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Despite the identification of severe acute respiratory syndrome-related coronavirus (SARSr-CoV) in Rhinolophus Chinese horseshoe bats (SARSr-Rh-BatCoV) in China, the evolutionary and possible recombination origin of SARSr-CoV remains undetermined. We carried out the first study to investigate the migration pattern and SARSr-Rh-BatCoV genome epidemiology in Chinese horseshoe bats during a 4-year period. Of 1,401 Chinese horseshoe bats from Hong Kong and Guangdong, China, that were sampled, SARSr-Rh-BatCoV was detected in alimentary specimens from 130 (9.3%) bats, with peak activity during spring. A tagging exercise of 511 bats showed migration distances from 1.86 to 17 km. Bats carrying SARSr-Rh-BatCoV appeared healthy, with viral clearance occurring between 2 weeks and 4 months. However, lower body weights were observed in bats positive for SARSr-Rh-BatCoV, but not Rh-BatCoV HKU2. Complete genome sequencing of 10 SARSr-Rh-BatCoV strains showed frequent recombination between different strains. Moreover, recombination was detected between SARSr-Rh-BatCoV Rp3 from Guangxi, China, and Rf1 from Hubei, China, in the possible generation of civet SARSr-CoV SZ3, with a breakpoint at the nsp16/spike region. Molecular clock analysis showed that SARSr-CoVs were newly emerged viruses with the time of the most recent common ancestor (tMRCA) at 1972, which diverged between civet and bat strains in 1995. The present data suggest that SARSr-Rh-BatCoV causes acute, self-limiting infection in horseshoe bats, which serve as a reservoir for recombination between strains from different geographical locations within reachable foraging range. Civet SARSr-CoV is likely a recombinant virus arising from SARSr-CoV strains closely related to SARSr-Rh-BatCoV Rp3 and Rf1. Such frequent recombination, coupled with rapid evolution especially in ORF7b/ORF8 region, in these animals may have accounted for the cross-species transmission and emergence of SARS.
Coronaviruses can infect a wide variety of animals, causing respiratory, enteric, hepatic, and neurological diseases with different degrees of severity. On the basis of genotypic and serological characteristics, coronaviruses were classified into three distinct groups (2, 20, 54). Among coronaviruses that infect humans, human coronavirus 229E (HCoV-229E) and human coronavirus NL63 (HCoV-NL63) belong to group 1 coronaviruses and human coronavirus OC43 (HCoV-OC43), and human coronavirus HKU1 (HCoV-HKU1) belong to group 2 coronaviruses, whereas severe acute respiratory syndrome-related coronavirus (SARSr-CoV) has been classified as a group 2b coronavirus, distantly related to group 2a, and the recently discovered group 2c and 2d coronaviruses (6, 8, 10, 18, 31, 38, 43, 46, 49, 50). Recently, the Coronavirus Study Group of the International Committee for Taxonomy of Viruses has proposed renaming the traditional group 1, 2, and 3 coronaviruses Alphacoronavirus, Betacoronavirus, and Gammacoronavirus, respectively (http://talk.ictvonline.org/media/p/1230.aspx ).
Among all coronaviruses, SARSr-CoV has caused the most severe disease in humans, with over 700 fatalities since the SARS epidemic in 2003. Although the identification of SARSr-CoV in Himalayan palm civets and raccoon dogs in live animal markets in southern China suggested that wild animals could be the origin of SARS (11), the presence of the virus in only market or farmed civets, but not civets in the wild, and the rapid evolution of SARSr-CoV genomes in market civets suggested that these caged animals were only intermediate hosts (24, 39, 42, 52). Since bats are commonly found and served in wild animal markets and restaurants in Guangdong, China (47), we have previously carried out a study of bats from the region and identified a SARSr-CoV in Rhinolophus Chinese horseshoe bats (SARSr-Rh-BatCoV) (21). Similar viruses have also been found in three other species of horseshoe bats in mainland China (25), supporting the hypothesis that horseshoe bats are a reservoir of SARSr-CoV. Recently, viruses closely related to SARSr-Rh-BatCoV in China were also reported in Chaerophon bats from Africa, although only partial RNA-dependent RNA polymerase (RdRp) sequences were available (41). In addition, more than 10 previously unrecognized coronaviruses of huge diversity have since been identified in bats from China and other countries (1, 3, 5, 9, 22, 27, 32, 33, 40, 46, 51), suggesting that bats play an important role in the ecology and evolution of coronaviruses.
As a result of the unique mechanism of viral replication, coronaviruses have a high frequency of recombination (20). Such a high recombination rate, coupled with the infidelity of the polymerases of RNA viruses, may allow them to adapt to new hosts and ecological niches (12, 48). Recombination in coronaviruses was first recognized between different strains of murine hepatitis virus (MHV) and subsequently in other coronaviruses, such as infectious bronchitis virus, between MHV and bovine coronavirus, and between feline coronavirus type I and canine coronavirus generating feline coronavirus type II (12, 16, 17, 23). Recently, by complete genome analysis of 22 strains of HCoV-HKU1, we have also documented for the first time that natural recombination events in a human coronavirus can give rise to three different genotypes (48).
Although previous studies have attempted to study the possible evolutionary and recombination origin of SARSr-CoV, no definite conclusion can be made on whether the viruses from bats are the direct ancestor of SARSr-CoV in civets and humans, given the paucity of available strains and genome sequences. To better define the epidemiology and evolution of SARSr-Rh-BatCoV in China and their role as a recombination origin of SARSr-CoV in civets, we carried out a 4-year study on coronaviruses in Chinese horseshoe bats in Hong Kong and Guangdong Province of southern China. Bat tagging was also performed to study the migration pattern of bats and viral persistence. The complete genomes of 10 strains of SARSr-Rh-BatCoV obtained at different time were sequenced and compared to previously sequenced genomes. With the availability of this larger set of genome sequences for more accurate analysis, recombination and molecular clock analyses were performed to elucidate the evolutionary origin and time of interspecies transmission of SARSr-CoV.
Sample collection and bat tagging.Sample collection was approved by and performed in collaboration with the Department of Agriculture, Fisheries and Conservation (AFCD) of the Hong Kong Special Administrative Region (HKSAR) and Guangzhou Center for Disease Control and Prevention, Guangzhou, China. Chinese horseshoe bats (Rhinolophus sinicus) were captured from various locations in Hong Kong and in Guangdong Province of southern China over a 4-year period (April 2004 to March 2008). Respiratory and alimentary specimens of the bars were collected using procedures described previously (21, 53). All specimens were placed in viral transport medium before transportation to the laboratory for RNA extraction. To assess the migration range and chronicity of coronavirus infections, 511 bats from Hong Kong were also tagged during sample collection before release. Tagged bats, when identified in subsequent site visits, were recaptured and recorded for sample collection before release.
RNA extraction.Viral RNA was extracted from the respiratory and alimentary specimens using QIAamp viral RNA minikit (QIAgen, Hilden, Germany). The RNA was eluted in 50 μl of AVE buffer and was used as the template for reverse transcription-PCR (RT-PCR).
RT-PCR for coronaviruses and DNA sequencing.Coronavirus screening was performed by amplifying a 440-bp fragment of the RdRp gene of coronaviruses using conserved primers (5′-GGTTGGGACTATCCTAAGTGTGA-3′ and 5′-CCATCATCAGATAGAATCATCATA-3′) designed by multiple alignments of the nucleotide sequences of available RdRp genes of known coronaviruses (49). Reverse transcription was performed using the SuperScript III kit (Invitrogen, San Diego, CA). The PCR mixture (25 μl) contained cDNA, PCR buffer (10 mM Tris-HCl [pH 8.3], 50 mM KCl, 3 mM MgCl2, and 0.01% gelatin), 200 μM (each) deoxynucleoside triphosphates (dNTPs), and 1.0 U Taq polymerase (Applied Biosystems, Foster City, CA). The mixtures were amplified in 40 cycles of PCR, with 1 cycle consisting of 1 min at 94°C, 1 min at 48°C, and 1 min at 72°C, and a final extension step of 10 min at 72°C in an automated thermal cycler (Applied Biosystems, Foster City, CA). Standard precautions were taken to avoid PCR contamination, and no false-positive result was observed for the negative controls.
The PCR products were gel purified using the QIAquick gel extraction kit (QIAgen, Hilden, Germany). Both strands of the PCR products were sequenced twice with an ABI Prism 3700 DNA analyzer (Applied Biosystems, Foster City, CA) using the two PCR primers. The sequences of the PCR products were compared with known sequences of the RdRp genes of coronaviruses in the GenBank database.
All cDNAs positive for SARSr-Rh-BatCoV were subjected to Rh-BatCoV HKU2 screening using Rh-BatCoV HKU2-specific primers (5′-GGAGTATGCAGCGTTGGGTTA-3′ and 5′-GACACATAGCGCTCAAGCAAA-3′), and all cDNAs positive for Rh-BatCoV HKU2 were subjected to SARSr-Rh-BatCoV screening using SARSr-Rh-BatCoV-specific primers (5′-CAAGTGGGGTAAGGCTAGACTTT-3′ and 5′-AACATATTATGCCAGCCACCATA-3′) using the PCR conditions described above.
Statistical analysis.Comparison of the body weights of bats in different groups was performed using Student's t test and covariate analysis (SPSS version 11.5). A P of <0.05 was regarded as statistically significant.
Complete genome sequencing of SARSr-Rh-BatCoV.Ten complete genomes of SARSr-Rh-BatCoV detected in the present study were amplified and sequenced using the RNA extracted from an alimentary specimen as the template. The RNA was converted to cDNA by a combined random priming and oligo(dT) priming strategy. The cDNA was amplified by degenerate primers as described previously (21). A total of 57 sets of primers, available on request, were used for PCR. The 5′ end of the viral genome was confirmed by rapid amplification of cDNA ends (RACE) using the 5′/3′ RACE kit (Roche, Germany). Sequences were assembled and manually edited to produce the final sequences.
Genome analysis.The nucleotide sequences of the genomes and the deduced amino acid sequences of the open reading frames (ORFs) were compared to those of other coronaviruses using the CoVDB coronavirus database (14). Phylogenetic tree construction was performed using the neighbor-joining method with ClustalX 1.83.
Bootscan analysis.Sliding window analysis was used to detect possible recombination, using nucleotide alignment of the available genome sequences of different SARSr-Rh-BatCoV strains and civet SARSr-CoV SZ3 generated by ClustalX version 1.83 and edited manually. Bootscan analysis was performed using Simplot version 3.5.1 (26) (F84 model; window size, 1,500 bp; step size, 300 bp) with selected strains, including SARSr-Rh-BatCoV Rf1 and civet SARSr-CoV SZ3, as the query sequence.
Estimation of synonymous and nonsynonymous substitution rates.The number of synonymous substitutions per synonymous site, Ks, and the number of nonsynonymous substitutions per nonsynonymous site, Ka, for each coding region were calculated for all available SARSr-Rh-BatCoV, civet SARSr-CoV, and human SARSr-CoV genomes using the Nei-Gojobori method (Jukes-Cantor) in MEGA 3.1 (19). Identical genes were excluded from analysis.
Estimation of divergence dates.The time of the most recent common ancestor (tMRCA) and the time of divergence were estimated on the basis of an alignment of ORF1 sequences, using the uncorrelated exponentially distributed relaxed clock model (UCED) in BEAST version 1.4 (7). Under this model, the rates were allowed to vary at each branch drawn independently from an exponential distribution. The sampling dates of all strains were collected from the literature or from the present study and were used as calibration points. Depending on the data set, Markov chain Monte Carlo (MCMC) sample chains were run for 1 × 108 states, sampling every 1,000 generations under the GTR nucleotide substitution model, determined by MODELTEST and allowing γ-rate heterogeneity for all data sets. A constant population coalescent prior was assumed for all data sets. The median and the highest posterior density regions at 95% (HPD) were calculated for each of these parameters from two identical but independent MCMC chains using TRACER 1.3 (http://beast.bio.ed.ac.uk ). The tree was annotated by TreeAnnotator, a program of BEAST and displayed by FigTree (http://tree.bio.ed.ac.uk/software/figtree/ ).
Nucleotide sequence accession numbers.The nucleotide sequences of the 10 genomes of SARSr-Rh-BatCoV have been lodged within the GenBank sequence database under accession no. GQ153539 to GQ153548.
Epidemiology of SARSr-Rh-BatCoV in Chinese horseshoe bats.A total of 1,398 respiratory specimens and 1,648 alimentary specimens from 1,337 and 64 Chinese horseshoe bats were obtained from Hong Kong and in Guangdong Province in southern China, respectively, over the 4-year study period. RT-PCR of a 440-bp fragment in the RdRp genes of coronaviruses was positive for SARSr-Rh-BatCoV in respiratory specimens from 2 of the 1,337 Chinese horseshoe bats from Hong Kong and in alimentary specimens from 126 (9.4%) of the 1,337 Chinese horseshoe bats from Hong Kong and 4 (6.3%) of the 64 Chinese horseshoe bats from Guangdong, China, with ≥99% nucleotide identities to SARSr-Rh-BatCoV (GenBank accession no. DQ022305) (21). Another previously described group 1 coronavirus, Rhinolophus bat coronavirus HKU2 (Rh-BatCoV HKU2), coinfecting Chinese horseshoe bats, was identified in alimentary specimens from 62 (4.6%) bats from Hong Kong and from 7 (10.9%) bats from Guangdong, China, with ≥99% nucleotide identities to Rh-BatCoV HKU2 (GenBank accession no. DQ249235) (22). Seventeen bats from Hong Kong and three bats from Guangdong, China, were coinfected by SARSr-Rh-BatCoV and Rh-BatCoV HKU2. The 126 bats from Hong Kong positive for SARSr-Rh-BatCoV were from 15 of the 27 sampling locations in Hong Kong, with bats from seven locations harboring both viruses (Fig. 1). Peak activity of both SARSr-Rh-BatCoV and Rh-BatCoV HKU2 was observed in the spring (see Fig. S1 in the supplemental material). However, the prevalence of SARSr-Rh-BatCoV was higher than that of Rh-BatCoV HKU2 during the spring of 2005 and 2007, while the prevalence of Rh-BatCoV HKU2 was higher than that of SARSr-Rh-BatCoV in the spring of 2006.
A total of 511 Chinese horseshoe bats from 11 sites were tagged, with 152 (29.7%) recapturing episodes from six sites during subsequent visits (Fig. 2). A total of 113 tagged bats were recaptured, with 84 bats recaptured once, 21 recaptured twice, 6 recaptured three times, and 2 recaptured four times after tagging. The time interval between tagging and recapture of the same bat ranged from 2 weeks to 21 months. Migration between water tunnels at short distances was most common (Fig. 1). The longest distance of migration was approximately 17 km within 3 months from tagging to recapture (October 2006 to January 2007), while the shortest distance between two habitats was 1.86 km. Sixteen bats were positive for SARSr-Rh-BatCoV, and 23 were positive for Rh-BatCoV HKU2 at the time of tagging, with one bat being positive for both viruses. Among these 38 bats, 10 bats were recaptured, but all were subsequently negative for coronaviruses within a period of 4 to 16 months (Fig. 2). Twenty-three and nine bats initially negative for coronaviruses were subsequently positive for SARSr-Rh-BatCoV and Rh-BatCoV HKU2, respectively, among which seven bats were positive for both viruses. However, only one of these bats was positive for coronavirus at more than one episode, which carried SARSr-Rh-BatCoV at first and both SARSr-Rh-BatCoV and Rh-BatCoV HKU2 during the next visit at the same site 2 weeks later (Fig. 2). With the longest shedding period of 2 weeks and the shortest documented clearance time of 4 months in bats positive for SARSr-Rh-BatCoV, it is estimated that viral clearance occurred between 2 weeks and 4 months.
No disease association was observed in bats positive for SARSr-Rh-BatCoV or Rh-BatCoV HKU2. However, lower body weights were observed in bats positive for SARSr-Rh-BatCoV (body weight [mean ± standard deviation {SD}], 10.9 g ± 1.4 g) than those negative for coronaviruses (body weight [mean ± SD], 11.6 ± 2.2 g) (P < 0.0001 by Student's t test). A similar phenomenon was not observed when bats positive for Rh-BatCoV HKU2 (body weight [mean ± SD], 11.5 ± 1.5 g) were used for comparison (P = 0.783 by Student's t test). To control for the confounding effect of age and possible lower body weights after hibernation during which SARSr-Rh-BatCoV showed the highest detection rate, covariate analysis was performed using only data from the peak season (during March) with forearm lengths (which correlate with age) as a possible cofactor. The results showed that the SARSr-Rh-BatCoV carriage state is an independent factor in association with lower body weight (P = 0.002). Similarly, no significant difference in body weight was observed when similar analysis was performed on bats positive for Rh-BatCoV HKU2 despite its similar seasonality, suggesting that this phenomenon is specific to SARSr-Rh-BatCoV.
Complete genome comparison of SARSr-Rh-BatCoV genomes.In addition to the eight previously described genomes of SARSr-Rh-BatCoVs, complete genome sequence data of 10 additional strains of SARSr-Rh-BatCoV were obtained by assembly of the sequences of the RT-PCR products obtained directly from 10 individual specimens collected at different times. Eight strains were detected in bats from Hong Kong, while two strains were from bats from Guangdong, China (see Table S1 in the supplemental material). Their genome sizes were 29,677 to 29,716 nucleotides, with a G+C content of 41%, comparable to the previously reported genomes. The eight Hong Kong strains were more closely related to each other with an overall nucleotide identity of 99.9%, while the two strains from Guangdong, China, had 98.5% nucleotide identity to the Hong Kong strains. Except for strain HKU3-8 from Guangdong, China, all SARSr-Rh-BatCoV strains share the same genome organization, containing the putative transcription regulatory sequence (TRS) motif, 5′-ACGAAC-3′, at the 3′ end of the 5′ leader sequence and preceding each ORF except ORF7b.
Similar to previous findings, analysis of the full-length sequences of all currently available SARSr-Rh-BatCoV genomes showed that the major differences between SARSr-Rh-BatCoV genomes and civet/human SARSr-CoV genomes were observed in spike (mainly S1 domain), ORF3, and ORF8 regions, which were also the most variable regions among human SARSr-CoV genomes (21, 25, 34). All genomes possessed 87% nucleotide identities to civet and human SARSr-CoV, except for SARSr-Rh-BatCoV strain Rp3, which possessed 91% and 92% nucleotide identities to civet and human strains, respectively. The higher overall sequence similarity of strain Rp3 to civet and human strains is mainly due to the higher sequence homology within the ORF1 region. At nsp2, nsp3, nsp12, and nsp14 regions, strain Rp3 possessed the highest amino acid identities among all SARSr-Rh-BatCoV strains to the corresponding regions in civet SARSr-CoV SZ3 (see Table S2 in the supplemental material). Interestingly, the sequences of our SARSr-Rh-BatCoV strains from Hong Kong and Guangdong, China, possessed higher (98%) amino acid identities in the nps1 region to civet SARSr-CoV than other strains from China (92 to 93%). On the other hand, at ORF3a and ORF8, the sequences of strains Rf1 and 273/04 possessed the highest amino acid identities to those of civet SARSr-CoV (see Table S2 in the supplemental material).
ORF8 represents the most variable region within the SARSr-CoV genomes. In contrast to human SARSr-CoV which contains a 29-bp deletion at ORF8 region which resulted in two overlapping ORFs, ORF8a and ORF8b, all SARSr-Rh-BatCoV genomes except that of strain HKU3-8 contain a single long ORF8, similar to civet SARSr-CoV. This 29-bp deletion present only in human strains has been shown to disrupt the functional expression of the ORF8 region (30). Strain HKU3-8 has a short deletion at the ORF8 region that breaks this ORF into three small ones (see Fig. S2 in the supplemental material). This 26-bp deletion was only 14 bp downstream of the 29-bp deletion in human SARSr-CoV, suggesting that this region is a frequent site for deletions. As a result of the frequent deletions observed within this region, the ORF8 region of SARSr-Rh-BatCoV possessed very low (35 to 37%) amino acid identities to that of civet SARSr-CoV, except for two strains, Rf1 and 273/04, which possessed 80% amino acid identities (see Table S2 in the supplemental material).
Phylogenetic analysis.Phylogenetic trees were constructed using the nucleotide sequences of the nonstructural protein 3 (nsp3), RdRp, spike (S), ORF3a, envelope protein (E), membrane protein (M), ORF8, and nucleocapsid protein (N) genes of SARSr-CoV (see Fig. S3 in the supplemental material). In general, SARSr-Rh-BatCoV strains from the same geographical area are more closely related to each other. Among all SARSr-Rh-BatCoV genomes, strain Rp3 from Guangxi Province, China, is most closely related to human and civet strains in the ORF1 region, as exemplified by their close clustering in the nsp3 and RdRp trees. However, from the S gene onwards, strain Rp3 was more closely related to other SARSr-Rh-BatCoVs than to human and civet strains. Moreover, from ORF3a to ORF8, clustering of human and civet SARSr-CoVs with strains Rf1 and 273/04 from Hubei Province, China, was observed. This suggested that civet SARSr-CoV may have arisen from recombination between strains from different geographical locations that were related to present strains from Guangxi and Hubei provinces in China.
Recombination analysis.To detect recombination between genomes of different strains of SARSr-Rh-BatCoV or civet SARSr-CoV, sliding window analysis was conducted. Results showed frequent recombination events among the bat viruses in China. When civet SARSr-CoV SZ3 was used as the query sequence with SARSr-Rh-BatCoV strains Rm1, Rf1, and Rp3 as the potential parents, a recombination breakpoint at the nsp16/S intergenic region was identified (Fig. 3). Upstream of this breakpoint before position 21300, high bootstrap support for clustering of civet SARSr-CoV SZ3 with SARSr-Rh-BatCoV strain Rp3 was observed. However, an abrupt change in clustering occurred after position 21300, with high bootstrap support for clustering of civet SARSr-CoV SZ3 with SARSr-Rh-BatCoV strain Rf1. This is in line with results from phylogenetic analysis, where the ORF1 sequences of civet and human SARSr-CoV strains clustered with SARSr-Rh-BatCoV Rp3, but the sequences from ORF3a to ORF8 clustered with SARSr-Rh-BatCoV strains Rf1 and 273/04.
Apart from this recombination event, other putative recombination events were also observed when SARSr-Rh-BatCoV strains were used as the query sequence. When SARSr-Rh-BatCoV strain Rf1 was used as the query sequence with SARSr-Rh-BatCoV HKU3-1, 279/04, and civet SARSr-CoV SZ3 as the potential parents, putative recombination events were observed throughout the genome, as shown by frequent shuffle of clustering with the three putative parent strains (Fig. 4A). The most notable site occurred at around position 20700, corresponding to nsp16. From positions 16400 to 20700, high bootstrap support for clustering with strain 279/04 was observed. From position 20700 onwards and especially toward the 3′end of the genome, high bootstrap support for clustering with civet SARSr-CoV SZ3 was observed. These bootscan results were also supported by the shifting of positions upon phylogenetic analysis (Fig. 4B). From positions 16400 to 20700 (corresponding to helicase to nsp16), SARSr-Rh-BatCoV Rf1 was clustered with strain 279/04 with a bootstrap value of 1,000 away from civet SARSr-CoV. From positions 20700 to 25000 (corresponding to nsp16 to S2), it exhibited a distant relationship with both other SARSr-Rh-BatCoVs and civet SARSr-CoV. However, from position 25000 (S2) onwards, it was more closely related to civet and human SARSr-CoV strains than to other SARSr-Rh-BatCoV. Similar results were also observed when similar analysis was performed using strains 273/04, 279/04, and Rm1 as the query sequence with corresponding strains as the potential parents.
On the other hand, when SARSr-Rh-BatCoV strain Rf1 was used as the query sequence with strains 273/04, Rm1, and Rp3 as the potential parents, a single recombination breakpoint from position 18300 to 19900 corresponding to nsp14/15 (Fig. 5A) was observed. Before position 18300 and after position 19900, high bootstrap support for clustering between strains Rf1 and 273/04 was observed, whereas between these two positions, an abrupt shift in phylogenetic signals occurred, with high bootstrap support for clustering with strain Rm1. In fact, from phylogenetic analysis of other regions of the whole genome, strain Rf1 is closely related to 273/04, and SARSr-Rh-BatCoV Rm1 is closely related to SARSr-Rh-BatCoV 279/04, except in this breakpoint region, where discordance of phylogenetic positions was observed (Fig. 5B). These results suggest that SARSr-Rh-BatCoV strains from different bat species may undergo frequent recombination.
Estimation of synonymous and nonsynonymous substitution rates.Using all available SARSr-Rh-BatCoV genome sequences for analysis, except for strain HKU3-8 not used for ORF8 analysis, the Ka/Ks ratios for the various coding regions, compared to those of civet SARSr-CoV and human SARSr-CoV, are shown in Table 1. Notably, the Ka/Ks ratio for the S of SARSr-Rh-BatCoV is only 0.054, compared to that of civet SARSr-CoV (1.5) and human SARSr-CoV (1.0), suggesting that the spike gene of SARSr-Rh-BatCoV is unlikely under positive selection. Moreover, the Ka/Ks ratio for ORF3a, E, and M of SARSr-Rh-BatCoV were also markedly lower than that for civet and/or human SARSr-CoV. On the other hand, the highest Ka/Ks ratios were observed at ORF7b (0.546) and ORF8 (0.554), suggesting that this region is under strong positive selection.
Estimation of divergence dates.Using the uncorrelated relaxed clock model on ORF1ab, the date of the most recent common ancestor (MRCA) of all SARSr-CoVs was estimated to be 1972.39 (HPDs, 1935.28 to 1990.63), approximately 31 years before the SARS epidemic (Fig. 6). The date of divergence between human or civet SARSr-CoV and the closest SARSr-Rh-BatCoV was estimated to be 1995.10 (HPDs, 1986.53 to 2000.13), approximately 8 years before the SARS epidemic. Moreover, the MRCA date of human and civet SARSr-CoV was estimated to be 2001.36 (HPDs, 1999.16 to 2002.14). The estimated mean substitution rate of the ORF1ab data set under the UCED model was 2.82 ×10−3 substitution per site per year. This estimate is comparable to a previous estimation using fewer SARSr-Rh-BatCoV genome sequences (2.79 × 10−3 substitution per site per year) and the estimate in other RNA viruses (13, 15).
This is the first report on the migration pattern of horseshoe bats in China and its relation to the epidemiology of coronaviruses. In this study, SARSr-Rh-BatCoV was found among 9.4% and 6.3% of alimentary specimens from Chinese horseshoe bats from Hong Kong and Guangdong, China, respectively, with some bats coinfected with a group 1 coronavirus, Rh-BatCoV HKU2. Both viruses showed peak activity during spring, with an apparent alternate biennial activity. Mating and feeding activity soon after hibernation in spring may have facilitated the spread of the virus within the same roost and from roost to roost. Although no disease association could be observed, lower body weights were observed for bats positive for SARSr-Rh-BatCoV (but not for bats positive for Rh-BatCoV HKU2) than those negative for coronaviruses. The results of a tagging exercise showed that long-distance migration of Chinese horseshoe bats is uncommon, with the longest distance being 17 km from a northern location in fall to an eastern location in winter, compatible with data from other Rhinolophus species which may migrate up to 30 km for hibernation (28, 29). Nevertheless, such migration distances are sufficient for migration between Hong Kong and many areas in Shenzhen, China, including the wild animal markets where the first civet SARSr-CoV was identified (Fig. 1). Such foraging ranges could have allowed for mixing of different SARSr-Rh-BatCoV strains of different geographical origins. Except for one bat which carried SARSr-Rh-BatCoV for at least 2 weeks, all bats positive for coronavirus were cleared of the same virus during recapture. Moreover, tagged individuals positive for SARSr-Rh-BatCoV were healthy during subsequent recapture, evidencing survival after the viral infection as reported for European bat lyssavirus in meridional serotine bats from Spain (44). The present findings suggest that SARSr-Rh-BatCoV causes an acute, self-limiting infection associated with weight loss in Chinese horseshoe bats, with viral clearance occurring between 2 weeks to 4 months. This is compatible with our previous finding that the presence of neutralizing antibody in their sera correlated with a lower viral load in alimentary specimens (21).
The present study revealed that recombination events are common between different SARSr-Rh-BatCoV strains from different species of bats and geographical locations, which may account for the emergence of a civet SARSr-CoV capable of cross-species transmission from bats to civets and from civets to humans. Genome sequence comparison of SARSr-Rh-BatCoVs from horseshoe bats and human/civet SARSr-CoV showed that they shared only 87 to 92% nucleotide identity. Therefore, genetic events, such as mutation and/or recombination, would have occurred during the evolution of these SARSr-CoVs before the possible emergence of direct progenitors of SARSr-CoV capable of infecting palm civets and subsequently humans. Reconstruction of chimeric recombinant SARSr-CoV using the S sequences of various SARSr-Rh-BatCoV found in horseshoe bats will reveal if any particular part of the S sequence is important for infection of civets and/or humans (37). In the present study, frequent recombination events were identified among SARSr-CoVs. Moreover, civet SARSr-CoV strain SZ3 was shown to be a potential recombinant of SARSr-Rh-BatCoV strain Rp3 from Guangxi Province, China, and SARSr-Rh-BatCoV strain Rf1 from Hubei Province, China, by both phylogenetic and bootscan analyses, with the recombination breakpoint identified at the nsp16/S intergenic region. This suggests that civet SARSr-CoV may have either evolved from an ancestor that is a direct recombinant between strains Rp3 and Rf1 or is a direct recombinant of lineages closely related to Rp3 and Rf1 that are yet to be identified. This finding is in line with a previous study showing that SARSr-Rh-BatCoV Rp3 was a potential recombinant of SARSr-Rh-BatCoV and an unidentified lineage closely related to civet SARSr-CoV (13). However, civet SARSr-CoV was not used as the query sequence for analysis in this study, and no conclusion was drawn on the origin of civet strains. In addition, we also detected other potential recombination events when different SARSr-Rh-BatCoV strains were used as the query sequence for analysis, with the most notable recombination sites found to be located at nsp16 and nsp14/15. We have previously described the first evidence of natural recombination in a human coronavirus, HCoV-HKU1, that led to the generation of different genotypes, which also represents the first report to describe a distribution of the recombination spots in the entire genome of field isolates of a coronavirus (46). Interestingly, in this report, the most significant recombination event was also observed at nsp16. Therefore, the 3′ region of ORF1 and the ORF1/S junction are likely frequent sites of natural recombination in coronaviruses. Alternatively, the apparent hot spot of recombination may be due to the nonviability of the recombinant viruses that cross over at other points. In fact, recombination at the junction of the nonstructural and structural genes essentially defines the evolution of the nidoviruses. In wildlife food markets and restaurants in southern China, a huge variety of animals of different geographical origins are often caged in a crowded environment, which may allow cross-species viral transmission and recombination events (47). Further surveillance studies of different horseshoe bat species from different provinces of China and genome analysis of their SARSr-Rh-BatCoV strains may reveal further evidence for the recombination origin of SARSr-CoV.
With the availability of a larger set of SARSr-Rh-BatCoV genome sequences for analysis, the present study also attempts to more accurately estimate the time of emergence of SARSr-CoV in civets, which was shown to be only 8 years before the SARS epidemic in 2003. Despite the identification of SARSr-Rh-BatCoV in various horseshoe bat species from China, there has not been sufficient evidence to determine if bats are the host for the direct ancestor of civet and human SARSr-CoV. On the basis of the considerable phylogenetic distance between bat and civet/human strains, one study concluded that the current bat strains are unlikely to be the direct ancestor (34). A subsequent study using molecular dating analysis showed that the estimated date of interspecies transmission event from bats to an amplifying host, such as the civet, was 17 years (HPD, 2 to 39 years) before the SARS epidemic (45). It was therefore concluded that there may be a yet unidentified intermediate host between bats and civets or unidentified SARSr-Rh-BatCoV strains that are even more closely related to civet SARSr-CoV. Another study estimated the date of interspecies jumping to be much more recent, approximately 4.08 years (HPD, 1.45 to 8.84 years) before the SARS epidemic (13). Results from the present study are more in line with the latter estimate, with the date of divergence between human/civet and bat strains estimated to be 8 years (HPDs, 2.9 to 16.5 years) before the SARS epidemic. The discrepancies between the different estimated results may be due to the choice of different gene sequences and different number of strains for analysis. The study that concluded a much older date of divergence used only the helicase domain sequence for their analysis, whereas the other study, similar to this study but with fewer strains, used the complete ORF1 sequence for analysis. The availability of sequences of more strains collected over a longer period of time may further improve the accuracy of such estimation. On the other hand, all three studies supported that SARSr-CoV are likely a newly emerged subgroup of Betacoronavirus, with the median date of their MRCA estimated to be from 1961 to 1982 (13, 45). The emergence of diverse virus strains in the different Rhinolophus species within a few decades suggested that this novel group of coronaviruses is rapidly evolving and may easily cross the species barrier.
Comparison of all available complete genome sequences of SARSr-Rh-BatCoVs showed that their genome sequences were closely related with the same genome organization, except for strain HKU3-8 from Guangdong, China, with three small ORFs at ORF8 region instead of a single ORF. In particular, strains from the same geographical location were highly similar. Upon phylogenetic analysis of the individual ORFs, the strains from Hong Kong and Guangdong, Guangxi, and Hubei, China, formed separate clusters for most of the time, although the strains from Hong Kong and Guangdong, China, are more closely related, probably due to the close geographical proximity. However, given the frequent recombination and short genetic distance among the different bat strains, no distinct subgroups can be classified. Similar to previous studies, the most variable regions in the SARSr-Rh-BatCoV genomes were located in S, ORF3, and ORF8 (21, 25). In particular, the frequent deletions in ORF8 region in SARSr-Rh-BatCoV, together with the previously reported 29-bp deletion in human SARSr-CoV, suggested that this is a frequent site for deletions in SARSr-CoV. Moreover, the relatively high Ka/Ks ratios observed at ORF7b and ORF8 further supported that it is a region subject to rapid evolution under strong positive selection. The human SARSr-CoV 7b protein is an integral membrane protein localized in the Golgi apparatus and contributes to virus-induced apoptosis (35, 36). The human SARSr-CoV ORF8a enhances viral replication and induces apoptosis through a mitochondrion-dependent pathway, whereas the longer ORF8 protein of civet and early human isolates is a cleaved protein stable in the endoplasmic reticulum (4). Further studies are required to understand the biological significance of the high mutation rate in this part of the SARSr-CoV genomes.
We thank Alan Chi-Kong Wong, Siu-Fai Leung, Chik-Chuen Lay, Ping-Man So, and K. F. Chan (Hong Kong Special Administrative Region [HKSAR] Department of Agriculture, Fisheries and Conservation [AFCD]) and Hong Kong Police Force for facilitation and support; Cynthia S. M. Chan and Joseph W. K. So from AFCD; and King-Shun Lo (Laboratory Animal Unit) and Cassius Chan for their excellent technical assistance and collection of animal specimens. We are grateful to the generous support of Carol Yu, Richard Yu, Hui Hoy, and Hui Ming in the genomic sequencing platform.
This work was partly supported by the Research Grant Council Grant (HKU 7687/07M); University Development Fund and Outstanding Young Researcher Award, The University of Hong Kong; The Tung Wah Group of Hospitals Fund for Research in Infectious Diseases; the HKSAR Research Fund for the Control of Infectious Diseases (04050232) of the Health, Welfare and Food Bureau; the Providence Foundation Limited in memory of the late Lui Hac Minh; and Consultancy Service for Enhancing Laboratory Surveillance of Emerging Infectious Disease for the HKSAR Department of Health.
- Received 21 October 2009.
- Accepted 3 January 2010.
- Copyright © 2010 American Society for Microbiology


