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URL: http://www.bpb.de/internationales/weltweit/sicherheitspolitische-presseschau/247165/the-daily-caller
Pfad: Internationales / Weltweit / Sicherheitspolitische Presseschau / Medien / T / The Daily Caller
»http://dailycallernewsfoundation.org«
"Here's How Impeachment Obsession Is Allowing Big Tech To Build A China-Like Surveillance State"
https://dailycaller.com/2019/12/30/big-tech-surveillance-china-system/
Chris White warnt, dass Tech-Unternehmen und Behörden in den USA politisch weitgehend unbehelligt ein massives Überwachungsnetzwerk nach chinesischem Vorbild aufbauen könnten, da der Kongress durch das laufende Amtsenthebungsverfahren gegen den Präsidenten gelähmt sei. "Lawmakers are too busy wrestling with matters related to President Donald Trump's impeachment to address issues related to the government's deployment of facial recognition technology. Big tech is selling such technological know-how to police agencies and embedding it in smartphones while lawmakers remain distracted, Politico reported Monday. Other issues are also taking precedent, namely the death of one lawmaker who led efforts to regulate artificial intelligence. 'Unfortunately impeachment has sucked all the energy out of the room,' Rep. Stephen Lynch of Massachusetts, who chairs the House Oversight National Security Subcommittee, said in a statement, according to Politico."
"'Hispanic Invasion': Alleged El Paso Shooter's Manifesto Is An Anti-Immigrant Call To Arms"
https://dailycaller.com/2019/08/04/el-paso-shooter-manifesto/
Jason Hopkins berichtet über den Inhalt des Manifests des mutmaßlichen Täters von El Paso. "The nearly 2,400-word manifesto allegedly outlines the shooter's reasons for the launching the attack, breaking down the 'political' and 'economic' motivations. The entire diatribe argues that U.S. leaders — both Republicans and Democrats — are guilty of allowing unfettered immigration into the country and that violent action is needed to preserve the white population. (...) The author expressed explicit support to the Christchurch shooter, who killed around 50 people at a mosque in Christchurch, New Zealand, in March. The author also gained inspiration for his Saturday attack by reading 'The Great Replacement,' a book by a white nationalist French author who claims European elites are conspiring to replace their native populations with non-European immigrants. The manifesto ticks off a number of far-right grievances, however, the author also devoted a significant portion to overpopulation and its contribution to environmental degradation."
"Xi Jinping Tightens His Grip On China With Power Not Seen In Decades"
http://dailycaller.com/2017/10/20/xi-jinping-tightens-his-grip-on-china-with-power-not-seen-in-decades/
Pickrell beschreibt Chinas Präsident Xi Jinping als einen der mächtigsten Staatschefs in der jüngeren Geschichte des Landes. "Moving into his second term, the Chinese president is expected to turn his grand visions for the revival of China as a great, global power into action. China is likely to speak louder and forcefully in international affairs. The Trump administration may be facing a different, perhaps a much more assertive, China going forward, as the Chinese president stressed that no one 'should expect China to swallow anything that undermines its interests.' The Economist recently referred to Xi as 'the world's most powerful man,' and he may be. Regardless, Xi is one of the most powerful Chinese leaders since Chairman Mao Zedong, who ruled over the country with an iron fist for several decades."
"Hidden Camera Catches CNN Producer Saying Trump 'Probably Right' About Russia 'Witch Hunt'"
http://dailycaller.com/2017/06/27/cnn-producer-calls-trump-russia-story-mostly-bullsht-says-ceo-encourage
d-russia-coverage-video/
Ein mit versteckter Kamera gefilmter CNN-Produzent hat die Russland-Berichte seines Senders größtenteils als "Bullshit" bezeichnet. Die Berichte würden vor allem wegen der Einschaltquoten veröffentlicht werden, so der Produzent. "When asked by the Project Veritas reporter, 'But honestly, you think the whole Russia shit is just like, bullshit?' Bonifield replies, 'Could be bullshit. I mean, it's mostly bullshit right now. Like we, don't have any big giant proof.' (...) At another point, the Project Veritas journalist asks, 'Then why is CNN constantly like, Russia this, Russia that?' 'Because it's ratings,' Bonifield answers. 'It's a business, people are like the media has an ethical phssssss … All the nice cutesy little ethics that used to get talked about in journalism school you're just like, that's adorable. That's adorable. This is a business,' Bonifield also says. 'Trump is good for business right now.'"
"Turkey Turns To Russia's Most Advanced Air Defense System Instead Of Buying From NATO"
http://dailycallernewsfoundation.org/2017/04/24/turkey-turns-to-russias-most-advanced-air-defense-system-
instead-of-buying-from-nato/
Das NATO-Mitglied Türkei ist Berichten zufolge dabei, die Verhandlungen zum Kauf moderner russischer Luftabwehrsysteme vom Typ S-400 abzuschließen. "Turkey's decision to go with the S-400 was supposedly the result of NATO's failure to present a 'financially effective' alternative offer, according to TASS. (...) The S-400 is considered one of the most advanced air defense systems in the world. It is capable of shooting down both aircraft and ballistic missiles. (...) The missile defense deal presents potential challenges for NATO from an integration standpoint (NATO allies generally use standardized Western military equipment across their ranks). It also poses a geopolitical dilemma given Turkey's important role in the counter-Islamic State fight and their border with Syria."


Agent Justin Wells will join the group via Zoom technology. Alternately, Guild members can view the workshop from their own computers. Check the Member's Room (www.stlwritersguild.org) for details.
As an added bonus, Justin is offering a giveaway for those in attendance. Two winners will receive a 30-minute consultation at a later date to discuss their book and ask more personalized questions relating to their goals as a writer and about the writing industry.
Finding that one agent who is the right fit, and then trying to get them to take you on, can be tough. At the St. Louis Writers Guild's Jan. 6 workshop, literary agent Justin Wells will discuss a lot of things that you need to know in order to land your dream agent, and maintain a healthy relationship with that agent once you do.
The "Landing an Agent (query letters, relationships, and more!)" workshop will be held from 10 a.m. to noon in the Dogwood Room at The Lodge Des Peres, 1050 Des Peres Rd, St. Louis, MO 63131. Free to Guild members, $5 for nonmembers.
Justin is a Literary Agent with the Corvisiero Literary Agency. He will be graduating with a B.S. in Mass Communications with a focus in Public Relations. Justin loves being able to utilize his skills in public relations to assist the agency and his own clients through his work as a literary agent. His main focus is young adult, middle grade, and new adult, though he has been known to branch out and explore other books that catch his eye.


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Kangaroo River may refer to:

 Kangaroo River (Clarence Valley), a watercourse with its source in the Clarence Valley LGA, New South Wales, Australia
 Kangaroo River (Shoalhaven), a tributary of the Shoalhaven River in the Southern Highlands region of New South Wales, Australia
 Kangaroo River (Queensland), a watercourse in Queensland, Australia

See also
 Kangaroo (disambiguation)

redpajama_set_name: RedPajamaWikipedia


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Anatomy of the Piano sold out its Edinburgh runs in both 2013 and 2014 and has toured to Australia twice to great acclaim. In a performance that's part recital, part fantasy, part lecture ?piano man Pickvance' uncovers the physical, emotional and spiritual anatomy of the instrument. Along the way he sheds light on his own story as a musician, from not getting the Christmas present he wanted as a boy (he got a piano instead of a space rocket) to the codependency he discovered between the piano and pianist, each equally temperamental, each largely redundant without the other. Piano stories, piano solos, songs and dissection of the piano are accompanied by eccentric hand-drawn projections to create a quirkily subtle show of virtuosity, dissection and charmingly surreal humour. Just where exactly is the heart of the piano? Will is the man to tell you as he takes a trip through its body and soul using elements of classical, jazz, rock and pop with original songs and verse interspersed with the odd classic and a gently humorous presentation. Anatomy of the Piano for Beginners will delight anyone who has ever enjoyed the sound of the instrument in whatever genre of music - and charm any who haven't into compliance! Anatomy of the Piano - (for beginners) website.
Performers Will Pickvance.
Anatomy of the Piano - (for beginners)
Anatomy of the Piano - (for beginners) (One person show) production archive for QTIX code T1572123145. Details of all Anatomy of the Piano - (for beginners) archived productions can be found under the QTIX code: S525043496
17 Dec 16 Half Moon Young People's Theatre
Inner London
22 May 16 Theatre Royal
Bath & North East Somerset
31 May 15 General

