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Genetic Diversity and Evolution
Comparative Analysis of 22 Coronavirus HKU1 Genomes Reveals a Novel Genotype and Evidence of Natural Recombination in Coronavirus HKU1
Patrick C. Y. Woo, Susanna K. P. Lau, Cyril C. Y. Yip, Yi Huang, Hoi-Wah Tsoi, Kwok-Hung Chan, Kwok-Yung Yuen
Patrick C. Y. Woo
Susanna K. P. Lau
Cyril C. Y. Yip
Yi Huang
Hoi-Wah Tsoi
Kwok-Hung Chan
Kwok-Yung Yuen
DOI: 10.1128/JVI.00509-06
We sequenced and compared the complete genomes of 22 strains of coronavirus HKU1 (CoV HKU1) obtained from nasopharyngeal aspirates of patients with respiratory tract infections over a 2-year period. Phylogenetic analysis of 24 putative proteins and polypeptides showed that the 22 CoV HKU1 strains fell into three clusters (genotype A, 13 strains; genotype B, 3 strains and genotype C, 6 strains). However, different phylogenetic relationships among the three clusters were observed in different regions of their genomes. From nsp4 to nsp6, the genotype A strains were clustered with the genotype B strains. For nsp7 and nsp8 and from nsp10 to nsp16, the genotype A strains were clustered with the genotype C strains. From hemagglutinin esterase (HE) to nucleocapsid (N), the genotype B strains were clustered closely with the genotype C strains. Bootscan analysis showed possible recombination between genotypes B and C from nucleotide positions 11500 to 13000, corresponding to the nsp6-nsp7 junction, giving rise to genotype A, and between genotypes A and B from nucleotide positions 21500 to 22500, corresponding to the nsp16-HE junction, giving rise to genotype C. Multiple alignments further narrowed the sites of crossover to a 143-bp region between nucleotide positions 11750 and 11892 and a 29-bp region between nucleotide positions 21502 and 21530. Genome analysis also revealed various numbers of tandem copies of a perfect 30-base acidic tandem repeat (ATR) which encodes NDDEDVVTGD and various numbers and sequences of imperfect repeats in the N terminus of nsp3 inside the acidic domain upstream of papain-like protease 1 among the 22 genomes. All 10 CoV HKU1 strains with incomplete imperfect repeats (1.4 and 4.4) belonged to genotype A. The present study represents the first evidence for natural recombination in coronavirus associated with human infection. Analysis of a single gene is not sufficient for the genotyping of CoV HKU1 strains but requires amplification and sequencing of at least two gene loci, one from nsp10 to nsp16 (e.g., pol or helicase) and another from HE to N (e.g., spike or N). Further studies will delineate whether the ATR is useful for the molecular typing of CoV HKU1.
The recent severe acute respiratory syndrome (SARS) epidemic, the discovery of SARS coronavirus (CoV), and the identification of SARS CoV-like viruses from Himalayan palm civets and a raccoon dog from wild-animal live markets in mainland China have led to a boost in interest in the discovery of novel coronaviruses in both humans and animals (8, 23, 26, 28, 40, 42). In 2004, a novel group 1 human coronavirus (HCoV), NL63, was reported independently by two groups (6, 34). In 2005, we described the discovery, complete genome sequence, clinical features, and molecular epidemiology of a novel group 2 human coronavirus, HKU1 (genotype A) (17, 37-39, 41). This virus has also subsequently been found in patients with respiratory tract infections in other countries (1, 30, 33). Recently, we have also identified a SARS CoV-like virus in Chinese horseshoe bats and a novel group 1 coronavirus in large bent-winged bats, lesser bent-winged bats, and Japanese long-winged bats in the Hong Kong Special Administrative Region (16, 27). The discovery of SARS CoV-like viruses in horseshoe bats was confirmed by another group in other provinces in China (19).
As a result of the unique mechanism of viral replication, coronaviruses have a high frequency of recombination (15). Their tendency for recombination and high mutation rates may allow them to adapt to new hosts and ecological niches. However, no convincing evidence among human coronaviruses of genetic recombination that may have contributed to their ability to reinfect humans has been documented. In our study of the phylogeny of the RNA-dependent RNA polymerase (pol), spike (S), and nucleocapsid (N) genes of nine isolates of CoV HKU1 recovered from patients with pneumonia, it was discovered that the sequences of the S and N genes fell into two distinct genotypes, with seven strains belonging to genotype A and two belonging to genotype B (41). On the other hand, for the pol gene, one of the two "genotype B" strains as determined by its S and N sequences (from patient 8) was clustered with the other seven "genotype A" strains (41). Furthermore, the same phenomenon was also observed in our subsequent prospective study of CoV HKU1-associated respiratory tract infections (17). Based on these observations, we suspected that there is an additional CoV HKU1 genotype which has arisen from recombination between genotypes A and B of CoV HKU1.
To test this hypothesis, we performed complete genome sequencing on 21 additional strains of CoV HKU1 and compared their genomes to the CoV HKU1 genotype A strain (38). The sites of recombination were identified, and a novel CoV HKU1 genotype, genotype C, was defined.
CoV HKU1 strains.All 22 CoV HKU1 strains were isolated from patients with respiratory tract infections in Hong Kong in a 2-year period (March 2003 to February 2005) (Table 1) (17, 38, 41).
Characteristics of the 22 CoV HKU1 strains used in this study
RNA extraction.Viral RNA was extracted from the nasopharyngeal aspirates of the patients using a QIAamp viral RNA mini kit (QIAGEN, Hilden, Germany). The RNA pellet was resuspended in 10 μl of DNase-free, RNase-free double-distilled water and was used as the template for reverse transcription-PCR.
Complete genome sequencing and genome analysis.The complete genome sequence of the CoV HKU1 genotype A strain was described previously (GenBank accession no. NC_006577) (38). The complete genomes of the other 21 CoV HKU1 strains were amplified and sequenced using the RNA extracted from the nasopharyngeal aspirates of the corresponding patients as the template, by a strategy described previously (38). The RNA was converted to cDNA by a combined random-priming and oligo(dT) priming strategy. The 5′ ends of the viral genomes were confirmed by rapid amplification of cDNA ends using the 5′/3′ RACE kit (Roche, Germany). Sequences were assembled and manually edited to produce final sequences of the viral genomes. The 21 genomes were compared to that of the CoV HKU1 genotype A strain and were manually annotated.
Phylogenetic-tree construction.The nucleotide sequences for nsp1, nsp2, conserved portions of nsp3 (including papain-like protease 1 [PL1pro], a member of the Appr-1-p processing enzyme family [A1pp], papain-like protease 2 [PL2pro], and the hydrophobic domain [HD]), nsp4-nsp10, nsp12-nsp16, hemagglutinin esterase (HE), S, open reading frame 4 (ORF4), envelope (E), membrane (M), and N were extracted from the 22 CoV HKU1 genomes. Phylogenetic-tree construction was performed using the neighbor-joining method with ClustalX 1.83. The corresponding nucleotide sequences of human coronavirus OC43 (GenBank accession no. AY585229) were used as outgroups.
Bootscan analysis.To perform bootscan analysis, a nucleotide alignment of the genome sequences of one genotype A (38), one genotype B (patient 1 of reference 41), and one genotype C (patient 8 of reference 41) strain of CoV HKU1 and one HCoV OC43 strain (GenBank accession no. AY585229) was generated by ClustalX, version 1.83, and edited manually. Bootscan analysis was performed using Simplot version 3.5.1 (F84 model; window size, 1,000 bp; step, 200 bp) (20), with the genome sequence of HCoV OC43 as a query.
Nucleotide sequence accession numbers.The nucleotide sequences of the 21 additional genomes of CoV HKU1 strains (data not shown) have been lodged within the GenBank sequence database under accession no. AY884001, DQ339101, DQ415896, DQ415897, DQ415898, DQ415899, DQ415900, DQ415901, DQ415902, DQ415903, DQ415904, DQ415905, DQ415906, DQ415907, DQ415908, DQ415909, DQ415910, DQ415911, DQ415912, DQ415913, and DQ415914.
Complete genome sequence, genome organization, phylogenetic analysis, and genotypes.The sizes of the genomes of the 22 CoV HKU1 strains ranged from 29,295 to 30,097 nucleotides. The G+C contents of all 22 genomes are 32%. The overall genome organizations of the 22 CoV HKU1 strains were the same (Fig. 1A).
Genome organization and phylogenetic analysis of CoV HKU1. (A) Genome organization of CoV HKU1. The regions of the genome used for phylogenetic tree construction are labeled. (B) Phylogenetic analysis of nsp1, nsp2, conserved portions of nsp3 (PL1pro, A1pp, PL2pro, and HD), nsp4-nsp10, nsp12-nsp16, HE, S, ORF4, E, M, and N of the 22 CoV HKU1 genomes. The trees were constructed by the neighbor-joining method using Jukes-Cantor correction and bootstrap values calculated from 1,000 trees. Seven hundred forty, 1,821, 655, 333, 895, 1,263, 1,488, 909, 861, 276, 582, 330, 411, 2,783, 1,809, 1,563, 1,125, 900, 1,276, 4,126, 330, 253, 687, and 1,358 nucleotide positions in nsp1, nsp2, PL1pro, A1pp, PL2pro, HD, nsp4, nsp5, nsp6, nsp7, nsp8, nsp9, nsp10, nsp12, nsp13, nsp14, nsp15, nsp16, HE, S, ORF4, E, M, and N, respectively, were included in the analysis. The scale bar indicates the estimated number of substitutions per 50 or 100 nucleotides as indicated. The corresponding nucleotide sequences of HCoV OC43 were used as the outgroups. A, genotype A; B, genotype B and C, genotype C.
Phylogenetic trees using the nucleotide sequences of genes for putative proteins and polypeptides (nsp1, nsp2, conserved portions of nsp3 [PL1pro, A1pp, PL2pro, and HD], nsp4-nsp10, nsp12-nsp16, HE, S, ORF4, E, M, and N) of the 22 CoV HKU1 strains were constructed and are shown in Fig. 1B. In 18 of the 24 trees, the 22 CoV HKU1 strains fell clearly into three clusters, named genotype A (13 strains), genotype B (3 strains), and genotype C (6 strains). The exceptions are the five trees constructed using nsp1, nsp2, PL1pro, PL2pro, and HD, in which the differences among the sequences were too small, and the nsp10 tree, in which two genotype A strains, N1 and N3, were clustered with genotype C strains.
The three genotypes exhibited different relationships to each other in different regions of their genomes. From nsp4 to nsp6, the genotype A strains were clustered with the genotype B strains, but for nsp7 and nsp8, the genotype A strains were clustered with the genotype C strains. From nsp10 to nsp16, the genotype A strains were clustered closely with the genotype C strains, with high bootstrap values, but from HE to N, the genotype B strains were clustered closely with the genotype C strains, with bootstrap values of 1,000 in all cases. No association was observed between the genotypes and the time of detection or the age, sex, clinical disease, presence of underlying disease, or outcome of the patients (Table 1).
