The Protectors (1972) Synopsis The Protectors (1972) was an action thriller television series produced by ITV that premiered in September of 1972. The Protectors (1972) was a British action thriller series created by Gerry Anderson (Thunderbirds) about an international group of rich troubleshooters. Contessa di Contini (played by Nyree Dawn Porter), Paul Buchet (played by […]

Wormwood Synopsis Wormwood was a children’s television series produced by Network Ten and ABC1 that premiered in October of 2007. Made as a joint Canadian-Australian production, Wormwood was a TV series about a quirky town where a group of kids investigated strange events. The series was based on the stories written by Paul Jennings. Wormwood […]

A Sharp Intake Of Breath Synopsis A Sharp Intake Of Breath was a situation comedy television series produced by ITV that premiered in February of 1978. Created by Ronnie Taylor A Sharp Intake Of Breath followed the story of Peter Barnes (played by David Jason – Only Fools And Horses), an ordinary man who made […]

Joshua Summers, CEO of linear television supply-side platform clypd, discusses the major advanced targeting trends expected to infiltrate traditional TV ad buying within the next year.

Pandemic gives TV viewership boost, but will be short-lived   April 28, 2020 (New York, NY) – As stay-at-home orders remain in effect due to COVID-19, TV viewership and time […]

From better campsite cooking to car maintenance to smartphone-free communication, here are 12 gadgets to up your car camping game.

The post Up Your Car Camping Game with These 12 Clever Gadgets appeared first on Vagabondish.

It's not the brightest headlamp on the market, but it's more than adequate for cooking, reading, or finding a pee tree in the dark. Plus, it's comfortable, easy-to-use, and USB rechargeable.

The post BioLite HeadLamp 200 Is the Featherweight Head Lamp You’ll (Probably) Forget You’re Wearing appeared first on Vagabondish.

People are always curious about work-from-home (WFH), remote working setups. So, I posted a Basecamp message asking our employees to share a photo of their home office, desk, table, whatever. Here’s what came in. First, the ask: And the answers, in the order they came in: Andy Didorosi, Marketing Justin White, Programmer Jonas Downey, Designer… keep reading

David and I spent nearly 2-hours giving a livestream tour of our very own Basecamp account. We wanted to show you how Basecamp uses Basecamp to run projects, communicate internally, share announcements, know what everyone’s working on, build software, keep up socially, and a whole bunch more. Our entire company runs on Basecamp, and this… keep reading

In this livesteam, David and I answer audience questions about how to work remotely. At Basecamp we’ve been working remotely for nearly 20 years, so we have a lot of experience to share. This nearly 2-hour video goes into great detail on a wide variety of topics. Highly recommended if you’re trying to figure out… keep reading

We’ve been teaching people how to do remote work well for the better part of two decades. We wrote a whole book about the topic in 2013, called REMOTE: Office Not Required. Basecamp has been a remote company since day one, and our software is sold as an all-in-one toolkit for remote work. Yeah, we’re… keep reading

Video: Merlin Mann - "Time & Attention Talk (improvised)"

Audio (mp3): "Merlin Mann - 'Rutgers Time & Attention Talk'"

This is a talk I did at Rutgers earlier this month. I kinda like it, but for a weird reason. Something something, perfect storm of technology Ragnarok, and yadda yadda, I had to start the talk 20 minutes late with no slides. Nothing.

So, I riffed.

And, I ended up talking about a lot of the new stuff you can expect to see in the Inbox Zero book—work culture, managing expectations, the 3 deadly qualities of email, and one surprising reason email's not as much fun as Project Runway.

Some people liked it. I think. I liked it. I hope you do, too.

Here's the slides I would have shown. ;-)

Many thanks, again, to my great pal, Dr. Donald Schaffner, for bringing me in for this visit. I had a great time and met some fantastic, passionate people. Much appreciated.

update 2010-04-27_13-50-00

Apologies—my friends at Rutgers (inexplicably) have placed this video under lock and key. Fortunately, I have a lock-picker called Firefox. Samizdat video available soon...

update 2010-04-27_14-42-24

Yay, fixed! Many thanks to my hero, Jesse Schibilia.

