Wolbachia are maternally transmitted intracellular bacteria that invade insect populations by manipulating their reproduction and immunity and thus limiting the spread of numerous human pathogens. Experimental Wolbachia infections can reduce Plasmodium numbers in Anopheles mosquitoes in the laboratory, however, natural Wolbachia infections in field anophelines have never been reported. Here we show evidence of Wolbachia infections in Anopheles gambiae in Burkina Faso, West Africa. Sequencing of the 16S rRNA gene identified Wolbachia sequences in both female and male germlines across two seasons, and determined that these sequences are vertically transmitted from mother to offspring. Whole-genome sequencing of positive samples suggests that the genetic material identified in An. gambiae belongs to a novel Wolbachia strain, related to but distinct from strains infecting other arthropods. The evidence of Wolbachia infections in natural Anopheles populations promotes further investigations on the possible use of natural Wolbachia– Anopheles associations to limit malaria transmission.
Wolbachia bacteria live within the cells of many insect species, manipulating their hosts’ reproduction and immune responses. Here, the authors show that these microbes also infect wild populations of malaria-spreading Anopheles mosquitoes, supporting a potential use of Wolbachia to limit malaria transmission.
Malaria-transmitting Anopheles mosquitoes are the deadliest animals on this planet, causing the death of more than 600,000 people each year and endangering the lives of half of the world’s population 1. Current insecticide-based control strategies to stop malaria transmission by targeting the mosquito vector are limited by the rapid spread of insecticide resistance 2. In addition, these interventions target only indoor feeding and resting populations, with the use of insecticide-treated bednets and the application of indoor residual sprays, respectively. For decades, the use of Wolbachia endosymbionts has been proposed as an alternative to chemical strategies because of the ability of these bacteria to rapidly invade insect populations via cytoplasmic incompatibility 3, and successful Wolbachia invasions in field settings have been demonstrated in the case of the dengue and yellow fever vector Aedes aegypti 4. Recent proof that Wolbachia infections of Anopheles vectors limit the development of the Plasmodium parasites that cause malaria 5 6 7 8 makes these bacteria a particularly attractive tool for the control of both endo- and exophagic populations of malaria-transmitting anophelines. Long-standing limitations concerning the introduction of Wolbachia into laboratory colonies of Anopheles mosquitoes have been recently overcome 5; however, the usefulness of this system for the control of Anopheles populations has been undermined by the apparent absence of natural infections. Indeed, while Wolbachia strains have been detected in many insects 9 10, attempts to identify these bacteria in field Anopheles have failed, promoting the belief that these mosquitoes are not natural hosts for Wolbachia 11 12 13.
In this study, we report evidence of natural Wolbachia infections in two incipient species of the major malaria vector, Anopheles gambiae. We isolate Wolbachia-specific 16S rRNA sequences from the reproductive organs and carcasses of adult mosquitoes and from larval carcasses in three different villages in Burkina Faso, West Africa. Whole-genome shotgun sequencing of two positive samples reveals a previously uncharacterized Wolbachia strain that is maternally transmitted in laboratory settings. These results open new avenues for exploiting Wolbachia infections for field applications targeting the malaria mosquito.
We collected mating couples from natural An. gambiae mating swarms in Burkina Faso, West Africa, to identify the microbial populations of the male and female reproductive tracts. We analysed two reproductively isolated populations of An. gambiae, the M and S molecular forms, which do not interbreed in the field 14 and are now classified as two separate species ( An. coluzzii and An. gambiae, respectively 15). For simplicity, in the text and in the tables, we will refer to these two species as M and S forms. Our collections included 81 couples captured in three different villages: two villages in Vallée du Kou (VK5 and VK7), where the M form is prevalent, and one village in Soumousso, primarily populated with S form ( Fig. 1, Supplementary Table 1).
An initial high-throughput sequencing of the 16S rRNA gene (variable region V4, average 23,589, s.d. 16,070 assembled reads per sample) amplified from ovaries and testes dissected from 30 mating couples produced a molecular fingerprint of the bacterial population of these reproductive tissues. Surprisingly, this analysis identified one sample, derived from the testes of an S form male collected in Soumousso, infected with Wolbachia. This sample contained 21.8% of reads (5,412 out of 24,800) matching the V4 regions of the Wolbachia 16S rRNA gene, with a percentage identity ranging between 95.3 and 97.6%. This percentage is fully consistent with the overall diversity of known Wolbachia V4 sequences (average identity 95.9%, s.d. 1.75%, median 96.0% as estimated from available sequences, see Methods), while it is incompatible with any other sequenced bacterial 16S gene (closest matches at <89% identity). Phylogenetic analysis rooted the Wolbachia sequences into one of the two main subtrees of the genus, further confirming their taxonomic placement ( Fig. 2).
The identification of Wolbachia sequences in the testes of an An. gambiae male prompted us to analyse the remaining mosquito specimens collected from the same mating swarms in the three villages. To this aim, a more sensitive PCR-based amplification and sequencing of a 16S rRNA gene segment comprising three variable regions (V6, V7 and V8) was utilized to specifically detect Wolbachia in ovaries and testes dissected from the remaining 51 mating couples, using previously validated primers 16. The mosquito carcasses were also examined using the same method. Out of the 102 mosquitoes analysed, 11 were positive for Wolbachia 16S rRNA, leading to a frequency of infection of 10.8% similarly distributed between males and females (5 males and 6 females) ( Supplementary Table 1). Interestingly, Wolbachia sequences were PCR amplified from either the reproductive tissues or from the carcasses, but never from both, with the exception of one male, where 16S sequences were identified by high-throughput sequencing in the testes and by PCR in the carcass. Although sequences corresponding to these endosymbionts were more prevalent in M mosquitoes, the difference in frequency between the two species was not statistically significant (12.8% in M samples compared with 4.2% in S samples). We also amplified DNA from fourth instar larvae collected from the same three villages (100 specimens from Soumousso, 80 specimens from VK5 and 70 specimens from VK7) using the same primer sets described above. Wolbachia sequences were found in five larval samples (three from Soumousso, one from VK5 and one from VK7), suggesting occurrence of maternal transmission ( Supplementary Table 2).
Analysis of the amplified regions determined the presence of at least two distinct clusters of Wolbachia sequences ( Supplementary Fig. 1). One cluster was identical to the reference strains wAlbB isolated from A. albopictus 17, while the second was closely related to several wPip strains isolated from Culex mosquitoes and Drosophila species 18. The wAlbB-like cluster was detected only in reproductive tissues (ovaries and testes), while wPip-like sequences were more widespread and were found in ovaries, carcasses and whole larvae ( Supplementary Fig. 1). Although these Wolbachia ribosomal sequences are not host specific, the identification of two distinct clusters suggests the occurrence of independent Wolbachia infections in An. gambiae, as observed in other hosts 19.
We next analysed the distribution of the Wolbachia sequences among the different villages. A larger number of positive individuals were isolated from mating swarms in VK5, where 7 out of the 36 collected mosquitoes showed evidence of infection (19.4%). The other 4 positive samples were found in VK7 (3 samples out of 42, 7.1%) and Soumousso (1 sample out of 24, 4.2%) ( Supplementary Table 1). Strikingly, six of the seven positive samples isolated from VK5 had been collected in just two of the seven mating swarms analysed in that village ( Supplementary Table 1). If confirmed, such clustering of Wolbachia-positive individuals in specific swarms would suggest that ecological and environmental factors might play a key role in the establishment of Wolbachia infections in the An. gambiae host.
To expand the 16S rRNA-based analysis and better characterize the Wolbachia strain found in An. gambiae, we performed whole-genome shotgun (WGS) metagenomic sequencing of two of the Wolbachia-positive samples from mosquito ovaries ( Supplementary Table 3). Sequences corresponding to the An. gambiae genome were screened out, and a pipeline for detecting Wolbachia-specific sequences based on the unique marker approach was applied 20 (see Methods). A total of 571 reads uniquely attributable to Wolbachia were detected ( Fig. 3a,b, Supplementary Fig. 3a,b) with 86±3.9 and 87.0±4.4% average identity for the two samples analysed, which is in line with the sequence divergence observed for Wolbachia strains in different supergroups ( Supplementary Fig. 2). These sequences matched 134 Wolbachia genes belonging to different functional categories ( Fig. 3c, Supplementary Fig. 3c). The majority of reads (32.2% on average across the two samples) matched genes from metabolic pathways, while a relevant number of reads (13.3%) corresponded to Wolbachia-specific transposases; this is in line with the observation that transposases abundantly populate the genome of Wolbachia strains from many insects including mosquitoes (for example, transposases correspond to 8.32% of all genes annotated in the wPip genome 18). Alignment of our reads to eight fully or partially sequenced Wolbachia reference genomes ( Supplementary Table 4) demonstrated that the strain identified here belongs to a potential Anopheles-specific phylogenetic supergroup, distinct from the arthropod-associated and evolutionarily related supergroups A and B ( Fig. 3d, Supplementary Fig. 3D). We henceforth call this strain wAnga.