The putative transcription regulatory sequence motif, 5′-AAUCUAAAC-3′ (as in mouse hepatitis virus [MHV] and bovine coronavirus) (22) or, alternatively, 5′-UAAAUCUAAAC-3′, that was found at the 3′ end of the leader sequence and precedes each translated ORF except ORF5 described to occur in the genome of the CoV HKU1 genotype A strain, was also present in all of the other 21 CoV HKU1 genomes. On the other hand, the sequence of the putative internal ribosomal entry site (IRES) (32) for the ORF of the envelope protein in the genomes of all three CoV HKU1 genotype B strains and all six genotype C CoV HKU1 strains were UUUUAUCGCUUGG, instead of AUUUAUUGUUUGG in all 13 CoV HKU1 genotype A strains, although both sequences were similar to the IRES element, UUUUAUUCUUUUU, in MHV (10).
The 22 genomes differed in their numbers of tandem copies of the 30-base acidic tandem repeat (ATR) in the N terminus of nsp3 inside the acidic domain upstream of PL1pro (Tables 1 and 2). All 22 genomes had tandem copies of a perfect 30-base repeat which encodes NDDEDVVTGD and various numbers of imperfect repeats. The median number of tandem copies of the perfect 30-base repeat was 11.5 (range, 2 to 17), and the median number of imperfect repeats was 2 (range, 1 to 4). All of the 10 CoV HKU1 strains with incomplete imperfect repeats (1.4 and 4.4) belonged to genotype A.
Amino acid sequences of acidic tandem repeats of the 22 CoV HKU1 strainsa
Bootscan analysis.Bootscan analysis showed that from the 5′ end of the genome to position 12000, there could be a number of possible recombination sites in the genomes of the three genotypes (Fig. 2). Right upstream to position 11500, high bootstrap support for clustering between the CoV HKU1 genotype A strain and the CoV HKU1 genotype B strain was observed. From positions 13000 to 21500, high bootstrap support for clustering between the CoV HKU1 genotype A strain and the CoV HKU1 genotype C strain was observed. From position 22500 to the 3′ end of the genome, high bootstrap support for clustering between the CoV HKU1 genotype B strain and CoV HKU1 genotype C strain was observed. These findings indicate that recombination has possibly taken place between nucleotide positions 11500 and 13000, corresponding to the nsp6-nsp7 junction, and between nucleotide positions 21500 and 22500, corresponding to the nsp16-HE junction.
Bootscan analysis of the CoV HKU1 genomes. Bootscanning was conducted with Simplot version 3.5.1 (F84 model; window size, 1,000 bp; step, 200 bp) on a gapless nucleotide alignment, generated with ClustalX, with the genome sequence of HCoV OC43 (GenBank accession no. AY585229) as the query sequence. The dashed line denotes CoV HKU1 genotype A (NC_006577), the solid line denotes CoV HKU1 genotype B (AY884001), and the dotted line denotes CoV HKU1 genotype C (DQ339101).
Comparative sequence analysis of the nsp6-nsp7 junction and nsp13-HE gene junction.Since both phylogenetic trees and bootscan analysis showed that there was a possible recombination site at the nsp6-nsp7 junction and the nsp16-HE gene junction, multiple alignments among the nucleotide sequences of the 22 genomes were performed to ascertain the exact sites of recombination.
Upstream of nucleotide position 11750 of the CoV HKU1 genotype A genome (227 bases before the end of nsp6), there was high nucleotide identity between the sequences of the CoV HKU1 genotype A and genotype B strains, whereas downstream to nucleotide position 11892 of the CoV HKU1 genotype A genome (85 bases before the end of nsp6), there was high nucleotide identity between the sequences of the CoV HKU1 genotype A and genotype C strains (Fig. 3). This indicates that the site of crossover was probably within a 143-bp region between nucleotide positions 11750 and 11892.
Comparative sequence analysis of the nsp6-nsp7 junction. Multiple alignment of the nucleotide sequences of CoV HKU1 genotypes A, B, and C. In CoV HKU1 genotype B and CoV HKU1 genotype C, only the nucleotides differing from those in CoV HKU1 genotype A are depicted. The nucleotides in CoV HKU1 genotype C that are the same as those in CoV HKU1 genotype A but different from those in CoV HKU1 genotype B are highlighted in black, and those in CoV HKU1 genotype B that are the same as those in CoV HKU1 genotype A but different from those in CoV HKU1 genotype C are highlighted in gray. The putative template switching region is underlined and bold. The first (TCA) and last (CAG) codons of nsp7 are also underlined. The arrows denote positions with nucleotide polymorphism (at position 11414, N16, N17, N20, N21, and N22 [genotype C] were T instead of C; at 11422, N16 and N20 [genotype C] were C instead of T; at 11449, N25 [genotype B] was C instead of T; at 11528, N16, N17, N20, and N22 [genotype C] were C instead of T; at 11740, N6, N7, N9, N10, N11, N13, N14, N18, N23, and N24 [genotype A] were T instead of C; at 12095, N6 and N7 [genotype A] were T instead of C; at 12140, N23 and N24 [genotype A] were C instead of T; at 12367, N15 and N25 [genotype C] were A instead of G; and at 12400, N6, N7, N9, N10, N11, N13, N14, N18, N23, and N24 [genotype A] were C instead of T).
Upstream of nucleotide position 21502 of the CoV HKU1 genotype A genome (249 bases before the stop codon of ORF1ab), there was high nucleotide identity between the sequences of the CoV HKU1 genotype A and genotype C strains, whereas downstream of nucleotide position 21530 of the CoV HKU1 genotype A genome (221 bases before the stop codon of ORF1ab), there was high nucleotide identity between the sequences of the CoV HKU1 genotype B and genotype C strains, including a 13-bp insertion just downstream of the stop codon of ORF1ab (Fig. 4). This indicates that the site of crossover was probably within a 29-bp region between nucleotide positions 21502 and 21530.
Comparative sequence analysis of the nsp16-HE gene junction. Multiple alignments of the nucleotide sequences of CoV HKU1 genotypes A, B, and C. In CoV HKU1 genotype A and CoV HKU1 genotype B, only the nucleotides differing from those in CoV HKU1 genotype C are depicted. The nucleotides in CoV HKU1 genotype C that are the same as those in CoV HKU1 genotype A but different from those in CoV HKU1 genotype B are highlighted in gray, and those in CoV HKU1 genotype C that are the same as those in CoV HKU1 genotype B but different from those in CoV HKU1 genotype A are highlighted in black. The putative template switching region is underlined and bold. The stop codon of ORF1ab (TAG) and the start codon of the HE gene (ATG) are also underlined. The arrows denote positions with nucleotide polymorphisms (at 21297, N6, N7, N9, N10, N11, N13, N14, N23, and N24 [genotype A] were T instead of G; at 21429, N6 [genotype A] was C instead of T; at 21576, N15 [genotype B] was C instead of T; at 21908, N15 and N25 [genotype B] were G instead of A; and at 21949, N14 was T instead of C).
This is the first time that evidence for natural recombination is documented for coronavirus associated with human infection. Coronaviruses are unique in having a high frequency of homologous RNA recombination, as a result of random template switching during RNA replication, thought to be mediated by a "copy choice" mechanism (2, 4, 13, 14, 21, 35). In feline coronavirus (FCoV), it has been documented that FCoV type II strains originated from a double recombination between FCoV type I and canine coronavirus, and the site of recombination has been pinpointed to a region of about 50 nucleotides in the M gene by multiple alignment (9). As for recombination between different strains of MHV, in vitro studies have shown variations in both sites and rates of recombination, with the S gene having a frequency threefold that of the polymerase gene (7, 21). In the present study, by comparing the sequences of 22 complete genomes of CoV HKU1 strains, we documented that major recombination has occurred among the three CoV HKU1 genotypes. Both phylogenetic and bootscan analysis showed that the nucleotide sequences of the six genotype C strains were almost identical to those of the 13 genotype A strains from nsp10 to nsp16 (Fig. 1B and 4). Interestingly, the topologies of the phylogenetic trees changed dramatically starting from the HE gene. From HE to N, the nucleotide sequences of the six genotype C strains were almost identical to those of the three genotype B strains (Fig. 1B). This is also in line with results of bootscan analysis, suggesting recombination between genotypes A and B, giving rise to genotype C (Fig. 2). Multiple alignments of the nucleotide sequences of the nsp16-HE regions of the three genotypes confirmed our suspicion, and results localized the site of recombination to a stretch of 29 nucleotides in nsp16, just upstream to the stop codon of ORF1ab (Fig. 4). This is in keeping with the finding that the putative IRES of all the genotype B and genotype C strains were the same but different from those of the genotype A strains, as it is located downstream of HE. In addition to the recombination site in nsp16, there was another one at the end of nsp6, also evidenced by a shift in clustering in the phylogenetic trees, bootscan analysis, and multiple-alignment results (Fig. 1B, 2 and 3). In contrast to the nsp16 recombination site, recombination has occurred between genotypes B and C in this region, giving rise to genotype A. Furthermore, as shown in bootscan and phylogenetic analyses, additional recombination events might have occurred in ORF1ab upstream of nsp5 (Fig. 1B and 2). However, due to the relatively small variations in the sequences among the three genotypes, these putative recombination sites were difficult to ascertain with multiple alignments.
A novel genotype, genotype C, of CoV HKU1 is defined. It has been well known that recombination is an important mechanism for the generation and evolution of virus genotypes (12, 29, 31). In our previous study, we showed that seven of the nine CoV HKU1 strains were of genotype A and one of the nine strains was of genotype B by pol, S gene, and N gene sequence analysis (41). In the present study, we showed that the latter half of the genomes of the six genotype C strains probably represents a result of recombination between genotypes A and B. Analysis of the complete genomes of more CoV HKU1 strains from other countries will reveal the relative prevalences of the different genotypes in different localities. From the results of the present study, no association was observed between the genotypes and clinical characteristics of the patients. Furthermore, amplification and sequencing of a single gene is not sufficient to define the genotype of CoV HKU1. It would require amplification and sequencing of at least two gene loci, one from nsp10 to nsp16 (e.g., pol or helicase) and another from HE to N (e.g., S or N).