redpajama_set_name: RedPajamaCommonCrawl


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Genetics and Molecular Biology
Global Genome Transcription Profiling of Bifidobacterium bifidum PRL2010 under In Vitro Conditions and Identification of Reference Genes for Quantitative Real-Time PCR
Francesca Turroni, Elena Foroni, Barbara Montanini, Alice Viappiani, Francesco Strati, Sabrina Duranti, Alberto Ferrarini, Massimo Delledonne, Douwe van Sinderen, Marco Ventura
Francesca Turroni
Laboratory of Probiogenomics, Department of Genetics, Biology of Microorganisms, Anthropology and Evolution, University of Parma, Parma, Italy
Elena Foroni
Barbara Montanini
Department of Biochemistry and Molecular Biology, University of Parma, Parma, Italy
Alice Viappiani
Francesco Strati
Sabrina Duranti
Alberto Ferrarini
Dipartimento di Biotecnologie, Università degli Studi di Verona, Verona, Italy
Massimo Delledonne
Douwe van Sinderen
Alimentary Pharmabiotic Centre and Department of Microbiology, Bioscience Institute, National University of Ireland, Western Road, Cork, Ireland
For correspondence: marco.ventura@unipr.it
Bifidobacteria have attracted significant scientific attention due to their perceived role as health-promoting microorganisms, although the genetics of the bacterial group is still underexplored. In this study, we investigated the transcriptome of Bifidobacterium bifidum PRL2010 during in vitro growth by microarray technology. When B. bifidum PRL2010 was grown in liquid broth, 425 of the 1,644 PRL2010 genes represented on the array were expressed in at least one of the three investigated growth phases, i.e., the lag, exponential, and stationary phases. These transcriptional analyses identified a core in vitro transcriptome encompassing 150 genes that are expressed in all phases. A proportion of these genes were further investigated as potential reference genes by quantitative real-time reverse transcription-PCR (qRT-PCR) assays. Their expression stability was evaluated under different growth conditions, which included cultivation on different carbon sources, exposure to environmental stresses (thermal, acidic, and osmotic), and growth phases. Our analyses validated six reference genes suitable for normalizing mRNA expression levels in qRT-PCR experiments applied to bifidobacteria.
Bifidobacteria represent one of the principal members of the gut microbiota of infants (12) and are thought to positively influence the health status of their hosts (9). Thus, in recent years, a very significant body of research has been directed to exploring the biology of this group of microorganisms (for recent reviews, see references 3 and 22). However, the genetic basis of these purported health-promoting effects is poorly, if at all, understood (16). Thirty-eight species of bifidobacteria have been described so far (22), and several bifidobacterial genomes have been sequenced (for reviews, see references 7, 25, 26, and 33). The advent of genomics has allowed the determination of complete genetic blueprints of many bifidobacteria and has provided genetic evidence that underpins the specific adaptation of these bacteria to the human gut (for reviews, see references 7, 10, 19, 25, 26, and 33). In this context, the decoding of the genome sequence of Bifidobacterium bifidum PRL2010, a microbial isolate from an infant gut, led to the identification of an arsenal of enzymes necessary for the metabolism of host glycans, such as mucins and human milk oligosaccharides (HMO) (19), which opened avenues for further scientific exploration of the microorganism as a representative component of the infant gut microbiota.
Global transcriptional profiling has already been applied to explore changes in the transcriptome of B. bifidum PRL2010 cultivated under different growth conditions related to the available carbon source, i.e., mucin oligosaccharides, HMO, and lactose (19). Similarly, transcriptome analyses have been employed to monitor differential transcription based on carbohydrate metabolism in Bifidobacterium dentium Bd1 (36), and responses to various stressful conditions in Bifidobacterium breve UCC2003, such as heat stress and osmotic shock (39). In contrast, whole-transcriptome analyses have not yet been published to investigate changes in the transcription profile of bifidobacteria at different growth phases. Such analyses are important, not only to provide insights into the molecular mechanisms underpinning adaptation of bifidobacteria to the different growth phases, but also to generate a valuable database of housekeeping genes. The genes in this category are assumed to be constitutively and uniformly expressed (18) and are important as reference genes to normalize the level of transcription in quantitative real-time reverse transcription-PCR (qRT-PCR) protocols (23, 24). The application of the qRT-PCR technique contributes very significantly to the understanding of complex biological processes that allow adaptation of bifidobacteria to the human gut (20). However, the validity of reference genes under specific experimental conditions must be determined before the application of quantitative mRNA expression studies. To provide a genetic baseline for differential expression studies in bifidobacteria, we investigated the transcriptome of B. bifidum PRL2010 at different growth phases under in vitro conditions. This analysis led to the identification of a set of PRL2010 housekeeping genes confirmed by qRT-PCR.
Growth conditions.B. bifidum PRL2010 was routinely cultivated in an anaerobic atmosphere (2.99% H2, 17.01% CO2, and 80% N2) in a chamber (Concept 400; Ruskin) at 37°C for 32 h in Man-Rogosa-Sharp (MRS) medium (Scharlau Chemie, Barcelona, Spain), supplemented with 0.05% (wt/vol) l-cysteine hydrochloride. Growth curves were performed at least in triplicate in PRL2010 batch cultures. Samples were taken at regular intervals from the cultures to measure the optical density at 600 nm (OD600) and to determine the CFU/ml and pH values until 32 h following inoculation. Aliquots of 20 ml for the lag, early exponential, late exponential, and stationary phases were centrifuged for 10 min at 10,000 × g and 4°C. The pellets were then immediately frozen in liquid nitrogen and subjected to RNA extraction. We had to employ this extensive centrifugation step due to the severe difficulty in pelleting B. bifidum PRL2010 cells. The mRNA turnover was checked by qRT-PCR using log cell fractions collected at different times of centrifugation, i.e., 2 min, 5 min, and 10 min, targeting molecular chaperone genes, such as groEL and dnaK. No obvious change in the expression level between these different centrifugation times was noticed (data not shown).
RNA isolation.RNA was isolated according to the protocol described previously (37). The quality and integrity of the RNA was checked by Experion (Bio-Rad) analysis. As shown in Fig. S2 in the supplemental material, all RNA samples appeared to be of good quality with no obvious degradation.
Microarray, description, labeling, and hybridizations.Microarray analysis was performed with an oligonucleotide array based on the B. bifidum PRL2010 genome: a total of 39,249 oligonucleotide probes 35 bp in length were designed on 1,644 open reading frames (ORFs) using OligoArray 2.1 software (13). The oligonucleotides were synthesized in triplicate on a 2,000 by 40,000 CombiMatrix array (Mulkiteo). Replicates were distributed on the chip at random, nonadjacent positions. A set of 74 negative-control probes designed based on phage and plant sequences were also included on the chip.
Reverse transcription and amplification of 500 ng of total RNA were performed with a MessageAmp II-Bacteria kit (Ambion, Austin, TX) according to the manufacturer's instructions. Five micrograms of RNA was then labeled with a ULS labeling kit for Combimatrix arrays with Cy5 (Kreatech, The Netherlands). Hybridization of labeled DNA to B. bifidum PRL2010 arrays was performed according to CombiMatrix protocols (19).
Microarray data acquisition and treatment.Fluorescence scanning was performed on an InnoScan 710 microarray scanner (Innopsys, France). Signal intensities for each spot were determined using GenePix Pro 7 software (Molecular Devices). The signal background was calculated as the mean of negative controls plus 2 times the standard deviation (1). The absolute expression pattern was determined from at least three independent biological samples. A gene was considered to be subject to active transcription when it was detected in all three experiments and, consistent with previously published studies, when its hybridization signal exceeded the background standard deviation by 3-fold (4). This definition allows comparative transcription analysis of the different growth phases, although it provides only qualitative data.
Expression profiles of actively transcribed genes were analyzed using the EPCLUST module (http://www.bioinf.ebc.ee/EP/EP/). Signal intensities (s) were first transformed into log2 values (v) [vi,j = log2 (si,j/Sj), where si,j is the signal intensity of gene j in the experiment i, and Sj is the average signal intensity of gene j in all experiments]. The EPCLUST K-means tool (default parameters) was used to find clusters of coregulated genes. The presence of enriched cluster orthologue gene (COG) functional categories was investigated in each of the eight clusters identified. This analysis highlighted the fact that one to four COG functional categories were enriched more than 2-fold with respect to the entire genome. The EPCLUST hierarchical-clustering tool (default parameters) was used to construct a heat map of genes belonging to these functional categories.
qRT-PCR.qRT-PCR primers (see Table S1 in the supplemental material) were used to amplify the reference genes indicated in Table S1. The criteria for primer design were based on a desired melting temperature (Tm) value between 58 and 60°C and an amplicon size of approximately 100 bp. qRT-PCR was performed using the CFX96 system (Bio-Rad, CA). PCR products were detected with SYBR green fluorescent dye and amplified according to the following protocol: one cycle of 95°C for 3 min, followed by 39 cycles of 95°C for 5 s and 66°C for 20 s. The melting curve was 65°C to 95°C with increments of 0.5°C/s.
Each PCR mixture contained the following: 12.5 μl 2× SYBR SuperMix Green (Bio-Rad, CA), 1 μl of cDNA dilution, and each of the forward and reverse primers at 0.5 μM; nuclease-free water was added to obtain a final volume of 20 μl. In each run, negative controls (no cDNA) for each primer set were included.
Data on the expression levels of the housekeeping genes were obtained in the form of crossing point (CP) values based on the "second derivative maximum" method as computed by CFX96 software (Bio-Rad, CA). Further data analysis was performed with CP raw data using the Excel-based application BestKeeper tool program (11).
Microarray data accession number.The microarray data have been deposited in the Gene Expression Omnibus database (http://www.ncbi.nlm.nih.gov/geo/) with the associated accession number GSE30832.
In vitro cultivation.Previous studies have shown that the fermentation abilities of B. bifidum PRL2010 are limited to a relatively small number of carbohydrates, including complex sugars, such as human milk oligosaccharides and mucin, or disaccharides, such as lactose (19). However, here, we decided to study the fermentation behavior of PRL2010 on glucose, as it represents the carbon source present in most of the commercially available synthetic growth media. For this reason, B. bifidum PRL2010 was grown in MRS plus l-cysteine hydrochloride (see Materials and Methods) supplemented with 2% glucose as the unique carbon source. Growth was evaluated by constant monitoring of the OD600 for 32 h using a microplate reader, as well as by determining the CFU/ml and pH values at different time points. As expected, the growth curve of B. bifidum PRL2010 obtained in this medium consists of a lag phase, an exponential phase, and a stationary phase (Fig. 1). The viable count of the culture decreases dramatically after 24 h, and the acidification of the medium by PRL2010 rapidly increases following 6 h of incubation to reach a minimal pH value of 4.2, as previously described for other bifidobacterial cultures (6) (Fig. 1).
Growth curve of B. bifidum PRL2010 in MRS broth. Growth was monitored as a function of time by measuring the OD600, viable colony counts (circles), and pH values. The time points (T1 to T4) analyzed by microarray expression and qRT-PCR are indicated by arrows.
Analysis of the in vitro transcription profile.mRNA was isolated from B. bifidum PRL2010 cells collected from a liquid culture at four time points following inoculation (Fig. 1). In total, 12 hybridization assays were carried out, consisting of a triplicate for each of the four time points (lag, early exponential, late exponential, and stationary phases). Genes were scored as being transcribed if their signal was identified in all hybridization experiments while also fulfilling additional signal strength criteria as outlined in Materials and Methods. Overall, 425 genes, representing 26% of the total identified PRL2010 gene arsenal, were shown to meet these criteria and were thus considered to be expressed under in vitro conditions. This repertoire of transcriptionally active genes was shown to be differently distributed in the different growth phases (Fig. 2a). Functional classification of these PRL2010 growth phase-specific genes according to the COG families is shown in Fig. 2b. Sixteen genes were specifically transcribed in the lag phase, i.e., they were not detected in the early/late exponential and stationary phases. These genes were predicted to be involved in protein translation, lipid and nucleotide transport, and metabolism. Similarly, of the 283 genes expressed in the early exponential phase, 12 were specifically expressed in that growth phase and were predicted to be involved in protein translation; signal transduction; amino acid, nucleotide, inorganic ion, and carbohydrate transport and metabolism; and energy production (Fig. 2a). In both the lag and exponential phases, PRL2010 cells appeared to be metabolically active, as was also indicated by the rapid decrease in the pH. Finally, 21 and 62 of the 273 ORFs and 274 ORFs, respectively, were shown to be specifically transcribed during the late exponential and stationary phases. These genes are predicted to be involved in mechanisms commonly found in metabolically active cells, such as those that specify proteins involved in amino acid and carbohydrate transport and metabolism, as well as in typical processes occurring in senescent cells, like repair of misfolded proteins by molecular chaperones and of DNA damage. Moreover, among the genes whose transcription was shown to be upregulated at these time points is the F1F0 ATPase-encoding operon, whose function is to counteract acid stress (15, 27). Notably, during stationary phase, transcription of genes whose products are presumed glycolytic enzymes, such as glyceraldehyde-3-phosphate dehydrogenase, was increased, which would be expected to lead to increased production of reducing equivalents and energy-rich intermediates that protect cells against the deleterious consequences of this late growth phase (e.g., low pH and DNA damage). Interestingly, several genes that are known to provide protection against stressful conditions (dnaK, dnaJ, grpE, clpB, clpC, hsp20, groEL/groES, and hspR) in bifidobacteria (for a review, see reference 28) increase their transcription levels following the transition from lag to exponential phase (see Fig. S1 in the supplemental material). However, a large body of published data supports the notion that molecular-chaperone-encoding genes are also expressed during "normal" growth (i.e., under nonstressful conditions) of bifidobacteria in laboratory media, perhaps simply to assist the proper folding process of nascent proteins (28–32, 38, 39). Also, the increased production of certain glycolytic enzymes in response to stressful conditions has previously been reported for Bifidobacterium longum (14). Overall, we noticed a reduction in the transcription of genes belonging to categories that are involved in the maintenance of an active metabolic state during growth and, in particular, the transition from lag to stationary phase. Compared to their abundance as a functional category in the genome, the transcriptomes of all in vitro growth phases were somewhat enriched in functions related to transport and metabolism of carbohydrates (9.8%, 10%, 11%, and 10% of the transcribed genes in lag phase, early and late exponential phase, and stationary phase, respectively, compared to 9% of the genes in the genome) and translation and ribosomal biogenesis (24%, 22%, 14%, and 12% of the transcripts compared to 11% of the genome), as well as posttranslational modification, protein turnover, and chaperones (5% of the transcripts in the tested growth phases and 7% in stationary phase compared to 3% of the genes in the genome) (Fig. 2).