The presence of Wolbachia DNA in the reproductive tissues of female adults prompted the question of whether these bacteria are inherited from mother to offspring, an essential prerequisite for their spread through a population. To obtain irrefutable proof of vertical transmission and estimate its efficiency, semi-gravid females were collected from houses in VK5 two seasons after the initial collections. The same sets of Wolbachia 16S fragments were identified at a frequency of 21% (19 out of 91 females), and the progenies of the 5 Wolbachia-positive females that laid eggs were then analysed for infection. Occurrence of maternal transmission was detected in all progenies, with an average transmission frequency of 68% (ranging from 56 to 100%) ( Fig. 4). Taken together, these data confirm the presence of Wolbachia sequences over the course of 2 years and indicate occurrence of vertical transmission from mother to offspring, as normally observed in Wolbachia infections.
The identification of genomic sequences from a novel Wolbachia strain in two incipient species of An. gambiae over the course of different seasons suggests that anopheline mosquitoes naturally harbour these bacteria, prompting renewed efforts to exploit Wolbachia to block malaria transmission. Past attempts to identify Wolbachia in these mosquitoes may have failed due to possible methodological limitations in the detection systems used, including non-optimal DNA amplification and extraction methods, and size of the sampled mosquito population 11 12 13. In addition, the newly identified wAnga strain appears to be highly divergent from Wolbachia strains isolated in other insects. Indeed, our attempts to amplify by PCR two Wolbachia-specific genes commonly used in phylogenetic analyses, the wolbachia surface protein wsp and the fructose-biphosphate aldolase fbpA, were unsuccessful despite numerous attempts (see Supplementary Table 5 for primer sets used), suggesting a low degree of sequence conservation. Interestingly, in some Wolbachia strains infecting C. pipiens, the wspB gene is disrupted by the insertion of an IS256 transposon 21, which belongs to the same family of transposons identified in wAnga. Similar transposon insertions into wsp may have occurred in the Wolbachia strain infecting An. gambiae, compromising the amplification of this gene.
Although examples of horizontal gene transfer (HGT) between Wolbachia and insect hosts are widespread 22 23, two major observations strongly argue against the possibility of HGT of Wolbachia sequences into the An. gambiae genome: (1) with one exception, evidence of infection was identified in reproductive tissues (ovaries and testes) from nine females and males but not in the carcasses dissected from the same individuals, or vice versa, ruling against a possible transfer into the mosquito chromosomes; (2) the average coverage of Wolbachia in our WGS samples was lower than 0.05 × , which is incompatible with the three orders of magnitude higher coverage of the An. gambiae genome in the same samples (>75 × ). Alternatively, HGT may have occurred into a bacterium or eukaryotic microorganism that infects the Anopheles germline and is maternally inherited from mother to progeny (based on the evidence of vertical transmission of 16S sequences). Regardless of their origin, the Wolbachia sequences identified here may still be sufficient to induce Wolbachia-like reproductive phenotypes, such as bidirectional cytoplasmic incompatibility, that would impact future field deployments of experimental Wolbachia infections 4.
The unexpected discovery in the mosquito germline of maternally inherited Wolbachia organisms will prompt further studies of the ecological, environmental and genetic determinants of susceptibility of Anopheles mosquitoes to Wolbachia infections. It will also spark critical investigation into whether natural Wolbachia–Anopheles associations limit the development of Plasmodium parasites in the mosquito host, thus aiding the design of novel effective bacterial infection strategies to control malaria transmission.
Mosquito samples were initially collected during August–September 2011 in three different sites near Bobo-Dioulasso, Burkina Faso. The village of Soumousso (11°00′N; 4°02′W) is located 55 km North-East of Bobo-Dioulasso. It is characterized by wooded savannah and by temporary breeding sites, more favourable to S form ( An. gambiae) 14. The two other collection sites are located in Vallée du Kou, a large rice-growing area situated 30 km North-West of Bobo-Dioulasso. The village of VK5 (11°23′N; 4°24′W) is completely surrounded by rice fields, while VK7 (11°24′N; 04°24′W) is characterized by rice fields to the South and by Savannah to the North. Because of the irrigation system, rice fields form permanent mosquito breeding sites in which the M form ( An. coluzzii) thrives 14. Nonetheless, few transient breeding sites could be found in depressions and ponds within the villages. Male and female adult mosquitoes were collected in copula 24 from an average of six mating swarms per collection day. Fourth instar larvae were also collected from temporary and permanent water pools from each site. A schematic representation of the villages and swarm locations is provided in Fig. 1. In August 2013, blood-fed An. gambiae females were collected inside the houses in VK5 and allowed to individually oviposit in the insectary.
Genomic DNA was extracted from dissected reproductive tissues (testes and ovaries) using DNeasy kit (Qiagen), and from carcasses using NucleoSpin 96 Tissue kit (Macherey-Nagel). In 2013, to estimate Wolbachia maternal transmission, DNA was extracted using DNeasy kit (Qiagen) from whole females that were allowed to oviposit, and from their progenies. For M and S genotyping, DNA was extracted from a leg using a fast extraction method. In brief, individual legs were incubated in 40 μl of grinding buffer (10 mM Tris-HCl pH 8.2, 1 mM EDTA, 25 mM NaCl) with 0.2 mg ml −1 proteinase K for 45 min at 37 °C, then 5 min at 95 °C to inactivate the enzyme. DNA extracts (1 μl) were then subjected to PCR amplification targeting the locus S200 × 6.1 using specific primers (FWD: 5′-TCGCCTTAGACCTTGCGTTA-3′; and REV: 5′-CGCTTCAAGAATTCGAGATAC-3′) 25. M and S genotyping was also used on larvae and adult carcasses. Larval DNA was used to determine the sex using Y-specific primers (F: 5′-CAAAACGACAGCAGTTCC-3′; and R: 5′-TAAACCAAGTCCGTCGCT-3′).
The 16S rRNA gene data set consisted of Illumina MiSeq sequences targeting the V4 variable region. Detailed protocols used for 16S amplification and sequencing were previously described 26. In brief, genomic DNA from testes and ovaries was subjected to 16S rRNA amplifications using primers incorporating the Illumina adapters and a sample barcode sequence, allowing directional sequencing covering the variable region V4 (15F: 5′-GTGCCAGCMGCCGCGGTAA-3′; and 806R: 5′-GGACTACHVGGGTWTCTAAT-3′). PCR mixtures contained 10 μl of diluted template (1:50), 10 μl of HotMasterMix with the HotMaster Taq DNA Polymerase (5 Prime) and 5 μl of primer mix (2 μM of each primer). The cycling conditions consisted of an initial denaturation at 94 °C for 3 min, followed by 30 cycles of denaturation at 94 °C for 45 s, annealing at 50 °C for 60 s, extension at 72 °C for 5 min and a final extension at 72 °C for 10 min. Amplicons were quantified on the Caliper LabChipGX (PerkinElmer, Waltham, MA), pooled in equimolar concentrations, size selected (375–425 bp) on the Pippin Prep (Sage Sciences, Beverly, MA) to reduce nonspecific amplification products from host DNA and a final library size and quantification was done on an Agilent Bioanalyzer 2100 DNA 1000 chips (Agilent Technologies, Santa Clara, CA). Sequencing was performed on the Illumina MiSeq v2 platform, according to the manufacturer’s specifications with addition of 5% PhiX, generating paired-end reads of 175 bp in length in each direction. The overlapping paired-end reads were stitched together (~\n97 bp overlap) and size selected to reduce nonspecific amplification products from host DNA (225–275 bp).
We first applied the QIIME pipeline version 1.6 (ref. 27) to the 16S rRNA data set that detected in the testis sample G23656 (male SMS5.1 in Supplementary Fig. 1) a total of 19.9% of reads assigned to Wolbachia. To specifically investigate and validate the prediction about the presence of Wolbachia in the sample, we implemented an additional pipeline. We first integrated all 16S rRNA sequences assigned to Wolbachia included in the SILVA 28 and NCBI repositories; we manually inspected those sequences with a >3% nucleotide divergence from any other Wolbachia 16S, which let us exclude five sequences that were mislabeled with close endosymbionts such as Bartonella, Rickettsia and Francisella. We obtained a set of 2,064 Wolbachia 16S sequences from which the 253-nt-long V4 region was extracted and clustered at 99% identity, generating 115 operational taxonomic units (OTUs) covering the diversity in the Wolbachia genus. The all-versus-all mapping of the 115 OTU sequence representatives provided a lower-bound estimate of the genus’ total diversity (average identity of 95.9%, s.d. 1.75%, median 96.0%).
We then performed the mapping of the 24,800 reads of sample G23656 against the full SILVA database, retaining only those sequences with full-length percentage identity of at least 95% with a Wolbachia 16S. The resulting 5,412 reads have a best hit other than Wolbachia at <89% identity, confirming that a fifth of the sample (21.82%) consists of V4 fragments from Wolbachia 16S rRNA. The percentage identity was in the 95.3–97.6% interval, which is fully consistent with the observed diversity in the V4 hypervariable region of the Wolbachia 16S rRNA genes (average 95.9%, s.d. 1.75% as reported above).
