ABSTRACT

Clostridium saccharoperbutylacetonicum is a mesophilic, anaerobic, butanol-producing bacterium, originally isolated from soil. It was recently reported that C. saccharoperbutylacetonicum possesses multiple cellulosomal elements and would potentially form the smallest cellulosome known in nature. Its genome contains only eight dockerin-bearing enzymes, and its unique scaffoldin bears two cohesins (Cohs), three X2 modules, and two carbohydrate-binding modules (CBMs). In this study, all of the cellulosome-related modules were cloned, expressed, and purified. The recombinant cohesins, dockerins, and CBMs were tested for binding activity using enzyme-linked immunosorbent assay (ELISA)-based techniques. All the enzymes were tested for their comparative enzymatic activity on seven different cellulosic and hemicellulosic substrates, thus revealing four cellulases, a xylanase, a mannanase, a xyloglucanase, and a lichenase. All dockerin-containing enzymes interacted similarly with the second cohesin (Coh2) module, whereas Coh1 was more restricted in its interaction pattern. In addition, the polysaccharide-binding properties of the CBMs within the scaffoldin were examined by two complementary assays, affinity electrophoresis and affinity pulldown. The scaffoldin of C. saccharoperbutylacetonicum exhibited high affinity for cellulosic and hemicellulosic substrates, specifically to microcrystalline cellulose and xyloglucan. Evidence that supports substrate-dependent in vivo secretion of cellulosomes is presented. The results of our analyses contribute to a better understanding of simple cellulosome systems by identifying the key players in this minimalistic system and the binding pattern of its cohesin-dockerin interaction. The knowledge gained by our study will assist further exploration of similar minimalistic cellulosomes and will contribute to the significance of specific sets of defined cellulosomal enzymes in the degradation of cellulosic biomass.

IMPORTANCE Cellulosome-producing bacteria are considered among the most important bacteria in both mesophilic and thermophilic environments, owing to their capacity to deconstruct recalcitrant plant-derived polysaccharides (and notably cellulose) into soluble saccharides for subsequent processing. In many ecosystems, the cellulosome-producing bacteria are particularly effective "first responders." The massive amounts of sugars produced are potentially amenable in industrial settings to further fermentation by appropriate microbes to biofuels, notably ethanol and butanol. Among the solvent-producing bacteria, Clostridium saccharoperbutylacetonicum has the smallest cellulosome system known thus far. The importance of investigating the building blocks of such a small, multifunctional nanomachine is crucial to understanding the fundamental activities of this efficient enzymatic complex.

Sleep pressure and sleep depth are key regulators of wake and sleep. Current methods of measuring these parameters in Drosophila melanogaster have low temporal resolution and/or require disrupting sleep. Here we report analysis tools for high-resolution, noninvasive measurement of sleep pressure and depth from movement data. Probability of initiating activity,...

The capacity of Listeria monocytogenes to adapt to environmental changes is facilitated by a large number of regulatory proteins encoded by its genome. Among these proteins are the uncharacterized LysR-type transcriptional regulators (LTTRs). LTTRs can work as positive and/or negative transcription regulators at both local and global genetic levels. Previously, our group determined by comparative genome analysis that one member of the LTTRs (NCBI accession no. WP_003734782) was present in pathogenic strains but absent from nonpathogenic strains. The goal of the present study was to assess the importance of this transcription factor in the virulence of L. monocytogenes strain F2365 and to identify its regulons. An L. monocytogenes strain lacking lysR (the F2365lysR strain) displayed significant reductions in cell invasion of and adhesion to Caco-2 cells. In plaque assays, the deletion of lysR resulted in a 42.86% decrease in plaque number and a 13.48% decrease in average plaque size. Furthermore, the deletion of lysR also attenuated the virulence of L. monocytogenes in mice following oral and intraperitoneal inoculation. The analysis of transcriptomics revealed that the transcript levels of 139 genes were upregulated, while 113 genes were downregulated in the F2365lysR strain compared to levels in the wild-type bacteria. lysR-repressed genes included ABC transporters, important for starch and sucrose metabolism as well as glycerolipid metabolism, flagellar assembly, quorum sensing, and glycolysis/gluconeogenesis. Conversely, lysR activated the expression of genes related to fructose and mannose metabolism, cationic antimicrobial peptide (CAMP) resistance, and beta-lactam resistance. These data suggested that lysR contributed to L. monocytogenes virulence by broad impact on multiple pathways of gene expression.