With the eviction date looming for campers to leave Oppenheimer Park, B.C.'s social development minister says everyone is still expected to be out of the park by Saturday.

The following article, BREAKING: President Trump’s Fiery Interview On Fox & Friends…”These are dirty politicians and dirty cops…They put our nation in danger with other nations, including Russia” [VIDEO], was first published on 100PercentFedUp.com.

This morning during a nearly one hour interview with Fox & Friends, President Trump addressed the decision by the DOJ to drop the case against the innocent General Michael Flynn. Trump ripped into the “dirty politicians and dirty cops” who went after General Michael Flynn. President Trump called the players involved in the horrible plot […]

Continue reading: BREAKING: President Trump’s Fiery Interview On Fox & Friends…”These are dirty politicians and dirty cops…They put our nation in danger with other nations, including Russia” [VIDEO] ...

Assembled α-synuclein in nerve cells and glial cells is the defining pathological feature of neurodegenerative diseases called synucleinopathies. Seeds of α-synuclein can induce the assembly of monomeric protein. Here, we used sucrose gradient centrifugation and transiently transfected HEK 293T cells to identify the species of α-synuclein from the brains of homozygous, symptomatic mice transgenic for human mutant A53T α-synuclein (line M83) that seed aggregation. The most potent fractions contained Sarkosyl-insoluble assemblies enriched in filaments. We also analyzed six cases of idiopathic Parkinson's disease (PD), one case of familial PD, and six cases of multiple system atrophy (MSA) for their ability to induce α-synuclein aggregation. The MSA samples were more potent than those of idiopathic PD in seeding aggregation. We found that following sucrose gradient centrifugation, the most seed-competent fractions from PD and MSA brains are those that contain Sarkosyl-insoluble α-synuclein. The fractions differed between PD and MSA, consistent with the presence of distinct conformers of assembled α-synuclein in these different samples. We conclude that α-synuclein filaments are the main driving force for amplification and propagation of pathology in synucleinopathies.

Heat shock factor 1 (HSF1) regulates cellular adaptation to challenges such as heat shock and oxidative and proteotoxic stresses. We have recently reported a previously unappreciated role for HSF1 in the regulation of energy metabolism in fat tissues; however, whether HSF1 is differentially expressed in adipose depots and how its levels are regulated in fat tissues remain unclear. Here, we show that HSF1 levels are higher in brown and subcutaneous fat tissues than in those in the visceral depot and that HSF1 is more abundant in differentiated, thermogenic adipocytes. Gene expression experiments indicated that HSF1 is transcriptionally regulated in fat by agents that modulate cAMP levels, by cold exposure, and by pharmacological stimulation of β-adrenergic signaling. An in silico promoter analysis helped identify a putative response element for activating transcription factor 3 (ATF3) at −258 to −250 base pairs from the HSF1 transcriptional start site, and electrophoretic mobility shift and ChIP assays confirmed ATF3 binding to this sequence. Furthermore, functional assays disclosed that ATF3 is necessary and sufficient for HSF1 regulation. Detailed gene expression analysis revealed that ATF3 is one of the most highly induced ATFs in thermogenic tissues of mice exposed to cold temperatures or treated with the β-adrenergic receptor agonist CL316,243 and that its expression is induced by modulators of cAMP levels in isolated adipocytes. To the best of our knowledge, our results show for the first time that HSF1 is transcriptionally controlled by ATF3 in response to classic stimuli that promote heat generation in thermogenic tissues.