The V4 sequences from sample G23656 were then clustered into OTUs at 99% identity, discarding those OTUs with <50 sequences. The resulting OTUs were merged with the Wolbachia OTUs from the SILVA and NCBI repository (generated as described above) and aligned with MUSCLE version 3.8.31 (ref. 29). A V4 sequence representative of the Rickettsia 16S was added as outgroup. A phylogenetic tree was then built using RAxML version 7.4.2 (ref. 30) with the GTRGAMMA model, bootstrapping (1,000 replicates), best maximum likelihood tree inference, and displayed with GraPhlAn (
DNA from testes and ovaries was rehydrated from the 96-well Qiasafe plate (Qiagen) using 30 μl of water. A total of 2 μl of DNA was used for Wolbachia PCR detection using primers specific for Wolbachia 16S rDNA (W-Specf 5′-CATACCTATTCGAAGGGATAG-3′, W-Specr 5′-AGCTTCGAGTGAAACCAATTC-3′) following standard procedures 16. Positive samples showed a 438-bp band that was purified with QIAquick Gel Extraction kit (Qiagen) and sequenced (Eurofins MWG Operon, Ebersberg, Germany). Sample DNA quality was assessed with PCR using primers for the RpS7 (AGAP010592) An. gambiae gene (FWD 5′-GGCGATCATCATCTACGTGC-3′; and REV 5′-GTAGCTGCTGCAAACTTCGG-3′). Similarly, a total of 2 μl of DNA was used for PCR detection of wsp and fbpA genes following standard procedures (see Supplementary Table 5 for primer sets used).
Shotgun metagenomic sequencing was performed on two mosquito samples from infected ovaries using the remainder of the DNA available after Wolbachia-specific PCR detection (1–2 ng). Due to this limiting condition, libraries were prepared with 1 ng DNA according to the Nextera XT protocol (Version Oct 2012). Briefly, the DNA was fragmented in 5 μl of Amplicon Tagment Mix and 10 μl of Tagment DNA buffer (Illumina, San Diego, CA, USA). Tagmentation reactions were completed by incubation for 5 min at 55 °C followed by neutralization with 5 μl of Neutralise Tagment Buffer for 5 min. Tagmented DNA was used as the template in a 50-μl limited-cycle PCR (12 cycles) and processed as described in the Nextera XT protocol. Amplified DNA was purified with AMPure XP beads and then normalized to 2nM. Sequencing was performed on a HiSeq2000 (Illumina, San Diego, CA, USA) employing one full lane per library with 101 bp paired-end reads.
Raw WGS results consisting of >400 M 101-nt-long paired-end reads ( Supplementary Table 3) were subject to quality control and sliding window trimming with a minimum resulting read length of 80 bp, and to sequencing artefact removal using PRINSEQ version 0.20.3 (ref. 31). As expected, An. gambiae DNA was quantitatively dominant in the read pool, and was removed by BowTie2 mapping 32 using the ‘very-sensitive’ preset option against the An. gambiae PEST reference genome (
The resulting read set was mapped against the seven available Wolbachia genomes and the high-quality draft assembly of the Wolbachia strain wAlbB ( Supplementary Table 4). To quantify the expected sequence divergence between Wolbachia strains and supergroups, we performed all-versus-all sequence mapping (with BLASTN) with all open reading frames (ORFs), considering as pairwise common ORFs those sequences with >80% identity over 50% of the ORF length ( Supplementary Fig. 2). Reads were uniquely attributable to Wolbachia on the basis of the concept of unique marker sequences 20, performed in four steps. First step: BowTie2 mapping against the eight Wolbachia reference genomes was performed to identify Wolbachia candidate reads. The mapping was performed with enhanced sensitivity (score-min L,-1.0,-1.0 -D 25 -R 5 -N 1 -L 12 -i S,2,0.25) to capture Wolbachia sequence divergence and host specificity as assessed by ORFs’ sequence comparison among available Wolbachia genomes ( Supplementary Fig. 3). The matches were also confirmed by BLASTN with word size of length 7. Second step: candidate reads coming from the small and large ribosomal units ( 16S rRNA and 23S rRNA genes) were screened out by sequence mapping against the comprehensive ribosomal sequences in the SILVA database release 111. Third step: the remaining ribosomal-free candidate reads were mapped against the full RefSeq genomic database version 60 to identify any non- Wolbachia-specific hits using BLASTN with word size of length 7. Fourth step: on the basis of the mapping results of steps 1 and 3, the final set of reads from the Wolbachia strain identified in An. gambiae ( wAnga) was compiled selecting all reads showing >80% identity over >95 nt to at least one Wolbachia strain, and no hits longer than 80 nt to other organisms. Reads hitting non- Wolbachia genomes with identities below 80% and at least one Wolbachia genome at >90% were also retained. As control, the same procedure was also applied to the two genera closest to Wolbachia according to the PhyloPhlAn tree of life 33, namely, Anaplasma (six reference genomes) and Ehrlichia (five reference genomes). No uniquely attributable reads were found in this analysis.
wAnga reads mapping to all seven Wolbachia genomes were then retained for phylogenetic analysis. The homologous sequences were extracted and aligned to each wAnga reads with MUSCLE version 3.8.31 (ref. 29) and the alignments edited to remove leading and ending gaps. Sequence-specific phylogenetic trees were built using RAxML version 7.4.2 (ref. 30) with the GTRGAMMA model, bootstrapping (1,000 replicates) and best maximum likelihood tree inference. Sequence-specific phylogenetic distances were computed inferring the patristic distances within each tree, and reported with box plots in Fig. 3b and Supplementary Fig. 3B. A final phylogenetic tree was also built using RAxML (GTRGAMMA model, 1,000 bootstrapping replicates) on the concatenated alignments ( Fig. 3d, Supplementary Fig. 3D).
F.C., E.A.L., F.B., J.P., N.S. and A.D. designed the experiments. F.B., J.P., W.R.S. and P.M. performed the experiments. F.C., N.S., F.B., J.P. and E.A.L. analysed the data. N.S. performed the sequencing analysis. R.K.D. and A.D. provided samples for the analysis. F.C., N.S., F.B., J.P., E.A.L. and A.D. wrote the manuscript. F.B., N.S. and J.P. contributed equally to this study.
Accession codes: Nucleotide sequences of PCR-amplified fragments of Wolbachia 16S rRNA genes have been deposited in the GenBank nucleotide database under accession codes
How to cite this article: Baldini, F. et al. Evidence of natural Wolbachia infections in field populations of Anopheles gambiae. Nat. Commun. 5:3985 doi: 10.1038/ncomms4985 (2014).
Supplementary Figures 1-3, Supplementary Tables 1-5 and Supplementary References
We thank S. Ilboudo, H. Guel, D. D. Soma, S. Sawadogo and E. Bilgo for help with sample collection and dissection, V. De Sanctis and R. Bertorelli (NGS Facility at the Centre for Integrative Biology and LaBSSAH, University of Trento) for WGS sequencing and D.E. Neafsey and L. Waldron for helpful discussions. We thank the Broad sequencing platform for 16S sequencing. This work was funded by a European Research Council FP7 ERC Starting Grant project ‘Anorep’ (grant ID: 260897), a Biotechnology and Biological Sciences Research Council grant (grant BB/I002898/1) and a Wellcome Trust grant (grant ID: 093553) to F.C., by the European Union FP7 grant (PCIG13-618833) and startup funding at the Centre for Integrative Biology (University of Trento) to N.S., by the EC FP7 Networks of Excellence EVIMalaR (242095) to E.A.L. and F.C., and by an ANR-11-BSV7-009-01 GIME to E.A.L. F.C. was supported by a NIH grant (grant ID: NIH 1R01AI104956-01A1). J.P. was supported by fellowships from FRM and the Fondation Des Treilles.
The maps describe the three villages (Soumousso, VK5 and VK7) where An. gambiae couples from different mating swarms (indicated by circles) and larvae from different breeding sites (indicated by squares) were collected. Swarm sites and larval breeding grounds are identified by numbers and letters, respectively (see Supplementary Table 1 and 2). Sites where Wolbachia-positive mosquitoes were found are highlighted in red. Maps were adapted with permission from originals courtesy of A. A. Millogo, IRSS/Centre Muraz, Bobo-Dioulasso, Burkina Faso.
The tree was built using V4 16S rRNA fragments from Wolbachia reference sequences available in public repositories (NCBI and SILVA) and from the sequences obtained in this study. The sequences are clustered at 99% identity and the cardinality of each OTU obtained is reported in a logarithmic scale as a bar chart external to the tree (green for reference sequences and red for the new sequences). Shadings highlight subtrees with high bootstrapping support (>90%) and host organisms are reported for those OTUs in which at least one-third of the sequences are consistently associated to the same host.
Ovaries from a Wolbachia-positive female (sample S1) were sequenced using WGS. ( a) Percentage (Perc.) identity of 395 short sequences uniquely attributable to Wolbachia from sample S1 versus eight sequenced strains. The Wolbachia reference strains and the supergroups are indicated with the corresponding percentage of assigned sequences. ( b) Distribution of phylogenetic distances between different Wolbachia supergroups, from phylogenies reconstructed separately on each of the short sequences universally conserved within Wolbachia genomes (light blue inset in a). ( c) Functional classification of Wolbachia loci identified by alignment to read sequences, based on NCBI annotated gene functions. ( d) Wolbachia phylogeny, comprising the new Wolbachia strain wAnga isolated in An. gambiae, reconstructed from the concatenated sequences of b. These analyses show that wAnga is different from all other strains sequenced so far. The same analyses for another PCR-positive Wolbachia sample are available in Supplementary Fig. 3.
A total of 14 Wolbachia-positive blood-fed An. gambiae females collected from houses in VK5 were allowed to lay eggs individually. The progeny of the five Wolbachia-positive females (W+) that laid eggs was screened for the presence of Wolbachia at the fourth larval developmental stage (L4). Red indicates Wolbachia-positive larvae and adults. The number of larvae screened is shown for each female ( n).


