the Banking Fraud Investigations Unit (BFIU) of the DCI, had received Sh80 million to influence the investigations, the multi-agency team has placed all investigations under its technical team.
"To avoid duplication, all NYS investigations to be undertaken under MAT. In this respect, EACC and BFIU shall henceforth share information regarding the ongoing NYS investigations. Further investigations to be conducted jointly," the reports recommends.
The team warned that parallel investigations by the BFIU could not yield the desired results.
"It was noted that parallel investigations are also being undertaken by the Banking Fraud Investigations Unit and this might negatively impact on the outcome. The BFIU shall henceforth be incorporated to the MAT technical team," it said.
The EACC had cleared former CS Anne Waiguru of any wrongdoing in the NYS scandal in a letter sent to Chief of Staff and Head of Public Service Joseph Kinyua.
However, it has reopened investigations following a damning revelation by Ms Josephine Kabura Irungu— in an affidavit filed in court on Monday — that Ms Waiguru was aware of what was going on at the NYS and condoned irregular payments.
Ms Irungu is expected to appear before the EACC on Thursday at 10am for grilling.
She alleged that Ms Waiguru received a share of the money in question and that the former CS played a key role in devising devious schemes to divert public attention and mislead investigative agencies from getting to the bottom of the scandal.
Ms Waiguru has denied the allegations, saying she has never met Ms Irungu and that the claims were meant to derail her political ambitions. Ms Waiguru has indicated that she will vie for the Nairobi governor's seat in next year's elections.
Besides Ms Irungu, eight other business people are in court over the NYS scandal, while 11 are facing the prospect of losing their property in a move initiated by the Asset Recovery Agency.
In its report, the multi-agency team said it had also initiated investigations into four more cases involving the NYS, amounting to Sh321.5 million.
The cases involve Sh302 million paid for consultancy services for the youth, women and people with disabilities programme, Sh15 million meant for the construction of NYS camps, Sh4 million for the Naro Moru retreat and Sh500,000 to repair a vehicle by one of the directors.
At the same time, President Kenyatta has ordered that all NYS projects be stopped until investigations into the use of funds have been concluded and suspects prosecuted.
"The President has said that all NYS work will resume in March after the conclusions of investigations into the scandals," said his spokesman, Mr Manoah Esipisu.
Meanwhile, the Independent Policing Oversight Authority yesterday said it had opened investigations against two police officers who were handling the NYS case.
The complaint was filed by Ms Charity Wambui, the mother of Mr Ben Gethi, one of the suspects.


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Blue Hills Residences - Sales Office
125 Berwick-Cranbourne Road
Cranbourne East, 3977
Looking for a new Lifestyle?? Look no further than the beautiful Blue Hills Residences. Everything you need on the one site, including: Workshop, 2 Heat Pools, Spa, Tennis Court, Bowling Green, Bus Service, Country Club with Restaurant, Shop, Hairdresser and much...
RETIREMENT… That is the first and last time you will hear that word from us. Blue Hills Residences Pty Ltd is the dream of Judy Clarkson(pictured) and her family; to purpose build a residential facility for people entering the time of their lives –...
Imagine doing everything you love right where you live. Enjoy a hit of tennis, a social bowl, a swim or even some art and craft; it's all just metres from your door. Be involved as much as you like, or simply join in when you...
Join the Community Making the move to Blue Hills is the easiest move you will ever make. Visit our new FOR SALE link in the top menu to see what great opportunities currently exist at Blue Hills Residences. You may also like...
About 3 weeks ago from Blue Hills Residence's Twitter via Facebook
Last month from Blue Hills Residence's Twitter via Facebook
betterhealth.vic.gov.au/heat betterhealth.vic.gov.au/heat
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New year with Deja Vu
About 2 months ago from Blue Hills Residence's Twitter via Facebook


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They'll get the wrong idea if you so much as talk to them, and they'll be happy to just get homemade cookies from you. So they don't have to taste good. — As long as he knows you tried your best, wouldn't that sway his mind?
-Hikigaya Hachiman, My Teen Romantic Comedy SNAFU
That quote from the very first episode of My Teen Romantic Comedy SNAFU was stuck in my head ever since I found out Yui Yuigahama's birthday was the 18th of June. While Hikki is talking about the simplicity of teenage boys, I feel it can definitely be turned around the other direction as well. As such, I decided to bake a cake for Yui. There's no secret as to how important SNAFU is to me, so I wanted to pay it back somehow.
Michael felt the same way, so he also made a cake (below) and we both decided to collaborate on the celebration by recording a cover of Hello Alone, the ED for SNAFU, both in it's original form and the ballade sung by Yui herself. We perform both in English and Japanese, with the English lyrics being written by the me (based on the original Japanese ones). We're no professional singers, we just did it for fun, so please enjoy our little single below;