IMPORTANCE Listeria monocytogenes is the causative agent of listeriosis, an infectious and fatal disease of animals and humans. In this study, we have shown that lysR contributes to Listeria pathogenesis and replication in cell lines. We also highlight the importance of lysR in regulating the transcription of genes involved in different pathways that might be essential for the growth and persistence of L. monocytogenes in the host or under nutrient limitation. Better understanding L. monocytogenes pathogenesis and the role of various virulence factors is necessary for further development of prevention and control strategies.

OBJECTIVE

To investigate factors related to glycemic management among members of a professional cycling team with type 1 diabetes over a 7-day Union Cycliste Internationale World Tour stage race.

RESEARCH DESIGN AND METHODS

An observational evaluation of possible factors related to glycemic management and performance in six male professional cyclists with type 1 diabetes (HbA_1c 6.4 ± 0.6%) during the 2019 Tour of California.

RESULTS

In-ride time spent in euglycemia (3.9–10.0 mmol/L glucose) was 63 ± 11%, with a low percentage of time spent in level 1 (3.0–3.9 mmol/L; 0 ± 1% of time) and level 2 (<3.0 mmol/L; 0 ± 0% of time) hypoglycemia over the 7-day race. Riders spent 25 ± 9% of time in level 1 (10.1–13.9 mmol/L) and 11 ± 9% in level 2 (>13.9 mmol/L) hyperglycemia during races. Bolus insulin use was uncommon during races, despite high carbohydrate intake (76 ± 23 g ⋅ h^–1). Overnight, the riders spent progressively more time in hypoglycemia from day 1 (6 ± 12% in level 1 and 0 ± 0% in level 2) to day 7 (12 ± 12% in level 1 and 2 ± 4% in level 2) (²[1] > 4.78, P < 0.05).

CONCLUSIONS

Professional cyclists with type 1 diabetes have excellent in-race glycemia, but significant hypoglycemia during recovery overnight, throughout a 7-day stage race.

Background:

Patients with multimorbidity often require services across different health care settings, yet team processes among settings are rarely implemented. We explored perceptions of specialists and family physicians collaborating in a telemedicine interprofessional consultation for patients with multimorbidity to better understand the value of bringing physicians together across the boundaries of health care settings.

Methods:

This was a descriptive qualitative, interview-based study. Physicians who had previously participated in the Telemedicine Interprofessional Model of Practice for Aging and Complex Treatments (Telemedicine IMPACT Plus [TIP] Program) were invited to participate and asked to describe their experience of being a member of the program. Interviews were conducted from March to May 2016. We conducted an iterative and interpretive process using both individual and team analysis to identify themes.

Results:

There were 15 participants, 9 specialists and 6 family physicians. Three themes emerged in the analysis: creating new perspectives on care for patients with multimorbidity by sharing knowledge, skills and attitudes; the shift from a consultant model to an interprofessional team model (allowing a window into the community, extending discussions beyond the medical model and focusing on the patient’s health in context); and opportunities for learners, including learning about interprofessional collaboration and gaining exposure to a real-world model for caring for people with multimorbidity in outpatient settings.

Interpretation:

Family physicians and specialists participating in a TIP Program believed the program improved their knowledge and skills, while also serving as an effective care delivery strategy. The findings also support that learners require more exposure to nontraditional consultant models in order to care for patients with multimorbidity effectively.