The origin and function of the ATR located inside the acidic domain upstream of PL1pro, unique to CoV HKU1, remain enigmatic. Significant variations were observed among the ATRs of CoV HKU1 strains. Only two pairs of CoV HKU1 strains (N1 and N3, N7, and N9) possessed the same nucleotide sequence in their ATRs. No relationship was found between the number of repeats and the genotype or virulence of the strains. We speculate that this "independent evolution" of the number of repeats was due to the random expansion or deletion of part of the repeat region during the process of viral replication as a result of inaccurate replication by the viral polymerase or recombination between the repeat regions of different CoV HKU1 strains, a phenomenon widely observed in tandem repeats of genomes in all domains of life (3, 24). On the other hand, the sequence of the imperfect repeats seemed to coevolve with the rest of the genomes, most notably that all 10 CoV HKU1 strains with incomplete imperfect repeats (NDDD) were of genotype A (Tables 1 and 2). This could be due to the deletion of part of a repeat in one genotype A strain and subsequent expansion or deletion of whole repeats in its descendants. Further studies will delineate whether this ATR is useful for the molecular typing of CoV HKU1 strains.
This high frequency of recombination has resulted in the generation of a high diversity of coronaviruses in different animals. Before the SARS epidemic in 2003, a total of 19 coronaviruses were known, including 2 human, 13 mammalian, and 4 avian coronaviruses. After the SARS epidemic, within a short period of 3 years, 20 additional novel coronaviruses were described (5, 6, 11, 16, 19, 25, 26, 34, 36, 38, 43). These include 3 human coronaviruses, 11 mammalian coronaviruses, and 6 avian coronaviruses. Notably, there was a recent discovery of at least eight different species of coronaviruses in bats in Hong Kong, including SARS CoV-like viruses and a probable novel subgroup, group 2c, of coronavirus (16, 43). The high frequency of recombination in such a high diversity of coronaviruses may easily result in the generation of novel coronavirus species or genotypes that can cross host species barriers, leading to major zoonotic outbreaks with disastrous consequences. The potential of generation of novel species leading to zoonotic outbreaks and major consequences is analogous to the situation of avian and human influenza epidemiology, although the mechanism of generation of novel types and variants is by reassortment, which is different from recombination in coronaviruses (18, 44). Amplification of conserved regions in coronaviruses using RNA extracted from various animal specimens will lead to the discovery of more coronaviruses and subsequent complete genome sequencing, and comparative genome analysis will reveal the intricate relationships among the various coronaviruses.
In memory of the late Ted Sun and with gratitude to his family for their generous support for research on emerging infections. This work is partly supported by a Research Grant Council grant (7616/05 M); the Research Fund for the Control of Infectious Diseases of the Health, Welfare, and Food Bureau of the Hong Kong Special Administrative Region Government; Commercial Radio's Fund; the Suen Chi Sun Charitable Foundation; and the William Benter Infectious Disease Fund.
Received 10 March 2006.
Accepted 25 April 2006.
↵*Corresponding author. Mailing address: Department of Microbiology, The University of Hong Kong, University Pathology Building, Queen Mary Hospital, Hong Kong. Phone: (852) 28554892. Fax: (852) 28551241. E-mail: hkumicro{at}hkucc.hku.hk.
↵† P.C.Y. Woo and S.K.P. Lau contributed equally to this paper.
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Marra, M. A., S. J. Jones, C. R. Astell, R. A. Holt, A. Brooks-Wilson, Y. S. Butterfield, J. Khattra, J. K. Asano, S. A. Barber, S. Y. Chan, A. Cloutier, S. M. Coughlin, D. Freeman, N. Girn, O. L. Griffith, S. R. Leach, M. Mayo, H. McDonald, S. B. Montgomery, P. K. Pandoh, A. S. Petrescu, A. G. Robertson, J. E. Schein, A. Siddiqui, D. E. Smailus, J. M. Stott, G. S. Yang, F. Plummer, A. Andonov, H. Artsob, N. Bastien, K. Bernard, T. F. Booth, D. Bowness, M. Czub, M. Drebot, L. Fernando, R. Flick, M. Garbutt, M. Gray, A. Grolla, S. Jones, H. Feldmann, A. Meyers, A. Kabani, Y. Li, S. Normand, U. Stroher, G. A. Tipples, S. Tyler, R. Vogrig, D. Ward, B. Watson, R. C. Brunham, M. Krajden, M. Petric, D. M. Skowronski, C. Upton, and R. L. Roper. 2003. The genome sequence of the SARS-associated coronavirus. Science 300:1399-1404.
Nakamura. Y., M. Leppert, P. O'Connell, R. Wolff, T. Holm, M. Culver, C. Martin, E. Fujimoto, M. Hoff, and E. Kumlin. 1987. Variable number of tandem repeat (VNTR) markers for human gene mapping. Science 235:1616-1622.
Pearks Wilkerson, A. J., E. C. Teeling, J. L. Troyer, G. K. Bar-Gal, M. Roelke, L. Marker, J. Pecon-Slattery, and S. J. O'Brien. 2004. Coronavirus outbreak in cheetahs: lessons for SARS. Curr. Biol. 14:R227-R228.
Peiris, J. S., S. T. Lai, L. L. Poon, Y. Guan, L. Y. Yam, W. Lim, J. Nicholls, W. K. Yee, W. W. Yan, M. T. Cheung, V. C. Cheng, K. H. Chan, D. N. Tsang, R. W. Yung, T. K. Ng, and K. Y. Yuen. 2003. Coronavirus as a possible cause of severe acute respiratory syndrome. Lancet 361:1319-1325.
Poon, L. L., D. K. Chu, K. H. Chan, O. K. Wong, T. M. Ellis, Y. H. Leung, S. K. Lau, P. C. Woo, K. Y. Suen, K. Y. Yuen, Y. Guan, and J. S. Peiris. 2005. Identification of a novel coronavirus in bats. J. Virol. 79:2001-2009.
Rota, P. A., M. S. Oberste, S. S. Monroe, W. A. Nix, R. Campagnoli, J. P. Icenogle, S. Penaranda, B. Bankamp, K. Maher, M. H. Chen, S. Tong, A. Tamin, L. Lowe, M. Frace, J. L. DeRisi, Q. Chen, D. Wang, D. D. Erdman, T. C. Peret, C. Burns, T. G. Ksiazek, P. E. Rollin, A. Sanchez, S. Liffick, B. Holloway, J. Limor, K. McCaustland, M. Olsen-Rasmussen, R. Fouchier, S. Gunther, A. D. Osterhaus, C. Drosten, M. A. Pallansch, L. J. Anderson, and W. J. Bellini. 2003. Characterization of a novel coronavirus associated with severe acute respiratory syndrome. Science 300:1394-1399.
Simmonds, P., and S. Midgley. 2005. Recombination in the genesis and evolution of hepatitis B virus genotypes. J. Virol. 79:15467-15476.
Sloots, T. P., P. McErlean, D. J. Speicher, K. E. Arden, M. D. Nissen, and I. M. Mackay. 2006. Evidence of human coronavirus HKU1 and human bocavirus in Australian children. J. Clin. Virol. 35:99-102.
Steininger, C., B. Schmied, M. Sarcletti, M. Geit, and E. Puchhammer-Stockl. 2005. Cytomegalovirus genotypes present in cerebrospinal fluid of HIV-infected patients. AIDS 19:273-278.
Thiel, V., and S. G. Siddell. 1994. Internal ribosome entry in the coding region of murine hepatitis virus mRNA 5. J. Gen. Virol. 75:3041-3046.
Vabret, A., J. Dina, S. Gouarin, J. Petitjean, S. Corbet, and F. Freymuth. 2006. Detection of the new human coronavirus HKU1: a report of 6 cases. Clin. Infect. Dis. 42:634-639.
van der Hoek, L., K. Pyrc, M. F. Jebbink, W. Vermeulen-Oost, R. J. Berkhout, K. C. Wolthers, P. M. Wertheim-van Dillen, J. Kaandorp, J. Spaargaren, and B. Berkhout. 2004. Identification of a new human coronavirus. Nat. Med. 10:368-373.
van der Most, R. G., L. Heijnen, W. J. Spaan, and R. J. de Groot. 1992. Homologous RNA recombination allows efficient introduction of site-specific mutations into the genome of coronavirus MHV-A59 via synthetic co-replicating RNAs. Nucleic Acids Res. 20:3375-3381.
Wise, A. G., M. Kiupel, and R. K. Maes. 2006. Molecular characterization of a novel coronavirus associated with epizootic catarrhal enteritis (ECE) in ferrets. Virology 349:164-174.
Woo, P. C., Y. Huang, S. K. Lau, H. W. Tsoi, and K. Y. Yuen. 2005. In silico analysis of ORF1ab in coronavirus HKU1 genome reveals a unique putative cleavage site of coronavirus HKU1 3C-like protease. Microbiol. Immunol. 49:899-908.
Woo, P. C., S. K. Lau, C. M. Chu, K. H. Chan, H. W. Tsoi, Y. Huang, B. H. Wong, R. W. Poon, J. J. Cai, W. K. Luk, L. L. Poon, S. S. Wong, Y. Guan, J. S. Peiris, and K. Y. Yuen. 2005. Characterization and complete genome sequence of a novel coronavirus, coronavirus HKU1, from patients with pneumonia. J. Virol. 79:884-895.
Woo, P. C., S. K. Lau, Y. Huang, H. W. Tsoi, K. H. Chan, and K. Y. Yuen. 2005. Phylogenetic and recombination analysis of coronavirus HKU1, a novel coronavirus from patients with pneumonia. Arch. Virol. 150:2299-2311.
Woo, P. C., S. K. Lau, H. W. Tsoi, K. H. Chan, B. H. Wong, X. Y. Che, V. K. Tam, S. C. Tam, V. C. Cheng, I. F. Hung, S. S. Wong, B. J. Zheng, Y. Guan, and K. Y. Yuen. 2004. Relative rates of non-pneumonic SARS coronavirus infection and SARS coronavirus pneumonia. Lancet 363:841-845.
Woo, P. C., S. K. Lau, H. W. Tsoi, Y. Huang, R. W. Poon, C. M. Chu, R. A. Lee, W. K. Luk, G. K. Wong, B. H. Wong, V. C. Cheng, B. S. Tang, A. K. Wu, R. W. Yung, H. Chen, Y. Guan, K. H. Chan, and K. Y. Yuen. 2005. Clinical and molecular epidemiological features of coronavirus HKU1-associated community-acquired pneumonia. J. Infect. Dis. 192:1898-1907.
Woo, P. C., S. K. Lau, B. H. Wong, H. W. Tsoi, A. M. Fung, K. H. Chan, V. K. Tam, J. S. Peiris, and K. Y. Yuen. 2004. Detection of specific antibodies to severe acute respiratory syndrome (SARS) coronavirus nucleocapsid protein for serodiagnosis of SARS coronavirus pneumonia. J. Clin. Microbiol. 42:2306-2309.
Woo, P. C., S. K. Lau, K. S. Li, R. W. Poon, B. H. Wong, H. W. Tsoi, B. C. Yip, Y. Huang, K. H. Chan, and K. Y. Yuen. 26 April 2006. Molecular diversity of coronaviruses in bats. Virology [Epub ahead of print].
Yuen, K. Y., P. K. Chan, M. Peiris, D. N. Tsang, T. L. Que, K. F. Shortridge, P. T. Cheung, W. K. To, E. T. Ho, R. Sung, and A. F. Cheng. 1998. Clinical features and rapid viral diagnosis of human disease associated with avian influenza A H5N1 virus. Lancet 351:467-471.
American Society for Microbiology
Journal of Virology Jun 2006, 80 (14) 7136-7145; DOI: 10.1128/JVI.00509-06
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"Time Spent Stitching" is the perfect stitchers alibi. This sampler is part of the premiere Traditions line introduced by Great Bear Canada. The pattern opens up to reveal a great big (17½" x 23½") color chart with easy-to-read symbols. A gift for tired eyes! Includes quotations you'll enjoy.
Item # GBC-112 stitched on aida or evenweave; DMC and Anchor codes.
My grandmother, Margaret Mellon, was one of those ordinary, extraordinary women. She was born in 1881 and lived deep into the next century. She lived, in fact, until she was nearly 100!
In between, she met and married my grandfather, moved to Maine and then back to New Brunswick, had nine children (two of whom she lost as babies), and helped run a farm. She also made things with her hands – everything from quilts to socks. She knit and wove and hooked and quilted.
My Aunt Mary tells me she and Grandma even knit socks for the boys off in the Navy. ("I used to feel terrible for the poor souls who had to wear them!" my aunt confesses now). The thought of those lovingly crafted, but probably scratchy and bulky, socks makes me smile now, as I count stitches and pull thread in my home in Hamilton, Ontario.


