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The identity of herpesviruses isolated in Europe from domestic pigeons (Columbid herpesvirus-1 - CoHV-1) as well as falcons and owls remains unknown. All these herpesviruses are antigenically and genetically related. The falcons and owls are thought to have become infected during the ingestion of pigeon meat thus suggesting the virus's capacity to infect a wide range of hosts. The aim of the conducted study was to detect the occurrence of CoHV-1 and estimating the similarities and differences in the DNA-dependent DNA polymerase gene of herpesviruses isolated from domestic pigeons, birds of prey and non-raptorial free-ranging birds in Poland.
The study has shown the presence of CoHV-1 in 20.4% (18/88) in the examined birds. In case of one CoHV-1, infected Peregrine Falcon (Falco peregrinus), neurological signs were observed. Nucleotide sequencing of the DNA-dependent DNA polymerase gene, showed a high similarity among Polish strains (100%), independently from the species of the affected birds. Only one compared CoHV-1 strain - KP 21/23 originating from Germany showed a slightly lower similarity at a level of 99.1%. Further analysis has shown the identity of DNA-dependent DNA polymerase of CoHV-1 strains and other herpesviruses present in poultry as well as other birds ranged from 35.4 to 44.9%. Interestingly CoHV-1 infection was also confirmed for the first time in four non-raptorial birds.
The current study has shown a high similarity of CoHV-1 strains and the possible transmission of herpesviruses between domestic rock pigeons and free-ranging birds including raptors and non-raptorial birds. Further studies focused on cloning and the analysis of the whole CoHV-1 genome which is needed to explain the role of the observed similarities and differences between field strains of columbid herpesviruses.
Keywords:Columbid herpesvirus-1; Phylogenetic analysis; Real-time PCR; Sequence analysis; DNA-dependent polymerase
Herpesviruses of domestic rock pigeons and free-ranging birds cause a variety of clinical signs. Due to latent properties of Herpesviridae they may also remain a subclinical infection that influences the immune status of birds [1,2]. Herpesviruses of free-ranging birds fall into seven different species including herpesviruses of pigeons, falcons, vultures, owls and Psittacid herpesvirus (PsHV-1) affecting parrots [2-6]. CoHV-1 isolated from pigeons and falcons, presents similar antigenic features thus its differentiation by serological assays is not possible [7,8]. CoHV-1 has been primarily classified as member of Betaherpesvirinae due to its pathology and growth features. However, it has been further reclassified as an Alphaherpesvirus because of its close relation to the Marek's disease virus in chickens (MDV) [9,10]. Meanwhile, MDV officially named as Gallid herpesvirus 2 (MDV-1), shares common genetic properties with other herpesviruses isolated in poultry including the infectious laryngotracheitis virus - ILTV (Gallid herpesvirus 1), MDV serotype 2 – MDV-2 (Gallid herpesvirus 3), and the previously mentioned PsHV-1 infecting parrots. Infection with CoHV-1 spreads by direct contact with the infected birds as well as by consumption of infected pigeon meat by birds of prey [1,2]. Therefore, pigeons are thought to be the main source of infection also for free-ranging birds [11]. The observed clinical signs in pigeons are called Smadel's disease [3,11] or fatal inclusion body hepatitis in falcons and owls [1,12-14]. Smadel's disease is present in young squabs between ten and sixteen weeks old [1,3]. The birds may not present any clinical signs except depression, anorexia or conjunctivitis. In case of inclusion body hepatitis in falcons, the clinical signs are non-specific and may also be manifested by weakness and anorexia. The specific lesions observed during Smadel's disease include upper respiratory tract inflammation and ulceration as well as hepatic and splenic necrosis [1,3]. In case of herpesvirus infection of falcons the lesions include small necrotic foci in liver, spleen, kidney, pharynx and bone marrow [14-16]. Neurological signs are also possible but rare [13]. The infection may also remain latent making pigeons susceptible to secondary infections with other viral pathogens [11,17]. The previously described infections with CoHV-1 were observed among Accipitridae family in Common Buzzards (Buteo buteo), Booted Eagles (Hieraaetus pennatus) and Cooper Hawks (Accipiter Cooperi)[13,14,16]. Owls (Strigidae), such as the great horned owl (Bubo virginianus), may also be affected by the same virus [4,5]. The study on CoHV-1 occurrence in free-ranging non-raptorial birds has not been previously described.
The study was done to explain the phylogenetic relationships of herpesviruses isolated from pigeons and free-ranging birds including birds of prey such as Peregrine Falcon (Falco peregrinus), Eurasian Kestrel (Falco tinnunculus), but also members of other families like Herring Gull (Larus argentatus) and Song Thrush (Turdus philomelos). The identity as well as transmission of herpesviruses isolated from free-ranging birds in Europe remains unclear. However since Alphaherpesviruses are characterised by their long survival in the environment, they pose a threat for all free-ranging birds, not only for raptors ingesting pigeon meat [18]. The study on CoHV-1 phylogenetic analysis among different free-ranging birds may provide additional information and evidence on the ubiquitous features of this virus.
Birds and samples
In total 88 dead birds were submitted by practitioners or bird rehabilitation centres during 2011–2012 for monitoring West Nile Virus (WNV) at the Department of Poultry Diseases at NVRI. During post-mortem, examination sections of brain were collected and examined for the WNV presence [19]. These brain sections were also used in this study for possible herpesvirus infection detection. The birds originated from different parts of Poland, predominantly form its east (Lublin) and south-west area (Wroclaw). The cause of death of the submitted birds was unknown. Due to the specific goal of the WNV surveillance project for its purposes only brains of free-ranging birds were submitted. Therefore, clinical history of birds from rehabilitation centres or practitioners was not available. However, in case of one female Peregrine Falcon (Falco peregrinus) submitted to our laboratory by dr Szymanski (Wild Animal Treatment and Rehabilitation Centre, Wroclaw) the clinical signs were observed. The bird was examined for the infection with AIV and aPMV-1 to exclude the influence of other associated infections.
Virus isolation
The 10% (w/v) brain homogenates of field CoHV-1 isolates were used for virus propagation in secondary chicken embryo fibroblasts (CEF) prepared from 11-day old SPF chicken embryos (Lohmann Tierzucht, Cuxhaven, Germany). The cells were grown at 37°C and 5% CO2 in a MEM medium (Gibco, Paisley, United Kingdom) supplemented with 5% calf bovine serum (Gibco, Paisley, United Kingdom) and with a 1% (w/v) mixture of antibiotics (Antibiotic-antimycotic, Gibco, Paisley, United Kingdom). All strains were passaged three times then submitted to DNA extraction and real-time PCR. The cytophatic effect visible as clumping cells forming syncytia was observed starting from the 2nd passage of the examined viruses. The cells were harvested 96 h p.i. when approximately 85% of the visible cytophatic effect (CPE) was observed. After triple freezing (−70°C), thawing (+20°C) the cell suspension was centrifuged at 600 × g, then the supernatant was aliquoted and stored at −70°C as stock solutions.
Standard strains
Standard CoHV-1 strain GJW1126 originating from rock pigeon (Columbia livia) (Accession number: JX892997) and GJW1132 (Accession number: JX893002) from Peregrine Falcon (Falco peregrinus) were used (Department of Poultry Viral Diseases, National Veterinary Research Institute, Poland). These strains were used as a positive control for further real-time PCR development. The set of control strains represented by Marek's disease virus (MDV) strain 31_07 (Department of Poultry Viral Diseases, NVRI, Poland), turkey herpesvirus (HVT) FC126 strain (Mérial, France), pigeon circovirus POL1 (PiCoV) (Department of Poultry Viral Diseases, National Veterinary Research Institute, Poland) and Fowl adenovirus 1 strain (FadV-1) (Charles River Laboratories International, Wilmington, United States of America) were also applied. The DNA templates for further analysis were extracted from 200 μL stock solutions of strains.
DNA extraction
The total DNA for herpesvirus detection was extracted from 200 μL of virus stock solutions according to the procedure of the manufacturer for the QIAamp DNA Mini Kit (Qiagen, Hilden, Germany). The DNA was then stored and frozen at −80°C.
Primer design
The primers were designed in Primer Express software version 2.0.1 (Applied Biosystems, Foster City, California, USA) on the basis of the conserved DNA-dependant DNA polymerase gene of CoHV-1 PO1W124 strain (Genbank: GQ478232) and were as follows CoHV1: 5′-GATGGCGGCCTGCTGTTTGT-3′, CoHV2 5′-CGCCGTGGACGACTTGCGT-3′. The primers were synthesised by Genomed Co. (Warsaw, Poland).
Real-time PCR
The reaction was performed in 7500 Applied Biosystems system (Applied Biosystems, Foster City, California, USA). The reactions were set up on ice in 0.2 ml optical tubes with caps, using the Quantitect SYBR Green PCR Kit (Qiagen, Hilden, Germany). The reaction volume was 25 μL that contained: 12.5 μl of 2× QuantiTect SYBR Green PCR Master Mix, 40 pmol of each CoHV1 and CoHV2 primer, 1 μL of DNA template (~25 ng) and deionised water. All DNA templates were analysed in duplicates. The data were collected by Applied Biosystems software ver. 2.0.1 (Applied Biosystems, Foster City, California, USA). Reaction conditions were as follows: 50°C/2 min. (Uracil-DNA Glycosylase incubation), 95°C/15 min (initial denaturation), then 40 cycles of 95°C/1 min (exact denaturation) and 60°C/1 min (primer anealing and signal detection).