The CRISPR/Cas9 nucleases have been widely applied for genome editing in various organisms. Cas9 nucleases complexed with a guide RNA (Cas9–gRNA) find their targets by scanning and interrogating the genomic DNA for sequences complementary to the gRNA. Recognition of the DNA target sequence requires a short protospacer adjacent motif (PAM) located outside this sequence. Given that the efficiency of target location may depend on the strength of interactions that promote target recognition, here we sought to compare affinities of different Cas9 nucleases for their cognate PAM sequences. To this end, we measured affinities of Cas9 nucleases from Streptococcus pyogenes, Staphylococcus aureus, and Francisella novicida complexed with guide RNAs (gRNAs) (SpCas9–gRNA, SaCas9–gRNA, and FnCas9–gRNA, respectively) and of three engineered SpCas9–gRNA variants with altered PAM specificities for short, PAM-containing DNA probes. We used a “beacon” assay that measures the relative affinities of DNA probes by determining their ability to competitively affect the rate of Cas9–gRNA binding to fluorescently labeled target DNA derivatives called “Cas9 beacons.” We observed significant differences in the affinities for cognate PAM sequences among the studied Cas9 enzymes. The relative affinities of SpCas9–gRNA and its engineered variants for canonical and suboptimal PAMs correlated with previous findings on the efficiency of these PAM sequences in genome editing. These findings suggest that high affinity of a Cas9 nuclease for its cognate PAM promotes higher genome-editing efficiency.

This article is by L. James Rice.

Campfire Pro is a program that aims to be a comprehensive tool for worldbuilders, ranging from authors to game designers to TTRPG Gamemasters, which means it’s got many features to explore. With that in mind, it’s probably best to give a little overview of the product before delving into its details.

First off, between gaming, screenwriting, and writing in general, I’ve seen a lot of these sorts of programs over the years and been disappointed by them more often than not. Campfire’s user interface harkens back to many older programs, keeping things relatively simple, and if you’re looking flashy bells and whistles, they aren’t here. You can, however, beautify the background with a variety of themes as well as create your own. Where this program excels is in its most important aspect, functionality, while for me at least, the biggest downfall is a less than intuitive interface. This could just be my brain, results will vary, but don’t be surprised by a tiny learning curve and the occasional “good grief, did I really just do that?” This is more about little irritants than deal breakers, however, and a little tinkering tends to find answers.

Continue reading Review of Campfire Pro Writing Software at Mythic Scribes.

JP Segrest
Feb 1, 1992; 33:141-166
Reviews

JE Hixson
Mar 1, 1990; 31:545-548
Articles

Assembled α-synuclein in nerve cells and glial cells is the defining pathological feature of neurodegenerative diseases called synucleinopathies. Seeds of α-synuclein can induce the assembly of monomeric protein. Here, we used sucrose gradient centrifugation and transiently transfected HEK 293T cells to identify the species of α-synuclein from the brains of homozygous, symptomatic mice transgenic for human mutant A53T α-synuclein (line M83) that seed aggregation. The most potent fractions contained Sarkosyl-insoluble assemblies enriched in filaments. We also analyzed six cases of idiopathic Parkinson's disease (PD), one case of familial PD, and six cases of multiple system atrophy (MSA) for their ability to induce α-synuclein aggregation. The MSA samples were more potent than those of idiopathic PD in seeding aggregation. We found that following sucrose gradient centrifugation, the most seed-competent fractions from PD and MSA brains are those that contain Sarkosyl-insoluble α-synuclein. The fractions differed between PD and MSA, consistent with the presence of distinct conformers of assembled α-synuclein in these different samples. We conclude that α-synuclein filaments are the main driving force for amplification and propagation of pathology in synucleinopathies.