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About Me
- Jez Bragg
- Welcome to my blog which I hope to develop with some interesting material on ultra running both on the trails and road including reports on races and interesting training runs, views on kit and equipment as well as anything else I find of interest. I love running for adventure, opportunity and well being. Enjoy!
Thursday, 24 July 2014
Western States 100, 28 June 2014
This has done the rounds on Twitter & Facebook already, but just in case you missed it here's my write up on Western States.
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Georgia Residency Rules
Universities in Georgia, to generalize, are on the difficult side of the spectrum when it comes to earning in-state residency. But importantly, there are some harder and easier universities within the system and even the hardest is do-able for the right person.
The primary difficulty arises from Georgia's presumption that if you move to Georgia, then you're there primarily to attend college. If you can't overcome that presumption, then you're out of luck. For almost all universities in Georgia, that presumption is impossible to overcome if you go to school at all during your domicile period. There are some exceptions, but in general if you are prepared to take a gap-year and work for one year either before your freshman year or after, then it may be possible for you.
On the good side of things, you typically will not be required to be 100% financially independent; you can have some help. But you also must be prepared to work in Georgia and earn a not-insignificant amount of money.
Georgia Reciprocity Agreements:
Georgia is a member state of the Academic Common Market which is a limited regional reciprocity agreement among select Southern States. Favorable college tuition rates in nearby states are sometimes possible through reciprocity although many restrictions apply. There are also some special agreements between some Northern Florida and Southern Georgia counties and colleges-for those you will want to research your university in question.
See Tuition Reciprocity Agreements Explained
See Tuition Reciprocity Agreements Overpromise & Under-Deliver
University-Specific Overviews Below:
Georgia state regulations concerning in-state status are interpreted wildly differently for different universities. That being the case, it behooves you to figure out how those state rules are actually interpreted at the college in Georgia that you're interested in. Below please find links to in-state overviews for select universities in Arkansas. If your college is not highlighted, then please consider buying a University-Specific Nutshell Report or see all available university-specific in-state overviews.
Popular Universities in Georgia:
Armstrong Atlantic State University-Savannah
Georgia State University-Atlanta
Georgia Tech-Atlanta
University of Georgia-Athens
Other Universities in Georgia:
Albany State University-Albany
Clayton State University-Morrow
Columbus State University-Columbus
Dalton State College-Dalton
Fort Valley State University-Fort Valley
Georgia College and State University-Milledgeville
Georgia Gwinnett College-Lawrenceville
Georgia Regents U-Augusta
Georgia Regents U Medical College of Georgia-Augusta
Georgia Southern U-Statesboro
Georgia Southwestern State University Americus
Kennesaw State University-Kennesaw
Middle Georgia State College-Macon
Savannah State Georgia-Savannah
Southern Polytechnic State University-Marietta
University of North Georgia-Athens
University of West Georgia-Carrollton
Valdosta State University-Valdosta
Every College in Georgia Is Unique
Just as every state has its own in-state regulations, each college in the state has its own interpretations. Learn everything important about how to actually earn in-state tuition at the university in Georgia you're most interested in.


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Additional Dreamer Options
Here is where you will find a list of "Add Ons" for your package. Some parents find it helpful to understand that they have additional support and options available if they hit a bump in the road while working with their child to develop independent sleep strategies.
Additional Text Message Support:
Ten more text messages to tackle a particular struggle (going on a vacation, dropping a nap, etc.) with your sleep plan
Investment: $79
Additional Email Support:
One more week of unlimited e-mails to ensure that your questions are answered and your sleep plan is on track
Investment: $89
Additional Phone Call Support:
A 30 minute phone call to discuss your new situation/challenge and to get things back on track
Investment: $99
In-Home Travel Cost:
Packages include an in-home consultation within 20 miles of Ladera Ranch, CA; if you would like to have your consultation in your home (instead of over the phone or video chat) and you live 20-40 miles from Ladera Ranch, then an additional travel fee will be charged
Investment: $49
(Please e-mail or call for travel fee information if you are located more than 40 miles from Ladera Ranch, CA)


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Sunlight In The Middle
Mariann Kovats 6/24/2010 6:26:10 AMBeautiful capture, Brian! I love the sunlight from the middle! #1281759
Tammy Espino 6/24/2010 6:09:08 PMLovely~ #8715513
Kate Jackson 7/4/2010 12:39:18 AMFantastic detail in the white flower Brian. Love that sunlight in the middle!
Gail Vitikacs 7/12/2010 5:02:36 PMBreathtaking...so beautiful! Congratulations on this wonderful finalist! #8753477
Randy A. Myers 7/12/2010 5:59:16 PMCongratulations. Well done. #8754043
Kathy Cline 7/12/2010 6:47:04 PM
Sherran Andersen 7/12/2010 7:33:35 PMCongrats Brian... I love dogwood. #8754831
Sam Britt 7/12/2010 8:18:18 PMCongratulations on this very pretty finalist, Brian! #8755061
Carrie M. Groseclose 7/12/2010 11:16:11 PMHuge Congratulations Brian on this gorgeous Finalist. I love how the light is shining through! #8755724
Jeana Clark 7/13/2010 8:39:26 AM
Sharon Sawyer 7/13/2010 12:43:36 PMWhooo Hoooo!!! WTG, Brian! I love how you captured the light in the middle of the flower. Well done. HUGE congratulations on this well deserved finalist and all the best in the next round. #8758633
Tammy Espino 7/13/2010 2:41:53 PMCongrats on this gorgeous finalist!! #8759035
Mark Dodge 7/13/2010 3:01:02 PMStaggering clarity, Brian. Excellent work! Congratulations on your finalist here. Definitely deserves it :o) #8759139
Renata Gusciora 7/14/2010 7:19:02 PMCongratulations on your lovely finalist,Brian!!!:-) #8764000
Sam Britt 8/13/2010 5:47:29 PMCongratulations, Brian!! #8828267
Datha Y. Thompson 8/14/2010 8:42:37 AMCONGRATULATIONS again Brian!! Beautiful ;) #8831409
Merna L. Nobile 11/18/2010 7:57:39 PMI would personally like to congratulate you on your winning photograph for October, 2010 and wish you a Happy Thanksgiving. Merna Nobile #9049889
Kousa Dogwood Flower.
200-400mm VR Lens, ISO 500, f/5.6, 1/160s
Category: Flowers
Exif: F Number: 5.6, Exposure Bias Value: -0.67, ExposureTime: 625/100000 seconds, Flash: did not fire., ISO: 500, White balance: Auto white balance, FocalLength: 360.00 mm, Model: NIKON D700
Uploaded on 6/24/2010 5:27:57 AM
Camera: Nikon D700 Digital SLR Camera
Copyright for this gallery photo belongs solely to Photographer. Images may not be copied, downloaded, or used in any way without the expressed, written permission of the photographer.
Log in to follow or message this photographer or report this photo.


