Yadong Yu
Dec 1, 2020; 19:1997-2014
Research

Diana Samodova
Dec 1, 2020; 19:2139-2156
Technological Innovation and Resources

Post-transcriptional modifications of pre-mRNAs expand the diversity of proteomes in higher eukaryotes. In the brain, these modifications diversify the functional output of many critical neuronal signal molecules. In this study, we identified a brain-specific A-to-I RNA editing that changed glutamine to arginine (Q/R) at exon 20 and an alternative splicing of exon 4 in Tmem63b, which encodes a ubiquitously expressed osmosensitive cation channel. The channel isoforms lacking exon 4 occurred in ∼80% of Tmem63b mRNAs in the brain but were not detected in other tissues, suggesting a brain-specific splicing. We found that the Q/R editing was catalyzed by Adar2 (Adarb1) and required an editing site complementary sequence located in the proximal 5' end of intron 20. Moreover, the Q/R editing was almost exclusively identified in the splicing isoform lacking exon 4, indicating a coupling between the editing and the splicing. Elimination of the Q/R editing in brain-specific Adar2 knockout mice did not affect the splicing efficiency of exon 4. Furthermore, transfection with the splicing isoform containing exon 4 suppressed the Q/R editing in primary cultured cerebellar granule neurons. Thus, our study revealed a coupling between an RNA editing and a distant alternative splicing in the Tmem63b pre-mRNA, in which the splicing plays a dominant role. Finally, physiological analysis showed that the splicing and the editing coordinately regulate Ca2+ permeability and osmosensitivity of channel proteins, which may contribute to their functions in the brain.

Trypsin is the protease of choice in bottom-up proteomics. However, its application can be limited by the amino acid composition of target proteins and the pH of the digestion solution. In this study we characterize ProAlanase, a protease from the fungus Aspergillus niger that cleaves primarily on the C-terminal side of proline and alanine residues. ProAlanase achieves high proteolytic activity and specificity when digestion is carried out at acidic pH (1.5) for relatively short (2 h) time periods. To elucidate the potential of ProAlanase in proteomics applications, we conducted a series of investigations comprising comparative multi-enzymatic profiling of a human cell line proteome, histone PTM analysis, ancient bone protein identification, phosphosite mapping and de novo sequencing of a proline-rich protein and disulfide bond mapping in mAb. The results demonstrate that ProAlanase is highly suitable for proteomics analysis of the arginine- and lysine-rich histones, enabling high sequence coverage of multiple histone family members. It also facilitates an efficient digestion of bone collagen thanks to the cleavage at the C terminus of hydroxyproline which is highly prevalent in collagen. This allows to identify complementary proteins in ProAlanase- and trypsin-digested ancient bone samples, as well as to increase sequence coverage of noncollagenous proteins. Moreover, digestion with ProAlanase improves protein sequence coverage and phosphosite localization for the proline-rich protein Notch3 intracellular domain (N3ICD). Furthermore, we achieve a nearly complete coverage of N3ICD protein by de novo sequencing using the combination of ProAlanase and tryptic peptides. Finally, we demonstrate that ProAlanase is efficient in disulfide bond mapping, showing high coverage of disulfide-containing regions in a nonreduced mAb.

AAA+ ATPases constitute a large family of proteins that are involved in a plethora of cellular processes including DNA disassembly, protein degradation and protein complex disassembly. They typically form a hexametric ring-shaped structure with six subunits in a (pseudo) 6-fold symmetry. In a subset of AAA+ ATPases that facilitate protein unfolding and degradation, six subunits cooperate to translocate protein substrates through a central pore in the ring. The number and type of nucleotides in an AAA+ ATPase hexamer is inherently linked to the mechanism that underlies cooperation among subunits and couples ATP hydrolysis with substrate translocation. We conducted a native MS study of a monodispersed form of PAN, an archaeal proteasome AAA+ ATPase, to determine the number of nucleotides bound to each hexamer of the WT protein. We utilized ADP and its analogs (TNP-ADP and mant-ADP), and a nonhydrolyzable ATP analog (AMP-PNP) to study nucleotide site occupancy within the PAN hexamer in ADP- and ATP-binding states, respectively. Throughout all experiments we used a Walker A mutant (PAN^K217A) that is impaired in nucleotide binding as an internal standard to mitigate the effects of residual solvation on mass measurement accuracy and to serve as a reference protein to control for nonspecific nucleotide binding. This approach led to the unambiguous finding that a WT PAN hexamer carried – from expression host – six tightly bound ADP molecules that could be exchanged for ADP and ATP analogs. Although the Walker A mutant did not bind ADP analogs, it did bind AMP-PNP, albeit at multiple stoichiometries. We observed variable levels of hexamer dissociation and an appearance of multimeric species with the over-charged molecular ion distributions across repeated experiments. We posit that these phenomena originated during ESI process at the final stages of ESI droplet evolution.