The DNA template of standard GW1126 strain was used for amplification of DNA-dependent DNA polymerase. The pGW1126 plasmid was constructed by PCR product insertion (207 bp) into pGEM-T Easy vector (Promega, Fitchburg, Wisconsin, USA). The plasmid was cloned into DH5α cells (Invitrogen, San Diego, USA) with standard blue/white screening method. A single white colony with pGW1126 was used for the inoculation of 5 ml of liquid LB medium with ampicillin (100 μg/mL), and then the culture was incubated for 18 h at 37°C with shaking 225 rpm. The plasmid DNA was extracted from 5 ml of inoculated media using the Plasmid Mini Kit (Qiagen, Hilden, Germany) and used for the determination of real-time PCR sensitivity.
Real-time PCR specificity and sensitivity
For the specificity test, DNA templates extracted from MDV strain 31_07, HVT FC126 strain, PiCoV POL1 strain and FadV-1 were applied. The sensitivity of the real-time PCR was examined using 5 ten-fold dilutions (100 ng – 10 pg) of the entire GW1126 strain (200 ng/μL) as well as 5 ten-fold dilutions of pGJW1126 plasmid (from 108 to 101 copies). The detection limit was determined as the highest dilution that resulted in a fluorescent signal plotted as a curve detected as cycle threshold (CT) value.
DNA sequencing
The products obtained by real-time PCR were separated in a 1.5% agarose gel (Invitrogen, Green Island, New York, United States of America) then cut out and purified according to procedure of the QIAquick Gel Extraction Kit (Qiagen, Hilden, Germany). The amplicons were then sequenced on a GS FLX/Titanium sequencer (Roche, Branford, Connecticut, USA) by Genomed (Warsaw, Poland). Each product was sequenced in both forward and backward directions and then assembled into a single contig in Genious™ Software ver. 6.5.5 (Biomatters Ltd, Anzac Avenue, New Zealand). The sequences were aligned using Multiple alignment Genious™ algorithm with 93% cost matrix similarity and BOOTSTRAP analysis with 1000 repeats. To compare the obtained sequences 7 additional NCBI GeneBank entries of DNA-dependent DNA polymerase from CoHV-1 as well as other herpesviruses of poultry (MDV-1, MDV-2, HVT) and parrots (PsHV-1) were used. The results were also compared with the related human herpesvirus 1 (Accession number: JF810822) to show true phylogeny of the examined CoHV-1 Polish strains. The results were plotted as a similarity matrix and tree showing the phylogenetic distances and relationships between the analysed strains.
Birds and clinical signs
The cause of death for 88 birds used in this study was unknown since they were collected by practitioners for the monitoring study on WNV. In the case of one female Peregrine Falcon, clinical signs including keratitis and torticollis were observed (Figure 1). The Peregrine Falcon female from the rehabilitation centre was fed with pigeon meat, which further suggested the suspicion of herpesvirus infection, by ingestion. The bird died after the two weeks long treatment. The bird was also examined for possible infection with AIV, aPMV-1 and mechanical damage.
Figure 1. Clinical signs observed in young female of Peregrine Falcon (Falco peregrinus) showing torticollis and keratitis.
Virus isolation
The conducted virus isolation was successful in the case of 18 strains. The viruses formed cytophatic effect starting from the 2nd passage in CEF cell cultures.
Real-time PCR
The designed primers used in real-time PCR did not allow for the amplification of DNA templates extracted from MDV, HVT, PiCoV and FadV-1 strains. Therefore, we considered them specific only for the DNA-dependant DNA polymerase gene of CoHV-1. The method enabled detection of 100 pg of standard GW1126 strain or 102 copies of pGW1126 plasmid with the concentration determined as 1.32 μg/μL and was considered as very sensitive (Figure 2). Following the study, all 88 DNA templates from the propagated field strains were examined with the developed method. Eighteen samples (20.4%) were positive for CoHV-1 (Table 1). The highest cycle threshold (CT) values – 17.1 and 21.7 – were found in rock pigeons from the city of Lublin (GJW1130 and GJW1126). Lower values (CT ranging from 31.1 to 36.1) were found in free-ranging birds like Peregrine Falcon (Falco peregrinus) – strain GJW1122, Herring Gull (Larus argentatus) – strain GJW1124 or Tawny Owl (Strix aluco) – strain GJW1138.
Figure 2. Sensitivity determination of the developed real-time PCR assay. (A) Amplification in samples containing successive 10-fold dilutions of CoHV-1 standard isolate. Concentration of DNA in each sample was given. (B) Amplification in samples containing serial dilutions (108 – 1E8, 106 – 1E6, 104– 1E4, 102 – 1E2 and 101 – 1E1) copies of pGW1126 plasmid. ΔRn is an increment of fluorescent signal during successive cycles.
Table 1. GenBank accession numbers and origin of DNA sequences analysed in the study
The species distribution among the 88 examined birds has shown that the most frequently CoHV-1 was detected in Peregrine Falcons (2/2), rock pigeons (6/11) and Common Buzzards (2/11). Interestingly, CoHV-1 was identified for the first time in Herring Gull, Grey Heron (Ardea cinerea), Hooded Crow (Corvus cornix) and Song Thrush (Turdus philomelos). Seven out of seventeen examined species of birds were free from CoHV-1 infection (Table 2).
Table 2. Distribution of CoHV-1 among 88 birds received for the WNV surveillance program
DNA sequencing
The obtained PCR products are 207 bp long. The accession numbers of 18 new CoHV-1 sequences were shown in Table 1.
Phylogenetic analysis
The conducted Genious™ sequence alignment for the total 26 herpesvirus sequences showed a 100% identity within the group of Polish CoHV-1 strains. This similarity was independent from host species. These strains sequence was identical with the reference sequence of PO1 W124 strain originating from Australia. Minor differences within CoHV-1 sequence were found in KP 21/23 strain isolated in Germany since its identity to other examined CoHV-1 strains reached 99.1%. CoHV-1 strains showed an identity from 37.3 to 44.9% with three other compared Alphaherpesviruses of poultry (Gallid herpesvirus 2, Gallid herpesvirus 3 and Meleagrid herpesvirus 1). The identity of CoHV-1 strains to ILTV (Gallid herepesvirus 1) was lower and reached from 34.3 to 35.4%. Interestingly, PsHV-1 affecting parrots showed an identity of 37.3% with the examined CoHV-1 strains. Finally, the analysed CoHV-1 strains shared an identity from 32.7 to 36.3% with the related human herpesvirus 1 (Figure 3). On the basis of the multiple alignment, analysis and neighbour-joining (NJ) computational method, the phylogenetic tree was constructed (Figure 4). All CoHV-1 strains fall into a single group. The second group was represented by a single human herpesvirus 1 whilst the third less homogeneous group was composed of sequences of poultry herpesviruses. The BOOTSTRAP values exceeded 86 between the main created groups. Within the group of poultry herpesviruses, these values were lower and reached from 30 to 59, respectively. The conducted analysis confirmed the data obtained by similarity matrix data (Figure 3). In general, the sequences of the examined CoHV-1 strains are highly homogenous as this was found on the basis of similarity and phylogenetic analysis. The comparison with two other CoHV-1 homological sequences from GenBank has shown that herpesviruses of pigeons and free-ranging birds present a high similarity that is independent from the species or geographical origin.
Figure 3. Nucleotide sequence matrix similarity of 26 aligned sequences composed from 18 Polish field strains of CoHV-1 and 8 GenBank database accessible sequences. The similarity of each sequence is compared and given as percentage.
Figure 4. Phylogenetic tree constructed on the basis of the conducted alignment of the DNA-dependent DNA polymerase from Polish field CoHV-1 strains and herpesviruses derived from GenBank database. BOOTSTRAP values are indicated.
Columbid herpesvirus (CoHV-1) has been primary described in 1940 by Smadel et al. [3] as an agent affecting pigeons that caused visceral lesion such as necrosis especially in liver. Herpesvirus infection was frequently identified in young pigeons and free-ranging birds from different countries of Europe [12,16,20-22], Iraq [23], USA [1,13,15], Australia [4,14] and Canada [5]. The virus is antigenically related to CoHV-1 and was found in falcons and owls [1,4,5,12-16].
The current study has shown that eighteen samples (20.4%) were positive for the CoHV-1 presence. The applied real-time PCR method was highly sensitive which facilitated the collection of reliable results on the CoHV-1 proportion in Poland. The most frequent CoHV-1 was found in Falconidae, Strigidae and Accipitridae. In other reports from the United Kingdom conducted by Zesivanovitis et al. [24] the prevalence of CoHV-1 ranged from 4.9% within the Strigidae to 8.8% in the Accipititridae family. The same study showed CoHV-1 seroprevalence among free-ranging owls reaching 8.2%. The analytic method selection might explain this variation between the percentages of CoHV-1 positive birds since PCR-based techniques enable a more sensitive detection of viral pathogens in comparison to serological assays.