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Warm, bright sunlight, sumptuous green and magnificent, colourful flowers – spring has sprung and brought the Villeroy & Boch Easter collections along with it. From playful bunny motifs and colourful Easter eggs to artistically curved porcelain with delicate décor, the collections offer a huge range of tableware and decorative items.
Buy directly from the manufacturer and receive attractive offers on the Villeroy & Boch Online Shop.
Free Delivery on orders over 1250kr
Free & easy returns
Convenient & Safe Payments
Tableware Customer Care
Call us for free:
Telephone: 00800 6864 8110
Monday - Friday: 8am – 6pm
In light of current events, we can't always be reached by phone. But you can contact us by email at any time.
Enjoying breakfast or afternoon tea together on Easter Sunday has long been a tradition in many families. An Easter table decorated for the occasion is an essential part of this and is child's play to create with the Villeroy & Boch Easter collections. The Farmers Spring series with meadow flowers, hens and green accents gives your festive table real country flair. With flower décor and contrasting yellow edges, the Spring Awakening collection is less classic and more playful. Matching napkins pick up on each style and are the perfect finishing touch to the table.
Is your Easter table lacking colour? Does it need some extra decoration to welcome in the spring? Then discover the beautiful bowls, egg cups and vases in the Spring Fantasy Easter collection, with delicate flowers and cute bunnies. The Spring Decoration series also has adorable musical bunnies – both as tea light holders and as porcelain hanging decorations for your Easter bouquet. They are perfect teamed with the artistically-painted Mini Flower Bells and Spring Eggs.
Made from fine porcelain and coloured in soft pastel shades, the Mariefleur Spring Easter collection will capture the hearts of all Easter fans. The delicately-applied flower motifs in purple, pink and yellow cover the vases, bowls, decorative figures and porcelain hanging decorations in various shapes. Be it bunnies, birds or Easter eggs – you alone decide how you want to decorate your home at Easter. Combined with the fine Villeroy & Boch tableware and cutlery, you can conjure spring onto your Easter table.
The small bunny figures in the Bunny Family collection are a great surprise for the Easter nest. From female bunnies playing the violin or xylophone and male bunnies with drums to the entire bunny family as a choir, the decorative figures all look beautiful. They are ideal not just for children, but also for enthusiastic collectors. The bunnies in the Easter Decoration series have a more realistic look – with blue collars and bells. The Annual Easter Edition, on the other hand, offers Easter bunnies for tableware and decoration. Everyone will find what they are looking for in our Easter collections.