--- title: Portofolio permalink: portofolio/ profile: true portofolio: true --- {% for post in site.posts %}
{{ post.description }}


@(name: String, age: Int = 42) @* This template is a simple html template --- (this text btw. is a comment and is not rendered) *@
You are @age years old, @(if(age < 21) shouldn't you be in bed ?! else have a great evening ! asd)


--- layout: article category: meetups title: Computational Intelligence Unconference UK 2014 image: https://farm4.staticflickr.com/3881/14570525409_4f1246a63d.jpg description: Bringing people together to talk about Computational Intelligence and Intelligent Systems. ---
Yesterday I attended an unconference on computational Intelligence hosted by Daniel Lewis and the London Futurists group. The event was a lot like the Anticipating 2025 conference I attended a few months ago but with more of a focus on Artificial intelligence, Machine Learning and the Internet of Things.
Throughout the day several speakers took the stage in different rooms where attendees could informally pop into the ones which interested them the most. This was great as someone like myself who could attend all the tech-related talks, without going too deep into the more science or political related talks.
Deep Learning representations of images, sound and text
Piotr Mirowski gave a really interesting talk on the fundamental building blocks of deep learning algorithms and building an auto-encoder to learn from images, sound and text.
Superintelligence - how afraid should we be?
As always David Wood gave an eye-opening talk on the future of super intelligence and in particular the future intelligence of artificial intelligence.
This time his talk focused more on the dangers of AI far exceeding human intelligence and what we need to be researching and investing into today to make sure the future is bright.
Web Science, Internet Research & Development
Sepi Chakaveh gave a talk about SoFWIReD and how they help companies use future technologies.
One key theme explored was the size of 'Big Data' and how much of a problem it's going to be in the future as we spend more of our lives online, ever-more quantities of data is being stored and handling the data efficiently is the difference between Facebook and MySpace.
Internet of Things (IoT) and Machine Learning
Ajit Jaokar gave a brilliant talk on the convergence of IoT technologies and machine learning to "make dumb things smarter" and making "living things more robust"


{% trans 'Join to ' %} {{ community.service.title }} {% trans 'community' %} {% trans 'Require access'%}
{% blocktrans %}
To become member of this community, require access from its owner, who is responsible to accept or reject your request.
Fill in the fee and currency for your assistance, if you want to charge it. Otherwise, use the default values.
Also, you are able to send any message to the owner of the community.
{% endblocktrans %}
{% endblock content %}
{% block scripts %}
{% with 'app/scripts/bootstrap-table/dist/locale/'|add:bootstrap_table_locale_url as bootstrap_table_locale %}
{% endwith %}
{% endblock %}


A resolver follows a container pattern and is used throughout the framework to manage dependancies within classes. The main difference between this and containers found in other frameworks is that you can mount resolvers via traits.
use Cradle\Resolver\ResolverTrait;
class MyClass
{
use ResolverTrait;
}
Out of the box, resolvers can be used to instantiate classes like the following example.
use Cradle\Event\EventHandler;
$handler = $this->resolve(EventHandler::class, ...$args);
If you wanted to make this into a static class, where it returns the same instance every time we can do so like the following.
$this->addResolver(EventHandler::class, function(...$args) {
static $class = null;
if (is_null($class)) {
$class = new EventHandler(...$args);
}
return $class;
});
$this->resolveShared(EventHandler::class, ...$args);
What the example above also shows that there's nothing stopping us from out
right stubbing the EventHandler
class. The following shows how this can be
done.
$this->addResolver(EventHandler::class, function(...$args) {
return new EventHandlerStub(...$args);;
});
Resolvers can also resolve static methods like the following.
$this->resolveStatic(MyClass::class, 'aStaticMethod', ...$args);
$this->resolve('MyClass::aStaticMethod', ...$args);
In fact you can use Resolvers for anything that needs a resolution.
$this->addResolver('foo', function($number) {
return 'bar' . $number;
});
$this->resolve('foo', 1); //--> 'bar1'


The teaching material is produced in various formats for printing and on-screen reading.
The PDF files are based on LaTeX and have seldom technical failures that cannot be easily corrected. The HTML-based files, called "HTML" and "ipynb" below, apply MathJax for rendering LaTeX formulas and sometimes this technology gives rise to unexpected failures (e.g., incorrect rendering in a web page despite correct LaTeX syntax in the formula). Consult the corresponding PDF files if you find missing or incorrectly rendered formulas in HTML or ipython notebook files.
To understand why matter is stable, and thereby shed light on the limits of nuclear stability, is one of the overarching aims and intellectual challenges of basic research in nuclear physics. To relate the stability of matter to the underlying fundamental forces and particles of nature as manifested in nuclear matter, is central to present and planned rare isotope facilities.
Important properties of nuclear systems which can reveal information about these topics are for example masses, and thereby binding energies, and density distributions of nuclei. These are quantities which convey important information on the shell structure of nuclei, with their pertinent magic numbers and shell closures or the eventual disappearence of the latter away from the valley of stability.
During the last decade, the study of nuclear structure and the models used to describe atomic nuclei are experiencing a renaissance. This is driven by three technological revolutions: accelerators capable of producing and accelerating exotic nuclei far from stability; instrumentation capable of detecting the resulting reaction products and gamma radiation, often on an event-by-event basis, in situations where data rates may be many orders of magnitude less than has been traditional; and computing power adequate to analyze the resulting data, often on-line, and to carry out sophisticated theoretical calculations to understand these nuclei at the limits of stability and to unravel what they tell us about nuclei and their structural evolution.
The nuclear shell model plays a central role in guiding our analysis of this wealth of experimental data. The shell model provides an excellent link to the underlying nuclear forces and the pertinent laws of motion, allowing nuclear physicists to interpret complicated experiments in terms of various components of the nuclear Hamiltonian and to understand a swath of nuclei by following chains of isotopes and isotoones over wide ranges of nucleon numbers. The nuclear shell model allows us to see how the structure of nuclei changes and how the occupation of specific nucleonic orbits affects the interplay of residual interactions and configuration mixing. The computed expectation values and transition probabilities can be directly linked to experiment, with the potential to single out new phenomena and guide future experiments. Large-scale shell-model calculations represent also challenging computational and theoretical topics, spanning from efficient usage of high-performance computing facilities to consistent theories for deriving effective Hamiltonians and operators. Alltogether, these various facets of nuclear theory represent important elements in our endeavors to understand nuclei and their limits of stability.
However, the dimensionalities of interest for shell-model studies exceed quickly present computational capabilities of eigensystem solvers. In order to be able to describe nuclear systems with many more degrees of freedom as well as providing better effective operators, approximative many-body methods like Coupled Cluster (CC) theory or the In-Medium Similarity Renormalization Group (IMSRG) approach have lately gained wide interest and applicabilities in the nuclear many-body community.
It is the goal and motivation of this course to introduce and develop the nuclear structure tools needed to carry out forefront research using the shell model and many-body methods like CC theory and the IMSRG method as central tools, with applications to both structure and reaction theory studies, including continuum contributions and resonances. After completion, it is our hope that the participants have understood the overarching ideas behind central theoretical tools used to analyse nuclear structure experiments.
This three-week TALENT course on nuclear theory will focus on the Many-body methods for nuclear structure and reactions, focusing on nuclear shell model and/or coupled cluster theory and in-medium SRG with applications to structure and reactions. Via hands-on projects and series of exercise, the participants will have been exposed to the necessary tools and theoretical models used in modern nuclear theory.
We propose approximately forty-five hours of lectures over three weeks and a comparable amount of practical computer and exercise sessions, including the setting of individual problems and the organization of various individual projects. The course starts July 16 (with arrival on July 15) and ends (the course) on August 3. A three days workshop will be organized from August 4 to August 6. The mornings will consist of lectures and the afternoons will be devoted to exercises meant to shed light on the exposed theory, and the computational projects. These components will be coordinated to foster student engagement, maximize learning and create lasting value for the students. For the benefit of the TALENT series and of the community, material (courses, slides, problems and solutions, reports on students' projects) will be made publicly available using version control software like git and posted electronically on github.
As with previous TALENT courses, we envision the following features for the afternoon sessions:
The local organizers are
Thomas Papenbrock and Morten Hjorth-Jensen will also function as student advisors and coordinators.
The teachers are
Lectures are approximately 45 min each with a small break between each lecture. The morning sessions are scheduled to end around 1230pm. Every Friday we will have presentations from each group, where a summary of what has been achieved is presented.
Lectures and preparatory material on second quantization are all available at the Github address of the course, or go to URL: https://nucleartalent.github.io/ManyBody2018/doc/web/course.html"" for an easier read.
Furthermore, we strongly recommend that you read chapter 8 and 10 of Lecture Notes in Physics 936. This text contains also links to all codes we will discuss, in addition to the codes we have placed in the program folder of the course. If you cannot access the pdf file of the above text, you can reach chapters 8 and 10 via their respective arXiv versions, click here for chapter 8 and here for chapter 10.
Furthermore, for Coupled Cluster theory the review of Crawford and Schaefer III, An Introduction to Coupled Cluster Theory for Computational Chemists is highly recommended.
For nuclear structure problems, the book of Rick Casten is a highly recommended read. Similarly, Alex Brown's text on Nuclear Structure is a good companion read. The text of Jouni Suhonen is also an excellent read.
The course will be taught as an intensive course of duration of three weeks, with a total time of 45 h of lectures, 45 h of exercises, with the possibility to complete a final assignment if credits are needed.
The organization of a typical course day is as follows:
| Time | Activity |
|---|---|
| 9am-1230pm | Lectures, project relevant information and directed exercises |
| 1230pm-230pm | Lunch |
| 230pm-6pm | Computational projects, exercises and hands-on sessions |
| 6pm-7pm | Wrap-up of the day and eventual student presentations |
You are expected to have operating programming skills in in compiled programming languages like Fortran or C++ or alternatively an interpreted language like Python and knowledge of quantum mechanics at an intermediate level. Preparatory modules on second quantization, Wick's theorem, representation of Hamiltonians and calculations of Hamiltonian matrix elements, independent particle models and Hartree-Fock theory are provided at the website of the course. Students who have not studied the above topics are expected to gain this knowledge prior to attendance. Additional modules for self-teaching on Fortran and/or C++ or Python are also provided.