Political Opposition and Policy Alternatives in Zambia 31 October 2017 — 10:30AM TO 11:30AM Anonymous (not verified) 19 October 2017 Chatham House, London

In Zambia’s 2016 national election, the Patriotic Front (PF) was re-elected by a narrow margin. The PF’s Edgar Lungu secured 50.35 per cent of the vote according to the Electoral Commission of Zambia, narrowly avoiding a second round, while his main rival, Hakainde Hichilema, won 47.67 per cent. The UPND, led by Mr Hichilema, alleges electoral fraud and has challenged the result in the courts and through direct protests. Mr Hichilema was imprisoned for 100 days.

At this meeting, Hakainde Hichilema will discuss his UPND priorities, how to strengthen opposition parties and their role in Zambia’s democratic future.

Read transcript

North Dakota is the latest state to make a push for integrating Native American or other ethnic studies into school curricula.

Although Native American students make up a small percentage of Wyoming's student population, they are suspended more than their non-minority peers.

Freshman had a season-high 13 points off bench while contributing to Spartans' win over Eastern Michigan.

The president atoned for the federal government's role in forcing Native American children into boarding schools, where many were abused and more than 900 died

The National Hockey League announced earlier this week that Jake Allen, born and raised in Fredericton, has become the first goalie in NHL history to record a win against 33 franchises. In an interview, the 34-year-old said he was unaware he was breaking the record but called it "pretty neat."

Knee replacement doesn’t have to be ‘total’

More than 100,000 AutoCAD software users download free compatibility solution from SolidWorks Corporation

New Solar Thermal Power Plant Developed in SolidWorks 3D CAD Software Goes Online Near L.A.

Penn State Berks will welcome Piscataway Nation singers and dancers to honor and celebrate Indigenous and Native American Heritage Month. The event is free and open to the public and will take place on Monday, Nov. 4, at 12:15 p.m. in the Perkins Student Center Auditorium.

Roebuck will discuss his past and future projects, many of which were filmed in the Lehigh Valley.

The Delaware Department of Agriculture (DDA) is offering extensions and COVID credit exemptions for individuals who have exhausted all options in earning their continuing education credits needed to maintain their nutrient management or pesticide certifications.

The Delaware Department of Natural Resources and Environmental Control is working with the Delaware Department of Agriculture to execute a controlled burn at Brandywine Creek State Park in February or March. This prescribed burn will help manage nearly 30 acres of meadowland near the park’s nature center.

Some businesses overlook alternative data. That's probably a mistake.

The post Alternative data may feel risky: Here's why it's not appeared first on The Data Roundtable.

“Cyprus has gone from zero to over 330 licensed investment entities managing €9 billion in assets in just a decade,” according to Maria Panayiotou, president of the Cyprus Investment Fund Association (CIFA). Speaking at the 5th Fund and Asset Management Conference in Athens, Panayiotou attributed this growth to “relentless effort and strategic vision.” She added […]

Is it worth including activated charcoal in your terrarium’s potting mix? James Wong isn’t convinced by this pricey product

Title: Monkey Trials Raise Hope for Non-Addictive Opioid Alternative
Category: Health News
Created: 8/30/2018 12:00:00 AM
Last Editorial Review: 8/31/2018 12:00:00 AM

Title: AHA News: Native People Find Support, 'Sacred Space' Through This Nonprofit's Work
Category: Health News
Created: 8/23/2021 12:00:00 AM
Last Editorial Review: 8/24/2021 12:00:00 AM

Title: Alternative Medicine Popular Among Seniors, But Most Don't Tell Their Doctors About It
Category: Health News
Created: 7/27/2022 12:00:00 AM
Last Editorial Review: 7/27/2022 12:00:00 AM

The SARS-CoV-2 frameshifting element (FSE) has been intensely studied and explored as a therapeutic target for coronavirus diseases, including COVID-19. Besides the intriguing virology, this small RNA is known to adopt many length-dependent conformations, as verified by multiple experimental and computational approaches. However, the role these alternative conformations play in the frameshifting mechanism and how to quantify this structural abundance has been an ongoing challenge. Here, we show by DMS and dual-luciferase functional assays that previously predicted FSE mutants (using the RAG graph theory approach) suppress structural transitions and abolish frameshifting. Furthermore, correlated mutation analysis of DMS data by three programs (DREEM, DRACO, and DANCE-MaP) reveals important differences in their estimation of specific RNA conformations, suggesting caution in the interpretation of such complex conformational landscapes. Overall, the abolished frameshifting in three different mutants confirms that all alternative conformations play a role in the pathways of ribosomal transition.