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Yoe Bo, E15 1BG
- 020 8519 2020
- Unit 1, Gerry Raffles Square, E15 1BG (streetmap) (osm) (gmap) (bingmap) (streetview)
- yoebo.co.uk
A Chinese all you can eat buffet restaurant in Stratford near the theatre.
secretlondon went there with two others one Wednesday evening in March 2009. The only food on offer is the buffet which costs £12 a head (less for children and less at lunchtime, but £13 a head Friday and Saturday evenings). Chinese tea is £1.50 a head and tap water is 20p (and on the drinks list). They are licenced.
There is one large H-shaped buffet with some fruit in the middle. There wasn't much for vegetarians. They also had some desserts and other things on their own to the right. We didn't spot these until we were leaving.
The food was okay but it was still £12 a head for a Chinese buffet in outer London.
Last visited by secretlondon, March 2009.
OS X co-ord: 538808 OS Y co-ord: 184530 (Latitude: 51.542603 Longitude: 0.0002544)
Last edited 2014-02-12 11:06:37 (version 6; diff). List all versions.


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If you have questions about payment, product download, pricing, status of your order or general inquiries, please click below for further assistance.
If you have purchased one of our products and require technical support for an issue not covered in our FAQs, please click below for further assistance.
If you are a journalist and are interested in reviewing our products and RPG Maker or have other media enquiries, please click below to access our press section for the latest press releases, screenshots and logos.CLICK HERE TO CONTACT OUR PRESS OFFICE


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"In Turkey, the reign of Sulaiman was at the same time an age of victorious warfare, of widespread construction and of substantial legislative activity. Sulaiman bore the title of Kanuni, or law-maker, indicative of a revival of law studies and the existence of a special class of jurists in the states under his rule and above all at Constantinople. His legal code so suecessfully regulated the judicial machinery that it was said that Henry VIII of England sent a legal mission to Constantinople to study its workings. His Kanun-name is to the East what the Justinian Code is to the West and the Recopilacion de las Leyes to Spain. All the legal machinery established by Sulaiman in Hungary was the work of the jurist Abu'l-Su'ud; such a major achievement of legislation was it on the question of property that many of its detailed provisions remain in force to the present day. And the jurist Ibrahim Al-Halabi, author of a handbook on legal procedures, the multaka can be ranked alongside the most eminent western
jurists of the sixteenth century."
From Fernand Braudel, The Mediterranean and the Mediterranean World in the age of Philip II, Volume 2, pg. 683, Fontana/Collins, 1973.


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Without a full set of teeth, your oral health and appearance may decline. Sagging facial muscles can make you look older, and the inability to chew foods or maintain a healthy diet can cause overall health issues. Speaking may become difficult, causing a limit in social interactions. Also, missing teeth prompt remaining teeth to shift out of place in an effort to compensate. If you're missing many or all of your teeth, dentures may be the best treatment option. At Wager Evans Dental, we want to help improve your quality of life by replacing missing teeth and improving the health and function of your smile.
Partial Dentures
A partial denture replaces multiple teeth, not all teeth, on the upper or lower arch. Like a puzzle piece, partial dentures fill spaces so that your smile looks good and functions properly. Partials usually attach to neighboring teeth with inconspicuous clasps. The artificial teeth in a partial will be designed using impressions of your teeth and employing lifelike dental materials. Partials will also help prevent the misalignment that often accompanies tooth loss, keeping your smile whole, even, and healthy.
Full Dentures
Full dentures are a complete row of prosthetic teeth attached to a gum-colored base. A full denture can replace all upper and/or lower teeth. Your dentures will be designed based on Dr. Evans or Dr. Wager exact specifications. Once the dental lab has completed your prosthetic, we will check the fit and make any necessary adjustments. Once in place, your dentures will allow you to enjoy a wider range of foods and smile without hesitation. If you have any questions about either our full or partial dentures, then don't hesitate to contact Wager Evans Dental today.
Implant Dentures
Adhesive creams or natural suction are typically used to secure full dentures. Over time, the ridges on your jaw will wear down from friction, which can cause dentures to loosen, wobble, or slip out of place. In this situation, dentures must be relined and adjusted. To stabilize your dentures more securely, you might consider implant-retained dentures. With mini-dental implants, dentures snap onto small posts anchored in teeth sockets. You won't need to use adhesive to hold the dentures in place and implant dentures reduce the risk of slippage when eating or speaking. Mini-implants provide a solid, reliable, and attractive smile to enjoy.
What Kind of Denture Do You Need?
If you are interested in receiving dentures, then schedule a consultation with Dr. Evans and Dr. Wager. Click here to request an appointment, or call our office today at (775) 829-7700. Located in Reno, NV, we proudly welcome residents from South Meadows, Double Diamond, Caughlin Ranch, Donner Springs, Meadowood, Sommersett, Arrow Creek, and all surrounding communities.