Provides abstractions for the main objects, that handle the whole process of images going through all possibles phases.
The phases are:
Reading input
Sorting, detection, recognition, ...
Storing output


LMV Extractor Node.js Sample (local view)
This model was extracted using
http://extract.autodesk.io
© Copyright Autodesk. All rights reserved.


| Clients Name | Andrew Schlosser |
| Company Name | ALL HANDS LLC |
| New design or redesign? | New Design |
| Type of website? | Identity, marketing, shopping |
| Design goals | 5. Our goal with the website is to showcase ALL HANDS LLC as a unique, author-focused printing press and book manufacturer. Giving prospective clients a sense of our tastes and sensibilities, as well as our commitment to working closely with them are top priorities with this redesign. |
| Audience | issues | Concerns |
Companies: Computer literate. Usually using Macbooks and apple design products. 1280x1040 minimum monitor resolution. Prefers chrome browser. |
|
|
Authors: 7.2% of Writers & authors are White, making that the most common race or ethnicity in the occupation. Representing 4.23% of Writers & authors, Black is the second most common race or ethnicity in this occupation. The median age of Writers & authors is 42.5, and Male employees are generally 2.12 years older than than their Female counterparts. Independent artists, writers, and performers employs the largest share of Writers & authors at 14.6%, followed by Advertising, public relations & related services with 8.38% and Colleges, universities & professional schools, including junior colleges with 7.02%. |
|
|
Individuals On a gender basis, women are the more avid book readers, per the study, being 13% more likely than men to have read a book in the prior 12 months (77% vs. 68%). Non-Hispanic Whites (76%) outpace non-Hispanic Blacks (69%) and Hispanics (58%) in book-reading incidence, while there’s a clear age trend in play, with Millennials (18-28) being the most likely to read books (80%) and Boomers (65+) the least likely (67%). Typically on mobile based device. High resolution monitor. |
|
|


We've all heard horror stories about amphetamines, speed, whatever you want to call it. Anyone who's ever gone to public school or watched TV for more than twenty minutes at a time has seen some form of propaganda. Amphetamines are terrible for you, take them once and you'll become addicted, ect ect. Up until I was seventeen I pretty much believed all of it.
Up until then I'd been a screw up, but not because of drugs. Oh no, in fact I'm willing to bet I'd been the most sober screw up in the history of the world. 250 pounds, failing all classes, kicked out of school, and I'd never even smoked pot!
Anyway, I'm really only trying to set back story for my experiences on Adderall. All of that was in late 2007, early 2008. I'd been kicked out of school in my Junior year, and if I wanted to return, I would have to go to the public 'Alternative Center for Education'. Basically the school the district tossed the biggest screw ups in, then told the small staff to make it work.
So, I rejoined the school population for my senior year, after six months of doing nothing but playing video games and gathering fat. I had about 4 total credits, I should have had fifteen. At first things seemed to be alright, my grades were fairly decent, and I was keeping up with my required online courses to graduate. Then, slowly but surely, my resolve died down. I started failing again, sleeping through classes, skipping class, the usual.
It was at the end of first semester that I was first offered Adderall. At the time I wasn't interested, I didn't have the money, and was too scared to try it. However the crowd I was running with slowly started to wear me down, not with peer pressure, just general attitude. I decided to try it, and bought five 20mg instant release Adderall pills from a friend.
That night I went home and took one as soon as I got in the door. Within ten minutes I could feel it going to work. I felt strong, I felt fast, I could take on the world! I started working on my online classes, finishing one within four hours. Then I started going down, not a crash per say, but I could feel the energy leaving me. I made the decision to stay up all night and do more work. I took another pill, and finished two more classes that night. These things normally took me an average of 15 hours each, were suddenly child's play.
The next day I went to school and realized two things, one -- I was down to two pills already, and two -- I wanted more. (Keep in mind that word, 'wanted'. It's quite important.) However my friend couldn't get anymore, and I decided to let it go at first. I didn't have the money to support buying more at the time, and I did not want to become addicted to amphetamines.
As luck would have it though, I was offered a new supply. Instead of IR, I could get 20mg extended release (XR) for half the price I was paying earlier, and have them last twice as long. It was a great deal, and soon I was buying 10 at a time, every couple weeks or so. Whenever I needed some, he could get them.
It was during this period that my life started taking drastic changes, for the better. Due to the almost daily dosage of Adderall, I was suddenly motivated to do things I'd never done before. I was hitting the gym on a daily basis. Being social, making more friends in a month then I'd made in the past 4 years. I dropped forty pounds within three months, my grades shot up, and my online classes were being finished ahead of time.
I never built up a tolerance, and soon found that as long as I ate something on the Adderall (hard to remember to eat. I just don't need to), I didn't have any crash. Every time I took one of my little pills, I got everything done, no matter what. The only issues I ever suffered was a small amount of increased aggression. I learned to control it, however it took me some time.
It's been seven months since I first started taking Adderall, I'm now out of school, and my supply was cut off. I decided to get a prescription for Adderall, my family has great health insurance. I'm now paying a tiny amount a month, and have cut my dose down. I've found that I don't need the Adderall anymore, as soon as I got used to my new life, I've been able to function on my own. Now I save those little pills for days when I simply can't afford to sleep for whatever reason, and need to get work done while staying alert.
I believe Adderall can be a great tool if used correctly, look at me. Lost over 70 pounds, graduated high school by making up nearly 18 credits in a single year, create a whole new group of friends. I attribute it all to the Adderall. Yes, it can create addiction, hell, I might be a little addicted right now for all I know. However I believe when managed correctly, it has the potential to make a huge positive impact.
Experience Reports are the writings and opinions of the individual authors who submit them.
Some of the activities described are dangerous and/or illegal and none are recommended by Erowid.


Test the plugin by typing into fields below.
back to the article
Character Count plugin is brought to you by Css Globe, Easy front-end framework and supported by Css Templates by Templatica


the and herds was with The were swine production transmission for ASF from are herd number movements {background infected spread surveillance wild between African animals fever contact (). virus margin this which infection {border-style control into introduction pig clinical px;} #ffffff;} detection index through when considered distribution have these :â€". This case measures not small been density epidemiological more only that visits France French Number Vet al. has may two type virological within according cases different local model represent European among data dead due duration first free-range herds. introduced median pigs rate serological size time zones (Figure Although Figure Halasa However, Transmission affected also based boar both breeding could during each evaluated impact network one potential risk selected sites than therefore trade Dis. Emerg Zone additional all average being but described domestic epidemics evaluate herds, involved modeling per probability px; results zone {font-family Europe Table analyze areas boars consequences depending disease font-size found geographic infectious low-density over relatively same screening secondary seeder simulations used For Index Swine Transbound Union Veterinary al.. analysis any countries days direct double; epidemic high-density highest large movement obtained other outbreaks policies population previous provided randomly reaching route sites. subjected their using (days) -km ./tbed. All Eastern analyzed animal animals. area, assess border-color border-width clearly clinical, clinically commercial distance distances framework growing human implementation investigations its limited mainly outgoing parameters performed period rates related represented scenario scenarios set similar simulated site solid; structure susceptible systematically total types virus's where while {font-style {font-weight -day ./fvets.. Czech DTU-DADS-ASF EFSA France. Free-range Front Herds Indeed, Introduction Material Supplementary areas, article at: batch bold cases, contacts courier; days, densities derived designed despite detect developed did directed economic em; emergence exerted farm farrowing force further having higher huge implemented increasing incursion initial inspection inspections introduction, likely links location main major material model's mortality neighboring notification online original period. pigs. populations. potentially pt;} regarding reported revealed role serological, shared simulation since situation sizes slaughterhouse smaller specific throughout toward very via visits, without


As soon as possible :)
If you need to get in touch, contact me<EMAIL_ADDRESS>We'll do some crazy stuff - kaiquegarcia.dev


The Pansapia Creative is a collaboration designed to tell unique stories in interesting ways. Most of these stories will be set in the Pansapia shared universe and will eventually combine into a single narrative.
Follow Us


--- layout: interior title: "Contact Us" ---
Take Interstate 81 South to the I-81/I-64 Interchange at Staunton. Take Interstate 64 East to Waynesboro, Exit 94. Bear right off of the Exit onto Route 340 North/Rosser Avenue. Continue on Rosser Avenue to the stop light at Route 250/Main Street, approximately three (3) miles. Turn right. Go past Waynesboro High School (on the right). Continue on Route 250/Main Street, down the hill, to the stop light at Main Street & Wayne Avenue. Turn left onto Wayne Avenue. Law Firm is located one block ahead on the left (green/white/burgundy striped awning on front of building).
Take Interstate 64 West to Waynesboro, Exit 99. Turn left onto Route 250 West. Follow Route 250 into downtown Waynesboro, approximately five (5) miles. Turn right at the fourth light at Main Street & Wayne Avenue. Law Firm is located one block on the left (green/white/burgundy striped awning on front of building).
Take Interstate 81 North to I-81/I-64 Interchange at Staunton. Take Interstate 64 East to Waynesboro, Exit 94. Bear right onto Route 340/Rosser Avenue. Continue on Rosser Avenue to the stop light at Route 250/Main Street, approximately three (3) miles. Turn right. Go past Waynesboro High School (on the right). Continue on Route 250/Main Street, down the hill, to the stop light at Main Street & Wayne Avenue. Turn left onto Wayne Avenue. Law Firm is located one block ahead on the left.