text:
Berserker è una Space opera dello scrittore statunitense Fred Saberhagen. In Italia è stata edita parzialmente dalla collana fantascientifica Urania.

La parola Berserker, nell'antica Mitologia norrena, indicava quei guerrieri che in battaglia erano particolarmente pervasi da foga omicida. Nella saga di Saberhagen è un tipo di macchine, costruito millenni prima della nascita dell'uomo, che hanno sviluppato una propria intelligenza artificiale, giungendo alla decisione di sterminare dall'universo qualsiasi forma di vita biologica.

Opere del ciclo
Il mondo dei Berserker (Berserker, 1967), Urania n. 1152, Mondadori. Antologia di racconti.
Berserker! (Brother Assassin o Brother Berserker, 1969), Urania n. 1123, Mondadori. Testo online. Raccoglie i romanzi brevi:
Stone Man, in Worlds of Tomorrow, maggio 1967.
The Winged Helmet, in If, agosto 1967.
Brother Berserker, in If, novembre 1967.
Il pianeta Berserker (Berserker's Planet, maggio/giugno 1974 su If), Urania n. 1256, Mondadori.
L'uomo Berserker (Berserker Man, 1979), Urania n. 1133, Mondadori.
The Ultimate Enemy, 1979. Antologia di racconti.
Le guerre dei Berserker (Berserker Wars, 1981), Urania n. 1174, Mondadori. Testo online. Antologia di racconti:
L'elmo alato
La ballata delle stelle
Ultimi avvenimenti al radiante dei Templari
Ali nell'oscurità (anche Ali dall'ombra)
Il sorriso
Berserker Base: A Collaberative Novel, 1984, scritta con vari autori
What Makes Us Human, di Stephen R. Donaldson, su F&SF, agosto 1984.
With Friends Like These, di Connie Willis, su F&SF, febbraio 1985.
Itself Surprised, di Roger Zelazny, su Omni, agosto 1984.
Deathwomb, di Poul Anderson, su Analog, novembre 1983.
Pilots of the Twilight, di Edward Bryant, su Asimov's, dicembre 1984.
A Teardrop Falls, di Larry Niven, su Omni, giugno 1983.
Il trono dei Berserker (The Berserker Throne, 1985), Urania n. 1201, Mondadori.
Berserker: la morte azzurra (Berserker: Blue Death, 1985), Urania n. 1233, Mondadori.
The Berserker Attack, 1987. Antologia di racconti precedentemente editi.
Berserker Lies, 1991. Antologia con un racconto originale.
I Berserker uccidono (Berserker Kill, 1993), Urania Argento n. 6, Mondadori.
La furia dei Berserker (Berserker Fury, 1997), Urania n. 1349, Mondadori.
Berserker: il titano d'acciaio (Shiva in Steel, 1998), Urania n. 1381, Mondadori.
Berserkers: The Beginning, 1998 (raccolta omnibus di volumi precedenti)
Berserker's Star, 2003
Berserker Prime, 2004
Berserker Man: Mega Book (raccolta omnibus di volumi precedenti)
Berserker Death, 2005
Rogue Berserker, 2005