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|Tropical Tidbits
Click to enlarge
Top left: PNA Forecast
Top right: NAO Forecast
Bottom left: WPO Forecast
Bottom right: EPO Forecast
A quick refresher on the PNA, NAO, WPO and EPO...
The Pacific North American index involves what the atmosphere does in the northeast Pacific and the western coast of North America. When we see a stormy pattern in place over these regions, we call such a pattern a negative PNA, due to the below normal height anomalies in this region. In a similar sense, when high pressure dominates that same region, we call that a positive PNA. A negative PNA will bend the jet stream to give the storms to the Plains and the Deep South regions, frequently initiating high pressure system formations over the Central US. A Positive PNA will bring about an opposite response to high pressure (HP) over the West, and will have the stormy pattern evolve over the East US.
The North Atlantic Oscillation involves the presence of a high pressure system over Greenland (negative NAO) or the presence of a low pressure system over Greenland (positive NAO). In the negative NAO, the jet stream will buckle into the Northeast to allow storms and cold to thrive in that region. The positive NAO denies this region any of these benefits.
The WPO (West Pacific Oscillation) and EPO (East Pacific Oscillation) are very closely related. In the negative phase of the WPO, a strong ridge exists over the Bering Sea, which can allow for sustained cold weather in the Central and Eastern United States. The negative phase of the EPO gives similar results, though the ridge is positioned in the Gulf of Alaska instead. The positive phase of both the EPO and WPO see warm weather prevail in much of the US, as stormy weather replaces the ridges in each respective region.
The positive PNA and negative EPO have worked in tandem to indicate this strong ridge blossoming into the Gulf of Alaska and general northeast Pacific. In the upcoming couple of weeks, model guidance is indicating we see the ridge shift further offshore to the west, as the PNA dip from positive to negative shows. Over time, ensemble guidance is telling us that this ridge could keep shifting west, resulting in the EPO actually moving positive for the first time in a while. The positive EPO signal, however, is weak, and will have to be watched for a 'false positive', both literally and metaphorically. If this forecast does verify, we can expect a warm-up in the early days of March.
|Tropical Tidbits
You'll notice I haven't used tropical forcing in this post; that's because the ridge in the West is just so overpowering that the Madden Julian Oscillation can't do much of anything.
To summarize:
- A cold pattern is expected to round out February.
- A warm-up is expected for the last days of the month into early March.
- The first half of March should be characterized by a generally average to slightly below-average temperature pattern. There are hints of some stormy weather in early March.


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Have a heart
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The Unique Mind of Joseph Bismark
Who could ever achieve the greatness of Joseph Bismark? I never knew of this leader before, but after reading an article about him on the blog, "We Don't Love These Hogues," I can say he is truly an inspirational being. Raised in the mountains of the Philippines among monks, he learned at a young age what most of us would learn later in our life. Success can be achieved by everyone. When he returned back home in Malaysia, he applied that knowledge in the business industry and eventually worked his way to do wonders.
One of his best accomplishments was being the managing director for Qi Group. Applying his strong sense of spirituality into his work, he managed to do many things there, such as inspiring the creation of the Corporate Social Responsibility arm of the Qi Group. As for more modern news, the Qi Group opened a college in Malaysia called Quest International University Perak. At this astounding university, students are informed of the impacts of humans on the Earth which basically was a part of Bismark's spiritual belief. Joseph Bismark is very different from any other everyday businessman.
Most businessmen tend to be very arrogant and egoistic, but not Bismark. He is a humble being and even as a boss, he still respects his employees. Even when they are not doing exceptional work, he encourages them to work harder to achieve almost perfection with no harsh words. As stated above, he believes everyone can reach success.
He took his spirituality and combined it with the corporate world thus creating a powerful sense of brotherhood and humbleness. He brought uncountable businesses to follow his own beliefs and numerous people to follow in his footsteps. I believe Bismark is someone that not just businessmen, but everyone should hold in the highest esteem. He is a quintessential of how far a person can go in life by working to the best of their ability and just being optimistic.