Hepatocyte nuclear factor 4 alpha antisense 1 (HNF4A-AS1) is a long noncoding RNA (lncRNA) gene physically located next to the transcription factor HNF4A gene in the human genome. Its transcription products have been reported to inhibit the progression of hepatocellular carcinoma (HCC) and negatively regulate the expression of cytochrome P450s (CYPs), including CYP1A2, 2B6, 2C9, 2C19, 2E1, and 3A4. By altering CYP expression, lncRNA HNF4A-AS1 also contributes to the susceptibility of drug-induced liver injury. Thus, HNF4A-AS1 lncRNA is a promising target for controlling HCC and modulating drug metabolism. However, HNF4A-AS1 has four annotated alternative transcripts in the human genome browsers, and it is unclear which transcripts the small interfering RNAs or small hairpin RNAs used in the previous studies are silenced and which transcripts should be used as the target. In this study, four annotated and two newly identified transcripts were confirmed. These six transcripts showed different expression levels in different liver disease conditions, including metabolic dysfunction-associated steatotic liver disease, alcohol-associated liver disease, and obesity. The expression patterns of all HNF4A-AS1 transcripts were further investigated in liver cell growth from human embryonic stem cells to matured hepatocyte-like cells, HepaRG differentiation, and exposure to rifampicin treatment. Several HNF4A-AS1 transcripts highly displayed correlations with these situations. In addition, some of the HNF4A-AS1 transcripts also showed a strong correlation with CYP3A4 during HepaRG maturation and rifampicin exposure. Our findings provide valuable insights into the specific roles of HNF4A-AS1 transcripts, paving the way for more targeted therapeutic strategies for liver diseases and drug metabolism.

Previous studies demonstrated that sigma receptor (R) antagonists alone fail to alter cocaine self-administration despite blocking various other effects of cocaine. However, R antagonists when combined with dopamine transporter (DAT) inhibitors substantially decrease cocaine self-administration. To better understand the effects of this combination, the present study examined the effects of R antagonist and DAT inhibitor combinations in male rats discriminating cocaine (10 mg/kg, i.p.) from saline injections. The DAT inhibitors alone [(–)-2-β-carbomethoxy-3-β-(4-fluorophenyl)tropane 1,5-naphthalenedisulfonate monohydrate (WIN 35,428) and methylphenidate] at low (0.1-mg/kg) doses that were minimally active failed to shift the dose-effect function for discriminative-stimulus effects of cocaine to the left more than 2-fold. At 0.32 mg/kg the DAT inhibitors alone shifted the cocaine dose-effect function leftward 24- or 6.6-fold, respectively. The R antagonists (BD1008, BD1047, and BD1063) failed to fully substitute for cocaine, although BD1008 and BD1047 substituted partially. At 10 mg/kg, BD1008, BD1047, or BD1063 alone shifted the cocaine dose-effect function leftward less than 6.0-fold. In combination with 0.1 mg/kg WIN 35,428, the 10 mg/kg doses of R antagonists shifted the cocaine dose-effect function from 12.3- to 36.7-fold leftward, and with 0.32 mg/kg WIN 35,428 from 14.3- to 440-fold leftward. In combination with 0.1 mg/kg methylphenidate, those R antagonist doses shifted the cocaine dose-effect function from 5.5- to 55.0-fold leftward, and with 0.32 mg/kg methylphenidate from 10.5- to 48.1-fold leftward. The present results suggest that dual DAT/R inhibition produces agonist-like subjective effects that may promote decreases in self-administration obtained in previous studies.