Note

Voci correlate
Berserker

Collegamenti esterni

Cicli di fantascienza
Opere di Fred Saberhagen

redpajama_set_name: RedPajamaWikipedia


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text:
Michael Addelman
News and Media Relations Officer - Faculty of Biology, Medicine, and Health
Register for news releases (journalists only)
New research calls for better care for people who seek emergency help following self-harm
Research has found that people who go to A&E following self-harm receive varying quality of care and this has a significant impact on what they experience subsequently.
The study, which was codesigned and co-authored with people who have lived experience of self-harm and mental health services, found negative experiences were common, and revealed stigmatising comments about injuries from some hospital staff. Some participants reported being refused medical care or an anaesthetic because they had harmed themselves. This had a direct impact on their risk of repeat self-harm and suicide risk, as well as their general mental health.
The research, 'Relieved to be seen - patient and carer experiences of psychosocial assessment in the emergency department following self-harm: qualitative analysis of 102 free-text survey responses'*, was published today in the journal BMJ Open. It was funded by the National Institute for Health Research Greater Manchester Patient Safety Translational Research Centre (NIHR GM PSTRC). The Centre is a partnership between The University of Manchester and Salford Royal NHS Foundation Trust.
According to the research, the participants who received supportive assessments with healthcare staff reported feeling better, less suicidal and were less likely to repeat self- harm.
This research highlights the importance of learning from the experiences of individuals to help improve care for people who have harmed themselves. We involved patients and carers throughout the entire process and this enabled us to gain a greater insight into what patients want after they present to hospital having harmed themselves
Dr Leah Quinlivan
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Dr Leah Quinlivan, from The University of Manchester, a Research Fellow at the GM PSTRC's Mental Health theme, led the study, and said: "Our research looked at the care people received at A&E following self-harm. It's part of an overall programme of work that aims to improve patient safety in mental health services. This research highlights the importance of learning from the experiences of individuals to help improve care for people who have harmed themselves. We involved patients and carers throughout the entire process and this enabled us to gain a greater insight into what patients want after they present to hospital having harmed themselves. "
According to NICE guidelines anyone who arrives at A&E following a self-harm episode should receive a psychosocial assessment. The research has revealed how important these assessments are for people who harm themselves. For participants in the study, if the assessments were delivered correctly and with compassion, they reported being less likely to harm themselves in the future. The research also revealed that assessments can be helpful when staff work alongside the person affected by self-harm, and are empathetic to their emotional distress.
Participants in the research revealed what a negative experience looked like and this was when waiting times were unduly long, assessments overly standardised and a lack of compassion was shown by staff.
Elizabeth Monaghan, one of the co-authors with lived experience in this area said: "The research demonstrates the value of involving patients and carers throughout the research process. Our varied experiences of assessments in A&E during and after crises meant we were able to add helpful insights into the research design and findings.
"These insights shaped the research team's understanding of what factors are helpful for A&E staff to be able to carry out assessments more effectively and with compassion. Being part of the research process all the way through has given me an opportunity to use my experiences to improve care as well as giving me valuable skills in being part of a team carrying out research in this sensitive area."
Jenny Groves has 35 years of lived experience of self-harm and is the CEO and Founder of Battle Scars, a survivor led charity for people who have self-harmed. She, said: "This research shows the discrepancies in the procedures not just in different parts of the country but the huge differences in attitude and treatment even within the same emergency department.
"Ticking boxes, dismissing, judging and invalidating people who present at A&E following self-harm cause long-term damage and increase risk. Acceptance, compassion and a true person-centred approach, acknowledging the individual and their needs, can make a world of difference to someone who is struggling and resorting to self-harm to manage life."
Study at The University of Manchester opens in new window

redpajama_set_name: RedPajamaCommonCrawl


From the garden to your living room, keep a vibrant bouquet looking fresh with this simply luminous cylinder vase, designed for the heavy branches of plumeria. From Lenox.
A delicate, etched vine motif and a scalloped rim infuses your decor with gentle sophistication.
Sculpted fuschia pops with a tinge of pink against this distinctly shaped Flora vase. A quiet beauty in graceful Lenox porcelain.