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BEIJING -- China Construction Bank Corp. reported third-quarter net-profit results that missed market expectations, as the state-run bank and other lenders grapple with China's slowing economy and Beijing's efforts to liberalize financial markets.
The country's second-largest bank by assets reported Sunday that net profit for the quarter rose 9.4% to 56.8 billion yuan ($9.34 billion), lower than the average forecast of 57.69 billion yuan among four analysts polled by Thomson One Analytics.
The third-quarter profit growth for China Construction Bank, the first of the state-run banks to report quarterly earnings, marked a slowdown from its 13% growth in the first half of the year and the second-slowest growth in more than four years.
The profit gain was driven by a rise in net-interest income, which accounted for more than 70% of the bank's operating income. Third-quarter net-interest income rose 8.2% to 98.85 billion yuan.
Though they remain among the most profitable in the world, Chinese banks have been facing mounting challenges, including narrowing interest margins and rising bad loans, amid a slowing economy and Beijing's push to liberalize government-controlled interest rates.
China Construction Bank's net-interest margin, a measure of the profitability of lending, was 2.71% at the end of September, unchanged from the end of June, but was lower than the 2.74% the bank posted a year earlier.
An anticipated push by Beijing to further liberalize interest rates is expected to further hurt banks' margins, as funding costs rise and competition for customers increases.
In July, China's central bank scrapped a floor on lending rates, a move toward promised overhauls. On Friday, the central bank introduced a market-based loan prime rate in a bid to eventually replace official benchmark interest rates.
The regulatory changes come as growth in the Chinese economy, the world's second largest, has slowed from double-digit percentages in previous years to 7.8% year on year in the third quarter. That has triggered concern over rising bad loans, especially after Beijing's huge 2009 stimulus plan that was supercharged by bank lending.
Still, the ratio of nonperforming loans to overall loans at Chinese commercial banks stood at 0.96% at the end of the second quarter, unchanged from the first quarter.
China Construction Bank's bad-loan ratio declined to 0.98% by the end of September from 0.99% the pervious quarter. Its provision coverage ratio for bad loans was 267.9% at the end of the third quarter, down 3.41% from the end of last year.
The five biggest Chinese banks wrote off a total of 22.1 billion yuan of bad loans from their books in the first half of the year, in a bid to address a surge in bad assets and prepare for a fresh wave of defaults as the economy slows.
-- Write to Grace Zhu at firstname.lastname@example.com
Subscribe to WSJ: http://online.wsj.com?mod=djnwires


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One muted winter afternoon in 2006, Pete Yorn awoke from a sleeplike trance convinced that recording a duets album with Scarlett Johansson would save him. Unlike most people who have this thought, Yorn actually had the actress's number. The two were acquainted through Yorn's brother, an entertainment lawyer Johansson has worked with since she was a teenager. So Yorn texted her. Johansson hadn't yet recorded or even announced her own 2008 album, the slight but likable Anywhere I Lay My Head, a collection of Tom Waits covers. But it turned out to be an opportune moment for both.
Yorna singer-songwriter who broke out in 2001 with the CD musicfor themorningafterhad been touring nonstop for months behind his 2006 album, Nightcrawler, and was having a tough time reentering normal life. Johansson was weathering the transition from indie ingenue to grown-up movie star. Though she wasn't feeling as emotionally vulnerable as Yorn, they shared a common sense of being in between stages in life. Both were restless, had time on their hands, and were ready for a new challenge.
In the early weeks of January 2007, the pair met at an unassuming studio in Los Angeles and recorded Break Up, a collection of boy-meets-girl-girl-ruins-boy duets in the tradition of Serge Gainsbourg and Brigitte Bardot, Lee Hazlewood and Nancy Sinatra, and Lou Reed and Nico. Intuition spawned the idea and improvisation fueled the album's production; Yorn, 35, and Johansson, 24, felt the magic would come from the willfully haphazard way it was recorded.
Nearly three years, two solo albums (one his, one hers), a Woody Allen movie (hers), and a wedding (also hers) later, and Break Up is finally coming out (September 15 on Atco; the first single, Relator, was released in May, on vinyl and iTunes). The resulta delicate, elegiac album of jangly country-tinged songs about love gone wrongis like the perfect mix tape for anyone haunted by an ex.
This record was inspired by a dream?
PY: Sort of. I was home in Jersey for Christmas break after touring, and I just freaked out. I had all this anxiety, and I wasn't sleeping. One afternoon, I curled up on the bed, trying to pass out. I was jolted awake by one of those Oh my God, I left the oven on or I'm late for school feelings. Except it was I gotta make a duets record. This quick flurry of thoughts came together over a few seconds: Duets record, yeah! Guy-girl thing, yeah! Serge Gainsbourg, Brigitte Bardotthey're cool, I love them Brigitte Bardot Scarlett Johansson! Just a total chain reaction.
SJ: I don't even think he knew that I could sing. I think he asked me after I said I would do the album.
PY: I had a phone number for her from a few years earlier. I texted her not even knowing if it was still her number.
SJ: It didn't strike me as strange to get his message. I hadn't spoken to him in a long time, but we've always had a friendship where we catch each other every few months and it's like no time has passed. That said, we had never talked about music.
PY: She hit me back like, That sounds cool. Let's do it! Then I was like, um, now what? Before you knew it I was writing songs and we were in the studio making the tracks. Two months later, we had Scarlett come in for two afternoons.
Scarlett, at that point you hadn't made a record before. What gave you the confidence to step into a recording studio?
SJ: I've been singing for my whole life. When I was a kid I wanted to be on BroadwayI was a real jazz-hands kind of girl.
PY: Her voice is like mineit's not technically perfect, it's quirky.
SJ: It sort of happened by fluke that we could actually sing together.
PY: When Scarlett came in, she gave the album a huge presence without even trying. She played the part of the female perspective in the songsshe brought an aloofness I hadn't accounted for.
Did Pete send you the songs as he wrote them?
SJ: No, I had no idea what the songs were going to sound like when I got to the studio.
PY: The album cover is a photo my cousin took of me showing her a song right before we recorded it. If she wasn't singing and I wasn't singing, that's what we were doing, sitting there with the guitar, like, Okay, here's the song. Learn it.
SJ: A lot of the work we did was just collaborating on the harmonies. Finding them and figuring out who would sing which piece. I didn't expect to sing I Am the Cosmos by myself. But Pete preferred it with a female voice, so that became a bit of a solo.
PY: She ended up having a deeper voice, so I took a lot of the light, higher stuff.
SJ: Pete had mentioned the Serge and Brigitte recordings, and I had my own idea that it was Nico-ish, but it wasn't like we talked it all through. The songs are quite literal, so we weren't trying to find the meaning in them; the meaning is very much there.