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|Péter Király <kirun...@gmail.com>
|Re: Does any one know how to expand queries using Wordnet with Lucene 3.6?
|Fri, 15 Jun 2012 18:07:24 GMT
Hi, maybe not the one you are looking for, only a similar thing what I have create: http://drupalcode.org/project/xc.git/tree/refs/heads/6.x-1.x:/xc_wordnet. This is a Drupal module and a distinct Java class. The Java class creates an XML file, which can be imported into Solr, to build a Wordnet index (so I have two indexes: one for my content, and one for wordnet). The Drupal module provides synonyms as links, so if the user enters a term, below or upon the search results a list of synsets is shown up, as links. The user can click any synonym or a special "all synonyms" link, wich runs the refinement search. For me the suggested solution of using Wordnet in a SynonymFilter produced very-very strange results, because sometimes (and more frequently as toleratable) the synonyms go too far from the impliciltly intended meaning. For example Wiener -> Frankfurter -> hot dog is a real-world example. You can imagine that your result set will be fulfilled with unwanted hits when you search for Wien (capital of Austria). For me not to handling it automatically, but provide something the user to help him to refine the search is the optimal solution. Regards, Király Péter 2012/6/15 Robert Muir <email@example.com>: > See its test: http://svn.apache.org/repos/asf/lucene/dev/branches/lucene_solr_3_6/lucene/contrib/analyzers/common/src/test/org/apache/lucene/analysis/synonym/TestWordnetSynonymParser.java > > The idea is you parse the wordnet file with this thing, and create a > SynonymFilter that you use in your query Analyzer (The one you pass to > QueryParser). > > On Fri, Jun 15, 2012 at 6:27 AM, Kits89 <firstname.lastname@example.com> wrote: >> I've found the class WordnetSynonymParser in >> org.apache.lucene.analysis.synonym but there aren't examples of its usage >> neither in the API nor in google. Does any one have experience with it? >> >> Thank you! >> >> EDIT: I know that there used to be the class SynExpand, but with version 3.6 >> it disappeared... >> >> -- >> View this message in context: http://lucene.472066.n3.nabble.com/Does-any-one-know-how-to-expand-queries-using-Wordnet-with-Lucene-3-6-tp3989794.html >> Sent from the Lucene - Java Users mailing list archive at Nabble.com. >> >> --------------------------------------------------------------------- >> To unsubscribe, e-mail: firstname.lastname@example.com >> For additional commands, e-mail: firstname.lastname@example.com >> > > > > -- > lucidimagination.com > > --------------------------------------------------------------------- > To unsubscribe, e-mail: email@example.com > For additional commands, e-mail: email@example.com > --------------------------------------------------------------------- To unsubscribe, e-mail: email@example.com For additional commands, e-mail: firstname.lastname@example.com
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Blessings on Your First Holy Communion frame Girl Figurine
Item Number: 123410
Dimensions: 6.00 x 0.44 x 7.25 inches
Dimensions: 6.00 x 0.44 x 7.25 inches
A young girl preparing to receive the holy bread and wine for the very first time graces the front of this photo frame; the perfect way to showcase a 3.5" x 5" photo from her First Communion. Accented with golden paint, this gift becomes a treasured keepsake – a reminder of the day she received this blessed sacrament. Give this First Holy Communion gift to a special young girl you know who is making this important step in her religious life. Beautifully sculpted in fine bisque porcelain and meticulously hand painted. Approximately 7.25 inches high.
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Thinking about another build and was reading Greg Duleys article on Long Range Hunting which has got me interested. Especially since Berger has launched their 195gr.
I've lost his article but from what I remember there was a bit involved in case forming? However, what sparked my interest was the comment made on the LRH article about how brass for the Fatso was easier to make because of Norma's newly released cases (.338 Norma Mag??) .
Anyway, with regards to action, short or long? I seem to recall Greg built his Fatso on a Remington 700 short action with Wyatts extended magazine. I'm guessing that to run the 195gr you'd require a long action?
Any other information would be appreciated.