The CoHV-1 affected birds may show other non-specific signs like depression, diarrhoea and neurological signs, but this may be also related with other co-infections with adeno- (FadV), circo- (PiCV) and paramyxovirus aPMV-1 [17,25]. However, in the past two variants of neuropathogenic CoHV-1 strains occurred in rock pigeons from Iraq and Poland [22,26]. The specific lesions include hepatic inclusion bodies with focal necrosis in spleen and pancreas [1,4,5,12-14,16]. Meanwhile, there are many asymptomatic infections in pigeons older than 6 months. The reservoirs of CoHV-1 are latently infected pigeons and the environment, in which the virus may remain infectious for a long time. The latter is more convincing when taking into account the survival of other Alphaherpesviruses such as MDV which may remain infectious at least for 8 months in 22-25°C and for 3 years in 4°C [18]. Free-ranging pigeons are thought to transmit the virus to domestic pigeons [2]. In addition, the meat may be infectious for birds of prey that hunt these. We presume that Herring Gulls as scavengers may be infected by consumption of dead pigeons, or when they share food with pigeons in the cities. In our study, we have shown that also free-ranging non-raptorial birds may be infected with CoHV-1. We have also observed neurological signs in Peregrine Falcon female. The absence of WNV, AIV and aPMV-1 in addition to the presence of CoHV1 in the brain of this Peregrine Falcon with central nervous signs indicates a possible neuroinvasive form of herpesvirus infection, such as those described previously.
What is worth mentioning, the samples of brain are not commonly used for CoHV-1 isolation and detection. However, our study has shown that the proportion of CoHV-1 infection was even higher in comparison to previous reports [26]. The comparison of different detection methods is always very difficult because serological assays investigate the immunological response of the host to the pathogen whereas our real-time PCR method revealed the presence of DNA what does not determine the detection of fully functional virus and possible viremia. The complete investigation should include full necropsy and histological assay which we were unable to conduct. However, virus isolation in cell cultures confirmed the CoHV-1 presence. So far the methods of CoHV-1 detection included mainly histopathological examinations which is dedicated only to experienced pathologists, serum neutralisation tests, DNA endonuclease digestion and polymerase chain reaction (PCR) which in spite of their usefulness are laborious and time-consuming [7,9,10,13]. The uniplex-PCR technique facilitating the detection of all three most important viral pathogens of pigeon was further described by Frieck et al.[25]. The problem with CoHV-1 detection may also be caused by latent infection that is specific for Herpesviridae infecting pigeons and owls [1,4,5]. The present study describes the use of SYBR Green I – based real-time PCR technique, which provides fast and reliable results on the CoHV-1 infection of both pigeons and free-ranging birds. The design reason of this technique was to accelerate, simplify and increase the sensitivity of CoHV-1 detection. The main advantage of our technique is also the lower risk of false negative results as it is important due to the kind of used samples in this study. On the other hand, the main drawback of the used method was CoHV-1 detection after the culturing of CoHV-1 strains, thus our results lack quantification of the virus in the original brain samples. It would also be valuable to examine samples from other organs including liver and spleen in the future. It is well known that real-time PCR may not provide any information about the cause of the observed clinical signs. However, it is valuable in case of latent infections which cannot be identified by histological assays. The best combination would be the use of a histopathological assay together with our real-time PCR that may facilitate sensitive herpesvirus detection and provide valuable remarks on the potential death cause.
We presume that the obtained proportion of CoHV-1 in free-ranging birds may be slightly underestimated due to the kind of used samples. The results obtained in our study confirm previous findings that all recently reported herpesviruses from birds of prey belong to CoHV-1 [1,5,13,14,27]. Indeed, herpesviruses are not restricted to a specific host or tissue and may cross the barriers of the specific host. Our study has shown that the sequence identity of 18 CoHV-1 Polish strains was 100% independently from the species of the affected bird. The analysed 207 bp fragment was conserved among all examined Polish strains of CoHV-1. Consistently, CoHV-1 sequences identical to other examined herpesvirus strains have been found for the first time in four species of non-raptorial birds including: Herring Gull (Larus argentatus), Grey Heron (Ardea cinerea), Hooded Crow (Corvus cornix) and Song Thrush (Turdus philomelos). A similar identity (99.1-100%) was found by comparing examined CoHV-1 to two other foreign sequences originating from Germany and Australia. The identity of CoHV-1 sequences seems to be independent from species and geographical origin. This may be also caused by the low number of relevant CoHV-1 sequences accessible in the GenBank database. Interestingly, CoHV-1 strains shared at least 37.3% similarity with other related herpesviruses isolated from chickens and turkeys. In addition, the PsHV-1 that infects parrots has the same identity level, which may confirm the homology of the particular genes of Alphaherpesviruses. The related human herpesvirus 1 falls into a distinct group, but still its identity to CoHV-1 strain reached from 32.7 to 36.3% that implies on at least partially conservative character of the DNA-dependant DNA polymerase. The presented phylogenetic data were reliable in spite of three lower BOOTSTAP values estimated for Gallid herpesvirus 3, Psittacid herpesvirus 1 and Gallid herpesvirus 2 strains.
In concluding the conducted study, we have shown that the sequence of DNA-dependent DNA polymerase of 18 Polish isolates has shown 99.1% identity independently from their species and geographical origin. This may suggest a possible transmission of the virus between population of domestic and free-ranging birds in Poland. Interestingly the same isolates were found in domestic pigeons, raptors and non-raptorial birds as may be the evidence on close similarity between herpesviruses. Further studies including cloning of the whole CoHV-1 genome and its detailed analysis will hopefully explain the role of the observed similarities and differences in the genome.
NVRI: National veterinary research institute, Pulawy, Poland; CoHV-1: Columbid herpesvirus 1; PsHV-1: Psittacid herpesvirus 1; MDV-1: Marek's disease virus serotype 1 (Gallid herpesvirus 2); MDV-2: Marek's disease virus serotype 2 (Gallid herpesvirus 3); ITLV: Infectious laryngotracheitis virus 1 (Gallid herpesvirus 1); HVT: Herpesvirus of turkey (Meleagrid herpesvirus 1); WNV: West Nile Virus; AIV: Avian influenza virus; CEF: Chicken embryo fibroblasts; SPF: Specific pathogen free; MEM: Minimal essential medium; w/v: Weight by volume percentage; PCR: Polymerase chain reaction; h p. i.: Hours post-infection; μL: Microliter; pg: Pico gram (10-12 g); CT: Cycle threshold value; SYBR Green I: Real-time PCR assay kit containing SYBR Green I dye binding to double-strand structures of DNA; BOOTSTRAP: Statistical method in phylogenetics used to estimate an error during computation of relation between similar nucleotide sequences at phylogenetic tree
Competing interests
The authors declare that they have no competing interests.
Authors' contributions
Most of the experiments were conducted by WJG who conducted viral materials preparation, DNA extraction, real-time PCR development and sequence analysis. ESS assisted in experimental design of the study. SP provided infectious materials from falcons with detailed description and documentation. PW and ST provided materials for the study from herpesvirus infected pigeons. HM provided additional infectious materials from non-raptorial and raptor birds. ESS, SP and PW and HM participated in the coordination of the study. WJG wrote this manuscript and prepared figures for publication. The final manuscript was read and approved by all the authors.
The materials from free-ranging birds for the presented study were provided by the support of project number: W/110 entitled: "Occurrence of West Nile Virus in free-ranging birds in Poland" conducted by the Department of Poultry Viral Diseases at the National Veterinary Research Institute (NVRI).
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Get your strongest core ever with these six moves from the new DailyBurn Pilates Phase Two. Do them all in a row for an intense (but worth it) ab workout.
From key lime pie to chocolate chip cookie dough, these easy homemade energy bites recipes will conquer your cravings and make your taste buds smile.

subset: commoncrawl


text:
Meet Me at the Moon
By Gianna Marino
(Viking Books for Young Readers, Hardcover, 9780670013135, 40pp.)
Publication Date: March 29, 2012
List Price: $16.99*
* Individual store prices may vary.
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A heartwarming love story between mother and child
When Mama Elephant must leave Little One to ask the skies for rain, the young elephant is worried. Who will care for Little One? Who will sing Mama's special songs? When will she return?
Mama is very reassuring - Little One will hear her song on the wind and feel her love in the warmth of the sun, and, after the rains come, they will meet where the moon sets.
Exquisitely illustrated and supremely comforting, Meet Me at the Moon is a mother and child love story to be enjoyed again and again.


text:
Surveillance, Art and Photography in Europe post 9/11
Time: 7–8 October 2015
Place: Valand Academy, Gothenburg, Sweden
Detailed program will be announced in August 2015.
As surveillance has become a constitutive part of contemporary social life, new forms of discrimination and segregation have emerged, but also perhaps new means of empowerment and 'sousveillance'. How can art and visual theory contribute to the understanding of our current surveillance society? During a 2-day public research symposium, artists and scholars will present work on surveillance and its relation to representation, migration, community and control. Speakers include Peter Weibel, Hille Koskela, Marco Poloni, Shoshana Magnet, Jason Bowman, Ann-Sofi Sidén and Kari Andén-Papadopoulus.
The symposium is co-organized by Valand Academy of Art and the Hasselblad Foundation. It is a part of the project WATCHED! Surveillance, Art and Photography in Europe post 9/11 – an exhibition at The Hasselblad Center and a forthcoming book in spring 2016.


text:
Dogs & Cats Forever: Goldie is a boy with a 'golden' personality
FORT PIERCE - I've never met a human I didn't like. I'm a young Chi-Weenie who loves to cuddle. I'm sweet and handsome, and I would love an active family to play with. I get along with other dogs, too.
Meet me and take me home. I'm sure we're perfect for each other.
You can meet Goldie at Dogs & Cats Forever No Kill Shelter, 4600 Selvitz Rd., in Fort Pierce. For more information, please call 772-489-5454 or visit our web site at http://www.dogsandcatsforever.com. You can also follow us on Facebook or Petfinder
This story is contributed by a member of the community and is neither endorsed nor affiliated with TCPalm.