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Hey guys, so... has anyone got/played this?
I must admit, I first had a go and though, hmm I don't know. But I listen to a lot of music and well, I think i love it. its only a small game, I mean there's not much to do, but I can see me getting very addicted.
I love the fact that you upload your scores afterwards, so basically you are playing against loads of other peoples scores.
If anyone is interested in getting it, or has it let me know, we can sort something
edits:sorry misstyped AudioSurf


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Following a brand overhaul by Boccalatte, Never Sit Still were approached by MAAS (Museum of Applied Arts & Sciences) to create a motion system and guidelines based on their new visual identity.
We closely followed their comprehensive style guide to bring the brand elements to life, developing a motion system that would be flexible and easy to use for the in-house production team.
Motion templates were created for both the 'MAAS master brand' and the 'Recollect exhibition series'. The files we provided to MAAS included titles, end frames and supers that could be used with live-action video footage. PDF guideline documents were also created, explaining how to use the motion templates.


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It is better to frankly acknowledge that Donald Trump has fundamentally re-written the rules about what it means to be a Republican than to pretend that he still, somehow, represents "conservative" thought.
So argues R.R. Reno, in a Sunday Review piece in The New York Times. Republicans are now the "America First" party: Get over it and embrace it, he says.
Reno begins by enshrining his Reaganite conservative credentials. (Reno is the editor of the thoughtful social conservative magazine First Things):
Of all the people still trying to process Donald Trump's rise to the presidency, perhaps none are more confused than my generation of conservatives, who came of age under Mr. Reagan and drank deeply of that old orthodoxy. We are, by now, the establishment — the senators, governors, think-tank presidents and columnists who, until Mr. Trump came along, got to define what "Republican" and "conservative" meant. My cohort simply cannot accept that Mr. Trump has taken away that coveted role and revolutionized not just our party, but also the very terms of the American political divide.
But accept it they must. Unlike Republican candidates like Cruz, Trump won the nomination because he did "not adopt Ted Cruz's strategy of trying to revive the rotting flesh of Reaganism."
The problem, according to Reno, is that a blithe culture of globalism had spread to both parties, Democrats as well as Republicans. He reminds readers about Obama's promise, in his 2013 inaugural address, that innovation and mobility that would allow our nation to thrive in "this world without boundaries."
Obama, of course, thought of that as a good thing. Didn't everyone?
Here's the core of what Reno says he now accepts:
Rather, we need to become much more skeptical of post-national ways of thinking. For too long a globalist utopianism — Mr. Obama's happy, peaceful and inclusive world without boundaries — has tempted us to neglect one of the fundamental tasks of political leadership, which is to promote the kind of national solidarity that binds a country's leaders to its people.
Globalism poses a threat to the future of democracy because it disenfranchises the vast majority and empowers a technocratic elite. It's a telling paradox that the most ardent supporters of a "borderless world" live in gated communities and channel their children toward a narrow set of elite educational institutions with stiff admissions standards that do the work of "border control." The airport executive lounges are not open and inclusive.
John Q. Public is not stupid. He senses that he no longer counts. And he resents the condescension of globalist elites, which is why Mr. Trump's regular transgressions against elite-enforced political correctness evoke glee from his supporters.
There is much to say about, and criticize about, in Reno's view. Globalism should never be an excuse for elitism, or a sneering about traditional values that is too often exhibited by the cultural cognoscenti.
But this is the point at which the political project of our day gets exciting: Finding ways to show how "The Wealth of Nations" does indeed benefit the citizens of each nation. Liberal humanism may desperately be in need a new, catchy label for the hashtag era. We now need political leaders who draw the linkage between the philosophies of peace, trade, and the worth every of individual. That, indeed, is the very embodiment of the American national ideal.