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Air Science Purair Laminar Flow Cabinets
Have questions or need technical support? Contact Us to speak with a live representative.
Monday-Friday, 8:00 AM - 5:00 PM (EST)
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A $50 minimum applies to all orders.
Starting at $1,699.00
Have questions or need technical support? Contact Us to speak with a live representative.
Monday-Friday, 8:00 AM - 5:00 PM (EST)
Chat with a live representative
A $50 minimum applies to all orders.
Purair® FLOW™ Series laminar flow cabinets are designed to protect the work surface, products and materials from particulate contamination. Room air passes through a HEPA (High Efficiency Particulate Air) filter then uniformly by laminar flow through the cabinet interior to protect the work from unfiltered air. The airflow is oriented to exhaust airborne particulate introduced by the user.
The Purair® FLOW™ Series product line employs the Air Science exclusive Multiplex™ HEPA Filtration Technology to sustain the contamination-free environment.
Equipment Included with Purchase
- • Prefilter for Dust ($32 value)
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- • Integral fluorescent lighting ($175 value)
- • Cable slip ports (2 in rear wall) ($95 value)
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FLOW™ Benefits
- • Vertical laminar flow with HEPA/ULPA filtration protects materials inside the cabinet from particulates.
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- • Optional base stand and mobile cart with locking casters
Purair® FLOW™ Series cabinets offer a proven level of ISO Class 4 product performance at an economical cost. These cabinets are intended for use in non-hazardous applications where biological or biohazard byproducts are not generated and user protection is not required. A range of Air Sciecne innovations are integrated into the clean, simple, low-mainteance design, offering flexible access to the interior work area. Typical applications include:
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Performance specifications and construction meet or exceed OSHA, ANSI, and relevant international standards to assure operator safety.
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Each Air Science Purair® FLOW™ Cabinet includes features expressed through sound design and certified quality construction. Options and accessories add functional performance to meet specific applications.
- • Professional Quality. Air Science cabinets comply with current technical and safety regulations.
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Purair® FLOW™ Cabinets are available in 3 model sizes with various options.
- • Cabinets are shipped fully assembled with standard power cords; no installation required.
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- • Purair® FLOW™ cabinets are constructed of steel with a microbial powder finish coating.
2 year warranty (parts and labor) on mechanical and electrical and lifetime warranty (parts only) on the metal superstructure. Consumables are not covered.
|Airflow Pattern
|Vertical Downflow
|0.45 m/s to 90 fpm
(average airflow measured 6" from filter face; uniformity is ±20%)
|Disposable polyester fibers with 85% arrestance
|Main Filter
|HEPA efficiency: 99.99% at 0.3µm
|Screw Compression Clamp
|Cabinet Lighting
|Compact Fluoresecent Bulb Removed from Air Stream
|Side Window Construction
|Side Window Visible Opacity
|Side Window Color
|Construction Color
|White epoxy-coated steel frame with blue legs on cabinet sides.
|Pass-Through Ports
|ebmpapst™ centrigual fan
|120V, 60Hz
|Electrical Controls
|Main ON/OFF switch for fan and lighting. Solid-state fan speed
control with RFI filter maintains blower uniformity.
|Nominal Width
|24" / 610 mm
|36" / 910 mm
|48" / 1220 mm
|Internal Height
|24" / 610 mm
|Internal Depth
|24" / 610 mm
|External Dimensions
|24 x 24 x 35"
610 x 610 x 889 mm
|36 x 24 x 35"
914 x 610 x 889 mm
|48 x 24 x 35"
1219 x 610 x 889 mm
|Shipping Dimensions
|40 x 40 x 40"
1016 x 1016 x 1016 mm
|52 x 45 x 40"
1321 x 1143 x 1016 mm
|Net Weight
|72 lbs. / 33 kg
|99 lbs. / 45 kg
|138 lbs. / 63 kg
|Ship Weight
|129 lbs. / 59 kg
|157 lbs. / 71 kg
|195 lbs. / 88 kg
|Standards and Compliance (All Models)
|Quality Management Systems
ENERGY STAR® Partner
|Cabinet Performance
|Air Quality
|ISO 14644-1, Class 4
|Electrical Safety
CE Mark
RoHS Exempt under EEE Category 9