|
Vienna Vulkan Engine
|
Store texture data. More...
#include <VEMaterial.h>
Public Member Functions | |
| VETexture (std::string name, gli::texture_cube &texCube, VkImageCreateFlags flags=VK_IMAGE_CREATE_CUBE_COMPATIBLE_BIT, VkImageViewType viewType=VK_IMAGE_VIEW_TYPE_CUBE) | |
| VETexture constructor from a GLI cube map file. More... | |
| VETexture (std::string name, std::string &basedir, std::vector< std::string > texNames, VkImageCreateFlags flags=0, VkImageViewType viewtype=VK_IMAGE_VIEW_TYPE_2D) | |
| VETexture constructor from a list of textures. More... | |
| VETexture (std::string name) | |
| Empty constructor. | |
| ~VETexture () | |
| VETexture destructor - destroy the sampler, image view and image. | |
| Public Member Functions inherited from ve::VENamedClass | |
| VENamedClass (std::string name) | |
| Constructor. | |
| ~VENamedClass () | |
| Destructor. | |
| std::string | getName () |
Public Attributes | |
| VkImage | m_image = VK_NULL_HANDLE |
| image handle | |
| VkDescriptorImageInfo | m_imageInfo |
| Vulkan handles for the image. More... | |
| VmaAllocation | m_deviceAllocation = nullptr |
| VMA allocation info. | |
| VkExtent2D | m_extent = { 0,0 } |
| map extent | |
| VkFormat | m_format |
| texture format | |
Additional Inherited Members | |
| Protected Attributes inherited from ve::VENamedClass | |
| std::string | m_name |
| Name of this instance. | |
Store texture data.
Store an image and a sampler as shader resource.
| ve::VETexture::VETexture | ( | std::string | name, |
| gli::texture_cube & | texCube, | ||
| VkImageCreateFlags | flags = VK_IMAGE_CREATE_CUBE_COMPATIBLE_BIT , |
||
| VkImageViewType | viewType = VK_IMAGE_VIEW_TYPE_CUBE |
||
| ) |
VETexture constructor from a GLI cube map file.
Create a VETexture from a GLI cubemap file. This has been loaded using GLI from a ktx or dds file.
| [in] | name | The name of the mesh. |
| [in] | texCube | The GLI cubemap structure |
| [in] | flags | Create flags for the images (e.g. Cube compatible or array) |
| [in] | viewType | Type for the image views |
| ve::VETexture::VETexture | ( | std::string | name, |
| std::string & | basedir, | ||
| std::vector< std::string > | texNames, | ||
| VkImageCreateFlags | flags = 0 , |
||
| VkImageViewType | viewType = VK_IMAGE_VIEW_TYPE_2D |
||
| ) |
VETexture constructor from a list of textures.
Create a VETexture from a list of textures. The textures must lie in the same directory and are stored in a texture array. This can be used also as a 2D texture or a cube map.
| [in] | name | The name of the mesh. |
| [in] | basedir | Name of the directory the files are in. |
| [in] | texNames | List of filenames of the textures. |
| [in] | flags | Vulkan flags for creating the textures. |
| [in] | viewType | Vulkan view tape for the image view. |
| VkDescriptorImageInfo ve::VETexture::m_imageInfo |
Vulkan handles for the image.


|
|
Internet user privacy and confidentiality is of the utmost importance to all of us at The Natural Handyman. Our reputation and integrity is directly related to the trust we have with our clients and customers. Please find below a description of how The Natural Handyman strives to protect the privacy of Internet users, and maintain the Internet as a highly credible medium.
All users who simply visit The Natural Handyman website retain complete personal anonymity. We do not acquire or collect any personal information about you when you visit, nor do we require you to give us personal information to use our website. Your name, address, phone number, and email address all remain your private properties. The Natural Handyman does, however, collect non-identifiable personal information which is specific to how you use our website.
Our servers record non-personally identifying details such as, but not limited to, the server you are logged onto, the geographic location of the ISP you use, the type of browser you use, your IP address, the date and time, the duration of your visit and the pages viewed. This information is obtained solely for the purpose of determining the number of visits each of our pages receive, how much time our visitors spend viewing our articles and home many pages each of our visitors view. This information is pooled for statistical purposes.
There are instances when a user may provide specific personal information when filling out a form on our site, such as for contests or our newsletter. This information is kept in absolute confidence and is not shared with any other organization.
Furthermore we do not collect or sell the email addresses or demographic information of "casual visitors". A casual visitor is someone who visits pages on our website but does not fill out any forms.
The Natural Handyman uses cookies for collecting such general information as how many people visit our site, how long they stay, and how many pages they view. No personal information is obtained with cookies. Most web browsers automatically accept cookies, but you can usually change your browser to prevent that use if you desire.
However, if you block cookies from our website by changing the settings of your browser or using an internet security "firewall" program, you may not have access to portions of our content. These security programs can block important graphics and other content which are not injurious to you.

The Natural Handyman may use outside organizations to provide advertising for our site. The links on these ads may contain cookies that the advertiser uses to track the ad's effectiveness. Cookies received with banner ads are collected by the advertiser(s), and we do not have access or control over this information.
Though we try to maintain relationships with only the most reputable advertisers, we have no control over their policies or procedures and cannot accept responsibility for their actions. However, we welcome any questions or complaints regarding any ads posted on our website.
This site contains links to other websites or references to non-Internet informational sources. The Natural Handyman is not responsible for the privacy practices or the content of such Web sites or sources.
The Natural Handyman offers sweepstakes, contests or giveaways for the entertainment of our readers and the benefit of our sponsors. These sweepstakes may be internal (information is collected by us) or external (information is collected by the sponsor).
To enter our internal contests, we ask visitors for minimal personal contact information which is necessary to be able to identify and contact the winners. Entering our contests does not place the entrant on any permanent email or "spam" list, nor is this information given or sold to anyone. However, your name and city/town will be released for public announcement if you become a winner. The sponsor of the contest may be given the information submitted in your entry form.
We reserve the right to email any contest entrant once per contest concerning the selection of the winner(s) and a short note from the sponsor. Visit our contest rules for more specific information.
If the contest is external, meaning hosted on another website, we provide only a descriptive informational link to their website. All information regarding rules, privacy and information collection are the responsibility of the hosting website, and neither our rules nor privacy policies apply to them.
The Natural Handyman shares content with other websites and other websites share content with us. We do not, however, share any information given to us by visitors to our site.
The Natural Handyman has security measures in place to protect the loss, misuse, and alteration of the information under our control.
Our subscriber lists are secured from unauthorized viewing or "harvesting" within the limitations of the technology. We acknowledge, however, that there are no absolute privacy protections and thus cannot take responsibility for the malicious acts of others that are contrary to the policies listed in this document.
The Natural Handyman has created this privacy statement in order to demonstrate our firm commitment to privacy. If you have any questions pertaining to this privacy policy and its contents, please click HERE to send us a message. We reserve the right to change, clarify or extend this policy statement from time to time based on our needs, the needs of our visitors, changes in technology or changes in applicable law.
However, rest assured that:
You may also contact us regarding our privacy policy or any other matter at our corporate office at:
G George Ventures, Inc.
16 Hilltop Drive Canton,
CT 06019
860.906.6547 (Leave a message!!)


One-dimensional plots
Pre-processing
The pre-processing in a 1d-vector is based on the interpolation: function: findpeaks.interpolate.interpolate_line1d()
# Import library
from findpeaks import findpeaks
# Initialize
fp = findpeaks(method='topology', interpolate=10)
# Import example
X = fp.import_example("1dpeaks")
# Detect peaks
results = fp.fit(X)
# Plot
fp.plot()
Persistence
The persistence plot is called with the function: findpeaks.findpeaks.findpeaks.plot_persistence()
, and provides two plots.
The left is the detected peaks with the ranking of the peaks (1=best), and the right plot the homology-persitence plot. See section topology for more details.
# Plot
fp.plot_persistence()
Two-dimensional plots
Pre-processing plot
The pre-processing plot is developed for 2D arrays (images) only: function: findpeaks.findpeaks.findpeaks.plot_preprocessing()
Depending on the number of user defined pre-processing steps, the plot will add new subplots.
# Import library
from findpeaks import findpeaks
# Initialize
fp = findpeaks(method='topology', scale=True, denoise='fastnl', window=3, togray=True, imsize=(50,150))
# Import example
X = fp.import_example("2dpeaks")
# Detect peaks
results = fp.fit(X)
# Plot
fp.plot_preprocessing()
Plot
- The plot function
findpeaks.findpeaks.findpeaks.plot()
plots the 3 major steps: input data
final pre-processed image
peak detection.
# Plot
fp.plot()
Persistence plot
The persistence plot is called with the function: findpeaks.findpeaks.findpeaks.plot_persistence()
, and provides two plots.
The left is the detected peaks with the ranking of the peaks (1=best), and the right plot the homology-persitence plot. See section topology for more details.
# Plot
fp.plot_persistence()
3D-mesh
The mesh plot can easily be created using the function: findpeaks.findpeaks.findpeaks.plot_mesh()
.
It converts the two image into a 3d mesh plot.
# Plot
fp.plot_mesh()
# Rotate to make a top view
fp.plot_mesh(view=(90,0))


enum class Boundary
To be use as MsgCtrl.boundary
SUBSEQUENT
Middle packet of a message
LAST
Last packet of a message
FIRST
First packet of a message
SOLO
Solo message packet


kronos
[
class tree: kronos
] [
index: kronos
] [
all elements
]
Todo List
Packages:
undefined_package
DB
framework
kronos
PHPMailer
PHPUnit
Smarty
universibo
universibo_tests
Classes:
Krono
Files:
Krono.php
Procedural File: Krono.php
Source Location: /framework/Krono.php
Classes:
Krono
Class for compute some calculations on date and time
Page Details:
Class for compute some calculations on date and time
Tags:
link:
KronoClass Home at freshmeat
filesource:
Source Code for this file
link:
KronoClass Home at phpclasses
link:
Holosoft Home Page
version:
$Id v 0.6 2003/03/19 20.30.00(CET) marms Exp $
copyright:
2002-2003 Holosoft - Tommaso D'Argenio<EMAIL_ADDRESS>Documentation generated on Fri, 28 Nov 2003 13:09:37 +0100 by
phpDocumentor 1.2.3