The RESTORE-IMI 1 phase 3 trial demonstrated the efficacy and safety of imipenem-cilastatin (IMI) combined with relebactam (REL) for treating imipenem-nonsusceptible infections. The objective of this analysis was to compare the outcomes among patients meeting eligibility requirements based on central laboratory susceptibility versus local laboratory susceptibility. Patients with serious infections caused by imipenem-nonsusceptible, colistin-susceptible, and imipenem-REL-susceptible pathogens were randomized 2:1 to IMI-REL plus placebo or colistin plus IMI for 5 to 21 days. The primary endpoint was a favorable overall response. Key endpoints included the clinical response and all-cause mortality. We compared outcomes between the primary microbiological modified intent-to-treat (mMITT) population, where eligibility was based on central laboratory susceptibility testing, and the supplemental mMITT (SmMITT) population, where eligibility was based on local, site-level testing. The SmMITT (n = 41) and MITT (n = 31) populations had similar baseline characteristics, including sex, age, illness severity, and renal function. In both analysis populations, favorable overall response rates in the IMI-REL treatment group were >70%. Favorable clinical response rates at day 28 were 71.4% for IMI-REL and 40.0% for colistin plus IMI in the mMITT population, whereas they were 75.0% for IMI-REL and 53.8% for colistin plus IMI in the SmMITT population. Day 28 all-cause mortality rates were 9.5% for IMI-REL and 30.0% for colistin plus IMI in the mMITT population, whereas they were 10.7% for IMI-REL and 23.1% for colistin plus IMI in the SmMITT population. The outcomes in the SmMITT population were generally consistent with those in the mMITT population, suggesting that outcomes may be applicable to the real-world use of IMI-REL for treating infections caused by imipenem-nonsusceptible Gram-negative pathogens. (This study has been registered at ClinicalTrials.gov under identifier NCT02452047.)

When you view a property to rent, it pays to be as organised as possible. Our rental viewing checklist below should provide you with basic questions to ask and things to look out for at the property. Whether you are viewing a house or a flat, it could be your home for the foreseeable future, so be prepared and make a rational assessment of the property. staircaseAlthough you're going to be renting a house or a flat rather than buying it, remember that it will be your home for the foreseeable future. Of course you'll have an instant emotional reaction to any property, but you should balance this with a rational assessment of the potential of living in the property, including the local area and amenities, the condition of the property both inside and out and the costs that come with living in it. Obviously, your own priorities will vary depending on whether you're viewing houses, flats, studio flats or just rooms, but the checklist should act as a basic guide to help you keep all of your bases covered.

The decision to list your home for resale is a big one. No doubt it came about after much thought, weighing pros and cons, and series of long discussions. However, once you decide to put your home on the market, get ready for the next big discussion: interior design.

According to a study done by Professor Paul Anglin from the University of Guelph in Ontario, there are certain words used in home listings that have a magical effect on a home for sale. These are the most powerful words in a home for sale listing that help the owner sell quickly and/or get a higher asking price.

Writing a home for sale listing is not an easy task, especially for those who are doing it for the first time, but if you can manage to avoid the worst words in a home for sale listing, rest assured that you will be one step closer to selling your home in a timely manner.

Every home seller wants to sell it fast and reach as many buyers as possible. Here are some tips to make your listing stand out to potential buyers from the crowd.

If you make a mistake during the listing process, you might end up getting less than your home is worth or find that it takes significantly longer to sell your home than it should.

Every seller wants his homes to be sold smoothly and quickly with the highest possible price. However, there are still houses which have been listed on the market for months without any potential buyer because the sellers make one, or maybe more, of the following popular mistakes:

#GuildChat returns this Friday, April 24th at 11am PT / 2pm ET. Our topic is Listening to the Business. Right now there is a lot […]

The post #GuildChat for 04/24/20: Listening to the Business appeared first on e-Learning Feeds.

Fancy a 1v1 match on Shipment? Of course not. Duels to the death are played out and boring. Subscribing to the view that bigger is indeed always better, Call Of Duty: Modern Warfare has brought back its 10v10 Shipment playlist. After briefly appearing as an April Fool’s jab, Infinity Ward have decided to make 20-player […]

I'm a huge podcast tragic. From true crime to weird radio plays, I like shoving as much of them into my ear holes as possible. But despite my efforts, I realised that I don't listen to as many Australian podcasts as I'd like. Sure, I have a few favourites, but I could be doing more to support local creators. So I asked around the office to find out what the best Aussie podcasts are, and why I (and you) should be listening to them. More »

The Pulitzer Prize Board this week awarded its commentary award to The New York Times' Nikole Hannah-Jones for her essay launching the "1619 Project." This will accelerate a trend already underway: subjecting schoolchildren to a curriculum that blames slavery on capitalism and whose creator believes socialism offers the best path to racial equity.