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You can now invite anyone to meet at your favorite Starbucks.
Limited-time feature
29 year old man | North Olmsted, OH, USA | Seeking women 20-35 within 50 miles
Active within 24 hours
I was referred to this site by a friend. Just...
I was referred to this site by a friend. Just looking to meet new people possibly for a date. I used to go downtown a lot, but realized that the type of clientele that that area brings is honestly not the type of person I am trying to meet. Plus I am getting too old for that crap. I enjoy cars. I am currently employed as the Marketing Director for a multimillion dollar late model high performance domestic sport car auto yard. I enjoy modifying cars, the gym, boating, and enjoying good times with good friends. I graduated from Baldwin Wallace in 2009 with a bachelors degree in computer information systems, so I also enjoy technology and the latest advancements in electronics. I own my own house, and I enjoy modifying it as well. I am an honest guy - a little emotional at times, but 100% real. I think what makes me stand apart from others is that I will never be fake to impress someone or say things to people just because that's what they want to hear. I'd rather be completely upfront and honest than string things along and be fake. So why has it come to this site? Well, I believe love should never hurt, and if it does, then its not love. You should never have to look back and regret something if it was right. If you truly care about someone, you'll put their happiness before your own. Even if you don't fit in their plans, there's no point trying to be vengeful or spiteful. Stay true to yourself, and take value in knowing your morals and promises were exactly as you said. I don't settle, and I guess my biggest weakness is my pride. I'd rather not be in a relationship than be in the wrong one. I know my worth, and I'm not on here just to sleep around. People change. Life can change in the blink of an eye. Everything that seems so set in stone and forever can change in a second. So value your happiness. If you're happy today, be appreciative for it. If you were happy before, remember it. The ideal first date - that starts out with some drinks at a restaurant or bar, and maybe dinner. Or possibly a quiet cafe near a park. I know that sounds cliche, but honestly I would prefer to be stimulated by lively discussion rather than meet up at some crowded, overly loud bar where you cant hear where you are talking to. That disqualifies movies; I mean what good is a movie first date? All you do is sit next to each other and watch something on a screen. Then maybe a walk in that park, feeding ducks, romantic walk on the beach - you know, depending on how it goes. I'm not really expecting to find love here, but I've found you always find love in the most unexpected places.
Camping , Coffee and conversation , Business networking , Dining out , Fishing/Hunting , Movies/Videos , Museums and art , Music and concerts , Exploring new areas , Nightclubs/Dancing , Shopping/Antiques , Video games , Wine tasting
Auto racing / Motorcross, Basketball, Cycling, Football, Inline skating, Other types of exercise, Running, Swimming, Tennis / Racquet sports, VolleyBall, Walking / Hiking, Weights / Machines
I exercise 3-4 times per week.
I like Birds, Cats, Dogs, Exotic Pets, Fish, Horses, Other
Had a few dogs and cats growing up.
Middle of the Road
Baldwin-Wallace College, Berea, OH
I enjoy modifying cars, and current own an 09 BMW M6, 2010 Grand Cherokee, and a Supercharged Cobalt SS, among others.
I thoroughly enjoy john grisham and james patterson novels.
Z{{Cta}} {{#if HasMoreInfo}} ?{{CtaSecondary}} {{/if}}
|What he is looking for
|5'11" (180cms)
|3'0" (91cms) to 6'0" (182cms)
|Body type:
|Athletic and toned
|Slender, About average, Curvy, Athletic and toned, A few extra pounds
|No preference
|Dark brown
|No preference
|No Way
|No preference
|Social Drinker
|No preference
|Sales / Marketing
|No preference
I am currently employed as the Marketing Director for a multimillion dollar late model high performance domestic sport car auto yard. Been there 7 years, extremely happy.
|$50,001 to $75,000
|No preference
|Never Married
|No preference
|Have kids:
|No, Yes, and they live away from home
|Wants kids:
|No preference
|White / Caucasian
|No preference
|Christian / Other
|No preference
|Bachelors degree
|No preference


text:
Dr. Ravi Chandra, MD
General Surgery
- 1920 SW 20th Pl Ste 100
Ocala, FL 34471
- 3 Locations
- (352) 237-1212
Dr. Ravi Chandra graduated from the Gandhi Med Coll, Ntr Univ of Hlth Sci, Hyderabad, Ap, India in 1980. He works in Ocala, FL and 2 other locations and specializes in General Surgery and Vascular Surgery. Dr. Chandra is affiliated with Munroe HMA Hospital LLC, Ocallaghan Regional Medical Center and West Marion Community Hospital.
General Surgery, Vascular Surgery
Medical School: Gandhi Med Coll, Ntr Univ of Hlth Sci, Hyderabad, Ap, India; Graduated 1980
*This physician has multiple office locations. See Office Information for details.
1630 SE 18th St Ste 103
Ocala, FL 34471
1830 SE 18th Ave Ste 3
Ocala, FL 34471
1500 SE Magnolia Ext Ste 201
Ocala, FL 34471
2301 SE 3rd Ave Ste B
Ocala, FL 34471
2820 SE 3rd Ct Ste 100
Ocala, FL 34471


text:
|By PR Newswire
Article Rating:
|February 20, 2013 08:02 AM EST
NEW YORK, Feb. 20, 2013 /PRNewswire/ -- Reportlinker.com announces that a new market research report is available in its catalogue:
Cross Platform Mobile Development Tools: Market Analysis & Forecast
The market for cross platform mobile development tools is forecast to reach $8.2 billion by 2016. The promise of HTML5 as a write once run anywhere is a illusion but the technology is still indispensable in developing mobile apps. Developers are taking a number of cross platform development approaches and successful developers will match the right tools and approach to appropriate requirements and use cases. With the complexity of mobile app development continuing to grow, the tools vendors ability to reduce development time and increase application reach is generating significant opportunities. This new trend in mobile application development will also help fuel an more open and prosperous mobile app ecosystem.
This updated report provides expanded coverage of the cross platform mobile development tools market. Based on an objective survey of 228 mobile developers and 9 vendor interviews, the report provide insights into:
- Size of the Market Opportunity for Web & Non Web Cross Platform Mobile Development Tools