Player 1: {{ form.player1(size=40) }} Player 2: {{ form.player2(size=40) }} Player 3: {{ form.player3(size=40) }} Player 4: {{ form.player4(size=40) }}


--- layout: default link-title: Blog title: Blog - Java Design Patterns description: Java Design Patterns blogs are easy to understand guides and tutorials into design pattern implementation. permalink: /blog/ icon: fa-align-justify page-index: 4 ---


BREAKING: Photo: Governor Bello casts his vote - The Nation Newspaper
By Samuel Oamen
The Gubernatorial candidate of All progressive Congress in the 2019 Gubernatorial election in kogi state, Governor, Yahaya Bello, has voted.
Gov. Bello voted at his Agasa, Okene LG polling unit, along side his wife at exactly 8.50am.
Details Shortly…


GitUser App is an angular website that allows users to enter a GitHub username into a search input, submit it, and see names, nicknames, locations, email account, followers, people they are following, biography and descriptions of that person's public repositories.


{% comment %} * * Make a picture responsive. * * Usage: {% include components/responsive_picture.html file="mypicture.jpg" alt="my title" %} * {% endcomment %} {%- assign file = include.file -%} {%- assign alt = include.alt -%}


Company profile - Canadian Company Capabilities
Third-Party Information Liability Disclaimer
Some of the information on this Web page has been provided by external sources. The Government of Canada is not responsible for the accuracy, reliability or currency of the information supplied by external sources. Users wishing to rely upon this information should consult directly with the source of the information. Content provided by external sources is not subject to official languages, privacy and accessibility requirements.
Rousseau Métal Inc.
Mailing Address:
105 de Gaspé Ave WSAINT-JEAN-PORT-JOLI, Quebec
G0R 3G0
Location Address:
105 de Gaspé Ave WSAINT-JEAN-PORT-JOLI, Quebec
G0R 3G0
Tel.:<PHONE_NUMBER>
Tel.:<PHONE_NUMBER>
Fax:<PHONE_NUMBER>
Logo
Company Profile
Rousseau Metal was founded in 1950. Through the years, Rousseau has gained a solid reputation in its field as a company known for innovative products of premium quality, advanced manufacturing methods and exceptional customer service. Rousseau pioneered the modular drawer in shelving solution.
Rousseau designs, manufactures and markets modular drawers, shelving and workstation systems for industrial and commercial use, offered through distributors as well as strategically located companies in high potential markets in North America.
Through the years, Rousseau has acquired a name for itself in North America with clients in industrial sectors such as; Automotive, Wholesalers, and the Government. They can count among their clientele: Alcan, Boeing, Bombardier, Canadair, Chrysler, Domtar, Esso, General Motors, Honda, Pratt & Whitney, Ultramar.
ISO 9001 Certification.
Member of the MHIA (Material Handling Industry of America
Award for the Rousseau workstation systems from the Design
Engineering Association
Full profile
Full profile
Contact information
Company description
Product / Service / Licensing
The new Rousseau cabinet comes in 5 different widths, each offered in 2 depths, for a total of 10 combinations, each available in 5 heights. Add to that the 10 drawer heights, and you have an incredible choice of possibilities. You're guaranteed to find the right size for your specific application.
Tested by an independent laboratory, the Rousseau cabinet proved to be the most durable on the market. The trials proved just how superior our drawer is, as it completed three times as many cycles as its closest competitor. This allows us to be 100% confident in offereing you a lifetime warranty on the drawer, which currently has a patent pending.
The new Rousseau cabinet, in addition to being sturdy, offers a refined and unique look, which will stand the test of time. A shining example of unwavering durability.
- The combination of shelving and Mini-Racking opens up a whole new avenue of possibilities.

- Vertical expansion is also a possibility thanks to our two-level shelving system.
Designed for intensive use, modular storage system will maximize the space in your warehouse, parts department, maintenance workshop, distribution center, etc.
our expertise ensures you peace of mind for all of your needs, present and future!
Whether you are installing a workbench for a production or assembly line, or a custom layout for your machine tool workshop, our line of products offers you an impressive selection of accessories that are sure to meet and surpass your expectations. Whether you are looking for basic work table with two legs and a top, or a specialized table, stationay or mobile, you will find a solution for each and every application. From shipping and receiving, to you foreman's office, the quality and variety of our product will more than satisfy you.
Moreover, the Rousseau multi-puropose stand presents an immense advantage by being so easy to reconfigure, without tools. It can be changed according to your evolving needs by simply adding accessories.
Rousseau offers you several work surface dimensions for each of the surfaces offered: painted steel, stainless steel, laminated wood, wood composite, laminated and dissipative plastic tops. Rousseau, the on-stop solution to simplify your life!
- By its modularity: it is compatible with the entire line of Rousseau products;
- By its flexibility: offered blank of with perforations, it can adapt to any type of tool;
- By its sturdiness: maximum support and stability, it prevents tools from falling, even when the rack is tilted;
- By its ergonomics: pratical, ergonomic handles make it easy to carry. Handling is made easier in the Spider shelving and on the multi-purpose stand by the 20o inclination of the adaptors.
- By its security: sturdy PVC extruded design, it can withstand blows and allows for coolant drainage, prolonging tool life.
The tool identification zone allows for quick retrieval of tools.
Is your need for storage space greater than you originally thought? Equipped with catwalk mezzanines and deck-over shelving, our two-level shelving system might be just what you're looking for. Ask for help from one of our specialists, they'll find the perfect solution, just for you!
In addition to the computer, Rousseau offers supports for monitors and keyboards. These are available in a wall-mounted version and for use with the multi-puropose frame, as well as the wall-mounted smart computer cabinet. All of these products will be very useful for your workspace.
No matter what your need is, Rousseau can supply a solution that is tailored to your situation. In addition, with Rousseau products, you will see the difference in the quality and the attention to detail.
Our specialists will suggest solutions to get around all of the obstacles in your path: construction support beams, columns, doors, ventilation conduits, etc.
Rousseau two-level shelving will maximize your storage by combining shelving, modular drawers and mini-racking and adapting it to your inventory. Store all of your parts in a small area and reduces waster time moving through your inventory to find parts.
Market profile
Geographic markets:
Export experience:
- United States
- Alabama
- Alaska
- Arizona
- Arkansas
- California
- Colorado
- Connecticut
- Delaware
- District of Columbia
- Florida
- Georgia
- Hawaii
- Idaho
- Illinois
- Indiana
- Iowa
- Kansas
- Kentucky
- Louisiana
- Maine
- Maryland
- Massachusetts
- Michigan
- Minnesota
- Mississippi
- Missouri
- Montana
- Nebraska
- Nevada
- New Hampshire
- New Jersey
- New Mexico
- New York
- North Carolina
- North Dakota
- Ohio
- Oklahoma
- Oregon
- Pennsylvania
- Rhode Island
- South Carolina
- South Dakota
- Tennessee
- Texas
- Utah
- Vermont
- Virginia
- Washington
- West Virginia
- Wisconsin
- Wyoming
Third-Party Information Liability Disclaimer
Some of the information on this Web page has been provided by external sources. The Government of Canada is not responsible for the accuracy, reliability or currency of the information supplied by external sources. Users wishing to rely upon this information should consult directly with the source of the information. Content provided by external sources is not subject to official languages, privacy and accessibility requirements.
Contacts
Contact information
Description
Company description
Products, services and licensing
Product / Service / Licensing
The new Rousseau cabinet comes in 5 different widths, each offered in 2 depths, for a total of 10 combinations, each available in 5 heights. Add to that the 10 drawer heights, and you have an incredible choice of possibilities. You're guaranteed to find the right size for your specific application.
Tested by an independent laboratory, the Rousseau cabinet proved to be the most durable on the market. The trials proved just how superior our drawer is, as it completed three times as many cycles as its closest competitor. This allows us to be 100% confident in offereing you a lifetime warranty on the drawer, which currently has a patent pending.
The new Rousseau cabinet, in addition to being sturdy, offers a refined and unique look, which will stand the test of time. A shining example of unwavering durability.
- The combination of shelving and Mini-Racking opens up a whole new avenue of possibilities.