Number and functional analysis according to COG categories of B. bifidum PRL2010 genes expressed in vitro by global transcription profiling. (a) Venn diagram displaying the numbers of genes expressed during the lag phase, early exponential phase, late exponential phase, and stationary phase. Inside each ring of the Venn diagram, the COG categories of the specifically upregulated genes for each of the tested growth phases are indicated. Each COG family is identified by a one-letter abbreviation: A, RNA processing and modification; B, chromatin structure and dynamics; C, energy production and conversion; D, cell cycle control and mitosis; E, amino acid metabolism and transport; F, nucleotide metabolism and transport; G, carbohydrate metabolism and transport; H, coenzyme metabolism; I, lipid metabolism; J, translation; K, transcription; L, replication and repair; M, cell wall/membrane/envelope biogenesis; N, cell motility; O, posttranslational modification, protein turnover, and chaperone functions; P, inorganic ion transport and metabolism; Q, secondary structure; T, signal transduction; U, intracellular trafficking and secretion; Y, nuclear structure; V, defense mechanisms; Z, cytoskeleton; R, general functional prediction only; S, function unknown. (b) Functional annotation of the in vitro-expressed genes of B. bifidum PRL2010 according to their COG categories. For each category, the black bar represents the percentage of genes in that category as detected in the sequenced genome of PRL2010 (19). The other bars show the percentages of genes transcribed during the lag, early exponential, late exponential, and stationary phases that belong to a particular category. The percentage was calculated as the percentage of transcribed genes belonging to the indicated COG category in respect to all transcribed genes.
Accordingly, when all upregulated genes were classified based on their transcription profiles, 8 gene groups were identified, where members of each group exhibit similar expression profiles (Fig. 3). These groups encompass 83 genes, which exhibit enrichment of particular COG families based on the growth phase relative to their abundance in the PRL2010 genome (Fig. 3). The lag- and early-exponential-phase-specific gene transcription pool was enriched in genes involved in functions pertaining to an active metabolic state (e.g., ribosomal biogenesis; energy production and conversion; and nucleotide, lipid, and coenzyme transport and metabolism) (Fig. 3). The late-exponential- and early-stationary-phase transcripts displayed a higher representation of genes that perform functions in protein turnover, protein folding, energy conversion, intracellular trafficking, cell division, and signal transduction mechanisms than those of the preceding phases (Fig. 3).
Correlation between gene expression groups and COG functional categories. From left to right, the hierarchical clustering of genes belonging to cluster-enriched COG categories is shown, along with the corresponding eight clusters of coregulated genes and the description of enriched COG categories. Color legend is to the right; showing increased (green) and decreased (red) transcription levels.
Notably, in contrast to what was noticed for other bifidobacterial strains (D. van Sinderen, M. Ventura, and F. Turroni, unpublished data), 1,219 genes, corresponding to 74% of the identified ORFs of the B. bifidum PRL2010 genome that were present on the array, did not appear to be expressed during any of the tested growth phases. Such apparent disagreement with other microarray data might be linked to the different cutoff criteria used, as well as to the different microarray platforms used, which may have different sensitivities with regard to hybridization signal generation and detection. A large part of these genes were clustered in particular genomic loci, for example, the DNA region encompassing the prophage locus Bbif-1 (34, 35) (see Fig. S1 in the supplemental material). Nontranscribed genes were also found in PRL2010 genomic regions predicted or shown to mediate host-microbe interaction, such as those that specify sortase-dependent pili (the pil1 and pil3 loci) and the biosynthesis and transport system for Tad (tight-adherence) pili, which have previously been characterized for PRL2010 (5, 19) and Bifidobacterium breve UCC2003 (10). In contrast, other DNA regions predicted to be involved in host interaction, such as the gene encoding the putative adhesion factor BopA and the pilus-like structure specified by the pil2 locus, are transcribed under such in vitro conditions (see Fig. S1 in the supplemental material).
Common genes expressed during in vitro growth.Transcription profiling analyses of PRL2010 cultivated under in vitro conditions revealed the existence of a core transcriptome, i.e., transcripts present in all three growth phases, consisting of 150 genes. Such genes are predicted to encode housekeeping functions, such as metabolism of lipids, amino acids, nucleotides, vitamins, and steroids, as well as glycolytic processes (Table 1). Furthermore, genes coding for hypothetical proteins and regulators are found among this core transcriptome of PRL2010.
Core transcriptome of B. bifidum PRL2010 grown under in vitro conditions
Comparing the core transcriptome of PRL2010 with the core genome contents of the genus Bifidobacterium (2) revealed a high level of correspondence between the two sets of genes (Table 1).
Validation of the identified reference genes.Based on the PRL2010 core transcriptome, 22 genes were selected as putative reference genes that could be used for mRNA level normalization in qRT-PCR protocols (Table 1) because they have homologs in all known bifidobacterial genomes and because of their COG assignments. To verify the suitability of these genes as internal reference (IRF) genes, their expression stability was investigated and compared to that of the very commonly used reference 16S rRNA gene. The PCR efficiencies and linearity of the housekeeping-specific real-time assays were defined using standard curves based on PRL2010 genomic-DNA samples. The evaluation of efficiency is considered an essential marker in gene quantification procedure. The amplification of putative IRF target genes obtained exhibited PCR efficiencies ranging from 96.4% to 100%. The expression stability of a tested housekeeping gene was analyzed as a function of different PRL2010 growth conditions, including exposure to stressful treatments, such as thermal, osmotic, and acid stress, and cultivation in the presence of different carbon sources (e.g., glucose and lactose) and different growth phases. Among the potential IRF genes tested, the most highly abundant transcript was shown to correspond to the 16S rRNA gene, whereas the pntA1 gene was found to generate the lowest level of transcripts (Fig. 4). The achieved crossing point (CP) data were further analyzed using the BestKeeper tool software (11), which allows identification of the "optimal" housekeeping reference genes (17) by calculation of CP data variation, CP data ranges, standard deviation (SD), and coefficient of variation (CV). As shown in Tables 2 and 3, the uvrD-rep, gluC, and pdxS genes displayed the highest level of expression stability (SD ≤ ±1 CP and CV of 2.5% CP). Notably, such data show that there is only a low level of variation associated with the expression of these three candidate genes under the different conditions tested (see Table S2 in the supplemental material). In fact, inspection of the CP data variations based on all the CP values derived from the different conditions analyzed (e.g., temperature shifts, treatment with NaCl, acid exposure, various carbon sources, and change in growth phases) displayed high expression stability for each of the three genes investigated.
Box plot overview of the CP values of B. bifidum PRL2010 cells cultivated under different growth conditions (growth phases; thermal, acidic, and osmotic stresses; and in the presence of different carbon sources). The box plots highlight the median 25th (white) and 75th (black) percentiles and expression range across the different reference genes.
Statistical analysis related to variation of CP values obtained from transcriptional behavior of various B. bifidum PRL2010 genesa
Data from BestKeeper correlation analysisa
Conclusions.Bifidobacteria have become a prominent group of microorganisms due to their perceived role as health-promoting bacteria in the human gut (33). However, despite a growing interest in the genomics, molecular biology, and genetics of this group of bacteria, surprisingly little is known about genome expression under in vivo or in vitro conditions (25). Microarray-based analysis allows insights into the metabolic states of bacteria under different culture conditions. The data described in this study show that, during in vitro growth, the lag, exponential, and stationary phases are characterized by a specific global transcription pattern. Analysis of the functional annotations of the transcribed genes revealed a shift from cell division-focused gene expression during the lag phase to a more pronounced carbohydrate metabolism-related gene repertoire in the exponential growth phase. Later in the growth cycle of B. bifidum PRL2010, cells modify their transcription patterns by reducing the total number of genes that are expressed, thereby sustaining the stationary growth phase.
Surprisingly, just 26% of the total number of genes present in the genome of B. bifidum PRL2010 were designated as being transcribed under in vitro conditions using our criteria, indicating that only a modest part of the overall genetic makeup of PRL2010 is active during growth in liquid broth. However, such a finding can be explained by assuming that all gut-expressed genes (e.g., those encoding mucin breakdown, as well as host-microbe interaction) remain transcriptionally silent during broth cultivation of B. bifidum PRL2010. Alternatively, the applied criteria may have been too stringent for the data sets obtained here, perhaps eliminating many genes that were in fact transcriptionally active. Nevertheless, if just 26% of the total gene repertoire of the microorganism is active under the in vitro conditions used here, this poses new questions related to the minimal transcriptome and the global regulation of genes when a bacterium is cultivated in a "nonnatural" environment. The core transcriptome of PRL2010 contains a large number of genes that are present in all bifidobacterial genomes so far sequenced. This highlights an interesting parallel between the core transcriptome and the minimal genome content of the genus Bifidobacterium, suggesting that the genetic requirements for the utilization of simple carbohydrate substrates are conserved and are always expressed in bifidobacteria.
Furthermore, apart from these gene categories, there are 346 nontranscribed genes belonging to the "unknown-function" category, and we cannot even speculate under what circumstances/conditions these genes may be expressed.
Our study facilitated the identification of a set of bifidobacterial housekeeping genes suitable as internal reference genes for the quantification of mRNA targets in bifidobacterial cells by quantitative gene expression analysis techniques, such as real-time RT-PCR. This technique is frequently applied to evaluate changes in gene expression under different growth conditions involving stress treatments, fermentation on different carbon sources, and protease treatments (for examples related to bifidobacteria, see references8, 21, and 36). However, the reference genes used so far for such gene transcript quantification by qRT-PCR approaches in bifidobacteria have not been subjected to solid validation. It is worth mentioning that the proposed reference genes, as suggested in this report, represent the optimal reference conditions for relative qRT-PCR analysis of gene expression for the growth conditions used in this study. However, since it is possible that specific but as yet untested environmental conditions affect bacterial gene expression, we have to accept that universal reference genes do not exist. Thus, a certain amount of caution should be applied in the selection of reference genes in qRT-PCR assays, including those proposed here, when using different growth conditions and/or different bifidobacterial species.
This study has focused on the B. bifidum PRL2010 strain, which we use as a model microorganism to study the genetics and physiology of intestinal commensal bifidobacteria. However, it is worth mentioning that the genetic behavior of the various members of the genus Bifidobacterium is likely to be different, and thus, the findings described here might not be applicable to all bifidobacteria. For these reasons, further research activities should be carried out to facilitate the determination of global transcription patterns of bifidobacteria under in vivo conditions, allowing comparative analysis with in vitro transcriptome data, which will be important in order to determine if bifidobacterial transcription in different gut sections corresponds to the transcription profile observed in a specific growth phase and thus to obtain molecular clues to gut colonization by bifidobacteria.
This work was financially supported by the Italian Award for Outstanding Young Researcher scheme "Incentivazione alla mobilità di studiosi stranieri e italiani residenti all'estero" and by a Marie Curie Reintegration Grant (MERG-CT-2005-03080) to M.V. and a FEMS Advanced Fellowship 2011 to F.T. D.V.S. is a member of the Alimentary Pharmabiotic Centre, which is a Centre for Science and Technology (CSET) funded by the Science Foundation Ireland (SFI) through the Irish government's National Development Plan (grant no. 02/CE/B124 and 07/CE/B1368).
We thank S. Ottonello for constructive and critical reading of the manuscript.
Received 28 July 2011.
Accepted 7 October 2011.
Accepted manuscript posted online 14 October 2011.
↵† Supplemental material for this article may be found at http://dx.doi.org/10.1128/AEM.06352-11.
Copyright © 2011, American Society for Microbiology. All Rights Reserved.
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Applied and Environmental Microbiology Dec 2011, 77 (24) 8578-8587; DOI: 10.1128/AEM.06352-11
You are going to email the following Global Genome Transcription Profiling of Bifidobacterium bifidum PRL2010 under In Vitro Conditions and Identification of Reference Genes for Quantitative Real-Time PCR