Mass spectrometry (MS)-based proteomics has great potential for overcoming the limitations of antibody-based immunoassays for antibody-independent, comprehensive, and quantitative proteomic analysis of single cells. Indeed, recent advances in nanoscale sample preparation have enabled effective processing of single cells. In particular, the concept of using boosting/carrier channels in isobaric labeling to increase the sensitivity in MS detection has also been increasingly used for quantitative proteomic analysis of small-sized samples including single cells. However, the full potential of such boosting/carrier approaches has not been significantly explored, nor has the resulting quantitation quality been carefully evaluated. Herein, we have further evaluated and optimized our recent boosting to amplify signal with isobaric labeling (BASIL) approach, originally developed for quantifying phosphorylation in small number of cells, for highly effective analysis of proteins in single cells. This improved BASIL (iBASIL) approach enables reliable quantitative single-cell proteomics analysis with greater proteome coverage by carefully controlling the boosting-to-sample ratio (e.g. in general <100x) and optimizing MS automatic gain control (AGC) and ion injection time settings in MS/MS analysis (e.g. 5E5 and 300 ms, respectively, which is significantly higher than that used in typical bulk analysis). By coupling with a nanodroplet-based single cell preparation (nanoPOTS) platform, iBASIL enabled identification of ~2500 proteins and precise quantification of ~1500 proteins in the analysis of 104 FACS-isolated single cells, with the resulting protein profiles robustly clustering the cells from three different acute myeloid leukemia cell lines. This study highlights the importance of carefully evaluating and optimizing the boosting ratios and MS data acquisition conditions for achieving robust, comprehensive proteomic analysis of single cells.

The CRISPR/Cas9 nucleases have been widely applied for genome editing in various organisms. Cas9 nucleases complexed with a guide RNA (Cas9–gRNA) find their targets by scanning and interrogating the genomic DNA for sequences complementary to the gRNA. Recognition of the DNA target sequence requires a short protospacer adjacent motif (PAM) located outside this sequence. Given that the efficiency of target location may depend on the strength of interactions that promote target recognition, here we sought to compare affinities of different Cas9 nucleases for their cognate PAM sequences. To this end, we measured affinities of Cas9 nucleases from Streptococcus pyogenes, Staphylococcus aureus, and Francisella novicida complexed with guide RNAs (gRNAs) (SpCas9–gRNA, SaCas9–gRNA, and FnCas9–gRNA, respectively) and of three engineered SpCas9–gRNA variants with altered PAM specificities for short, PAM-containing DNA probes. We used a “beacon” assay that measures the relative affinities of DNA probes by determining their ability to competitively affect the rate of Cas9–gRNA binding to fluorescently labeled target DNA derivatives called “Cas9 beacons.” We observed significant differences in the affinities for cognate PAM sequences among the studied Cas9 enzymes. The relative affinities of SpCas9–gRNA and its engineered variants for canonical and suboptimal PAMs correlated with previous findings on the efficiency of these PAM sequences in genome editing. These findings suggest that high affinity of a Cas9 nuclease for its cognate PAM promotes higher genome-editing efficiency.

• The Hobbit Review
• Christmas Movies
• What We Watched: Sunshine/Silver Lining's Playbook/The Grey/Argo

• Zero Dark Thirty Review
• Our Top 10 Most Anticipated Films of 2013
• What We Watched: Jiro: Dreams of Sushi, Take This Waltz, Futurama, The Great Happiness Space, Indie Game: The Movie, Somewhere Between, Lincoln & Chasing Ghosts

• The Hunger Games: Catching Fire review
• The Wolverine Reviews
• What We Watched: jOBS, Steve Jobs: The Lost Interview, WWE For All Mankind: Life and Career of Mick Foley and The Lakota 38

ssm euromicron GmbH, a system house subsidiary of euromicron AG, is involved in a project to provide the technical equipment for the new building for the Institute of Geosciences at Kiel’s Christian Albrechts University. The seven-story new building is part of a campus-wide modernization initiative that is one of the largest public high-rise projects in the federal state of Schleswig-Holstein.

Hydel, despite trailing their rivals for two straight days, easily retained their title

The world's two largest economies are both partners and rivals, deeply intertwined but also with divergent interests. How will these contradictions resolve themselves following COVID-19?