Long before NLP became a hot field in AI, people devised rules and machines to manipulate language

This is a video from Russia's Zapad-2017 military exercises where a KA-52 Alligator helicopter accidentally fired two missiles towards a group of journalists covering the event (allegedly as a result of a short-circuit in the helicopter's electric system according to the Russian military *eyebarrelroll*). Three of the journalists were injured in the incident (amazingly not critically either), including the one seen strolling along the highway to the danger zone the video. Man, those missiles come QUICK. If you think you're gonna outrun one of those things like a tarantula in Animal Crossing, think again. I mean sure The Flash could do it, but he could also pleasure himself infinitely in the time it took you to read this article. "Huh?" How his penis doesn't catch fire is beyond me, there must be other superpowers at play too. Keep going for a couple versions of the video (the second with slow-mos), as well as a news report which includes the copter's gun view.

Failed to get friends list and several more Pokemon GO friends list errors are once again impacting players on iOS and Android.

Touhou hits the western hemisphere. Yo-kai watches will soon compete with Apple watches. Monster Hunting gets more cute. And Square Enix announces that they might...

While Chris destroys Anna Marie's right joycon (press R to pay respects), the rest of the cast discusses their now playing, the news of the week, and there are still far too many games releasing every week. Please save us.

"Fortnite" Party Royale had another big show on May 8. Here's everything you need to know about the Dillion Francis, Steve Aoki and deadmau5 concert that just took place.

Contagion was a film released in 2011 about a fictional pandemic of a virus called MEV-1 which kills between 25 and 30 per cent of those it infected. Here is our review of the film originally published in September 2011, now that it's on Netflix

Use our interactive checklist to help fill up your museum with all the dinosaur fossils in Animal Crossing: New Horizons.

Thousands of cyclists took over streets in the centre of the Slovenian capital Ljubljana on Friday evening to protest against the government of Prime Minister Janez Jansa and the restrictions it has imposed to fight the coronavirus.

Couple bought the property months before the former Beatle's 1980 assassination.

Economic reconstruction is a chance to speed up decarbonisation, and the pandemic has shown a different kind of politics is possible

We’re already being swamped with ideas about “reforms” needed to recover from the pandemic crisis. But the word reform is like gift wrap – a handy cover for any offering, thought-through or otherwise.

Perhaps we should ditch the word entirely, and with it the forest of feelpinions about what governments “must” do to advance an author’s previously-held ideological positioning in the post-corona world.

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Former Los Angeles Lakers player Shannon Brown was arrested last week for allegedly firing a gun at two people who entered his Georgia home that was listed for sale.

A cyclist has died after being involved in a collision with a van.

Read our live updates on coronavirus HERE Coronavirus: The symptoms

A motorcyclist has died after a crash on a busy road in north-east London.

A 35-year-old motorcyclist has died after a crash in east London.

The National Union of Journalists gathered for a special virtual tribute

Donate at virginmoneygiving.com/fund/FoodforLondonNOW

A motorcyclist has died after colliding with a lamppost in east London.

Police in Barnsley have launched an investigation after a three-year-old girl had spit wiped on her face by a teenage cyclist.

A police motorcyclist has died after a collision with a car while responding to an incident in Sheffield, South Yorkshire Police said.

If the pandemic lasts another three months, news brands could suffer a £50 million loss

A man has died after his motorbike crashed into the central reservation of a major London road.

Whirlpool has announced they will recall thousands more washing machines which could risk causing fires in people's homes.

A new survey suggests people think the behaviour of drivers and cyclists has deteriorated since the start of the coronavirus pandemic.

The Supreme Court is now holding hearings over a teleconference, making the audio publicly available. Jerry Goldman, the founder of the Oyez Project, offers some guidance on how to listen to it.

The author of The Miniaturist on the beguiling blend of tradition and modernity (and pancakes) in a city that provided the inspiration for her 17th-century-set debut novel and Waterstones Book of the Year 2014

• Top 10 free things to do in Amsterdam
  
• A new Amsterdam – and a new Rijksmuseum

Amsterdam is classically romantic but is also funky, forward-thinking and citizen-friendly. In the old centre, around the southern canal belt, there are these beautiful 17th-century merchants’ houses that 21st-century Amsterdammers still live in. I’ve always thought it wears its historical cloak quite casually and doesn’t just dwell in the past.

The Rijksmuseum is stunning and I love it as a fascinating, cool, accessible museum, as well as for the part it played in inspiring The Miniaturist. I came across Petronella Oortman’s doll’s house there by chance. It’s an exact scale replica of her real home, and Oortman spent a fortune having it created. I thought at the time it was an interesting story, but I didn’t think I was going to write a novel about it. I’m in its debt, really.

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