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← Life in the Time of Corona
A Short Trip to the Italian Lakes →
Posted on June 27, 2020 by twobbs
Italy began lifting its lockdown restrictions in mid-May, and in early June the ban on traveling between Italian regions was removed. That didn't mean you could immediately go anywhere – flights to Sicily, where we had originally planned to go, won't resume until the end of the month. But the uneven end of the lockdown provided a golden opportunity to visit popular sites at a time when not not many foreign tourists would be around.
So as soon as we could, we were on the road again.
When we arrived in Venice, on a Wednesday evening, the city was so empty as to be almost desolate. We have been to Venice in low season before, trading off the shorter days and sometimes bad weather of late winter to avoid the huge high season crowds. But this was different. Smaller hotels were open, but many of the larger ones were not. Many shops and restaurants were open, but a surprising number remained closed. The Basilica of San Marco was closed to tourist visits, although you could still attend mass or visit the loggia upstairs. And the Piazza San Marco, normally teeming with tourists, shoppers and vendors, was practically empty. The Cafe Florian, the iconic coffee bar in the middle of the Piazza, was closed when we arrived. Even the pigeons seemed to have temporarily deserted the place.
Fortunately, things improved over the course of our 4-day visit. By Friday, the Cafe Florian had reopened (although their coffee was still too expensive to actually consider buying). By Saturday, a few souvenir vendors had returned to the Piazza, but they were the licensed kiosks, not the guys selling "irregular" merchandise spread out on the street. Even a few pigeons had ventured back. And more shops and restaurants opened their doors.
The number of tourists increased each day, too, and by Saturday there was a line to get in to the Doge's Palace (although it didn't look very long). Most of the tourists were Italian, including many families, taking the opportunity, like we were, to rediscover their own cities as they were in a quieter era.
Venice, like most parts of Italy, does not require people to wear masks outdoors, and the social distancing requirement is only 1 meter (about 3 feet). You are required to wear masks in indoor public spaces – hotel lobbies, churches, museums, stores. They are required in bars and restaurants, but you can take them off as soon as you are seated and, given the fine weather, we took most of our meals outside. You are also required to wear them on public transportation, although on the 40-minute trip to Burano, those of us sitting outside took our masks off.
Aside from that tiny act of civil disobedience on the boat, compliance with the masking rules was quite high. We saw one guy take his mask off inside a church, admiring a Tintoretto, when the custodian, a young woman, came over and politely asked him to put it back on. "Later," he said. When she asked a second time, still politely, the guy looked around at the other visitors giving him the stinkeye, and realized he was at risk of making a "brutta figura," which literally means "cutting a bad figure," but actually means "embarrassing yourself by acting like a jerk." And no Italian wants to do that. He put the mask back on.
I do wonder, though, how Venice and other big cities are going to enforce the social distancing requirements on public transportation, which means leaving every other seat vacant, when there are more visitors.
Carpaccio: Panel from Legend of St. Ursula (restoration finally complete!)
We visited the Accademia, Venice's largest art museum where we were able to spend time with the works of Bellini, Veronese and Carpaccio in uncrowded rooms. We also visited the Peggy Guggenheim museum. We don't always like museums devoted to modern art, but Guggenheim selected her pieces according to her personal taste, and makes a convincing case for her selections. Even Jackson Pollock's strange abstractions look more organized when paired with Peggy's wrought-iron windows. And Peggy's placement of modern abstract pieces next to contemporary African art is also very interesting – she knows exactly where the inspiration for the European works is coming from, and she wants you to know it too.
Magritte: La voce dell'aria (at the Guggenheim)
We took a guided tour of the Casa Bartoli, a 16th C structure which, much renovated, was a private residence until the owners died a few years ago. The house was elegant, but comfortable, and had killer views of the Grand Canal and the church of Santa Maria della Salute across the way. (The church, whose name means Our Lady of Good Health, was built by the city in gratitude for surviving the second coming of the Black Death in the 17th C – a story that had unexpected resonance in these pandemic times.)
Casa Bortoli interior
Casa Bortoli: Fantastic chandelier
The Salute Church from Casa Bortoli
We even took a gondola ride, for the first time in many years. Without the big cruise ships in the lagoon, you could venture out into the canal in a gondola without fear of being swamped.
Mostly, though, we enjoyed just walking the streets of Venice, being able to travel side by side instead of single file for once. Most of the restaurants we have enjoyed in the past were open, and doing good business, but they had time to talk to us. So did our gondolier, the folks running our hotel, and shopkeepers. They all wanted tourists back – just maybe not so many of them.
The Cinque Terre are five spectacularly sited villages along the Ligurian coast. Although there are some roads, the towns are connected mainly by hiking trails and a rail line. We had never been to this part of Italy before, since we had heard that it was often crowded during high season and the hiking season is short (it's not really a place you can visit during the winter).
The villages are as beautiful as advertised. From our base in Monterosso, we enjoyed hiking to Vernazza and then Corniglia, and we visited a fourth town (Manarola) by train. We even got in a little beach time. All in all, though, I'm not sure we would have enjoyed it so much had it been more crowded.
We have been to Lucca before, some years ago, but again, it seemed like a good time to revisit this popular city. Lucca has a very old cathedral (started in the 11th C) and very fine city walls, which you can walk around. But the highlight of this trip was our visit to Villa Reale di Marlia, a few miles outside the city.
Villa Reale was the country residence of Elisa Baciocchi, the sister of Napoleon Bonaparte, who was named Princess of Lucca after her brother conquered much of northern Italy. Elisa restored the 17th C manor house, and completely redesigned the gardens in the style of English country houses (including very large lawns, unusual in Italy). After Napoleon was deposed, the house was sold to a succession of new owners, who maintained the magnificent gardens but allowed the manor house to fall into disrepair. The house has recently been restored by an anonymous Swiss couple who fell in love with the place and clearly spared no expense to restore the house to its original Napoleonic splendor. It is spectacular, and we highly recommend a visit here to anyone who happens to be in the vicinity of Lucca.
Travel in the post Covid era
How has travel changed in the post Covid era?
The first and most obvious difference is that we took our car instead of the train. Neither of us minds wearing a mask during a visit to a church or museum, or on a short trip on a bus or train. But wearing one continuously for 5 or 6 hours seems a bit much. I can't even imagine a transAtlantic flight under those conditions.
Going to a restaurant where all the waiters are masked is a little weird, but you get used to it pretty quickly.
One less obvious casualty of the pandemic is the hotel buffet breakfast. The hotel we stayed at in Venice used to offer breakfast, but their breakfast room was so tiny they could not comply with the new social distancing requirements. So they no longer offer breakfast, and have adjusted their room rates accordingly. Given that the hotel was situated within 5 minutes walk of at least half a dozen cafes, including an excellent pastry shop literally outside the door, this was no hardship.
The other two hotels we stayed at continued to offer a buffet style breakfast. You could see everything on offer, but you had to ask a hotel staff member to get it for you and put it on your plate. In some ways, that's a good thing – you will probably be too embarrassed to ask for that second piece of pastry you didn't need anyway. But some might be less willing to try an unfamiliar local specialty – deviled eggs in Turkey, or spinach with pine nuts in Spain, which would be too bad.
The Peggy Guggenheim museum in Venice is currently operating an an advance reservation basis – you go to the website and select a date and an entry time. I think this is a great idea for small museums, which can sometimes be unpleasantly crowded at peak times. The Galeria Borghese in Rome has been doing this for several years now. It requires a bit of advance planning, but it makes your visit to these smaller museums much more enjoyable. I hope it's a permanent change.