text:
Areas of freezing fog early. Cloudy early with peeks of sunshine expected late. High near 40F. Winds N at 5 to 10 mph..
Partly cloudy skies this evening will become overcast overnight. Low 23F. Winds light and variable.
Legislators go over some thorny issues
Area legislators discussed various "misunderstandings" experienced between the Oklahoma Legislature and Governor Brad Henry during this year's legislative session during the Woodward Chamber of Commerce's final eggs and Issues conference Friday morning.
Sen. Owen Laughlin and Reps. Gus Blackwell, Jeff Hickman and Rob Johnson participated in the conference which was held at Northwestern Electric's Medallion Room.
All four legislators agreed that one of the biggest misunderstandings has been over the state's general appropriations budget.
Blackwell, R-Goodwell, said that in the past the Senate was known as the place where all good bills go to die, but that was not the case this year. He said the Senate worked amazingly well, especially considering the 24-24 tie in that chamber, and that instead the Governor's desk was treacherous ground for bills, especially for the state budget.
Johnson, R-Kingfisher, explained that for once the legislature had the budget ready for approval before the April 1 deadline, but after returning from a month long vacation the governor vetoed it.
Hickman, R-Dacoma, said Henry only expressed problems with about 10 percent of the budget, but he vetoed 100 percent of the budget even though he had line item veto authority.
The legislature then tried to pass portions from the budget Henry proposed in his State of the State address, he said, but those were vetoed as well.
"He doesn't want to play well with others," Johnson said of the governor's behavior.
Hickman added that he believes "the governor has been completely unreasonable."
Johnson said Henry has refused to compromise and continues to stick to his original budget plan, which according to Johnson and Hickman, was based on old numbers and flawed projections.
"We can't do much if the governor isn't willing to work with us," Johnson said.
And with only nine days left in the session, it does not look like they have enough time to get everything accomplished that they need to, the legislators said.
Blackwell said it is likely that the legislature will have to go into a special session before they can finally pass a budget.
"We will have a budget," he promised. "When, I'm just not sure."
Johnson claimed the delay was an attempt by the governor to get the legislature to pass the budget that he wants, which will spend more money than he said the state can afford. He also claimed that the governor was also trying to take some of the heat off himself by making it look like the legislature failed in accomplishing what they needed to in the regular session.
Hickman agreed saying "it's about spreading the blame."
Laughlin, R-Woodward, said another big misunderstanding revolved around lawsuit reform, which also ended in a vetoed bill.
He discussed how the governor claimed to be for tort reform, but then vetoed the tort reform bill which the legislature had worked hard to develop.
Johnson said that like the budget, the lawsuit reform bill was "a compromise piece of legislation," which many within both parties and even within the professions which the legislation would have affected worked together to develop.
He said by his veto the governor showed that he does not really want tort reform, because "if he did, he could have had it."
Hickman explained why lawsuit reform is so important, saying that it would help to fight against the "crisis" of rising health care costs.
Lawsuit reform is also important, Laughlin said, because it can help bring higher paying jobs to Oklahoma.
Johnson said that because it is so important, the legislature will probably send the bill back again, although he is not confident that it will do any better next year.
While the legislature does not have the votes to override Henry's vetoes on the budget or on lawsuit reform, Laughlin said they are working hard to get the votes to override his veto on a bill that would bar the use of public funds for abortions.
Hickman reminded everyone that the governor did not veto everything.
He noted how Henry had signed a bill which put over $200 million into the teacher's retirement system and allowed for an additional $60 million to be added each year for the next several years, so that by 2026 the system will be 80 percent funded, where at it is only 49 percent funded today.
Another important bill that Henry approved was an immigration reform bill which focuses on immigrants' work eligibility, helps prevent illegal immigrants from obtaining benefits and also allows state and local law enforcement to enforce federal immigration laws, Hickman said.
Laughlin said another successful bill will provide ad valorem tax breaks to help the new assisted living center which is being built in Woodward.
Other issues that were dealt with during this year's legislative session were also discussed during a question and answer period which ended the Eggs and Issues conference.
These other issues included the governor's plan to place three-year-olds in the classroom, the performance audit planned for the Department of Corrections, and even more about lawsuit reform.
Also during the question and answer period, Mayor Bill Fanning publicly thanked Blackwell, Hickman, Johnson, and Laughlin for how well they supported Woodward throughout the session.


Hedi Slimane photographed Sasha Pivovarova for an editorial spread titled Tough Love for Vogue Japan August 2011, styled by Mika Mizutani. Characteristic of Hedi Slimane, Sasha Pivovarova was shot in mostly black and white imagery donning soon-to-be-released Fall 2011 collections from the likes of Louis Vuitton, Azzedine Alaia, Gucci, Rick Owens, Ann Demeulemeester, Balenciaga and Haider Ackermann among others.
I don't know if I will ever get bored with Hedi Slimane's style and his mostly blonde subjects like Sasha Pivovarova here. Till then, while we wait for his return to fashion design, let's indulge for now, shall we?
This entry was posted in Editorials and tagged Ann Demeulemeester, Azzedine Alaia, Balenciaga, Gucci, Haider Ackermann, Hedi Slimane, Louis Vuitton, Rick Owens, Sasha Pivovarova, Vogue by fashionslop. Bookmark the permalink.


Free download old songs for mohamed abdelwahab. ( Yes but it' s been a tradition here since 1999. Ignatz Jerry Miller Andres Huicochea Kartways you almost certainly just hear fun Thank you for making the sincere effort to idp ilsaf13 embarrasing ยิ นดี.
Mawazine ( Arabic: موازين , translit. mawāzīn, meaning " rhythms" ) is a Moroccan International music festival held annually in Rabat, Morocco, featuring many international and local music artists. The festival is presided over by Mounir Majidi, the personal secretary of the Moroccan King Mohammed VI and founder and president of Maroc Culture, the cultural foundation that yla and Majnun ( Arabic: مجنون ليلى  ), also Leili o Majnun ( Persian: ليلى و مجنون  ), is a narrative poem composed in 584/ 1188 by the Persian poet Niẓāmi Ganjavi based on a semi- historical Arab story about the 7th century Bedouin poet Qays ibn Al- Mulawwah and his ladylove Layla bint Mahdi ( or Layla al- Aamiriya). Nizami also wrote Khosrow and Shirin in the 12th.


The .357 SIG is a necked down 10mm, or a a necked down .40 S&W depending on how you want to look at it. This round was created back in 1994 not so very long ago, and was designed by SIG-Sauer. (thats where the name comes from). The .357 Sig is a very fast round much faster than the 380, 9mm, 40, 10mm,45 ACP, and even the large 357 Cartridge. The sig has an average speed of 1379 fps if you round speed of all know ammo.
This round is hard to find and very expensive compared to the 9mm, and the 40 S & W. This is most likely due to the fact that it's not produced in the numbers that the other cartridges are. If it becomes a more popular round in the future, chances are the price will drop when there are more ammo manufacturers producing the bullet.
The average gr is 122 and this number again is created by averaging all known ammo for the cartridge, we are not even sure if they make a 122 gr bullet it's just the average. It has on average 515 lb of energy at the muzzle. This means the Sig 357 has more energy at muzzle than the 40, 45 gap, 45 auto, 9mm and of course the smaller rounds. Data shows this as a very stout and fast cartridge flying flatter and further than its parent case the 40 with ease according to our ballistic calculations. Overall power ranking however sets it between the 9mm and 40 S & W for a final tally.
*Casing image above is an artist rendering and not a real photo of .357 SIG Ballistics cartridge. While we have went to great lengths to make sure that it's as accurate as possible this rendering should not be used to generate specs for casings.