- Vertical expansion is also a possibility thanks to our two-level shelving system.
Designed for intensive use, modular storage system will maximize the space in your warehouse, parts department, maintenance workshop, distribution center, etc.
our expertise ensures you peace of mind for all of your needs, present and future!
Whether you are installing a workbench for a production or assembly line, or a custom layout for your machine tool workshop, our line of products offers you an impressive selection of accessories that are sure to meet and surpass your expectations. Whether you are looking for basic work table with two legs and a top, or a specialized table, stationay or mobile, you will find a solution for each and every application. From shipping and receiving, to you foreman's office, the quality and variety of our product will more than satisfy you.
Moreover, the Rousseau multi-puropose stand presents an immense advantage by being so easy to reconfigure, without tools. It can be changed according to your evolving needs by simply adding accessories.
Rousseau offers you several work surface dimensions for each of the surfaces offered: painted steel, stainless steel, laminated wood, wood composite, laminated and dissipative plastic tops. Rousseau, the on-stop solution to simplify your life!
- By its modularity: it is compatible with the entire line of Rousseau products;
- By its flexibility: offered blank of with perforations, it can adapt to any type of tool;
- By its sturdiness: maximum support and stability, it prevents tools from falling, even when the rack is tilted;
- By its ergonomics: pratical, ergonomic handles make it easy to carry. Handling is made easier in the Spider shelving and on the multi-purpose stand by the 20o inclination of the adaptors.
- By its security: sturdy PVC extruded design, it can withstand blows and allows for coolant drainage, prolonging tool life.
The tool identification zone allows for quick retrieval of tools.
Is your need for storage space greater than you originally thought? Equipped with catwalk mezzanines and deck-over shelving, our two-level shelving system might be just what you're looking for. Ask for help from one of our specialists, they'll find the perfect solution, just for you!
In addition to the computer, Rousseau offers supports for monitors and keyboards. These are available in a wall-mounted version and for use with the multi-puropose frame, as well as the wall-mounted smart computer cabinet. All of these products will be very useful for your workspace.
No matter what your need is, Rousseau can supply a solution that is tailored to your situation. In addition, with Rousseau products, you will see the difference in the quality and the attention to detail.
Our specialists will suggest solutions to get around all of the obstacles in your path: construction support beams, columns, doors, ventilation conduits, etc.
Rousseau two-level shelving will maximize your storage by combining shelving, modular drawers and mini-racking and adapting it to your inventory. Store all of your parts in a small area and reduces waster time moving through your inventory to find parts.
Market
Market profile
Geographic markets:
Export experience:
- United States
- Alabama
- Alaska
- Arizona
- Arkansas
- California
- Colorado
- Connecticut
- Delaware
- District of Columbia
- Florida
- Georgia
- Hawaii
- Idaho
- Illinois
- Indiana
- Iowa
- Kansas
- Kentucky
- Louisiana
- Maine
- Maryland
- Massachusetts
- Michigan
- Minnesota
- Mississippi
- Missouri
- Montana
- Nebraska
- Nevada
- New Hampshire
- New Jersey
- New Mexico
- New York
- North Carolina
- North Dakota
- Ohio
- Oklahoma
- Oregon
- Pennsylvania
- Rhode Island
- South Carolina
- South Dakota
- Tennessee
- Texas
- Utah
- Vermont
- Virginia
- Washington
- West Virginia
- Wisconsin
- Wyoming


jQuery conditionals plugin 2007/08/22
jQuery provides a ton of ways to select sets of elements but pretty much no way to pare down a selection in an arbitrary way. Fortunately for me it's easy to extend jQuery, so I wrote a plugin that allows you to apply a function to your set of elements to pare the set down before applying an action. Source first, talk later:
jQuery.fn.if = function(arg) { var elems = []; var all = this.get(); var ii = all.length; for (var i = 0; i < ii; ++i) { if (arg.apply(all[i])) { elems.push(all[i]); } } return jQuery(elems); };
jquery.if.js (version 0.1)
The plugin takes as its argument a function that must return a boolean value. This function will be applied to every argument in the current set. Those that the function finds true will be returned in the final subset.
The use-case that inspired this quickie plugin is this: I wanted an action to happen to form fields that were empty, which previously required a bit of work. If I had more work to do with each match or a more complex matching requirement, this could get ugly.
$('input.promptable').focus(function() { var to = $(this); if ('' == to.val()) { to.val(this.id); to.addClass('prompt'); } });
With this plugin, the chaining action jQuery is known for is not interrupted. I can select all input
elements with the promptable
class and pare down that selection to only those whose value is empty. On the set returned there I can set values and add classes without breaking the chain.
$('input.promptable').focus(function() { $(this).if(function() { return '' == $(this).val(); }).val(this.id).addClass('prompt'); });
jquery.if.js (version 0.1)
Comments (2)
Hey Richard,
Just wanted to let you know that the functionality you are looking for is built into the .filter() method. Filter can take either a selector or a function. Those jQuery core methods can be hard to find sometimes and it is just so easy to quickly write a plugin for what you need. :) Anyways, here are the docs for the filter method: http://jquery.bassistance.de/api-browser/#filterFunction http://jquery.bassistance.de/api-browser/#filterString
— Brandon Aaron — 2007/08/22 10:42 am
D'oh! You're right, they're sometimes quite hard to find and I never would have called it "filter". Thanks for the tip, Brandon.
— Richard Crowley — 2007/08/22 10:48 am


@(The form loads template from the server without default content and creates a new content according to HTML editor. The existing content will be replaced.)
@(Template)
@(HTML)
@(Please select widgets which want to export. Only templated widgets can be exported.)
@(Data)
@(Copy the data above and share them with another Total.js CMS.)
@(Existing widgets)
@(Please paste export of widgets into the field below:)


Enum OculusApi.BoneId
Bone definition for the Oculus API
Namespace: XRTK.Oculus
Assembly: XRTK.Oculus.dll
Syntax
public enum BoneId
Fields
| Name | Description |
|---|---|
| Hand_End | |
| Hand_ForearmStub | |
| Hand_Index1 | |
| Hand_Index2 | |
| Hand_Index3 | |
| Hand_IndexTip | |
| Hand_MaxSkinnable | |
| Hand_Middle1 | |
| Hand_Middle2 | |
| Hand_Middle3 | |
| Hand_MiddleTip | |
| Hand_Pinky0 | |
| Hand_Pinky1 | |
| Hand_Pinky2 | |
| Hand_Pinky3 | |
| Hand_PinkyTip | |
| Hand_Ring1 | |
| Hand_Ring2 | |
| Hand_Ring3 | |
| Hand_RingTip | |
| Hand_Start | |
| Hand_Thumb0 | |
| Hand_Thumb1 | |
| Hand_Thumb2 | |
| Hand_Thumb3 | |
| Hand_ThumbTip | |
| Hand_WristRoot | |
| Invalid | |
| Max |


Module: RSpec::Core::SharedExampleGroup
- Included in:
- ExampleGroup
- Defined in:
- lib/rspec/core/shared_example_group.rb
Overview
Shared example groups let you define common context and/or common examples that you wish to use in multiple example groups.
When defined, the shared group block is stored for later evaluation.
It can later be included in an example group either explicitly
(using include_examples
, include_context
or it_behaves_like
)
or implicitly (via matching metadata).
Named shared example groups are scoped based on where they are defined. Shared groups defined in an example group are available for inclusion in that example group or any child example groups, but not in any parent or sibling example groups. Shared example groups defined at the top level can be included from any example group.
Defined Under Namespace
Modules: TopLevelDSL
Instance Method Summary collapse
-
#shared_examples(name, *args, &block) ⇒ void
(also: #shared_context, #shared_examples_for)
Stores the block for later use.
Instance Method Details
#shared_examples(name, &block) ⇒ void #shared_examples(name, metadata, &block) ⇒ void Also known as: ,
Stores the block for later use. The block will be evaluated
in the context of an example group via include_examples
,
include_context
, or it_behaves_like
.
88 89 90 91 92 93 94 95 96 97 |
# File 'lib/rspec/core/shared_example_group.rb', line 88 def shared_examples(name, *args, &block) top_level = self == ExampleGroup if top_level && RSpec::Support.thread_local_data[:in_example_group] raise "Creating isolated shared examples from within a context is " \ "not allowed. Remove `RSpec.` prefix or move this to a " \ "top-level scope." end RSpec.world.shared_example_group_registry.add(self, name, *args, &block) end |


Automation Never Tasted So Good!
Hi-Efficiency is raising the bar with customized solutions to fit and meet your needs. The automated bar is designed for beer service along with mix drinks and non-alcoholic beverages. This one-of-a kind patented technology is designed to make your life a little easier.
We combine the art of cocktails with the experience of automation.
Powerful APP
Our innovative app combines an eye-catching, sleek design with the smoothest user experience available. Available for iOS and Android devices.


Next: software-type, Previous: machine-type, Up: Environment Dictionary
machine-version
<no arguments> ⇒ description
description—a string or nil.
Returns a string that identifies the version of the computer hardware on which Common Lisp is running, or nil if no such value can be computed.
(machine-version) ⇒ "KL-10, microcode 9"
The machine version, and the implementation.


Barillia
About
Menu
Contact us
Nestled in the heart of Lutsk, Barillia provides an inviting pub-style experience, offering authentic casual cuisine using fresh locally sourced ingredients.
Weekdays 11 am - late
Weekends 11 am - 2 am
Kriviy Val Street, 31
Lutsk, Volyn region, Ukraine