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The Deep Blue Sea, Maltings Arts Theatre, St Albans, 8-10 & 15-17 March
This should be good. A flat in Ladbroke Grove, West London. 1952. When Hester Collyer is found by her neighbours in the aftermath of a failed suicide attempt, the story of her tempestuous affair with a former RAF pilot and the breakdown of her marriage to a High Court judge begins to emerge. With it comes a portrait of need, loneliness and long-repressed passion. This is Terence Rattigan's classic. 8pm.
Liz Earle, Spirella Ballroom, Letchworth, Weds 7 March
Liz Earle MBE is coming to Letchworth to help us all to be radiant before, during and after the menopause. Sounds good to me! Liz is known for her passion for beauty, natural health and
wellbeing, as well as her considerable charity and campaign work. She co-found the Liz Earle Beauty Co., now owned by Walgreen Boots Alliance. This is a David's Bookshop event at the Spirella Ballroom.
Liz Earle Spirella Ballroom, Letchworth SG6 5ET
Writing and getting published, Bibliomaniac, Harpenden, Weds 7 March
Bibliomaniac is always up to something good in Harpenden. She organises brilliant events that are usually packed out. Ian Ridley, Kerensa Jennings, Jennie Ensor and Jessica Duchen will be talking about their novels, their writing and their road to publication. Free drink & goody bag included in the £10 ticket price. 8-10pm.
Bibliomaniac The Harpenden Arms, 188 High Street, AL5 2TR
Zoe Folbigg, The Note, David's Bookshop, Thurs 8 March
Zoë Folbigg plucked up the courage to give a note to the handsome man on the Hitchin to Kings Cross commuter train and her bravery paid off. They are now married and have two children. Zoë then wrote her debut novel The Note, a book I loved so much I read it in one go. She'll be talking about being bold and writing a bestseller in David's Bookshop on Thursday. 7.30pm.
Zoë Folbigg at David's Bookshop, Eastcheap, Letchworth, SG6 3DE
Community tree planting, Heartwood Forest, Sun 11 March
Do you know Heartwood Forest? It's (almost) a brand new forest, planted from scratch. On Sun 11 March, the last trees are being planted and everyone is invited to join in. Trees are provided! And you'll also be shown how to plant them, so novices are welcome. Wear wellies or boots. Park in the main car park and there will be shuttle buses to and from the planting site. Refreshments will be on sale. Free! Booking not necessary.
Heartwood Forest
Orchid Festival, Kew Gardens, London, until 11 March
Things are going totally tropical this month at London's gorgeous Kew Gardens. Their annual orchid festival takes in Insta-friendly displays of the vibrant Thai blooms, Thai street food and musical performances, workshops on how to grow these notoriously tricky flowers and Thai afternoon tea at The Botanical restaurant. King prawn, lemongrass and coriander tartlet followed by Thai sticky coconut cream rice with mango salsa? Yes please.
Nicholas Hytner, Kings Place, London, Sun 11 March
Photo: Johan Persson
Nicholas Hytner, former artistic director of the National Theatre, talks about his experiences gained working with many of the leading actors, musicians and designers in the UK. Expect behind the scenes stories from many of his productions, including The History Boys and One Man, Two Guvnors. Chaired by none other than David Hare. 2pm.
Jewish Book Week Kings Place, 90 York Way, Kings Cross, London N1 9AG
Adam Kay, This is Going to Hurt, Old Town Hall, Weds 14 March
The show of the book. Comedian Adam Kay shares entries from diaries as a junior doctor in this hour-long show of stand-up and music. The Guardian described it as 'electrifying'. Adam will also have a chat and sign books after the performance. 8pm.
Adam Kay, Old Town Hall, High Street, Hemel Hempstead, HP1 3AE
Wirra Wirra wine dinner at Loft, St Albans, Weds 14 March
Sam Temme from award winning winery Wirra Wirra in McLaren Vale in Australia will be presenting a range of their wines including the 12th Man Chardonnay, the brand new Ashton Hills Pinot Noir & Woodhenge Shiraz. Learn the stories behind the names of the Catapult Shiraz and Dead Ringer. This will be matched by a menu designed by Nick Male, chef and owner of the recently opened Loft restaurant in St Albans. £55.
Wirra Wirra at Loft 23B George St, St Albans AL3 4ES
Tipis4Hire open weekend, Rothamsted Manor, Harpenden, Sat 17 – Sun 18 March
Tipis4Hire is a local company and they'll be open for business (well, you can have a good snoop) at Rothamsted Manor for the whole weekend. I went to a similar event last year and it was great. Music, food, and drink and, I do believe there was sunshine. If you don't know it, Rothamsted Manor is a relatively new venue (in an old manor house) and it's stunning. Perfect for weddings, obviously, but also more than suitable for those big birthdays. The gardens are wonderful and there are loads of interesting walks all around the estate.
Tipis4Hire at Rothamsted Manor West Common, Harpenden, AL5 2BG
Julius Caesar, National Theatre Live, Broadway, Thurs 22 March
Nicholas Hytner's production. Ben Whishaw and Michelle Fairley play Brutus and Cassius, David Calder plays Caesar and David Morrissey is Mark Antony. Broadcast live from The Bridge Theatre, London. 6.45pm
Julius Caesar, Broadway, Letchworth SG6 3DD
The Harpenden Big Read Quiz, Bennet's Club, Weds 21 March
How well do you know your crime (fiction)? This is a night of killer quiz questions with rounds on crime fiction, Harpenden and many more fun categories. It's a celebration of the launch of Leigh Russell's brand new thriller, Class Murder, published by Harpenden's No Exit Press. Leigh will be there! Ticket price (£10) includes one free drink and a goody bag. 7.30pm.
Big Read Quiz Bennet's Club 21 Leyton Road Harpenden AL5 2HU
Gangsta Granny LIVE on stage! Hertford Theatre, Thurs 22 – Sat 24 March
I can hear the cheers already. This production of David Walliams' book is from the producers of Horrible Histories so if I were you I would book tickets quickly!
Gangsta Granny Hertford Theatre, The Wash, Hertford SG14 1PS
St Albans South Signal Box Big Weekend, Sat 24 – Sun 25 March
Bells and whistles and levers. This is a free event, complete with signalling demonstrations and refreshments. 2-5pm.
South Signal Box, Ridgmont Road, St Albans, AL1 3AJ
Chilean Supper Club, White Hart, Welwyn, Thurs 29 March
I went to the January Chilean supper club at the White Hart and it was a wonderful evening. Home cooked food (this isn't a lavish tasting menu) and with lots of wine tasting and drinking. The food was delicious. To give you an idea, the menu included: empanadas, beef flank, and sausages with chimichurri. Pudding was cake made from dulce de leche. YUM! Not sure what's on the menu this time, but the wine comes from James at Wine2Trade in Hertford. He is very knowledgeable, and told us about the wines in an interesting (and succinct) way. You could buy the wine we tasted on the night. This is great to book with friends; it's a really good way of catching up and you don't waste any time pouring over menus or paying the bill. Buy tickets well in advance for this one because they usually sell out. At £45 it's phenomenal value. Be warned, you can drink up to six glasses of wine (nobody forced me, but…) so you might want to forget about your car, book a room and enjoy a leisurely breakfast the next morning.
Argentinian Supper Club
Bawden's Beasts, The Higgins, Bedford
Over the border to the Higgins in Bedford. This exhibition opened in January and it's on for a while, but I just wanted to let you know, particularly if you are an Edward Bawden fan. Creepy crawlies join the ranks of jungle giants and other beasts to sell products, narrate stories or decorate walls. More than sixty art works featuring animals are on show. Entry is free.
Bawden's Beasts The Higgins Bedford, Castle Lane, Bedford, MK40 3X
Ocean Liners: Speed and Style, V & A Museum, London, until July 2018
I went to see this V&A exhibition last week and it's glorious! It's also totally immersive. One minute you're sipping a cocktail on deck as you power through the waves to New York in the 50s, , the next, you're wondering whether you could ever squeeze into the Dior suit worn by Marlene Dietrich on a transatlantic crossing. Kate Winslett and Leonardo di Caprio even make an appearance. For a truly elegant and glamorous experience, I recommend a trip to the V&A.
Ocean Liners: Speed and Style
Kate Terry, A System of Parallels, Broadway Gallery, Letchworth, until Sun 22 April
This is a wonderful new exhibition by British artist Kate Terry and I urge everyone to go and see this. Kate Terry lives in London and she has created a thread installation for the gallery, that has to be experienced. There are also drawings and sculptures and lots of Farrow and Ball paint. Uplifting and colourful.
Broadway Gallery The Arcade, Letchworth, SG6 3EW
Barbara Hepworth: Finding Form, River & Rowing Museum, Henley, until 3 June
Now you can see Barbara Hepworth's work without making a trip to Cornwall. Although I went to St Ives last October and sitting in Barbara Hepworth's garden in the sun was complete bliss. This retrospective at the Rowing Museum covers the sculptor's work from the 1930s to the 1960s, borrowing pieces from the Tate and National Portrait Gallery. A must-see.
rrm.co.uk
Fly the Link trainer, De Havilland Museum, London Colney
You've always wanted to do this, haven't you? Get into the LINK cockpit and fly? Produced to help to train pilots, it was used extensively during World War Two. You'll have an instructor by your side who will guide you through the trainee pilot test.
The de Havilland Aircraft Museum Salisbury Hall, London Colney, AL2 1BU
Book for: Courtney Pine, Saffron Hall, Saffron Walden, Thurs 12 April
The king of British jazz sax! Courtney Pine will be playing music from his new album, Black Notes From The Deep. Be quick, this is bound to sell out. The words 'living legend' aren't an overstatement.
Courtney Pine, Saffron Hall, Audley End Road, Saffron Walden CB11 4UH
This MonthWhat's On