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H.15149 - Bonita Isles single family lakefront home, located steps away from the amenities. Impact double pane windows throughout. Best selling Salerno floor plan with open kitchen/great room/dining room. One of the few homes with direct, lighted waterfall view. Large L-shaped covered lanai is the largest in the neighborhood. High tray ceilings in great room and master. Kitchen upgrades include center island, backsplash, undercabinet lighting, pullouts, and stainless appliances. Hunter Douglas blinds and plantation shutters. Tile throughout - no carpet! Full length hall mirror in master. Added shelving in garage with insulated door. Amenities include resort-style pool, fitness center, pickleball/tennis, kids play area, and event area. Lawns are maintained by the HOA. No CDD!
BN06 - North Bonita East of US41
Architectural, No Commercial, No RV
BBQ - Picnic, Clubhouse, Community Park, Community Pool, Exercise Room, Internet Access, Pickleball, Play Area, Sidewalk, Streetlight, Tennis Court, Underground Utility
Cable Prewire, Custom Mirrors, Foyer, Smoke Detectors, Tray Ceiling
The source of this real property information is the copyrighted and proprietary database compilation of the Southwest Florida MLS organizations Copyright 2020 Southwest Florida MLS organizations.. All rights reserved. The accuracy of this information is not warranted or guaranteed. This information should be independently verified if any person intends to engage in a transaction in reliance upon it.
The data relating to real estate for sale on this Website come in part from the Broker Reciprocity Program (BR Program) of M.L.S. of Naples, Inc. Properties listed with brokerage firms other than Equity Realty are marked with the BR Program Icon or the BR House Icon and detailed information about them includes the name of the Listing Brokers. The properties displayed may not be all the properties available through the BR Program.


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Safety Data Sheets (SDSs) contain all the data a downstream user needs to work with a hazardous chemical. Required by law to be provided by the manufacturers of each chemical, SDSs cover everything from basic identification, hazards and safe handling processes to toxicological information, exposure control information, and substance stability and reactivity. The newer, more consistent format (designed in 2012) is comprised of 16 sections and is meant to be much easier to understand all the way around. If you have any questions or would like more information on how to use your SDSs, check out this page on the OSHA website, where they cover every section in detail.This entry was posted on Monday, March 28th, 2016 By
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With all of the courses available today it is difficult to understand which one is the best. This simple list explains the steps you will need to take to become and EBC accredited English language teacher. If you still have questions, contact us.
1. Contact
Get in touch with us to find out more about TEFL, TESOL, accreditation and jobs. EBC's experienced TEFL consultants are standing by to give you personalised guidance and advice to help you with your research. We will respond to your enquiries within 24 hours or less.
2. Meet and Talk
Finalise your schedule with your dedicated EBC student advisor who will assist and guide you from this moment through to getting your job teaching English abroad. You will get plain, realistic and practical advice about: teaching English abroad, costs, pay, travel, visas, documents, health insurance, housing and more.
3. Apply
Fill out a simple form to apply for your chosen course. We will process your application within 2 working days. You will be sent a confirmation. If you have applied for our CertTESOL course, you will be sent instructions about the Trinity College pre-acceptance process.
4. Pre-acceptance interview
The pre-acceptance interview makes sure that you are happy with your chosen course and that EBC can justify your enrolment on an EBC course as a quality student. The pre-acceptance interview is for classroom-based course students.
5. Enrol and guarantee your course place
To enrol and guarantee your course place you must either send a deposit or the full course payment. You may do this through our secure inline payment system or by bank transfer. Once your payment is received, your place is guaranteed.
6. Travel
Book your travel tickets and get ready to go. If you need accommodation, we can provide it. We can give non-legal advice about visas and other bureaucracy. If you want a student visa for Spain, we can help to arrange for it. Visas are given by governments not by EBC.
7. Train
Take your four-week classroom-based course or your online course. When you pass the course, you will be a certified TEFL teacher.
8. Teach
On the day you pass, you may ask EBC to send your CV to English language schools using our lifetime, proactive, worldwide job placement service. You should be interviewing and teaching within a few days of finishing your classroom-based course.
Join our growing EBC family
To become an English teacher all that is required is some of your time to take one of our EBC training courses. Once you graduate, EBC gives you lifetime, worldwide career support that will get you working quickly and will always be there for you when you want a change.