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This is Cladun: This is an RPG's English trailer.
This is where I first learned about the game's built-in screenshot function. Handy!
Just an example of the game's strange sense of humor.
It's gushing!
Inside one of the earlier dungeons.
One of the earliest new characters you meet not only parties with you, but also runs a convenient item shop.
The Magic Circle. Don't try to make sense of it from this screenshot, it'll just make your brain hurt.


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By Amy Reid
Lab tables cluttered with artifacts, scales, digital calipers, coffee cups, Excel spreadsheets and to-do lists that seem to get longer-not shorter. The analysis phase of an archaeological project can look more like chaos than science, but in reality there is a method — or methodology rather — to the madness. This post aims to describe the various analyses we have conducted and are planning to perform on the data recovered from our 2014 excavations at the Spring Lake Site. You will also get a sneak peak at some of the results available to date.
Like most systematic archaeological projects, my colleagues and I identified a series of research questions and objectives before the excavations even began. All of our work planning, excavating, sampling and processing (washing, sorting and cataloging) the artifacts was performed with 3 research domains in mind:
- chronology and culture historical reconstruction;
- geoarchaeology, site formation, and environmental reconstructions; and
- better defining the Calf Creek horizon at Spring Lake, including its technological characteristics and how subsistence-related behaviors changed during this brief interval of big game (bison) hunting at the end of the Early Archaic, ca. 5750 to 6000 years ago.
The analyses planned to address these questions cannot begin until there is 1) a complete catalog of artifacts, ecofacts and samples collected, and 2) a preliminary list of Analytical Units (AUs). AUs are a set of discrete, intact deposits of sediment that represent recognizable periods in the occupational history of the site. They are chronological in nature, and therefore rely on the law of superposition, cross-dating principles, typology, and context. Ultimately, the AUs will be a series of excavated proveniences that can be associated with a given time period that will provide the basis for all detailed, context-specific analyses that will be conducted. This step is vital for assessing behavioral changes through time. I refer to them here as preliminary because eventually the AUs will be updated and refined once we have absolute dates from bone and charcoal.
The first step in the process of defining AUs, was to take a closer look at the time-diagnostic projectile points. I went through the entire collection of points and point fragments and conservatively assigned types to the specimens that fit within existing type definitions. The typological outliers and the specimens that I could not identify with confidence (or as I have typed them, "Elton Points") were set aside to be examined by Elton Prewitt, an independent researcher and expert in Texas projectile point typology (Figures 4 and 5). Elton has graciously assisted us with typing projectile points from previous projects in and around the Spring Lake site and his help is much appreciated for the sake of consistency within our ongoing studies at this important site.
In all, twenty six different temporally diagnostic projectile point types were found during our 2014 excavations at Spring Lake, including: Wilson and Dalton of the Paleoindian (n=5); Gower, Uvalde, Bandy, Merrell, and Martindale of the Early Archaic (n= 13); Bell (Calf Creek) from the Calf Creek component (n=6); Baird (aka Taylor or Early Triangular), Nolan, Travis, Desmuke and Pandale from the Middle Archaic (n=25); Bulverde, Castroville, Ellis, Edgewood, Ensor, Fairland, Marcos, Montell, Morhiss, Pedernales, Lange, Marshall and Darl of the Late Archaic (n=59); Scallorn and Perdiz arrow points of the Late Prehistoric (n=7) (Figures 6-10).
My next step involved discussions with Jacob Hooge our resident Geoarchaeologist about which excavation units showed signs of post depositional disturbances. In other words, he gave me a list of excavated units and levels that were considered to have mixed deposits that I should therefore avoid when defining the AUs. Then, the typed projectile points that were recovered from within unit levels that were determined to have good contextual integrity were evaluated stratigraphically. Unit-levels that showed evidence for more than one time period were considered mixed or compressed and were not considered reliable for analysis. Alternately, unit-levels that did not contain any time-diagnostic information at all did not receive an analytical unit designation since it would be impossible to know whether these contain mixed or intact deposits.
Currently, the analytical units for our SLDR project include a series of lots (proveniences) that correspond with all of the general Prehistoric time periods in the existing Central Texas chronology from the Toyah phase of the Late Prehistoric at the top to the Late Paleoindian at the bottom (Figures 11 and12).
Importantly, we encountered intact Calf Creek components in both excavation blocks at similar elevations. These, alongside the Calf Creek component recorded during our 2012 data recovery excavations at Spring Lake (Lohse et al 2012) are believed to be the only intact Calf Creek components with datable materials yet documented in Central Texas!
Chronometric dating of Carbon (C14) and bone will help refine these AUs making it possible to observe a more precise scale of resolution that corresponds to intervals or phases defined for Central Texas by Prewitt 1981, 1983, 2005), Johnson and Goode (1994) and Lohse (2012) (see table). In particular, we hope to advance our understanding of the Calf Creek interval at Spring Lake. The Calf Creek horizon currently believed to have occurred during the terminus of the Early Archaic at Spring Lake. We have sent off 14 samples of carbon from the above described AUs to undergo AMS Radiocarbon dating and are patiently awaiting the results.
The faunal bone from our preliminary AUs will be identified by a faunal analyst (TBD) and then studies on subsistence at Spring Lake will be possible. Following identification, we plan to submit a selection of bison bone to be dated which will allow us to reconstruct periods of intensive focus on bison exploitation.
Another special study we are excited about is protein residue. A selection of lithic tools from our 2014 excavations at Spring Lake have been submitted for protein residue analysis, which will identify presence of animal and/or plant proteins on the tools.
We have also focused our attention on a feature that appears to be an intact cooking feature – Feature 6 (Figures 13 and 14). Feature 6 consists of a high concentration of burned rock below and surrounding an area full of burned clay. Interestingly, a dart point was found from within the feature with a potlid still in place. This point has been preliminarily typed as an Ensor, suggesting the feature may have been used during the Late Archaic time period. However, multiple lines of evidence will ultimately be used to date Feature 6 including both AMS radiocarbon dating and thermoluminescence dating. Archeobotanical analyses are also planned for this feature by way of flotation. Thanks to the help of Leslie Bush, we have already begun the flotation process for sediment samples from our AUs (Figure 15).
Check back for more posts on the results of these analyses as well as special studies on ceramics, bone and shell beads, and the possibility of prehistoric dwellings at Spring Lake.
References Cited:
Johnson, LeRoy, Jr., and Glenn T. Goode
1994 A New Try at Dating and Characterizing Holocene Climates, as well as Archeological Periods, on the Eastern Edwards Plateau. Bulletin of the Texas Archeological Society 65:1–51.
Lohse, Jon C., Amy E. Reid, David M. Yelacic, and Cinda L. Timperley
2013 Data Recovery and Analysis at the Texas State University Ticket Kiosk Project, Located at 41HY160, Spring Lake, Hays County, Texas. Center for Archaeological Studies: San Marcos
Prewitt, Elton
1974 Preliminary Archeological Investigations in the Rio Grande Delta Area of Texas. Bulletin of the Texas Archeological Society 45:55–65.
1981 Culture Chronology in Central Texas. Bulletin of the Texas Archeological Society 52:65–89.
1985 From Circleville to Toyah: Comments on Central Texas Chronology. Bulletin of the Texas Archeological Society 54:210-238.
Amy Reid is the Collections Manager at the Center for Archaeological Studies, and director of the PAST Program. She earned her Masters degree from Texas State University, and teaches a course in Archaeological Curation during the Fall semester.