- Developer Demand, Skill Set, and Requirements
- Competitive Differentiators
- Market Structure, Growth Drivers and Adoption Barriers
Table of Contents
1.0 Introduction
2.0 Mobile Application Development Technology
2.1 Mobile Website Development
2.2 Mobile Web Applications
2.2.1 Native Applications
2.2.2 Hybrid Applications
3.0 Market Structure
3.1 Front End
3.1.1 Hand coded applications
3.1.2 Mobile web application frameworks
3.1.3 Native wrappers
3.1.4 Native Widget Platforms
3.1.5 Recompilers & Translators
3.2 Market Leaders Participation in Cross Platform Mobile Development Ecosystem
3.2.1 OEM and OS vendors
3.2.2 Browser vendors
3.3 Back-end
3.3.1 Pure play backend as a service vendors
3.3.2 Cross-platform tools vendors
3.3.3 Carriers
4.0 Business Models
5.0 Market Drivers and Barriers
5.1 Drivers
5.1.1 4G Networks
5.1.2 Native OS fragmentation
5.1.3 Significant development cost reduction
5.1.4 Enterprises & brands building consumer facing apps
5.1.5 Web is evolutionary, flexible and open
5.2 Barriers
5.2.1 Challenges with HTML5
5.2.2 Varying platform support
5.2.3 Varying browser rendering engines
5.2.4 Challenges of moving from desktop web development to mobile
6.0 Changing Market Dynamics
6.1 Discovery
6.2 Emerging new open source OS's focused on supporting web apps
6.3 New development techniques to manage multiple screens
7.0 Developer Survey
7.1 Demographics
7.2 Mobile Developer Experience
7.3 HTML5 and Development Approaches
7.4 Tools Used by Developers
7.4.1 Web frameworks
7.4.2 Cross platform development platforms
7.4.3 Game engines
7.4.4 Back end services
7.5 Value of Cross Platform Development Tools
7.5 Platforms Targeted
8.0 Forecasts
8.1 Regional Forecasts
8.2 Segmentations by Tool Types
9.0 Conclusions & Recommendations
9.1 Robust Mobile HTML5 Apps Will Evolve Slowly and Unevenly
9.2 Line Between Native and Web Will Become Very Grey
9.3 Better Linking of Front and Back-End Will Be a Growing Trend
9.4 Future Influence of Carriers and OEMS
9.4.1 Carriers will have to partner to participate
9.4.2 How big an influence will Microsoft be in driving fragmentation ?
9.5 Recommendations For Tools Vendors
9.5.1 Partner with the right carriers
9.5.2 Make tools easy to use
9.5.3 The Enterprise is fertile ground
10.0 Vendor Profiles
10.1 AppMobi – Vendor Profile
10.1.1 Overview
10.1.2 Products
10.1.3 Business Model
10.1.4 Competitive Advantages
10.1.5 Competitive Challenges
10.2 Adobe – Vendor Profile
10.2.1 Overview
10.2.2 Products
10.2.3 Business Model
10.2.4 Competitive Advantages
10.2.5 Competitive Challenges
10.3 Sencha Inc.
10.3.1 Overview
10.3.2 Products
10.3.4 Business Model
10.3.5 Competitive Advantages
10.3.6 Competitive Challenges
10.4 Motorola Solutions – Vendor Profile
10.4.1 Overview
10.4.2 Products
10.4.3 Business Model
10.4.4 Competitive Advantages
10.4.5 Competitive Challenges
10.5 Xamarin– Vendor Profile
10.5.1 Overview
10.5.2 Products
10.5.3 Business Model
10.5.4 Competitive Advantages
10.5.5 Competitive Challenges
List of Figures
Figure 1 – Cross Platform Mobile Development Tools Market Opportunity
Figure 2 – Mobile Application Types
Figure 3 – Application Framework Architecture
Figure 4 – Native Wrapper Architecture
Figure 5 – Native Web App Architecture
Figure 6 – Survey Respondents Reduction in Development Time Using Cross Platform Tools
Figure 7 – Mobile Developers With Experience in Programming Languages
Figure 8 – US Mobile Ad Spending 2010-2016
Figure 9 – Android vs. iOS Device Sales June 2010-July 2012
Figure 10 – Survey Respondents Segmented by Geographic Region
Figure 11 – Survey Respondents Segmented by Revenues of Employer
Figure 12 – Types of Applications Developed
Figure 13 – Types of Games Developed
Figure 14 – Mobile Development Experience
Figure 15 – General Software Development Experience
Figure 16 – Developer Commitment to HTML5
Figure 17 – Approach to Cross Platform Development
Figure 18 – Usage of Cross Platform Development Tools
Figure 19 – Web Frameworks Currently Using
Figure 20 – Web Frameworks Developers are Interested in Using
Figure 21 – Web Frameworks Tried but Abandoned
Figure 22 – Cross Platform Mobile Development Platforms Currently Using
Figure 23 – Cross Platform Development Platforms Interested in Using
Figure 24 – Cross Platform Mobile Development Platforms Tried but Abandoned
Figure 25 – Gaming Engines Currently Using
Figure 26 – Gaming Engines Interested in Using
Figure 27 – Gaming Engines Abandoned
Figure 28 – Back End Services Currently Using
Figure 29 – Back-End Service Interested in Using
Figure 30 – Back-End Services Tried but Abandoned
Figure 31 – Reduction in Development Time Using Cross Platform Tools
Figure 32 – Investment in Cross Platform Development Tools
Figure 33 – Cross Platform Tools and Training Spending Change Expected
Figure 34 – Amount of Spending Changes Forecast
Figure 35 –Tool Features Rankings
Figure 36 –Tool Features Ranking, Top 2 & Bottom 2
Figure 37 – Mobile Platforms Targeted
Figure 38 – Mobile Platforms Targeted in Next 12 Months
Figure 39 – Total Mobile Developers
Figure 40 – Mobile Developers by Development Approach
Figure 41 – Total Developers Using Cross Platform Development Tools
Figure 42 – Developers Using Cross Platform Tools Segmented by Paying Developers
Figure 43 – Average Spend per Paying Developer per Year
Figure 44 – Cross Platform Tools Revenues
Figure 45 – Cross Platform Tools Users Segmented by Region
Figure 46 – Cross Platform Tool Revenues Segmented by Region
Figure 47 – Total Developers Segmented by Tool Types
Figure 48 – Developers Using Frameworks Segmented by Types of Frameworks
Figure 49 – Developers Using Platforms Segmented by Types of Platforms
List of Tables
Table 1– Mobile Web Frameworks
Table 2 – Native Wrappers
Table 3 – Recompilers and Translators
Table 4 – Sample Pure Play BaaS Providers
Table 5 – BaaS Offerings From Tools Vendors
Table 6 – Browser Rendering Engines
Table 7 – Emerging Open Standard Mobile Operating Systems
Table 8 – Emerging Open Standardized Mobile Operating Systems
Table 9 – Mobile Developer Experience Segmented by Language
Table 10 – Language Experience Segmented by Experience Developing for Mobile Devices
Table 11 – Application Type Segmented by Commitment to HTML5
Table 12 – Perceived Limitations of HTML5
Table 13 – Perceived Benefits of HTML5
Table 14 – HTML5 Benefits and Limitations Segmented by Application Type
Table 15 – Development Approach Segmented by Mobile Development Experience
To order this report:
Software_and_Consulting Industry: Cross Platform Mobile Development Tools: Market Analysis & Forecast
Contact Clare: [email protected]
Intl:+1 339 368 6001
SOURCE Reportlinker
"We have a tagline - "Power in the API Economy." What that means is everything that is built in applications and connected applications is done through APIs," explained Roberto Medrano, Executive Vice President at Akana, in this SYS-CON.tv interview at 16th Cloud Expo, held June 9-11, 2015, at the Javits Center in New York City.
Jul. 1, 2015 05:00 PM EDT Reads: 863
The Internet of Things is not only adding billions of sensors and billions of terabytes to the Internet. It is also forcing a fundamental change in the way we envision Information Technology. For the first time, more data is being created by devices at the edge of the Internet rather than from centralized systems. What does this mean for today's IT professional? In this Power Panel at @ThingsExpo, moderated by Conference Chair Roger Strukhoff, panelists addressed this very serious issue of profound change in the industry.
Jul. 1, 2015 03:30 PM EDT Reads: 1,077
Explosive growth in connected devices. Enormous amounts of data for collection and analysis. Critical use of data for split-second decision making and actionable information. All three are factors in making the Internet of Things a reality. Yet, any one factor would have an IT organization pondering its infrastructure strategy. How should your organization enhance its IT framework to enable an Internet of Things implementation? In his session at @ThingsExpo, James Kirkland, Red Hat's Chief Architect for the Internet of Things and Intelligent Systems, described how to revolutionize your archit...
Jul. 1, 2015 02:21 PM EDT Reads: 663
It is one thing to build single industrial IoT applications, but what will it take to build the Smart Cities and truly society-changing applications of the future? The technology won't be the problem, it will be the number of parties that need to work together and be aligned in their motivation to succeed. In his session at @ThingsExpo, Jason Mondanaro, Director, Product Management at Metanga, discussed how you can plan to cooperate, partner, and form lasting all-star teams to change the world and it starts with business models and monetization strategies.
Jul. 1, 2015 11:30 AM EDT Reads: 2,256
To many people, IoT is a buzzword whose value is not understood. Many people think IoT is all about wearables and home automation. In his session at @ThingsExpo, Mike Kavis, Vice President & Principal Cloud Architect at Cloud Technology Partners, discussed some incredible game-changing use cases and how they are transforming industries like agriculture, manufacturing, health care, and smart cities. He will discuss cool technologies like smart dust, robotics, smart labels, and much more. Prepare to be blown away with a glimpse of the future.
Jul. 1, 2015 11:30 AM EDT Reads: 803
Internet of Things is moving from being a hype to a reality. Experts estimate that internet connected cars will grow to 152 million, while over 100 million internet connected wireless light bulbs and lamps will be operational by 2020. These and many other intriguing statistics highlight the importance of Internet powered devices and how market penetration is going to multiply many times over in the next few years.
Jul. 1, 2015 10:30 AM EDT Reads: 2,146
Internet of Things (IoT) will be a hybrid ecosystem of diverse devices and sensors collaborating with operational and enterprise systems to create the next big application. In their session at @ThingsExpo, Bramh Gupta, founder and CEO of robomq.io, and Fred Yatzeck, principal architect leading product development at robomq.io, discussed how choosing the right middleware and integration strategy from the get-go will enable IoT solution developers to adapt and grow with the industry, while at the same time reduce Time to Market (TTM) by using plug and play capabilities offered by a robust IoT ...
Jul. 1, 2015 09:45 AM EDT Reads: 1,953
Today air travel is a minefield of delays, hassles and customer disappointment. Airlines struggle to revitalize the experience. GE and M2Mi will demonstrate practical examples of how IoT solutions are helping airlines bring back personalization, reduce trip time and improve reliability. In their session at @ThingsExpo, Shyam Varan Nath, Principal Architect with GE, and Dr. Sarah Cooper, M2Mi's VP Business Development and Engineering, will explore the IoT cloud-based platform technologies driving this change including privacy controls, data transparency and integration of real time context wi...
Jul. 1, 2015 08:00 AM EDT Reads: 948
SYS-CON Events announced today that BMC will exhibit at SYS-CON's 16th International Cloud Expo®, which will take place on June 9-11, 2015, at the Javits Center in New York City, NY. BMC delivers software solutions that help IT transform digital enterprises for the ultimate competitive business advantage. BMC has worked with thousands of leading companies to create and deliver powerful IT management services. From mainframe to cloud to mobile, BMC pairs high-speed digital innovation with robust IT industrialization – allowing customers to provide amazing user experiences with optimized IT per...
Jun. 29, 2015 12:15 PM EDT Reads: 2,709
There will be 150 billion connected devices by 2020. New digital businesses have already disrupted value chains across every industry. APIs are at the center of the digital business. You need to understand what assets you have that can be exposed digitally, what their digital value chain is, and how to create an effective business model around that value chain to compete in this economy. No enterprise can be complacent and not engage in the digital economy. Learn how to be the disruptor and not the disruptee.
Jun. 29, 2015 11:00 AM EDT Reads: 2,200
The Internet of Things is not only adding billions of sensors and billions of terabytes to the Internet. It is also forcing a fundamental change in the way we envision Information Technology. For the first time, more data is being created by devices at the edge of the Internet rather than from centralized systems. What does this mean for today's IT professional? In this Power Panel at @ThingsExpo, moderated by Conference Chair Roger Strukhoff, panelists will addresses this very serious issue of profound change in the industry.