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Nurag – stożkowata wieża wzniesiona nad kamiennym ołtarzem budowane na Sardynii przez Nuragijczyków. Najstarsze datowane są na XV wiek p.n.e.

Wieża zbudowana jest z kamieni ułożonych bez użycia zaprawy. Na zewnętrznej powierzchni znajduje się wiele wypustek i małych okien. Nuragi były budowane na wzniesieniach, często powstawały przy nich wioski, które korzystały z nuragów jako fortyfikacji obronnych.

Jedna z najlepiej zachowanych nuragów – Su Nuraxi – znajduje się w pobliżu miejscowości Barumini. Zachowany kompleks został wpisany w 1997 roku na listę światowego dziedzictwa UNESCO.

Bibliografia

Przypisy

Historia architektury starożytnej
Sardynia
Stanowiska archeologiczne we Włoszech


Tune in to watch the episode of "World's Greatest!..." Lumber Treater featuring a local Madison County Virginia business, Madison Wood, next week on national television! This episode will air on DirecTV Channel 305 and Dish Network on channel 250. Check out the website's television schedule page (http://worldsgreatesttelevision.com/tv_schedule) to see upcoming episode information and specific channel and network information. Please note, a third party is responsible for the listing of "World's Greatest!..." on interactive television guides. This third party does not always list "World's Greatest!..." by name. Thus, you may find "World's Greatest!..." is listed as "paid programming".
Madison has implemented it's newest program to train our employees on best practices for our yard. Today we learned about avoiding fork lift damage by understanding the lengths/widths of the different pack sizes. The guys were asked to guess how far the forks would stick out from under each unit. Here at Madison we know it is important for us to continue to help grow that next generation of leaders. We go into detail on topics like "choosing quality over quantity" so our guys understand how important each step of the treatment process is for our end users. Our promise is to produce the best stick of wood out there and we plan to do just that!
Good news! We can come to your yard and teach a #MADskillz training on best practices directly with your team. You can learn how to efficiently maintain your yard to avoid cull piles and waste from one of our reps at Madison Wood. We can even bring along a representative from SPIB (Southern Pine Inspection Bureau) to teach about the different grades that are available and common characteristics of treated lumber. Talk to your Account Manager today to schedule a #MADskillz training for the Fall!
Save time with our automated user-friendly approach to inventory management. MADTRAK accurately keeps records through barcode technology in real-time without the hassle and miscellaneous fees you may encounter with alternative programs. Our 12 digit barcode is uniquely designed to sync with current hardware inventory programs. Units can now be scanned in or out directly from the yard. Look for the MADTRAK barcode on the side of the white Madison Wood unit tags!
Tired of counting your inventory by hand?
Learn more about Madison Wood!


As lovers of sport, we all seem to have our favorite event. It might be The Super Bowl, Daytona 500, Masters, or The World Serious. If we're slightly crazy about sport, maybe it's more than one event. One of my favorite games to watch throughout the year is The Rose Bowl. Each New Year's Day you'll find me on the coach at 2:00 in the afternoon. One of the unique aspects of the game is when it begins. Being played in Southern California the game often begins under bright sunshine. Due to its afternoon start time, the Rose Bowl always concludes after dark under the lights. It always tickles me to watch highlights after the game. The first half is in bright sunlight and the second half in lit darkness. Our first playoff game was played in similar circumstances. We had a unique start time of 4:15, which meant we began with the sun still shining. By the time we ended, the lights had fully taken over. It turned into a great setting for our first playoff game.
Warming up our team was different in preparation for the playoff game. We began preparation for the game behind the end zone nearest the stadium entrance. During the first few minutes our kids where run through some stretching. About halfway through the stretches one of our coaches noticed we were in plain sight of our opponent. They were resting on the exterior of the fence, before entering the field to begin their warm-up. So, we took some extra time with stretching, slowing everything down. When our opponent for this playoff game entered the field, they walked down behind the opposite end zone to begin their warm-up process. After they became securely out of view, our preparation continued in full. Coach ran the kids through some new plays from the week of practice. Something new took place to end warm-ups, coach had our kids rest by cutting warm-ups short, having players sit and watch the game before us, conserving energy. Before long, it was time to play.
The playoff game saw my sideline role transition again. It had been a sunny day and was going to lead into a dry and cold night for football. Which meant we wouldn't have the need to dry off footballs. Normally without ball boy duty, my responsibility would be flashing a place card, helping rely plays in from the sideline. Playing a team for the second time required mixing up our sideline activities. In changing things up, I was given a different place card from the one in earlier games. It was meant to be a decoy for any onlookers. A couple coaches had different place cards, so no one could tell which place card actually pertained to the play. There was truly no way of knowing if the plan was necessary. Like anything else, it probably didn't go exactly how we had hoped. There were plays where mine forgot to be flashed, or too many coaches were gathered and my card couldn't be seen. It worked on a handful of plays though. The decoy role lead to a relaxing sideline experience for me. Probably the most fun game from my perspective, because there wasn't busy time pressure, it allowed me to take it all in.
Shadows crept across the field with the progression of our game. From the sideline, it almost felt like waiting in anticipation for the lights to take hold. We were involved in a competitive game for the first time in a while. Our opponent was moving the ball against our defense on the ground. Our linebackers were successfully getting into their backfield poised to make a tackle. But, those tackles were consistently being missed. Their running backs would slip through our players' arms, breaking away with runs to the outside. Our defense seemed to be allowing more yards than before. It took well into the second half before we found some true containment of their running game. However, on the offensive side of the ball, things seemed to go relatively well.
Our offense seemed to shine with the bright lights. It kept us in the game, while the defense sputtered. The running game worked well. We ran off pretty large chunks of yard during this game. With our offensive line creating good holes for running backs to run through. Our quarterback continued the growth that has steadily progressed throughout the year. There seemed to be unmistakable confidence in his body language. Watching our running backs during this game became exciting. At times, the frustration of our opposing defense was evident. It's been rare for me to view a run from behind this year. When one of our backs has broken a run to the outside along the sideline, it seems my position hasn't allowed for a good view. That wasn't the case during our first playoff game.
One of the best running plays left me viewing it from behind. As our back scampered down the near sideling, I watched in awe. It was probably our best athlete, as he broke off about a 30-yard run. Watching him work within a few yards of the boundary, as he slipped a couple tackles and juked past a defender. When the defender finally got a hold of him, our back was thrown to the ground. Frustration seemed to grip the tackler in that moment. But, then I witnessed something outstanding. His father is one of the coaches, he made a woo hoo exclamation, followed by encouraging his son to just get up. Our running back, who had just been unexplainably thrown to the ground, simply got up and jogged back to the huddle. He could have been visibly mad, but seemed to realize his play was doing the talking. His opponent had clearly reached a point of frustration following the run. Nothing more needed to be said.
The first playoff game was successful. We were in a competitive game for much of the time, but won going away. After the game, when both teams gathered on the field, one of our coaches talked about how rare it was to play football under the lights. It's true most of these kids won't experience high school football. This may be the only football game they ever play at night. If that is the case, hopefully they enjoyed the experience. It didn't seem to simply be a football game played at night. The game was played in the uniqueness of day transitioning into night. As the lights took over for long shadows, our team took hold of the game. They continued playing together and our defense began stopping the opponents running game. In the end, we cruised to another victory. As the clean crisp fall air became wintery cold, our undefeated season remained in tacked. One more week of practice before playing for a championship.