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I must admit, I got a little misty when I saw this picture of Tricia and Davis. Fine, I had a tear or two but how could you not? To me, this photo oozes love, respect, friendship, all the good stuff that marriage is supposed to mean. How is it possible to capture it all in one photo? I have no freaking clue but Scobey Photography did it and did it well. And if you love this picture then you will need a tissue for the rest of the wedding.
their official website.
AND you can connect with us on FACEBOOK and join the conversation on TWITTER. We would love it if you stopped by. Please.
What else do you think love looks like? Got an image that embodies everything that love is? Well send it our way so we can share it!
Make it de Lovely,


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4 April 2017 12:46PM • 0 comments
HARARE - The clueless and desperate Zanu PF administration has come up with a raft of new taxes, further strangling its overtaxed citizens.
President Robert Mugabe's government last Thursday imposed a new 10 percent tax on tobacco sales, resulting in angry farmers threatening to withhold their crop for the current marketing season.
The farmers' response to the new tax is likely to disrupt the much-anticipated 2017 tobacco marketing season.
Impoverished Zimbabweans rank among the most heavily-taxed in the region given that they are already reeling from a wave of other additional taxes recently imposed on them, including the recent shock hike in traffic fines, the taxing of commuter omnibus operators, driving schools, hairdressers and cross-border traders.
This is against the background of government recently giving the green light for the assumption of the Health Levy — a five percent tax on airtime and data.
In addition to that, the long-suffering citizens have been paying the Aids Levy while their salaries are significantly taxed at least 20 percent through pay-as-you-earn (Paye).
Recently, Mugabe's desperate administration imposed a 15 percent value added tax (Vat) on meat and potatoes, which they later reversed following fierce outrage by consumers.
Motorists are also levied through toll fees and vehicle licences and authorities are planning to introduce urban tolling.
It appears government is trying to raise funds to meet its obligations, including paying bonuses to civil servants.
Government last month agreed to pay bonuses to its bloated workforce, following threats of industrial action by union leaders.
The cocktail of taxes is not good for the economy and reminds people of the disastrous hyperinflation of 2008.
Meanwhile, bond notes — a surrogate currency introduced late last year — continued to lose value against the United States dollar, which is now scarce on the open market. Apart from having to contend with its depleted coffers, the government is also struggling to contain rising public discontent over the worsening shortages of cash and plummeting standards of living.
Surprisingly, while government collects substantial amounts from these taxes and fees — including fines at the numerous police roadblocks — an insignificant amount is channelled towards its intended purpose with the remainder sieving through into the unorthodox but institutionalised corruption network.
Following the controversial 2013 elections, the country has seen a steady reversal of its growth trajectory.
The agricultural and manufacturing sectors in Zimbabwe are in a near-comatose state.
On the other hand, health delivery services and standards in the education sector have plummeted to alarming levels. If only the money collected was used to fund the productive as well as service sectors, the better.
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vulgar, racist, tribalistic, sexist, discriminatory and hurtful
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- Editor
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