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Chinese Water Dragon
Reptile Community - Here's a handsome, young water dragon living in a nice little paradise. Build pictures for this cool enclosure can be seen here: http://www.reptile-community.com/smf/index.php?topic=25516.0;topicseen
Reptile Forums UK - A member shares some pictures of a very nice looking CWD and its very cool viv.@ Reptile Forums UK


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The Career Path of a Nephrologist
Nephrology is projected to be a growing field, as the aging population grows in addition to there being more cases of kidney problems. According to the National Kidney Foundation, 33% of all adults are at risk of kidney disease. Kidney function begins to decline at the age of 50, leading to an increase in chronic kidney disease in those over 60 years of age. The National Kidney Foundation goes on to state that the average adult is more likely to have complications with kidney disease than they are to experience diabetes or breast cancer.
The Characteristics of a Successful Nephrologist
Like most positions in the medical field, success depends on many factors. Education, drive, bedside manner, and intelligence are all important factors in being a good nephrologist. You will have to develop a relationship with your patient and be able to help answer their questions. Correctly diagnosing a situation and helping a patient is frequently reported as the most rewarding part of the job.
As a patient, everyone has a favorite physician that they've worked with. Think about the characteristics that they had that made them good at their job. They were patient and compassionate, paid attention to the details, and used critical thinking skills and problem-solving skills to not only diagnose the situation but to also help the patient. Subjects like internal medicine and physiology, combined with a lot of math, are a big part of the skillset needed to be a successful nephrologist.
If you are a student, you may want to explore adding a third-year study that includes transplantation, critical care, or other subjects that can set you apart and possibly increase your salary range.
Workplaces for Nephrologists
If you choose to go into the field of nephrology, you have a wide variety of places that you can find employment. These places include:
- Clinics
- Hospitals
- Medical Centers
- Private Practice
- Universities
You may also work in academic settings, outpatient clinics, healthcare organizations, solo practices, hospitals, office-based group practices, and other locations. You should be able to find a place to work that matches what you are looking for.
Regardless of where you work, the tasks will probably be the same for the most part. Your patients will be treated for kidney stones, dialysis, or even transplants. They may have chronic disorders or acute diseases impacting the function of their kidneys. You'll have to diagnose the problem and prescribe a plan of treatment for your patient.
Most nephrologists report working up to 50 hours a week, with 40 of those hours spent seeing patients. The time with patients averages around 20 minutes per patient. The remaining time is used for administrative duties, documentation, and organizational tasks.
Compensation for nephrologists depends on many variables including the practice's setting, the hiring situation, and the regional location. According to the Medscape Nephrologist Compensation Report (http://www.medscape.com/features/slideshow/compensation/2013/nephrology), nephrology ranks 14th for the highest income specialty. In addition, the pay gap between male and female nephrologists is 13%, less than many other medical specialties.
Research Opportunities in the Field of Nephrology
Research is an important part of Nephrology, so many doctors look to pursue this endeavor. Many nephrologists work in a research setting part-time, in addition to their clinic time.
The areas of research that are most important at this time are associated with subjects like making new organs available for transplant, discovering new therapies and treatments for acute renal failure, and improving the process for hospitals and surgical centers during kidney procedures and treatments.
Many research projects are underwritten by universities, non-profit foundations, and shareholders of private companies. Although government funding is available, currently 75% of research funding is provided by private institutions.
Opportunities Increasing in Nephrology
Unfortunately, the population of the United States is seeing an increase in Type II Diabetes, which in turn, is a factor in kidney disease. As more people develop kidney problems, the need for talented nephrologists will grow. It is critical that this increased demand is met with nephrologists that are knowledgeable and talented.


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Fences provide thе perfect boundary frоm оnе property tо another. Fences саn аlѕо provide homeowners with much-needed privacy. In addition to that, a fence can also provide protection for your property. Thеrе аrе several companies thаt manufacture аnd install fences. However, it is important to find a trusted, experienced and reliable fence company in NJ that fits all your needs and wants.
It is important to find a company that wіll оffеr a full service thаt wіll cover everything уоu need аnd tailor іt tо уоur specific requirements аnd thе individual demands оf thе site. In оrdеr tо mаkе thіѕ possible, a gооd company wіll provide a site survey tо assess уоur individual needs аnd thе possible solutions thеу саn оffеr. Frоm thіѕ initial survey, thеу wіll ensure уоur fences аrе manufactured tо thе highest possible standard аnd quality ensuring thаt thеу match уоur needs exactly. It іѕ best tо find a fence company in NJ thаt will give you a fair quote for the services and materials required for your specific fence project.
A trusted fence company will provide you with virtually аnу fencing requirement аnd always come up wіth аn imaginative аnd practical solution fоr thе client. Thе trick іѕ tо hаvе a clear idea оf what уоu want. A gооd design company wіll hеlр уоu develop thеѕе ideas once уоu commission them.
Thе designers аt a gооd fence company wіll mаkе sure thаt thе solution уоu аrе offered perfectly matches уоur individual needs. Thіѕ іѕ dоnе bу thе provision оf аn initial site survey. Thіѕ survey allows thеіr designers аnd engineers to assess thе area аnd ensure thаt the fence you choose meets уоur requirements. Along with meeting your specific needs, it is important that the fence contractor is providing you with true solutions thаt wіll work perfectly for you аnd last fоr a very long tіmе.
Thе durability оf a fence іѕ enhanced even further bу thе perfect matching оf thе design tо thе environment whеrе іt wіll bе installed. A reliable fence company in NJ wіll pride themselves оn thе care thаt іѕ taken bу thеіr design аnd engineering team tо ensure thаt thе fence wіll bе secure аnd long lasting. In addition tо thіѕ, thе installation itself ѕhоuld always bе carried оut wіth great care аnd professionalism. Anу problems thаt occur wіth installations оn site аrе referred back tо thе designers аnd engineers tо ensure thаt thе fence іѕ installed securely and tо thе highest possible standards.


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Archive for the 'Blanton' Category
Bye Week Update 8/27August 27, 2009
Fall Camp 8/19… With TAH-NOO-TAH!!!August 19, 2009
Wu Am I? Thick Killah & Homicidal Terra-hawkJuly 25, 2009
Welcome to another edition of Wu Am I? for the 2009 season. Once again- I am trying to identify the 36 players of impact for the 2009 squad. So without any further bullshit, I open chambers 23 & 24…
Eric Olsen #55
Offensive Guard
6'5″ 303 lbs.
Brooklyn, New York
Wu Name: Thick Killah
Although Eric was considered a 4 star prospect and the #2 ranked player in the state of New York, he didn't receive a whole lot of attention from the fanbase or the media when he chose Notre Dame over schools like Miami Fl., North Carolina, and Boston College. He stayed quiet during his freshman year while playing a little over 20 minutes (probably should have red-shirted), and became the fulltime starter at guard for the Irish during the middle of the 2007 season. In the spring of 2008, he opened the eyes of a conservative Irish fanbase, as he became very aggressive in practice. He was involved in more than a few fights, and was videotaped while yelling, "Fuck you defense! Fuck you!" I fell in love instantly.
2009 should be a much better year for Eric than in 2008- if he can stay healthy. One of the worst moments of the 2008 season was when Boston College defensive tackle and future first round pick by the Green Bay Packers, BJ Raji, basically threw Olsen back 5 yards into the ballcarrier (James Aldridge) and knocked said ballcarrier on his ass. Ouch! It should be noted that 1. Olsen was on two wounded legs, and 2. Raji was an animal and future NFL first rounder. I would bet against anyone who thinks that Olsen hasn't replayed that play in his mind a hundred times while chewing on glass as he works out. I think that play, and the help of the new offensive line coach, will turn Olsen into the animal that he really is in his heart. BTW- There are a few rumors going around about Olsen moving over to center to replace Wenger, and Trevor Robinson starting at what is currently Olsen's left guard spot. I'm not sure what will happen, but you can be sure that Olsen will be right in the middle of it… swinging.
Robert Blanton #12
6'1″ 185 lbs.
Matthews, North Carolina
Wu Name: Homicidal Terra-hawk
Robert Blanton is a special kid- and he will let you and anyone else within shouting distance know about it. When Blanton committed to Notre Dame in June of 2007, he passed up on a host of defensive powerhouses such as Georgia and Virginia Tech, to name just a few. You have to give this kid credit (and the whole incoming class of 2008 for that matter) for sticking to his commitment, even after the dismal 2007 season. Irish fans were treated to a very confident athlete that saw action very early in his career. The most memorable Blanton moment from 2008 was probably his INT return for TD against Purdue to spark what looked to be a somewhat flat Irish team. Personally, I liked the U$C pick. It gave me some hope in what turned out to be another ass-whooping in the Colosseum.
The loss of Darrin Walls last year really opened the door for Blanton to kick down and destroy. He showed up and played better than Lambert and at times better than McNeil. Don't think that Blanton is sweating his playing time with the return of Walls. He's probably telling anyone that will listen that he is going to run this shit. And perhaps he will. Notre Dame will be more talented and deeper in the secondary than at any point this decade, and Blanton could end up being the best overall corner on the roster. No matter how the playing time splits up between Walls, McNeil, Blanton , and even Gary Gray, I fully expect RJ to have a major impact and make some very big plays. I'm thinking pick 6 against Tommy Trojan. FU$C.
The MVP's & The GeneralApril 19, 2009
Armando Allen
Robert Blanton
Coach Weis
You can see more highlights and interviews at UND.com… HERE.
I'll have my own thoughts on the game and the gameday experience in a day or so. I just need a little more time to let this stuff marinate- it's not like I don't have some time between now and the Nevada game. Don't worry though- it will be up soon.
Bagpipe Monday Is On Delay… I Give You More InterviewsMarch 30, 2009
Bagpipe Monday will be out later today. Until then…
Corwin Brown
Raeshon McNeil
Robert Blanton
Harrison Smith
CornerbattleMarch 25, 2009
A look at the corners during todays practice
The Curious Case of Sergio BrownMarch 6, 2009
It's that time of year. Spring football is right around the corner and the message boards are a buzz with roster movement and depth chart speculation. People can't help themselves. It's fun, it's interesting, and its all we got right now.
I generally try to stay away from printing out any type of depth chart or starter projection-type material. In this era of football, more players are playing due to both sides of the ball using many different formations and packages. So instead of pinpointing 22 guys as "starters", I have instead pointed to 35-36 players of impact. These are players that will contribute heavily to the team. Sergio Brown is certainly one of those players, but could he be more?
If you ask around the Notre Dame locker room, "Who is the best athlete on the team?" More times than not, Sergio Brown would be the answer, and it has been for the last couple of years.
Last spring, Brown competed against Kyle McCarthy for the strong safety spot that opened up after the departure of 4 year starter Tom Zbikowski. McCarthy won that position battle and it bore fruit after Kyle led the team in tackles in 2008 with 110 and also had 2 interceptions. It was a very solid performance- that also had me yawning. Where were the big plays? Granted, Kyle did make some very big tackles that helped the team. However, it was done so quietly and 10-20 yards past the line of scrimmage, that it looked effortless.
After spring ball, the designated starters at defensive back were pretty much decided. These are the players that fell into the slots:
- CB- Darrin Walls
- CB- Terrail Lambert
- FS- David Bruton
- SS- Kyle McCarthy
- Nickel- Raeshon McNeil
- Dime- Gary Gray
Oh, how things change- and quickly at that. After Walls, the most talented defensive back on the roster, left school for personal reasons it opened up a huge hole. That hole got filled by Raeshon McNeil, but like a submarine taking on water, another hole opened up. The lineup soon looked like this:
- CB- Raeshon McNeil
- CB- Terrail Lambert
- FS- David Bruton
- SS- Kyle McCarthy
- Nickel- Sergio Brown
- Dime- Gary Gray
It didn't take long for this lineup to get tampered with either. Robert Blanton started kicking down the door from the day he stepped on campus and he soon got a lot of playing time. At first it was Gary Gray rotating in with Lambert and occasionally McNeil. Then, Blanton took over the role that Gray was occupying as the season wore on.
As we roll into spring football for 2009, we see the same type of battles that must take place to decide the participants. With Walls back, and the emergence of Robert Blanton the lineup will definitely be shook up- but is it the right shake up. Enter Sergio Brown.
After playing on special teams in his first two seasons and racking up a colossal 11 tackles, Brown stepped onto the stage in a huge way for the Irish at the Nickel position from game one last year. He was an effective blitzer and terrific in coverage. He showed great athleticism and playmaking ability and in my opinion- he proved himself to be able to contend for the SS spot once again. He deserves it, and keep in mind that this is his last year of eligibility. (That last part sucks balls).
I just feel that Brown brings more to the table than McCarthy. I like Kyle, but I also like safeties that make big plays. I'm not sure if Kyle can do it or can't do it, but I am sure that Sergio has more of the tools needed to make those big plays.
In today's game, we see more and more defenses having to move towards using the nickel package as somewhat of a base defense against spread teams. Spread teams are multiplying. So, the nickel is just as important as the SS for the most part. My ideal setup for the defensive backs in a nickel formation looks like this:
We can argue about the Blanton/McNeil battle at a later time. By now, the McCarthy brothers and fan club are ready to tear my ass apart, but we have to be honest with ourselves. You could at least pencil Kyle in as the Dime- but I keep hearing that the staff is very high on Jamoris Slaughter too. Uh, oh…
By now, if you're still reading this, you are probably wondering if I am moving towards some type of point or closing argument. Perhaps. But I think I have made it clear already. Sergio Brown is an exciting and athletic player and in my opinion- he should be starting at the SS spot. This puts our defensive backfield in better situations to make the big play. McCarthy led the team in tackles last year with 110, so I can understand that some people may be hesitant to accept this fact, but we should. A safety shouldn't be making that many tackles anyways.
This is the "Curious Case of Sergio Brown," but in essence it is also the " Curious Case of the Secondary." What do we do with what is the most talented defensive backfield the Irish have had in a very long time? In my opinion, we go with the playmakers and let them make plays, and somehow that all revolves around Mr. Brown.
Interviews Galore… #3, #7, #12, #23, #58December 13, 2008
Mike Floyd returns to action.
Golden Tate is glad Floyd is back.
Jimmy is happy Floyd is back too… duh.
R.J. talks trash?
Brian Smith is tougher than Charlie Weis.
The Ides Of CommitmentsJune 20, 2007
Robert Blanton is Irish. The 6'1″ 175 pound, 4 star prospect chose the Irish over Virginia Tech and…Skunkbear University. Lllloyd Carr- suck on that! Some have Blanton listed as free safety, but the Irish like him at cornerback. There are a lot of similarities between Robert and a former Irish player… Bobby Taylor. The size and speed this kid possesses is fantastic, but his hip movement and foot speed are what sets him apart as a CB as opposed to a FS.
This is the 15th Notre Dame commit for the 2008 incoming class. Our 15th commit last year was actually this year when Matt Romine chose ND at the Army AA Game. This class is filling up in a heartbeat. The Irish do have needs at WR, OL, and LB still to fill, but with every commit it adds pressure to those sitting on the fence to follow suit. (cough–Floyd–cough–Filer–cough–Page).
Click here to see video on Blanton.