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Home » eSports » The 3 best League of Legends teams of 2020
The 3 best League of Legends teams of 2020
2020-12-28 eSports Comments Off on The 3 best League of Legends teams of 2020
Here are the best rosters to hit Summoner's Rift this year.
As we approach the end of the year, many people will be thankful since 2020 was one of the most turbulent years in recent times. Many industries across the world had to quickly adjust to the dangerous circumstances of the COVID-19 pandemic—and the League of Legends esports scene was no exception.
It was hard for both teams and talent since not many had experienced League on the big stage like this before. Whether it was all online or in the studio without a crowd to cheer them on, several players admitted that it felt different to play out the season in this manner.
And yet there were some rosters that persevered through the difficulties and uncertainty. Some teams adapted and became even better iterations of themselves all to win.
Here are the three best League teams of 2020.
DAMWON Gaming
It should come as no surprise that the 2020 world champions are featured on this list. After their performance in the second half of the year, DAMWON etched their name into the history books as one of the most dominant rosters we've seen in a long time.
This team featured superstars like Nuguri, Canyon, and ShowMaker, all players who are considered one of the best in the world in their respective roles. Ghost and BeryL also made up one of the more underrated bottom lane duos at Worlds since the former became one of the best weakside AD carries in the world while the latter helped take control of the map with aggressive, decisive roams.
Although the team finished in fifth place and had an early exit from the 2020 Spring Split playoffs, the 2020 Summer Split was a completely different story. In fact, from the start of the summer, the team only lost two games on the way to their first LCK championship.
DAMWON's road through Worlds was just as destructive, with the Korean first seed only losing one game in the group stage to JD Gaming. Afterward, they swept past DRX with ease and beat G2 Esports in four games. In the finals, they only dropped a single match against Suning on their way to the World Championship.
DAMWON lost Nuguri in the offseason, though, and the talented top laner was a huge part of the team's identity. But that specific group of players will always be able to look back at 2020 and think "this was the year we brought the Summoner's Cup back to Korea."
Although Suning didn't have a star-filled roster like DAMWON, the whole team had a storyline that could get any League fan to root for them during the World Championship.
Throughout the tournament, Suning were an underdog in most of the matchups they faced, especially in the knockout stage. Whether it was JDG, Top Esports, or DAMWON, many people thought that the LPL third seed would get eliminated.
And yet their lineup shocked the world over and over. During the 2020 Summer Split, Suning lost against both JDG and TES—and were even swept by the latter in the playoffs. But during Worlds, they bested both of them in a best-of-five setting by only losing one game to each.
Whether it was jungler SofM showing off his flashy Lee Sin, rising ADC Huanfeng carrying with Jhin and Ezreal, or their budding young star top laner Bin placing the team in his backpack to pull them to victory, Suning's roster quickly became popular among fans thanks to their exciting and unconventional play.
This roster persevered through all of their shortcomings, transforming themselves from an 11th-place team in the spring to a Worlds finalist. Even though they didn't take down DAMWON, their 2020 run is still something to be proud of.
Next year, Suning will have to adjust its lineup due to the departure of veteran support, SwordArt, who's made the trip stateside to join TSM. The talented 23-year-old was a leader for this young roster, but this past year may have given the rest of Suning enough experience to help them stay at a high level.
This was an interesting year for G2 fans since they didn't look as dominant as they did in 2019. The European juggernaut was still the best team in the West this past year, however.
With a powerful core comprised of some of the best players in the region, the team stumbled a surprising amount of times, including playoff losses to MAD Lions and Fnatic and a third-place finish during the 2020 Summer Split regular season.
Even though the roster seemed to be going through a little bit of a rough patch, they knew how to turn it on when it mattered during the playoffs. G2 eventually worked their way back to the LEC finals, where they swept Fnatic for back-to-back championships.
Their Worlds 2020 run started off great, with the team tying Suning for first place in Group A with a 4-2 record. They also beat Gen.G in a convincing sweep but were eventually taken down by DAMWON in four games.
Many EU fans thought G2 were going to finally get their revenge for their finals loss against FunPlus Phoenix the year before, which is why this result was relatively disappointing. This early exit, however, might be exactly what the team needed to push them to another level.
In the offseason, the team parted ways with star mid laner Perkz. It was a decision that shocked the world, but sometimes, even the best teams need some changes to continue growing. With superstar ADC and former Fnatic player Rekkles now on the team, G2 is ready to break records and make history once again in 2021.
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Presidents of the Pacific Alliance bring to a close the III Business Summit, attended by over 700 participants | Alianza del Pacífico.
The President of Chile, Michelle Bachelet, and the presidents of Colombia, Juan Manuel Santos; of Mexico, Enrique Peña Nieto; and of Peru, Ollanta Humala, today brought to a close the Third Business Summit of the Pacific Alliance (PA), "Vision into the Future" in Frutillar, Los Lagos region, Chile.
In her introductory remarks, President Bachelet stressed the importance of strengthening innovation in the PA and the role that investment in young people's education should have, in order to meet this challenge. In this regard she mentioned that Chile has developed policies to achieve a productive transformation.
For his part, President Santos highlighted the importance of the creation of the Latin American Integrated Market (MILA) as part of the efforts to broaden and deepen capital markets among the four countries. In that regard, he underlined the fact that, for the PA, to boost its digital integration is today a necessity.
In his speech, President Peña Nieto mentioned the priority that must be given to trade integration. He noted that it is necessary to publicize the value of this regional integration initiative, which to date has attracted the attention of 49 observer countries, as well as provided benefits for the people.
Meanwhile, President Humala emphasized the need to invest in education, noting that Peru has been promoting a firm policy to achieve a high-quality school education. He also indicated that education is a key point in the cooperation program of the Pacific Alliance.
Finally, the Presidents agreed that the creation of the Venture Capital Fund and the progress toward the internationalization of SMEs, will allow a better projection of the PA to other regions. Similarly, they stressed that the PA is one of the most attention-grabbing business initiatives in the world.
The Minister of Foreign Affairs of Chile and the President of the Business Council of the Pacific Alliance (CEAP), Chilean chapter, conducted the opening of the event.
Later, the Foreign Minister of Chile, the Minister of Foreign Trade, Industry and Tourism of Colombia, the Secretary of Economy of Mexico, and the Minister of Foreign Trade and Tourism of Peru participated in the first panel of the morning.
Throughout the day, interventions by the Presidents of Argentina, Mauricio Macri, and of Costa Rica, Luis Guillermo Solis, as well as by the President-elect of Peru, Pedro Pablo Kuczynski were made. Similarly, the ministers of finance of Chile, Colombia, Mexico and Peru participated in a panel on financial integration and its challenges.
In the afternoon, participants were able to attend four panels that were held in parallel on the business opportunities for SMEs, entrepreneurship and innovation, trade facilitation and financial integration.
The III Business Summit of the Pacific Alliance (PA) attracted over 700 attendees, including top executives of companies and SMEs, and took place days after the IV Business Matchmaking event held in Santiago, which generated nearly USD 200 million in business prospects.