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Check Out These 3 Cafes The Next Time You're In Seoul!
Top 10 Items To Buy In A Korean Supermarket!
Geoje, a city located in the South Gyeongsang Province, is the second largest island in Korea. It is also easily accessible by bus or car from Busan, the second largest city in Korea. Geoje is a historically significant place, as it held the largest prisoner-of-war camp during the Korean War. About 170,000 POWs were housed in the prison war camp during the three-year war. In the last few decades Geoje has also grown to be a leader in the shipbuilding industries.
Probably one of the most famous attractions in Geoje is the Windy Hill, located at the southern part of Geoje island. Fans of Korean dramas and variety shows might feel like they have seen this place before. Other than appearing in shows, this place makes for a fantastic location to just sit down and enjoy the constant cool sea breeze, hence its name Windy Hill.
Visitors to Windy Hill often find themselves starting off from Hakdong, a village located about 15 minutes' bus ride away. Here at Hakdong, you can find affordable motels, restaurants and convenience stores easily.
Waiting for our bus from Handing where we stayed to Windy Hill.
Arrived at the foot of Windy Hill.
Restaurants and cafes scattered all around the dock.
If you do not have a car, you can choose to take the public bus which would bring you close to the foot of Windy Hill.
A common sight in Korea, elderly Koreans selling food in such makeshift tents.
The short climb up Windy Hill.
Here we are at Windy Hill!
A windmill is located nearby as well to harness energy from the constant wind.
Visitors come from all over Korea.
Perfect spot with a perfect view.
Coffee time at one of the many cafes nearby.
Heading back to the bus stop.
Most of the buses from Hakdong Village near Windy Hill runs to Windy Hill, but bus intervals can range anywhere from 15 minutes to 45 minutes. Another alternative, which we found much more practical was to take a cab from Hakdong Village to Windy Hill, which cost us no more than 8,000KRW.
Named as such due to the appearance of the black pebbles on the beach, Hakdong Mongdol Beach is one of the most popular beaches in the Gyeongsangnam-do province. The beach is about 40 minutes' drive away from the bus terminal, and there are plenty of motels and restaurants located in the area to cater to tourists. In summer, this place is a hive of activities with visitors coming from all over the country to soak in the summer fun.
Look at all the black pebbles.
In summer, this place turns into a mega playground.
Cyclists planning their route from the beach.
The row of seafood restaurants along the beach.
Our breakfast of seafood stew at one of the restaurants that were open in the morning.
The clean motel that we stayed in for the night. It directly faces the beach.
The main road of Hakdong.
For the history buffs, Geoje POW Camp provides visitors with a deep insight into the happenings of the Korean War. The outbreak of the Korean War in 1950 led to the capture of a large number of communist POWs by the ROK and US forces. The island of Geoje was then chosen to house these POWs due to its strategic location as an offshore island in the southernmost part of Korea. Upon the signing of the 1953 armistice that brought a temporary halt to the war, the camp was closed and then turned into an exhibition park in 1997.
The entrance to the POW Camp.
Ensuring that the war will never be forgotten.
Quite pricey for an admission ticket, 7,000KRW.
Tickets to the camp cost 7,000KRW per adult and 3,000KRW for children. The camp is open from 9am to 5pm from November to February, and 9am to 6pm from March to October each year.
A tank relic on display.
As you pass through the first building at the camp, you will be greeted by the key players on both sides of the Korean War.
Posters in different languages, including simplified and traditional Chinese.
A mockup of compound 62.
How POWs were transported to and from the camp.
The actual ground of the camp kitchen.
As well as actual ground for the bunkers.
The inside of the bunker.
Some structures like this have been preserved.
As we head out of the camp, we pass by a souvenir shop.
There you have it, 3 interesting places to check out in Geoje. Geoje is a fantastic place to check out, with plenty of beautiful sights and a deep history to boot. Getting to Geoje is also easy with the newly built bridge that connects Geoje to Busan. The next time you visit Busan, do consider checking out its sister city Geoje, which is only less than an hour away by bus!
Our botanic gardens in full colour.
The street where Chinese immigrants from Xiamen, Fujian used to congregate, hence its name Amoy(refers to Xiamen in colloquial speak) Street.
The unofficial ambassador of this colourful garden city - the ubiquitous bougainvillea plant.
Swimming with the world's busiest waterway as the backdrop.
Life is full of beauty, sometimes we just have to notice them.


text:
@interface AssignmentDetailViewController : UITableViewController <GMSourceFetcherDelegate, UIWebViewDelegate, MFMailComposeViewControllerDelegate> {
    GMAssignment *assignment;
    GMSourceFetcher *fetcher;
    NSString *details;
    UIWebView *sizingWebView;
    __unsafe_unretained UITabBarController *tabBarController;
    int webviewHeight;
}

@property (assign) UITabBarController *tabBarController;
@property (retain) GMAssignment *assignment;
@property (copy) NSString *details;

@end

redpajama_set_name: RedPajamaGithub


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The founders of Kombucha Culture are absolutely fascinating! Today we are chatting with Adi Ramon who not only brews delicious kombucha but is also a molecular biologist, scuba diver, adventure seeker, drummer and so much more! If you haven't experienced Kombucha Culture's Island Roots, I highly recommend it as one of my personal favorites! Here's more on the company's co-founder Adi.
Colombia – gorgeous country with very low tourism compared to neighboring countries.
Ocean conservancy, stopping the use of plastic, teaching kids & adults about delicious and nutritious plant-based foods.
Before you started Kombucha Culture, what were you doing?
I am a molecular biologist who worked in biotech for a decade.
I'm passionate about microbiome research and the role fermented foods play in keeping a healthy environment in the gut. And stay tuned for some exciting news from Kombucha Culture, coming soon…!


text:
Adtran in Huntsville, Alabama and Adva in Munich, Germany, announce merger in $931M deal.
Huntsville's Adtran and Munich-based Adva will merge, with the new company retaining Adtran's name and headquarters in Alabama and Germany.
MarketWatch valued the all-stock deal at $931.1 million.
The merger will "create a leading global, scaled provider of end-to-end fiber networking solutions for communications service provider, enterprise and government customers," according to a press release, combining Adtran's leadership in fiber access with Adva's leadership in data center interconnect, business ethernet and network solutions.
"We are in the early stages of an unprecedented investment cycle in fiber connectivity, especially in the U.S. and Europe, fueled by the demand for last-mile fiber access and middle-mile transport to provide high-speed connectivity to homes, businesses and future 5G infrastructure," said Adtran Chairman and CEO Thomas Stanton. "By joining forces, our combined firm's portfolio will better position us to capitalize on this highly compelling global opportunity. We expect the transaction will create significant long-term value for both companies' stakeholders by increasing our scale and improving our ability to serve as a trusted supplier for customers worldwide."
The merger will "leverage the complementary nature" of the two company's customers, said Brian Protiva, Adva CEO.


text:
Q: Use from json :how to recognize a json object as a string I have a jsonobject which contain a jsonobject.
String Example1 = {name1:value;name2:{ name21:value, name22:value}};
fromjson(Example1,some.class);

I want to use fromjson() method to transfer Example1 into JAVA class. But I want to let name2 be regarded as a String  not a JsonObject.
That is I want the value of name2 become a string { name21:value, name22:value}
What should I do?

redpajama_set_name: RedPajamaStackExchange


text:
06 October 2018 Venue Trillion Trophy Training Centre Attendance
Birmingham City U18
Burnley U18
PREVIEW: Birmingham City U18s v Burnley U18s
The youth team travel to Birmingham City this weekend in the U18 Professional Development League.
The Clarets head to the second City looking for a hat-rrick of victories, having defeated Barnsley in the league, followed up with a win in the Lancashire Youth Cup over Bolton Wanderers.
However, City are also in fine form, having won their last four league games.
And assistant Clarets coach John Townson admits his side are in for a tough test.
He said: "We have watched them, and they are a good side, however our recent performances and results have been good.
"So, it will be good to see how the lads get on, but we certainly won't take them lightly."
When the two sides met last season, Burnley ran out 4-3 winners in a seven-goal thriller at the Barnfield Training Centre.
And in the reverse fixture, the Blues edged out the Clarets 3-2 in Birmingham.
Burnley will be without Michael Fowler, Ethan Kershaw, Will Harris, who have extended their loan spells at Padiham, Barnoldswick Town and Nelson, respectively.
Second year scholar Richard Taylor will also miss the game afgter starting a loan with Colne.
However, Townson is looking forward to others seizing the opportunities vacated by loanees.
"It will be another mixed team, and as we have said in the past few weeks it brings opportunities for other players," added Townson.
Despite City only picking up a point in their first two games, the Midlands club have won their last four games, scoring 12 goals along the way.
However, Townson praises his side for their recent performances and hopes they can take it into the weekend.
"I've said previously that the lads deserve a lot of credit," he added. "There has been a lot going on with the lads out on loan and injuries, but we will have to do it again.
"There is no hiding place in football, so we will have to rise to it again this weekend."
The game, at Birmingham City's training ground, kicks off at midday on Saturday.
A report will be on the official website followiong the game.
Birmingham City U18s 1 Burnley U18s 3
Youths make it three wins out of three.
The U18s secured their third consecutive win in all competitions, with a 3-1 victory over Birmingham City.
The side, currently being led by assistant manager John Townson due to Danny Cadamarteri's departure, put an end to City's four-match winning streak.
Kai Smith put the Clarets ahead, before an own goal doubled Burnley's advantage.
Matty O'Neil added a third in the second-half, before the home side pulled one back late on.
Burnley started brightly, as Mitch George forced the Birmingham keeper' into action early on.
Minutes later, the Clarets took the lead, as George teed up Smith inside the box to finish.
With 30 minutes played, Townson's side had a second, as Kian Yari's cross was turned in by a City defender for an own goal.
Early into the second half, the visitors added a third, as O'Neil (below) finished off a good move from down the right side of the pitch.
The home side then pulled one back with 20 minutes to go.
However, after a shifting into a back five to hold out the Midlands' side pressure, Burnley held on and ensured they left with all three points.
The young Clarets are next in action on Saturday 13th October away to Nottingham Forest, 12:00pm kick off.