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Elastos solves the trilemma of all blockchain projects out there - the problem of security using the trusted runtime environment, scalability using the main chain sidechains solutions and decentralization using the peer to peer network.
Elastos is the safe and reliable internet of the future. Built utilizing the blockchain, this technological breakthrough provides the first completely safe environment on the web where decentralized applications are detached from the internet while also permitting full scalability to billions of users. Elastos enables the generation of wealth through ownership and exchange of your data and digital assets.
Elastos is not a blockchain project but rather a network operating system project powered by blockchain technology so in that sense, Elastos is not directly competing with any other blockchain projects. It can work together with them to form this new ecosystem where the decentralized applications run directly on the device instead of running on the blockchain along with decentralized peer to peer network to transfer assets in a completely closed sandboxed environment, thereby solving the trilemma of blockchain projects - security, scalability, and decentralization.
Inside elastOS, Elastos uses internet as the base-layer infrastructure and decentralized applications are forced to never directly connect to the internet and only interact with Elastos runtime(which in turn acts as a middle layer, then connecting to the internet). Within the elastos ecosystem, everything from devices to virtual machines to users to DApps have a DID(Decentralized ID) issued by the blockchain and everytime the application requests any sort of internet access, the ID has to first be verified using the blockchain which in turn eliminates many of the man-in-the-middle attacks, virus attacks, etc.
Elastos gets its hashpower from Bitcoin and some of the largest mining pools in the world including Bitmain, this secures the ELA token which is also used for DPoS node voting. By distributing load across multiple sidechains and replacing expensive PoW consensus in major blockchains like Ethereum with DPoS, Elastos is able to achieve far greater performance while maintaining a secure trustless environment.
Keep learning about Elastos in the following chapters:


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Sunday, August 21, 2016
LegalShield | Member Story of the Day
If you want peace of mind and answers to all your question about legal matters then today is the perfect time to get one and use it anytime you need legal help and advice. Here is a story that you may find helpful in making a decision.
Member Story of the Day - Big Pharma:
"Shortly after my beloved grandmother passed away, my Mom and my Uncle received a bill for $2,197.14 for a prescription drugs that were delivered, but, sadly, we lost my grandmother so quickly she never even opened the package.
The pharmaceutical company refused to take the drugs back, and the credit card company refused to credit the account. After several months of my Mom and her Brother arguing back and forth with the pharmaceutical and credit card companies, I finally got wind of the situation and told my Mom to just call LegalShield to see how they could help. She did call, and her almost 100-year old A.V. rated Provider Law Firm in California wrote a letter on her behalf to the pharmaceutical company, and within two weeks the entire debt was removed.
Thank you, LegalShield, for providing an awesome service and helping my family save a lot of money and eliminating the stress in an already sad and stressful time in our lives. It's resolutions like this that make me passionate about sharing this powerful membership."
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Lets talk and learn about life.Share your thoughts, ideas, opinions,experiences, points of view,and family matters and parenting ideas. A good outcome would come up by Connecting with other people from different walks of life around the world.


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It is January. It is cold and wet. We are all punishing ourselves with detox and starvation and resolutions that make us feel bad about ourselves.
It's not a good time.
So what I prescribe it an enormous injection of colour.
There now, that's going to make it all better, isn't it?
Alexander McQueen
Etoile Isabel Marant
Jil Sander
Jil Sander and Stella McCartney
Jonathan Saunders
Marc Jacobs
Miu Miu
Moschino Cheap and Chic
Nina Ricci and Juicy Couture
Pierre Hardy
Reed Krakoff
Roksanda Ilincic
Roland Mouret
See by Chloe
Stella McCartney
Theyskens' Theory
Vanessa Bruno Athé
(Images via Net a Porter)


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She told stories, flirted outrageously with boys and was constantly changing her hairstyle.
Anne Frank hid with her family in a secret room at her father Otto Frank's office in Amsterdam.
It could be the description of almost any young girl growing up in Europe. But this is how Eva Schloss remembers her childhood friend Anne Frank, who had she not died in a Nazi concentration camp, would have celebrated her 80th birthday this week.
Schloss described Frank, whose account of hiding from Jewish persecution in Nazi-occupied Amsterdam is one of the world's mostly widely-read books, as a spunky young schoolgirl with a passion for storytelling that often got her into trouble.
"She got her diary in 1942, so obviously her father knew she was interested in writing and I know she told stories," said Schloss.
"She talked a lot and she was called Mrs Quack Quack. Very often she used to write hundreds of lines [at school] of 'I'm not going to talk so much,' and so on -- but obviously she had a lot to tell."
In some ways the two friends lived parallel lives -- but tragically they had very different outcomes. Watch more about Schloss' story »
Schloss and Frank both came from Jewish families who fled to Holland to escape the wave of anti-Semitism spreading across Europe as the Nazis rose to power in Germany ahead of the Second World War.
But while Schloss was more of an introvert, Frank loved the limelight. Schloss said: "I was actually quite shy and she was the center of attention. We had steps where we sat, and she had a crowd of children around her.
"She was a big flirt -- she loved boys. She was always showing us who was her boyfriend at that particular time. She was always interested in her clothes. Her style, she always changed it. Sometimes she had curls, then she had straight hair."
Schloss says they were unaware of the full scale of what was going on around them as war escalated across Europe, placing their lives in increasing jeopardy.
"Our parents really protected us so there was no talk about the horrendous things which happened.
"You couldn't go out anymore after 8 o'clock, but for a 11 to 12 year old it didn't matter so much. Or not going to the cinema -- we were upset about those little things which we couldn't do, but we really didn't really take it seriously at that time."
Like Frank, Schloss was also forced into hiding when the Nazis took control of Holland.
Frank hid with her family in a secret room at her father Otto Frank's office. But Schloss and her family had to split up. Schloss stayed with her mother while her father and brother hid elsewhere. She and her mother moved around, staying in seven different hiding places over a two-year period.
Eventually both families were betrayed and were sent to concentration camps, where Frank died at the age of 15.
Schloss said: "My father and brother were betrayed by a Dutch nurse who was a double agent, and all four of us were arrested and taken to the headquarters to be interrogated.
"I didn't know anything, which was a good thing. So eventually they realized this and they gave up torturing me. Within two days we were put on a transport to Auschwitz."
Of her family, only Schloss and her mother survived Auschwitz, one of the most notorious concentration camps, located in southern Poland.
Today Schloss, who has just celebrated her own 80th birthday, has a husband, three daughters and five grandchildren.
Schloss says it took her decades to rebuild her life, with the help of Frank's father Otto, who also survived incarceration in a concentration camp.
She met Otto in August 1945, when he showed her Frank's diary.
Schloss said: "He read a few passages but he always burst into tears. It took me 20 years. I was really unhappy, but it was Otto who came to our apartment to talk to us, and he helped me a lot. He had lost everybody.
"Her book, she [Frank] made people aware of what happened. There are many messages. She believed in the goodness of mankind.
"People always ask me, what she would have done. I guess we will never know. But I guess she would have gone into politics -- she was a fighter. It's a pity, but also -- maybe her diary would have never been published."
CNN's Don Riddell contributed to this story.
All About Auschwitz-Birkenau • Anne Frank