Jun. 29, 2015 09:45 AM EDT Reads: 2,549
Business as usual for IT is evolving into a "Make or Buy" decision on a service-by-service conversation with input from the LOBs. How does your organization move forward with cloud? In his general session at 16th Cloud Expo, Paul Maravei, Regional Sales Manager, Hybrid Cloud and Managed Services at Cisco, discusses how Cisco and its partners offer a market-leading portfolio and ecosystem of cloud infrastructure and application services that allow you to uniquely and securely combine cloud business applications and services across multiple cloud delivery models.
Jun. 28, 2015 11:00 AM EDT Reads: 2,251
In his General Session at 16th Cloud Expo, David Shacochis, host of The Hybrid IT Files podcast and Vice President at CenturyLink, investigated three key trends of the "gigabit economy" though the story of a Fortune 500 communications company in transformation. Narrating how multi-modal hybrid IT, service automation, and agile delivery all intersect, he will cover the role of storytelling and empathy in achieving strategic alignment between the enterprise and its information technology.
Jun. 27, 2015 10:00 AM EDT Reads: 2,267
Buzzword alert: Microservices and IoT at a DevOps conference? What could possibly go wrong? In this Power Panel at DevOps Summit, moderated by Jason Bloomberg, the leading expert on architecting agility for the enterprise and president of Intellyx, panelists peeled away the buzz and discuss the important architectural principles behind implementing IoT solutions for the enterprise. As remote IoT devices and sensors become increasingly intelligent, they become part of our distributed cloud environment, and we must architect and code accordingly. At the very least, you'll have no problem fillin...
Jun. 26, 2015 12:00 PM EDT Reads: 2,257
Growth hacking is common for startups to make unheard-of progress in building their business. Career Hacks can help Geek Girls and those who support them (yes, that's you too, Dad!) to excel in this typically male-dominated world. Get ready to learn the facts: Is there a bias against women in the tech / developer communities? Why are women 50% of the workforce, but hold only 24% of the STEM or IT positions? Some beginnings of what to do about it! In her Opening Keynote at 16th Cloud Expo, Sandy Carter, IBM General Manager Cloud Ecosystem and Developers, and a Social Business Evangelist, d...
Jun. 26, 2015 10:00 AM EDT Reads: 2,136
Converging digital disruptions is creating a major sea change - Cisco calls this the Internet of Everything (IoE). IoE is the network connection of People, Process, Data and Things, fueled by Cloud, Mobile, Social, Analytics and Security, and it represents a $19Trillion value-at-stake over the next 10 years. In her keynote at @ThingsExpo, Manjula Talreja, VP of Cisco Consulting Services, discussed IoE and the enormous opportunities it provides to public and private firms alike. She will share what businesses must do to thrive in the IoE economy, citing examples from several industry sectors.
Jun. 25, 2015 02:00 PM EDT Reads: 2,040
In his keynote at 16th Cloud Expo, Rodney Rogers, CEO of Virtustream, discussed the evolution of the company from inception to its recent acquisition by EMC – including personal insights, lessons learned (and some WTF moments) along the way. Learn how Virtustream's unique approach of combining the economics and elasticity of the consumer cloud model with proper performance, application automation and security into a platform became a breakout success with enterprise customers and a natural fit for the EMC Federation.
Jun. 25, 2015 01:30 PM EDT Reads: 2,199
SYS-CON Events announced today that the "Second Containers & Microservices Conference" will take place November 3-5, 2015, at the Santa Clara Convention Center, Santa Clara, CA, and the "Third Containers & Microservices Conference" will take place June 7-9, 2016, at Javits Center in New York City. Containers and microservices have become topics of intense interest throughout the cloud developer and enterprise IT communities.
Jun. 22, 2015 02:15 PM EDT Reads: 2,844
SYS-CON Events announced today that the "First Containers & Microservices Conference" will take place June 9-11, 2015, at the Javits Center in New York City. The "Second Containers & Microservices Conference" will take place November 3-5, 2015, at Santa Clara Convention Center, Santa Clara, CA. Containers and microservices have become topics of intense interest throughout the cloud developer and enterprise IT communities.
Jun. 20, 2015 12:00 PM EDT Reads: 3,914
With major technology companies and startups seriously embracing IoT strategies, now is the perfect time to attend @ThingsExpo in Silicon Valley. Learn what is going on, contribute to the discussions, and ensure that your enterprise is as "IoT-Ready" as it can be! Internet of @ThingsExpo, taking place Nov 3-5, 2015, at the Santa Clara Convention Center in Santa Clara, CA, is co-located with 17th Cloud Expo and will feature technical sessions from a rock star conference faculty and the leading industry players in the world. The Internet of Things (IoT) is the most profound change in personal an...
Jun. 15, 2015 08:45 PM EDT Reads: 4,170


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Medium duty & school bus parts | Intercity touring coaches.
The parts division at Prevost has been supporting bus fleets throughout North America for over 90 years. With over 80,000 parts in our database, we offer a huge selection of original equipment and aftermarket products. We offer free ground freight from multiple locations throughout...
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1 of 1 people found the following review helpful
Sleep at last,
Verified Purchase(What is this?)
This review is from: PetSafe Anti-Bark Spray Collar - Citronella (Misc.)
This is a really good product and does what it advertises. My 14 year old Jack Russell has been keeping us awake at night for years. We have tried everything and have had no success, until this. The collar worked straight away. One bark, one spray and then silence. He did jump back at a hundred miles an hour after the first spray so stopped him mid bark. After a few nights he tried to gauge the volume of his bark and has now learnt to moan a bit instead of a full blown woof. Still 90% better than before. We don't need to put it on him hardly at all now and only put the collar on if he barks loudly. On the whole the best result we could have hoped for.
Review Details
Location: Somerset UK
Top Reviewer Ranking: 65,867


text:
Posted July 17, 2006 and updated on March 12, 2009
Why is My Super Ex-Girlfriend rated PG-13?
Official MPAA Rating: My Super Ex-Girlfriend is rated PG-13 for for sexual content, crude humor, language and brief nudity.
Please read our content details for this movie to help determine if it is suitable for members of your family. We also encourage you to check our full review and our movie information page.
Don't assume this superhero themed movie is kid material. The PG-13 rating is a result of the many adult-oriented sexual jokes, crude remarks, and nudity. (You will note the MPAA description doesn't even mention the violent content, giving parents a clue about the sexual focus.) Coarse language is frequent, and includes direct slang references for sexual activity. Violence typical of a superhero movie includes guns being fired at people, many physical altercations, and car accidents—one involving a pedestrian. Principal characters are seen drinking socially.


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Pronunciation: 钔
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WICHITA - The Common Threads Quilt Show 2012 will be held June 21-23 at the Century II Exhibition Hall in Wichita. This year marks the 15th year for the biennial quilt show presented by the Prairie Quilt Guild. The guild, which includes over 700 members from Wichita and surrounding areas, celebrates its 30th anniversary in 2012 and expects to host over 5,000 guests who will view over 500 quilts.
The College of Human Sciences at Oklahoma State University has awarded MaKenzie Sauder of Great Bend the Martha Johnston Nutritional Sciences Scholarship.
Great Bend piano instructor Darlene Boley recently received her 50-year membership pin from the Music Teachers National Association.
The public is invited to take a look at what some of the top photographers in the area are seeing through their lenses, with the Shafer Gallery's new show, "Exposure Time: Juried Photography Exhibition." The gallery, located in the Fine Arts Building at Barton Community College, will host an opening reception for the exhibit from 1-3 p.m. Sunday, May 6.
There are easier and cheaper ways to accredit a medical lab, but St. Rose Ambulatory & Surgery Center has never settled for one of those options, said Carla Herl, administrative lab director. St. Rose has always sought the inspections of the College of American Pathologists (CAP), which recently renewed the lab's accreditation.
Everything from restaurant equipment to medical supplies to office furniture will be included in the upcoming St. Rose Ambulatory & Surgery Center public auction. It is set for 10 a.m. Saturday, May 12 in the building just east of the Dominican Sisters Convent in the 3500 block of Broadway.
A St. John teenager was flown to Via Christi Medical Center's St. Francis Hospital in Wichita via air ambulance on Friday, after the car he was driving was hit by a construction crane at the U.S. 50 junction with U.S. 281, three miles south of St. John in Stafford County.
A two-vehicle injury accident occurred at 11:30 a.m. Thursday on NE 30 Road and North Washington Ave., three miles north of Great Bend.
LA CROSSE - Lloyd R. Rodie, 91, died May 2 at his home in La Crosse. Born Aug. 25, 1920, in rural Timken, he was the son of John and Carrie (Kaiser) Rodie. A lifelong resident of Rush County, he was a farmer and stockman before his retirement.
Shirley Ann Wright Ward, 81, died May 3, 2012 at St. Johns Nursing Home in Hays, Kansas.
Great Bend Fire Department
Great Bend's 19th annual Cinco de Mayo Festival was held Saturday in the courthouse square.
The Full Plate Diet weight-loss program is coming to Great Bend starting Tuesday. This is a new created by the doctors who wrote "The Full Plate Diet" from the Lifestyle Center of America.
Each year, thousands of volunteers in Barton County donate their time and energy to make our community a better place to live. National Volunteer Week was celebrated in April but I would still like to recognize our Extension and 4-H volunteers.
HOISINGTON - In December of 2010, Hoisington High School Principal Meg Wilson was approached by Sue Kidd, Kansas Character Education Initiative coordinator, at the National Character Education Conference in San Francisco to co-chair the Kansas state character education development committee in its infancy. The committee was charged with developing K-12 standards for schools to guide the process of integrating social and emotional character development for Kansas. The committee work began in January of 2011 and with diligent efforts throughout the past 14 months has reached a final product approved by the Kansas State Board of Education.