Tosylamide/Formaldehyde Resin is a polymer formed from the reaction of toluenesulfonamide and formaldehyde. Tosylamide/Formaldehyde Resin is used in nail enamel products. It is a soft resin material which, in conjunction with nitrocellulose, forms tough, shiny and durable films on nails.
Tosylamide/Formaldehyde Resin is a very unique resin that modifies the widely used nitrocellulose type polymers to form tough and shiny films, which have excellent wear and durability characteristics while maintaining high gloss of nail enamel throughout the wear period.
Tosylamide/Formaldehyde Resin is a synthetic resin, sometimes called Toluenesulfonamide/Formaldehyde Resin. It modifies the film-forming characteristics of Nitrocellulose type polymers. Nitrocellulose is extremely hard and brittle. Addition of Tosylamide/Formaldehyde Resin makes the film less brittle, tougher, and the resin also increases the gloss of the film.
The Food and Drug Administration (FDA) includes Tosylamide/Formaldehyde Resin on its list of indirect food additives. It is permited for use polymers having contact with food. The safety of Tosylamide/Formaldehyde Resin has been assessed by the Cosmetic Ingredient Review (CIR) Expert Panel. The CIR Expert Panel evaluated the scientific data and concluded that Tosylamide/Formaldehyde Resin was safe as a cosmetic ingredient. In 2004, as part of the scheduled re-evaluation of ingredients, the CIR Expert Panel considered available new data on Toluenesulfonamide/Formaldehyde Resin and reaffirmed the above conclusion.
CIR Safety Review: The CIR Expert Panel noted that Tosylamide/Formaldehyde Resin was practically nontoxic when given orally as a 70% solution in acetone and when applied to the skin as a 40% suspension in corn oil. Tosylamide/Formaldehyde Resin in the form of a finely ground powder was nonirritating to skin and slightly irritating to the eye.
The undiluted material was nonirritating to skin, slightly irritating to the eye, and practically nontoxic by dermal application. No significant treatment-related effects were observed in a subchronic oral study. Tosylamide/Formaldehyde Resin was negative in a bacterial test for mutagenicity. In clinical studies, 10% Tosylamide/Formaldehyde Resin in dimethyl phthalate was nonirritating and nonsensitizing to the skin.
Nail products containing up to 12% Tosylamide/Formaldehyde Resin were nonsensitizing, nonphotosensitizing and essentially nonirritating to human skin. Reports of nail polish dermatitis of allergic origin have been attributed to Tosylamide/Formaldehyde Resin and other liquid thermoplastic resins.
Tosylamide/Formaldehyde Resin may be used in cosmetics and personal care products marketed in Europe according to the general provisions of the Cosmetics Regulation of the European Union.
Tosylamide/formaldehyde Resin , also referred to as Toluenesulfonamide/Formaldehyde Resin, is a reaction product of toluenesulfonamide and formaldehyde. It is an amorphous resin and is used in conjunction with the polymer nitrocellulose in nail enamel products.
Tosylamide/Formaldehyde resin functions as a film former and plasticizer . It imparts high gloss and good flow properties and increases the hardness of nitrocellulose film. The purity and residual formaldehyde content are tightly controlled in resins used for in nail products with maximum levels of 0.5% formaldehyde and with typical levels at less than 0.2%.


I have recently noticed I am introducing more people to each other. It's enjoyable and something that falls into my "pay it forward" bucket.
But, through this process I have also learned how to properly execute an introduction so both people get the most out of the experience. The last thing you want to do is burn a contact by flooding their inbox with unwanted introductions, placing yet another time commitment on their already full plate.
It takes a bit of social intelligence to best set up the introduction. Here are a few thoughts as you go about connecting people in your life.
If someone you know asks you to introduce them to another contact, first determine what they are looking for so you can determine if it's actually a good idea, and something you are willing to associate yourself with.
If not, graciously decline or let them know it doesn't seem like a good fit for what they are requesting. Maybe offer someone else you think might work better.
Then, touch base with the requested person first and brief them with the idea of introducing a contact to them. Answer their inevitable question of "what do I get out of it?" If they are favorable, ask them if they mind if you email intro them to this other person.
Do not just throw an unwanted email into their email box without first checking with them to see if they are willing to respond and follow through with meeting the person.
If you get the green light, send an email with both in the TO: field.
Give the email a subject such as "Jane <> Joe" so it's obvious it's an introduction.
Give a short description of each person and why they are looking to meet the other, make it brief but pointed and include why the two people will add value to each other. The person's ask should be front and center.
Suggest a coffee meeting, phone call or chat over a beer or something that sets the expectations of the interaction.
End it by saying "I'll let you two take it from here" so you can step aside and allow the two people to communicate without them feeling like they need to keep you CC'd or included in future emails.
Although basic stuff, if not executed correctly email intro's can do more harm than good. Take note!
Note: I am writing this because I have seen a few intro's botched in my day, and if not careful, a person can burn social capital by annoying people they are connected with. Also, when you are on the receiving end of an intro, acknowledge and thank the person doing the introduction and place them in BC, so they are them removed from future communications.
During my latest Founders RAW conversation I was talking with Patrick Henley, founder and CEO of Amp Tab when he mentioned something quite profound.
Connecting with people – truly connecting with someone – is the key to business success.
It was a great moment in the conversation (will post the conversation when it goes live) but I want to riff on the concept of connecting with people and why it can have a huge impact on your success.
Some may view networking as cheesy, uncomfortable and a waste of time. I don't. And neither did Patrick. Networking is not an activity, it's an purposeful energy. Patrick says connecting with people has been the difference maker for him and his success in sales and business in general. He pointed to how connecting with someone on a deeper level, figuring out how he could help them in their pursuits tightens the relationship between him and the other person. Once tightened, those people will break down walls to help you.
So how do you really connect with people?
Shaking their hand. Looking them in the eye when they are talking to you. Remembering their name. Making them feel like they are the most important person in that room at that specific moment. Asking what you can do to help them. Asking if you can introduce them to someone they are looking to meet. Following up with them via email the next day to book a meeting or re-confirm what was talked about. These things show the other person you have their best intentions in mind, rather then trying to get something out of them.
Remember, people care more about themselves than others… take advantage of that and let them know you too care about what's important to them.
All too often people go about their days and nights looking to get something out of others. This is a terrible mistake, just as it is for the guy at the bar who obviously is trying too hard to take any girl home.
Once people feel you have their best intentions in mind, they let their guard down and feel safe to further communicate using all the different means we have available. They will then be willing to add you on LinkedIn, twitter, Facebook and others.
Do that year in and year out and before you know it you have a large network of people who not only feel comfortable with you but would be there when you need a favor (and you know that time will come sooner than you think) and willing to bust down walls to see you succeed.
This is the killer app for founders, creating a network of people you are truly connected with on a deeper level than just a digital click on a social network. If more and more people can hear your name in public and think "man, that person is great and I would do whatever I can to see them succeed" you are golden.
It means you are truly wealthy in the relationships you have built. And since success in business is based on the relationships we have created you will have no other option than to continue on your path towards success.


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C-based co-development environment rolls for FPGAs
January 12, 2004 Embedded Staff
SANTA CRUZ, Calif. — Startup Impulse Accelerated Technologies Inc. this week will roll out its CoDeveloper product, which offers a hardware/software co-development environment for FPGAs with embedded processors. The tool lets designers develop C-language algorithms that are compiled into FPGA hardware or embedded software.
CoDeveloper uses the vendor's Impulse C libraries to support multiple, parallel processes and includes a hardware compiler that generates synthesizable VHDL for portions of the design destined for hardware. The tool supports FPGAs with Altera Corp.'s Nios and Xilinx Inc.'s MicroBlaze embedded processors. It also generates hardware/software interfaces.
Impulse Accelerated Technologies (Kirkland, Wash.) released its first product, the CoValidator VHDL simulator. The company's technology is partially derived from the open-source Streams-C compiler licensed from Los Alamos National Laboratory.
Platform-based FPGAs with embedded processors are gaining momentum, said David Pellerin, co-founder, president and chief technical officer of Impulse. But HDL-based tools often aren't the best way to support them, he said.
"These platforms are very compelling for the acceleration of software algorithms," Pellerin said. "But the typical software engineer does not want to use low-level hardware design methods to express software algorithms. And hardware engineers don't want to deal with hardware/software interfaces."
Thus, Pellerin said, CoDeveloper is aimed at both software and hardware engineers. Hardware engineers will probably hand-tweak the VHDL code that the tool generates, while software engineers will not, he said.
By allowing designers to develop C-language algorithms, CoDeveloper enables a "10 to 1" productivity gain compared with HDL design entry, Pellerin said, because designers don't need to think in terms of hardware concepts such as register boundaries or clock cycles.
But there are trade-offs, he acknowledged. "Certainly, the VHDL designer can do a better job at hand coding for efficiency," Pellerin said. "We are certainly larger in area, but in terms of performance we're very close. A lot depends on the quality of the C code."
The Impulse C libraries leverage the "streams-based programming" approach provided by the Streams-C compiler. They add library functions to standard C that let users express parallel applications. Specifically, they support multiple processes that are independently synchronized and that communicate though data streams.
Also included with the Impulse C libraries are platform support packages for the Nios and MicroBlaze processors that make it possible to compile C code with multiple processes directly to the processor.
Pellerin said Impulse decided against using SystemC in favor of the Impulse C library approach. "SystemC is a large, complex language for high-level systems design," he said. "It's appropriate for ASICs, but for the embedded-systems programmer who wants to use C, SystemC is far too complex."
Once an algorithm is created, designers can simulate it using such standard tools as Microsoft Corp.'s Visual Studio or Metrowerks' CodeWarrior. During simulation, CoDeveloper monitors Impulse C applications and identifies communications bottlenecks, such as overused stream buffers.
A user then determines which functions are implemented in hardware and which in software. "We really believe that giving the user control over that is very important," Pellerin said. Users tag processes aimed at hardware with function calls. Processes that are not tagged for hardware are fed into the Altera Nios or Xilinx MicroBlaze compilers.
The hardware compiler then generates synthesizable VHDL, which can be fed into Altera or Xilinx design environments, or into third-party synthesis tools. CoValidator is included as a built-in VHDL simulator. In the future, Pellerin said, Impulse will add Verilog support to its tool.
Users can purchase either CoDeveloper for Nios or CoDeveloper for MicroBlaze. The former requires Altera's Quartus II and SOPC Builder software, along with a reference board. The latter requires Xilinx's Platform Studio and a reference board.
"We've had excellent support from both Altera and Xilinx in our launch of this product," Pellerin said.
CoDeveloper for either Nios or MicroBlaze is available now for $4,995 for a permanent license or for $1,995 for a one-year license. A universal version, supporting Nios, MicroBlaze and other processors to be announced, costs $8,995 for a permanent license.
Information about Impulse Accelerated Technologies Inc.'s CoDeveloper product, along with free 30-day evaluation licenses, can be found at the Impulse Web site..
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