Clinical reports suggest that the most effective strategies for managing opioid use disorder comprise a comprehensive treatment program of both pharmacological and nonpharmacological approaches. However, the conditions under which these combinations are most effective are not well characterized. This study examined whether the presence of an alternative reinforcer could alter the efficacy of Food and Drug Administration–approved opioid antagonist or agonist medications, as well as the nonopioid flumazenil, in decreasing oxycodone choice self-administration in nonhuman primates. Adult squirrel monkeys (n = 7; four females) responded under concurrent second-order fixed-ratio (FR)-3(FR5:S);TO45s schedules of reinforcement for intravenous oxycodone (0.1 mg/kg) or saline on one lever and 30% sweetened condensed milk or water on the other. Doses of naltrexone (0.00032–1.0 mg/kg), nalbuphine (0.32–10 mg/kg), buprenorphine (0.0032–0.032 mg/kg), methadone (0.32–1.0 mg/kg), or flumazenil (1–3.2 mg/kg) were administered intramuscularly prior to oxycodone self-administration sessions that occurred with either milk or water as the alternative. Naltrexone, a μ-opioid receptor antagonist, was >30-fold more potent when milk was available compared with water and abolished oxycodone intake (injections/session) while concomitantly increasing milk deliveries at the highest dose tested. Pretreatment with the low-efficacy μ-agonist nalbuphine was most effective in the presence of milk compared with water, decreasing oxycodone preference to <50% of control values. The higher efficacy μ-agonists, methadone and buprenorphine, and the benzodiazepine antagonist flumazenil did not appreciably alter the reinforcing potency of oxycodone under either condition. These results suggest that antagonist medications used in combination with alternative reinforcers may be an effective strategy to curtail opioid abuse–related behaviors.

BACKGROUND AND PURPOSE:

Angioplasty and stent placement have been described as a bailout technique in individuals with failed thrombectomy. We aimed to investigate Stent retriever AssIsted Lysis (SAIL) with tirofiban before angioplasty and stent placement.

May 29, 2024

The energy transition is driving demand for critical minerals and metals, such as lithium, nickel, cobalt, graphite, and rare earth elements. Sodium-based batteries could provide a more secure, affordable alternative to lithium-ion batteries and highly concentrated lithium supply chains.

"Check out Visual Studio optimizing their rating system to only include the ratings used," shared Fiorenzo R. Imagine the performance gain!

"This sounds about right," says Colin A.

"Wow! Must snap up some sweet Anker kit with this amazing offer; but less than four days to go!" exclaims Dave L., who then goes on to explain
"The actual WTF is this though. I sent this image to Anker with this email: But only 3days left? I hope this offer continues!
Anker replied: Thank you for your feedback! I understand that you appreciate the savings on the Anker SOLIX PS100 Portable Solar Panel and wish the offer could be extended beyond the current 3-day limit. Your suggestion is valuable and will be considered for future promotions to enhance customer satisfaction. If you have any other requests or need further assistance, please let me know.
I for one welcome our new AI overlords. "

Graham F. almost stashed this away for later. "Looks like Dropbox could use a few lessons in how to do Maths! Although maybe their definition of 'almost' differs from mine."

Finally Joshua found time to report a brand-new date-handling bug. "Teams is so buggy; this one just takes the cake. I had to check with the unix cal program to make sure I wasn't completely bonkers." For the readers, November 8 this year is supposed to be a Friday. I suppose things could change after the US election.

The nose, a portal to our bodies, filters air, scents, and medlinkpathogens/medlink. Within the nasal passage, the turbinates, bone structures resembling seashells, play a crucial role.

The WHO has issued a report urging the prioritization of health in climate action. The report, developed with over 100 organizations, calls for an end to fossil fuel reliance and investment in sustainable alternatives.

We’ll be straight with you. There’s no 1:1 Twitter replacement, but there are plenty of social apps that might be worth substituting into your obsessive timeline-checking routines if you’re done with Twitter for whatever reason (we can think of plenty). Twitter’s current situation — advertisers leaving, Nazis logging back on — presents an opportunity to […]

© 2024 TechCrunch. All rights reserved. For personal use only.

If Call of Duty: Black Ops 6 is proving too demanding for your PC, don't worry-Xbox Game Pass offers a fantastic selection of shooters that are engaging and easier on hardware. Here's a deeper look at six titles that provide everything

Feeling like X (formerly Twitter) just isn't your vibe anymore? You're not alone. But don't worry! There's a whole world of social media alternatives waiting to be explored. Here are 6 awesome platforms to check out, each with something unique to

Manchester : Manchester University Press, [2018]

London : Pluto Press, 2023.

Cambridge, Mass. ; London : MIT Press, [2009]

Manchester : Manchester University Press, [2019]

Covid disrupts lives of young women from North-East who’ve had to leave their jobs and go back home

Lily Gladstone picked up the Best Female Actor- Motion Picture award at the Golden Globes for her performance in Martin Scorsese's Killers of The Flower Moon.

We sample inventive menus across the lush, experimental township, which draws visitors from across the world

Chrome on Android will soon natively support third-party autofill services, providing a smoother user experience.

On Threads, people can post text and links and reply to or repost messages from others

Will a green line of credit help a net-zero transition in the auto sector?