Page 1478 of 2312
Brett and Kaley Williams of Parkville, Mo. announce the birth of their son Charles Joseph Williams on March 10, at St. Luke's Hospital in Kansas City. He weighed 8 lbs. 7 oz. and was 20 3/4 in. long.
Dale Burroughs of Ellinwood will celebrate his 80th birthday with an Open House from 2 to 4 p.m. on June 27, at the upstairs room of The Lone Wolf Restaurant, 108 East 1st Street, in Ellinwood. The event will be hosted by his children and their spouses David and Pam Burroughs of Wichita and Dana and Jerry Pfanenstiel of Scott City. He has five grandchildren and nine great-grandchildren.
Alfred "Al" Steinert will celebrate his 90th birthday on June 20, with a card shower.
Morris and Joyce Stukey will celebrate their 45th anniversary on June 20 with a card shower.
It seems like only yesterday when I raced my buddies down the red-carpeted ramp of the Pix Theater in Hoxie trying to nail down those good seats. You know the ones I'm talking about – those in the front row where tennis shoes could be heard latching into congealed soda from the earlier matinee.
Advancements in technology have arguably caused life to move at a much faster pace than it did even a decade ago. The speed at which change takes place today is phenomenal. While these advancements have brought about marvelous positive changes and benefits, they can inadvertently have equally dramatic and damaging negative effects.
Rayna Karst, an Emporia State University English major from Rush Center has been named to the dean's list for the College of Liberal Arts and Sciences. Nearly 500 students were named to dean's lists for the spring 2015 semester.
Hutchinson Community College has announced the honor rolls for the Spring 2015 semester, including grades through the end of the interterm session. 1,330 students have been honored with selection to the two honor roll listings, with 721 achieving the President's Honor Roll with a 4.0 grade point average for the semester.
Taylor Ann Latham of Great Bend has accepted membership in the National Society of Collegiate Scholars for her commitment to the ideals of scholarship, leadership, and service. NSCS is a member of the Association of College Honor Societies. Membership is by invitation only, based on grade point average and class standing. Taylor recently completed her freshman year on the Dean's Honor Roll with a 4.0 GPA at Kansas State University. She was elected as Vice President of College Republicans for her sophomore year and attended C-PAC in Washington, D.C. this spring. She is also organizing her second ...
WASHINGTON ─ Many individual states now rival the world's major energy producing countries, according to a ranking published today by API.
KANSAS CITY- Rookie left-hander Eduardo Rodriguez bounced back from a horrific start against Toronto and gave up just one run over 6 1/3 innings to lead Boston to a 7-3 win over the Royals on Friday night at sold-out (38,190) Kauffman Stadium.
The Great Bend Chiefs split a Friday night doubleheader with Hutchinson at the Great Bend Sports Complex. Great Bend fell in the opening game of the series by a score of 9-8. The Chiefs regrouped and delivered a 13-3 routing of the visitors in game two for the 1-1 split. Friday's second game was called in the fourth by virtue of the mercy rule. At 9-8 on the season Great Bend will face Buhler to start a five-game road swing on Tuesday, June 23. The doubleheader will begin at 4 p.m. The Chiefs return to the Great Bend ...
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Virgin Mobile Launches the Waterproof ZTE Reef
The water proof ZTE Reef washes to the shores of Virgin Mobile. As our love for mobile gadgets grows we begin to find ourselves with nicer and nicer hardware that is very fragile. While no one is quite sure where it all started, smart phone manufacturers are doing what they can to help us keep ...Tags: video recording, bluetooth, Virigin Mobile REEF, smart phone, RAM


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Make Your Move stars international dance sensation Derek Hough (Dancing With The Stars) along with Korean pop star BoA, in her first English language feature film.
She's ready for the summer!
Work it, girl.
Mariah Carey, her boobs and her new beau James Packer, aka the fourth richest man in Australia, took their love to Ibiza, Spain where they gallivanted on a yacht with her kids.
Bad Girl RiRi? More like Braless RiRi.


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iQualify offers a powerful, learner-centred education experience. It's tailored to fit your organisation and to inspire learner success. We get out of bed each day to build and evolve an outstanding world-class experience.
You, by us
iQualify can integrate with your technology and adopt your brand to provide a seamless experience for your learners.
Modern learning
Delivering a modern learning approach with iQualify is easy: it's digital, mobile and social from the ground up.
Digital learning tools
Keep your learners motivated to succeed with iQualify's digital learning tools - social notes, quizzes, notifications and more.
Data-driven insights
Boost retention and achievement through iQualify's data-driven insights. That's what we do.
iQualify is our platform, but it's your product. You can choose to just use the core platform, or adapt it to your organisation's needs. If you want iQualify to do something, it probably can – just ask us.
iQualify offers an engaging learner centric platform, developed for the modern, digital learner. And because it's web-based, you'll never need to upgrade it.
We'll work with you to create a seamless learner experience by integrating with your brand, identity and technology. Your learners will feel right at home.
Enhance your learners experience with our assessment tools, developed using our expert educational design knowledge.
If you already have content you love, but need a better way to deliver it to your learners we can help. Bring it to us and we'll work with you, bringing our expert knowledge and insights to ensure your content delivers learner success.
With our powerful analytics and insights you'll be able to increase learner engagement and improve your content.
Anyone can build software. But we're more than a tech startup, and iQualify is more than just software. Building on 70 years of evolution in educational design and learning expertise by Open Polytechnic, leaders in providing vocational distance learning, you can be sure that iQualify is the solution you need.
We're all about the custom touch, being flexible and responsive to your needs.
We're ready to work with you to create a solution based result, especially for you.
We'll work with you to deliver the content you need to reach your learner goals.
It's easy to get going with iQualify. And once you are, we'll be there to support you and keep everything running smoothly.
With iQualify, you can have your learning platform up and running incredibly quickly. To find out more, or to request a demo, just get in touch.Get in touch
At iQualify we're constantly looking to improve the user experience for our learners. To do this we monitor how learners are engaging with the platform and listen to feedback from our partners.
We understand that one of the most important functions of an assessed course is the assessment component. And (not-coincidentally) one of the core features of iQualify courses is that the majority of assessments are built into the course. This allows learners to practice and be assessed as they work through the material.


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BOSTON - There doesn't appear to be anything seriously wrong with Yoenis Cespedes' right hand.
Cespedes, who left Wednesday's game against the Reds in the seventh inning because of a right hand contusion, is in Thursday's lineup against the Astros, batting cleanup.
"And honestly he could have continued on [Wednesday]," John Farrell said of Cespedes before Thursday's game. "But [as a] precautionary [matter, we] just got him out of there and we don't anticipate any issues as he goes through BP today."
Cespedes looked fine during batting practice, even hitting one ball over the Monster while crushing a few out to center field.
According to NESN, Cespedes will wear padding that cushions the palm of his right hand when he bats as to not aggravate the injury.
Cespedes has only played in 10 games for the Sox since being traded over at the trade deadline in a deal that sent Jon Lester and Jonny Gomes to the A's, but he's already shown why he came to Boston as one of the more feared hitters in the game.
Take Sunday's and Tuesday's games for examples, when Cespedes hit his first two home runs as a member of the Sox.
Trailing 1-0 against the Angels on Sunday, Cespedes' three-run shot put them ahead in the eighth inning of an eventual 3-1 win. And on Tuesday, he hit a two-run homer in the eighth inning that gave Boston the lead in an eventual 3-2 win.
According to Elias, Cespedes becomes the first Red Sox since Reid Nichols to Aug. 23-24, 1992 at Seattle to hit a game-winning homer in the eighth inning or later in two straight games.
"He's got some home runs in timely moments," Farrell said. "There's a lot of bat speed. Obviously there's strength, we see that. There's an example of some fearlessness after [Reds reliever Jonathan] Brockton tries to knock him down and he comes back and takes him deep in the next pitch. I think he thrives in the moment, in big moments in the game. He doesn't fear those situations, I think he embraces them. And that's played out with the timeliness of a couple of home runs."
For Cespedes, the home runs were the first ones of his career that turned a deficit into a lead in the 8th inning or later.
With Cespedes batting fourth, here's how the rest of the Sox lineup looks:
Xander Bogaerts gets a night off, which puts Brock Holt at shortstop. Jackie Bradley will play center field and Will Middlebrooks gets the nod at third base.
Brock Holt, SS
Dustin Pedroia, 2B
David Ortiz, DH
Yoenis Cespedes, LF
Mike Napoli, 1B
Daniel Nava, RF
Will Middlebrooks, 3B
Jackie Bradley Jr., CF
Christian Vazquez, C
Allen Webster, SP
Cespedes back in Red Sox lineup after hurting hand
BOSTON - There doesn't appear to be anything seriously wrong with Yoenis Cespedes' right hand.


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Here's what we do:
We help lawyers surpass their client development goals. We provide a variety of online legal marketing services including:
- Building websites that attract meaningful attention and earn new clients.
- Developing and executing search engine marketing (SEM) campaigns (SEO & PPC).
- Implementing content marketing strategies that convert visitors into inquiries.