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The Sudanese government's seemingly lack of interest in the worsening crisis plaguing the Darfur region.
Paris Hilton, O.J., Britney, Lindsay - Pick One.


text:
Q: AWS S3 StringLike Condition preventing requests to bucket I have the following s3 IAM policy. It is intended to allow me to copy files from or put files into a bucket below from location temp/prod/tests within the bucket.
In the policy, I have added the StringLike condition, which I had hoped would allow the permissions in the policy to allow copying and puts when the object prefix contains temp/prod/tests.
{
    "Version": "2012-10-17",
    "Statement": [
        {
            "Sid": "",
            "Effect": "Allow",
            "Action": [
                "s3:ReplicateObject",
                "s3:PutObject",
                "s3:ListBucket",
                "s3:GetObjectAcl",
                "s3:GetObject",
                "s3:GetBucketLocation",
                "s3:GetBucketAcl",
                "s3:DeleteObject"
            ],
            "Resource": [
                "arn:aws:s3:::MYBUCKET/temp/prod/tests/*",
                "arn:aws:s3:::MYBUCKET"
            ],
            "Condition": {
                "StringLike": {
                    "s3:prefix": [
                        "temp/prod/tests/*",
                        "temp/prod/tests/"
                    ]
                }
            }
        }
    ]
}

My problem is that the condition prevents me from copying anything under temp/prod/tests/, or putting any new object in this bucket beneath this location.
 $ aws s3 cp --recursive s3://MYBUCKET/temp/prod/tests/ /tmp
download failed: s3://MYBUCKET/temp/prod/tests/testfiles/testfile to ../../../tmp/testfiles/testfile An error occurred (AccessDenied) when calling the GetObject operation: Access Denied

And
$ aws s3 cp /tmp/test s3://MYBUCKET/temp/prod/tests/
 An error occurred (AccessDenied) when calling the PutObject operation: Access Denied

If I remove the Condition, I am able to copy the files as expected.
I don't understand why the condition is not working, because as far as I can see, the requests I am making match the prefix of the condition.
Does anyone know why this is not working as I expect?

A: First of all, I think it is a good practice to split the rules according to resources. Some of the s3 actions require a bucket, some of them require an object. It's in the documentation to every service: https://docs.aws.amazon.com/service-authorization/latest/reference/list_amazons3.html
Furthermore, the conditions instead of proper resources make the policy even more confusing.
In theory, for uploading an object you need just PutObject, you don't even need any List action. But for various cmdline tools I am curious about how far you would get with something like:
{
    "Version": "2012-10-17",
    "Statement": [
        {
            "Sid": "rule1",
            "Effect": "Allow",
            "Action": [
                "s3:ListMultipartUploadParts",
                "s3:AbortMultipartUpload",
                "s3:PutObject",
                "s3:GetObject",
                "s3:DeleteObject"
            ],
            "Resource": [
                "arn:aws:s3:::MYBUCKET/temp/prod/tests/*",
            ]
        },
        {
            "Sid": "rule2",
            "Effect": "Allow",
            "Action": [
                "s3:ListBucket",
                "s3:ListBucketMultipartUploads"
            ],
            "Resource": [
                "arn:aws:s3:::MYBUCKET",
            ]
        }
    }
}

A: Most of the policy is derived from this blog post Writing IAM Policies: Grant Access to User-Specific Folders in an Amazon S3 Bucket
Following policy does as you mentioned in the question

It is intended to allow me to copy files from or put files into a bucket below from location temp/prod/tests within the bucket

PLUS all the actions within the folder temp/prod/tests/*. Those can be restricted further. Like you have few permissions asigned.
{
    "Version": "2012-10-17",
    "Statement": [
        {
            "Sid": "AllowGroupToSeeBucketListInTheConsole",
            "Effect": "Allow",
            "Action": [
                "s3:ListAllMyBuckets",
                "s3:GetBucketLocation"
            ],
            "Resource": "*"
        },
        {
            "Sid": "AllowListingOfUserFolder",
            "Action": [
                "s3:ListBucket"
            ],
            "Effect": "Allow",
            "Resource": [
                "arn:aws:s3::: MYBUCKET"
            ],
            "Condition": {
                "StringLike": {
                    "s3:prefix": [
                        "temp/prod/tests/*"
                    ]
                }
            }
        },
        {
            "Sid": "AllowAllS3ActionsInUserFolder",
            "Effect": "Allow",
            "Action": [
                "s3:*"
            ],
            "Resource": "arn:aws:s3::: MYBUCKET/temp/prod/tests/*"
        }
    ]
}

A: I think part of the confusion here is your expectation that s3:prefix will be present and testable during a CopyObject operation. It's present during a ListBucket operation, and I think that may be the only operation in which it's present. Condition keys for S3 are documented, but the documentation does not appear to include a matrix of which keys are present during which API operations.
Specifically, I believe that s3:prefix will be absent during an actual CopyObject operation and that means that IAM will treat this as values do not match, hence the conditional test fails and the CopyObject operation is denied.
AWS policy evaluation logic is reasonably straighforward and well-defined but the context in which AWS global condition context keys are present is not well-defined, or at least not well-documented. It's also quite difficult to determine exactly why a given API operation was denied after the fact (i.e. which part of the aggregated policies caused the failure), which makes it difficult to write and test complex policies.
Ideally, you'd know which keys are present on which operations, but that doesn't seem to be documented. One way to deal with this is to test (see what works and what does not). Another way is to use the ...IfExists condition check, but this is really designed for use with policy keys that are optional rather than that are not even relevant. When you use StringLikeIfExists, for example:

If the policy key is present in the context of the request, process the key as specified in the policy [i.e. perform a StringLike test]. If the key is not present, evaluate the condition element as true.

In the case of your policy, I'd suggest:

*

*use bucket resources with bucket actions and object resources with object actions (right now, you are mixing them together)

*limit your prefix conditions to the ListBucket operation

*no need to make GetObject or PutObject conditional, simply indicate the resource ARN for which these operations will be allowed (e.g. arn:aws:s3:::MYBUCKET/temp/prod/tests/*)

A: I found a solution that works with minimal change to the policy.
I added ForAllValues to the condition, and now I can copy any objects beneath temp/prod/tests/ or any of the subdirectories below temp/prod/tests/.
        "Condition": {
            "ForAllValues:StringLike": {
                "s3:prefix": [
                    "temp/prod/tests/*",
                    "temp/prod/tests/"
                ]
            }
        }


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With the hot weather still sticking around, the month has been full of restaurant openings, dinners with friends and lunches with my boys. This sunset was taken from the roof terrace of Kuti's Brasserie when I attended their re-opening!
We've had a month full of birthdays including my dads, friends and a few 1st birthday parties too! Joshua loved his first cupcake although he ended up covered in edible glitter.
Arran and I headed off on a date night in Winchester for cocktails and food. It was so lovely just being the two of us but we couldn't wait to get home to our squidge.
I headed off on my first ever press trip! I went to Leeds with Mayflower Theatre to watch the dress rehearsal of Calendar Girls The Musical and I loved it. Leeds is such a vibrant city and I wish we had more time to try all the great food available.
Joshua is ten months old! He's crawling, full of teeth and so very close to walking.
We hosted our first Southampton Bloggers Summer Picnic, the weather was touch and go but it stayed dry and we had such a fab afternoon.
We took Joshua to the farm for the first time and he slept through most of it! It turned out to be such a hot day so we were so thankful for shade filled flower gardens.
Joshua needs a mini. That is all... haha.
I finally tried Doughnut Time! My sister went to London and came back with the oh so sweet treats! I had No Share and it was bloody delicious.
What a lovely post, you've had a great fun month and shared some beautiful photos. Joshua has grown so much but still a little cutie. Hope September brings you lots of joy too.
I WANT DOUGHNUTS. And you are so right - Joshua does need a mini!