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The contract on work performance was signed in April 2012 between PJSC EMAlliance and OJSC Kharkov Boiler-Mechanical Works. According to contract relations EMAlliance is obliged to design, manufacture and supply supported water cooling system of high and low pressure, steam-to-steam heat-exchanger, bypass pipes of high and low pressure and low pressure convection superheater of second stage.
Equipment was designed by the specialists of Engineering Center of EMAlliance (Taganrog) and manufactured on the main manufacturing facility of the company – Taganrog Boiler-Making Works Krasny Kotelshchik.
Supplied parts will be mounted on the second unit of Tripolskaya TPP. The use of modified equipment will allow rejecting the use of supporting fuel at the plant and extend the resource of replaceable boiler elements.
Steam-to-steam heat-exchanger commissioning will be an innovation for plant operation. Equipment use will provide maintaining the temperature of secondary superheating within required load range increasing the reliability of Tripolskaya TPP operation.


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SingSnap - Online Karaoke Community
Register for free access to singing and recording thousands of karaoke songs, millions of recordings to listen to and entertainment that lasts a life time...
... SingSnap brought out a talent in me I never knew I had.
... The truth of the matter is that for the 3 minutes of a song, I can pretend that I am somebody that matters.
... it has the best features, staff and support who are there right away whenever I need assistance and most of all it boosts my confidence to sing.


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Data Wizards // SolutionsMaking data work for you
Growth | Innovation | Efficiency
Innovative use of data drives tangible business acceleration, fuels new business models and enables organisations to optimise their processes.
We develop data solutions that leverage the power of data with the collective human intelligence of the organisation. That's what we mean when we say:
The Power of Data, driven by People
Growth | Innovation | Efficiency
Innovative use of data enables tangible business acceleration, fuels new business models and enables organisations to optimise their processes.
We develop data solutions that leverage the power of data with the collective human intelligence of the organisation. That's what we mean when we say:
The Power of Data, driven by People
Our Services:
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The use of data as a catalyst for change is only possible when the required data is easily accessible in an environment where the right tools and applications are available.
Together we review the entire process from data inception through data transformation to storage, enabling you to decide on the optimal storage solution for your organisation.
Using our experience with data warehouses and data lakes, we can subsequently develop and build your new data infrastructure.
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Combining theoretical knowledge and practical experience in different industries and not-for-profit organisations, our team builds applications that meet your needs.
Using frameworks like Hadoop and Spark, platforms like Amazon Web Services, tools like Python and R and coupling this with concepts like artificial intelligence (AI), machine learning and deep learning, we are able to extract real value from your data.
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To derive the full benefits of a data centric approach you need continuous access to your ever changing data sets.
We provide you with the tools to analyse large and complex data sets and visualise the outcomes in reports and dashboards that drive better business decisions.
These tools are either developed exclusively for your organisation or use third party solutions like Google Cloud Platform or Microsoft Azure ,
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Our way of working is based on a 4D model. It is based on simple proven agile concepts and processes. This allows us to develop our applications in a structured manner while still enabling our customer to provide continuous input and maintaining track of the project's progress. The model is centered around 4 phases that can be performed individually or consecutively: Discover, Design, Develop and Deliver.
Identify and classify possibilities. Determine objectives.
Design based on objectives, desired business processes and interfaces.
An iterative process of developing, testing and reviewing.
Training, go live and ongoing support.
New or different ways of using data can offer opportunities for your organisation. There is however no 'one size fits all' solution. Discovery is about identifying how data can drive innovation and growth in your organisation.
This is done through meetings and workshops with the stakeholders in your organisation.
Once we have established the opportunities, these are prioritised and objectives are set. A clear understanding of these objectives is crucial for the next phase: Design.
Once the objectives have been defined, we create a detailed design for the application.
In addition to the various technical aspects, we also consider your organisation's desired process flows and user interfaces.
The initial design is validated through workshops and, if necessary, amended. This is an iterative process. Once we have agreement on the final design, we create a development plan.
The development of the application is split into multiple smaller projects. Each of these smaller projects is completed in short cycles where we plan, build, test and demonstrate the individual parts.
The feedback from these demonstration sessions is used to improve and finalise each of these projects.
Furthermore, this provides you with an opportunity to continuously monitor progress and, where possible, specify changes.
After succesfully completing the development phase we set up a rigorous user acceptance testing program. This is essential to ensure we deliver an application that meets your requirements and expectations.
Final delivery of the application takes place once acceptance testing is completed successfully,
If the application would need additional development in the future, we would follow a similar (but shorter) process.
Identify and classify possibilities. Determine objectives.
Design based on objectives, desired business processes and interfaces.
Iterative process of developing, testing and reviewing.
Training, go live and ongoing support.
Some of our clients:
WePowerPeople maakt werk van werkgeverschap.
De behoefte bij werkgevers in de mobiliteitsbranche aan ondersteuning op de thema's werkgeluk, verzuim, ontwikkeling, rendement, werkvermogen en productiviteit neemt verder toe.
WePowerPeople is van mens en werk en ondersteunt medewerkers en organisaties in de ontwikkeling. Voor welke uitdagingen staat je bedrijf? Welke talenten en kwaliteiten heb je nodig voor de ontwikkeling van de organisatie? Wat is de volgende stap in jouw carrière?
Dat is wat waar we graag met je over in gesprek gaan. Niet alleen omdat het ons een bestaan verschaft, het is wat ons drijft. Bij WePowerPeople, gaat het over mensen en hun potentieel, omdat mensen die in beweging komen méér bereiken!


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Wear what the World Team wears! Our Connect/Commit shoe tags were worn by the AKC/USA Agility Team at the 2014 World Agility Championship in Luxembourg!
This pair of hand stamped shoe tags makes a great motivational gift for the agility addict. This pair is stamped with "Connect" on one tag and "Commit" on the other. These words are part of the inspirational adage, Connect Commit Celebrate, which was developed by Nancy Gyes to be worn by the 2014 AKC/USA Agility Team. So now you can wear what the World Team wears!
The tags measure 3/8" by 1 1/2" and fit most running shoes. The holes are 5/32" and fit most shoestrings.
The tags are available in aluminum (shown here), copper, brass, or sterling silver.
The shoe tags will arrive in a gifty box.


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Speak to the team at Print on Collins or browse online for our full list of services. You won't be disappointed in the final outcome.


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Interest on the Notes will be payable in cash in arrears semi-annually through May 13, 2008. Thereafter, the principal amount of the debentures will accrete semi-annually at a rate of 2.75 percent per year to maturity.
RPM may redeem the notes, in whole or in part, on or after May 13, 2008. The company has also granted the initial purchaser of the notes an option to purchase up to an additional $49.5 million principal amount of notes, which would result in up to an additional $25 million gross proceeds to RPM.
The offering has been made only to qualified institutional buyers in accordance with Rule 144A under the Securities Act of 1933. RPM intends to use the net proceeds of the offering to repay borrowings outstanding under its credit facilities.
Neither the convertible notes nor the shares issuable upon conversion have been registered under the Securities Act of 1933 or any state securities laws and, until so registered, may not be offered or sold in the United States or any state absent registration or an applicable exemption from registration requirements.
RPM International Inc., a holding company, owns subsidiaries that are world leaders in specialty coatings serving both industrial and consumer markets. RPM's industrial products include roofing systems, sealants, corrosion control coatings, flooring coatings and specialty chemicals. RPM's consumer products are used by professionals and do-it-yourselfers for home maintenance and improvement, automotive and boat repair and maintenance, and by hobbyists. Industrial brands include Stonhard, Tremco, Carboline, Day-Glo, Euco and Dryvit. Consumer brands include Zinsser, Rust-Oleum, DAP, Varathane, Bondo and Testors.
This press release contains "forward-looking statements" relating to the business of the Company. These forward-looking statements, or other statements made by the Company, are made based on management's expectations and beliefs concerning future events impacting the Company and are subject to uncertainties and factors (including those specified below) which are difficult to predict and, in many instances, are beyond the control of the Company. As a result, actual results of the Company could differ materially from those expressed in or implied by any such forward-looking statements. These uncertainties and factors include (a) general economic conditions; (b) the price and supply of raw materials, particularly titanium dioxide, certain resins, aerosols and solvents; (c) continued growth in demand for the Company's products; (d) legal, environmental and litigation risks inherent in the Company's construction and chemicals businesses and risks related to insurance coverage inherent in the Company's disclosed litigation; (e) the effect of changes in interest rates; (f) the effect of fluctuations in currency exchange rates upon the Company's foreign operations; (g) the effect of non- currency risks of investing in and conducting operations in foreign countries, including those relating to domestic and international political, social, economic and regulatory factors; (h) risks and uncertainties associated with the Company's ongoing acquisition and divestiture activities; and other risks detailed in the Company's other reports and statements filed with the Securities and Exchange Commission, including the risk factors set forth in the Company's prospectus and prospectus supplement included as part of the Company's Registration Statement on Form S-3 (File No. 333-77028), as the same may be amended from time to time.
SOURCE RPM International, Inc.
CONTACT: Glenn R. Hasman, vice president finance and communications of RPM, 330-273-8820/
RPM International Inc. (NYSE: RPM) owns subsidiaries that are world leaders in coatings, sealants, building materials and related services. From homes to precious landmarks worldwide, their brands are trusted by consumers and professionals alike to protect, improve and beautify. Among its leading consumer brands are Rust-Oleum, DAP and Zinsser. Learn more about RPM brands >>
RPM is a compelling long-term investment.
The percent by which RPM's 10-year total return has bested the S&P 500. More reasons >>
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