Here's a longer list of services we provide:
- Website Design, Development and Marketing
- Search Engine Optimization (SEO)
- Paid Search Engine Marketing Management (SEM, PPC, Adwords)
- Conversion Rate Optimization (CRO)
- Analytics Implementation, Analysis and Reporting
- Online Reputation Management
- Content Development and Marketing
- Professional Photography
- Professional Copywriting
- Email Marketing
- Video Production and Marketing
- Data Visualization Marketing (Infographics)
- Social Media Marketing
Here's why we're different:
Here are some of the reasons our clients choose us:
- No long-term contracts.
- Transparency in what we do and why we're doing it.
- Meaningful, tangible results.
Most online legal marketing doesn't work.
There's plenty of blame to go around. Inexperienced consultants selling magic bullets. Desperate lawyers entranced by shiny objects. Gigantic legal web marketing conglomerates peddling websites, search engine optimization and social media cure-alls at every legal conference.
We founded AttorneySync to help lawyers cut through the noise.
We help lawyers attract more of the clients they want. In one year we helped one client increase his Google organic search traffic from 6,700 visitors to over 25,000 visitors which helped to generate over 1,000 web inquiries.
In a nutshell, we help lawyers develop web content that their target audience wants to find. We market that content in a way that motivates people to engage, share, link to and further publicize it online.
This process improves our clients' visibility in search results and elsewhere on the web.
Our clients have come to trust us to deliver meaningful results.
We are completely transparent as to our strategies and techniques. In other words, our clients know exactly what we are doing and why we are doing it throughout the entire process.
We work vigilantly to improve the results we deliver. Our team regularly updates our processes to remain current with the most recent innovations in internet marketing, web design, search engine optimization and paid search marketing.


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Call every Friday morning between 11 and Noon. If you can't be by the phone and the radio when the show is on (thanks for listening at work!) you can email an item in before the show at
Dial N Deal - August 1st 2014
by Doug Herendeen
posted Aug 1 2014 2:09PM
Marlboro Sherpa Jacket
2003 premium, tan/khaki, courdory, never worn,
Large, Sherpa collar, mint condition with tags - $25
JF, Altoona 943-9015
3 1/2 HP Motor
with horizontal shaft, on wheels - $40
RV How-To Books
four, various info in each - FREE
[Following for Dodge 3500 Series Pickup Trucks]
Louvered Tailgate
for easier hookup to 5th wheels - $40
Air Foil
- $20
Diamond Plate Tool Chest
Aluminum, 5 years old, excellent condition - $100
Ed, Altoona 943-3903
Hoover ProHeat Carpet Cleaner
shampoos rugs, used once, works well - $90
Elvis Presley Memorabillia Collection
- (reduced) $125
Avon Bottles Collection
- $50
Nancy, Altoona 932-4252
Wooden Rack Clothes Dryer
folds - $3
Hamilton Beach Wide-Slot Toaster
two slots, like new - $5
Shark Hand Vac
cordless, used - $2
Dirt Devil Hand Vac
used - $5
T-Fal Steamer
used - $4
Dumbbell Tree
with hand weights, 2 5-pound, 2 3-pound,
2 1-pound with tree holder - $7
Grace, Hollidaysburg 695-2975
Rustoleum Paint
two one-gallon cans, Hunter Green,
never used, unopened, 2-3 years old
- $20 per can
Jake, Newry 696-3377
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Leaf Cuff Drop EarringCA$62.00
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Unisex Stainless Steel Medical Tag ID Necklace
$10.00 Our Price
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Even if jewelry isn't your thing, sometimes just slipping on a simple chain adds a whole new dimension to your look. Let us show you the way to simple accessorizing with no fuss or muss, just a straight attractive solid stainless steel medical tag ID necklace to complement your image. This chain is guaranteed to bring out your swagger and easygoing style. Pair this cool necklace with jeans and a T-shirt, and you are certain to steal the show. Want to know more? • Made with stainless steel • Features a red medical ID tag • Measures 24" L • Designed in a classic ball chain motif • Lobster clasp closure • Hypoallergenic--suitable for all skin types Available in your choice of the following styles: • Rectangle Medical ID Necklace • Round Medical ID Necklace
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Matt Barkley holds off Geno Smith to lead season's first Heismanpundit/CBSSports.com Heisman Straw Poll
Originally written on Heisman Pundit | Last updated 9/5/12
USC quarterback Matt Barkley held off a surge from West Virginia quarterback Geno Smith to once again top the Heismanpundit/CBSSports.com Heisman Straw Poll, a survey of Heisman voter sentiment released every Tuesday. Barkley, the leader in the preseason edition of the straw poll, collected nine out of a possible 11 first-place votes and totaled 28 points after throwing for 372 yards and four [...]
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Flash Furniture Hercules Series Folding Chair
Flash Furniture Hercules Series Folding Chair
Weights & Dimensions
|Frame: Metal
|Seat Material: Metal
|Padding: No
|Outdoor Chair: Yes
|Country of Manufacture: China
About the ManufacturerFlash Furniture: Quality office furniture - at the speed of light. Flash Furniture takes pride in offering the highest quality office chairs and furniture at the best price that you will find anywhere. They proudly present an extensive selection of quality furniture for the commercial and home office. Flash Furniture offers a wide selection of office, hospitality, and restaurant furniture, available to ship within forty-eight hours – so you can get the support you need, right on time.
Flash Furniture Hercules Series Folding Chair
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Great product for great price!!!
Genet from Rexburg, ID – Verified Buyer
Was so excited to find these on sale in time for my Thanksgiving company! They are really nice and have an extra stabilizer bar on the back for strength. That's a quality that isn't readily noticeable. These ended up being the same price as the 'cheap' ones at Wallmart --- I scored again on Wayfair!!
18 gauge, not really, but a good chair by current standards, good value.
Jeff from Austin, TX – Verified Buyer
This review in no way reflects wayfairsupply, but the entire folding chair industry move to China and acceptable dump duties paid by the corporations which import the chairs. Let's start with the definition of 18 gauge tubing. By definition it would mean a wall thickness of 1.2mm. I knew this wasn't the case, so I removed the rubber feet and my first reading was 1.13mm. That is over the paint, and without a clean cut and smooth surface. There are tolerances for tubing, meaning +/- values allowed. The tubing manufacturers always make the tubing as close to the low end to increase their profits. Depending on what standard, be it ISO, ATSM, etc, there is a different value. In the USA, the manufacturer is going to need to use tube which meets certain standards. Leave the USA, and you can find standards with definitions as loose as "as agreed upon deviation". I have not proven it falls outside ISO standards, which are more generous than I would have imagined. The 1.13 still falls inside the allowable tolerances but if I were to make a significant effort to smooth the surface it would fail. Enough bashing, the paint is very resistant to chemical solvents in comparison with the finish on chairs from the Meco corporation. So far I am unable to remove it without abrasive using Kleen-Strip solvent. The chair is as sturdy as anything currently available. You would need an American made chair to find anything stronger, and you aren't going to find a new American made chair. It's a great price, but I would rather have a better chair over a low price point. Like I said at the beginning, wayfair supply did not start this. The manufacturers of the chairs order chairs made to certain standards, then give out the specifications of the chairs they had built (in China) to companies which are going to sell the chairs. Every website selling these chairs lists the specifications, and everyone believes them simply because they read it and it must be true. I won't go into providing an explanation of dump duties. If you are so inclined, searching for "dump duties folding chairs" will explain what they are and if you are so inclined you can follow the money trail to see who benefits. One last thing, from my experience the weak point on folding metal chairs is the rivet where the seat meets the front legs. I have seen failure of a rear cross bar but unusual forces were being applied, not as in use for normal seating.
Strong chairs!
Tonya from Nashville, TN – Verified Buyer
This is kind of weird, but I bought these for the chair dance classes at my dance studio. Our chairs kept breaking and bending so I went on a search for a "big booty chair". I couldn't find a chair with a large seat so I purchased these durable chairs instead. So far they have lasted several months with tons of booty and are still in great shape. Seriously.
Amy from Mulino, OR – Verified Buyer
I love these. The red is so pretty. Goes with every occasion and my decor. They are very sturdy and good quality paint. They are great for puzzles during holidays and just extra dining seating.
Great buy.
Angela from Pine Bluff, AR – Verified Buyer
These were bought for my place of employment. Used for our company party. Great buy. Shipped and delivered purfectly. Sturdy and well made product.
Great bargain and fast delivery
Natalie from Versailles, KY – Verified Buyer
These are great chairs for short term use and the delivery was super fast. I bought them in black and they seem to be fairly high quality. I haven't used them yet, so I don't know how they are going to hold up to weight, being banged around, and general wear and tear, so I didn't feel I could give them 5 stars yet. But, brand new, out of the box....they seem to be a great purchase.
SHARON from CA – Verified Buyer
Better than I had expected. Color is bright and the quality is better than I explected for the money! I would recomend these chairs! Pros: Nice looking, good color and easy to store.
just what i needed
Mark from Salem, NH – Verified Buyer
sturdy folding chairs. bought for when company is here, light and easy to store Pros: good price Cons: none
Very reasonable
Gina from Horseshoe Bay, TX – Verified Buyer
The "silk" plants were inexpensive and you can tell by looking at them. I'm glad I got them to put in my rentals.
Great chairs, terrific in red!
Laura from Madison, CT – Verified Buyer
We bought the reed chairs for our staff room for lunch and learns, they look great and function well!
OUR SKU # FFC2077Hercules Series Folding Chair is also sometimes listed under the following manufacturer numbers: HF3-MC-309AS-BGE-GG, HF3-MC-309AS-BK-GG, HF3-MC-309AS-GY-GG, HF3-MC-309AS-RED-GG