While Clemson and Alabama appear to be locks for the postseason bracket, Ohio State's loss opens up the bottom two spots to a few different tiers of teams.

Accelerate, the 5-2 favorite in the final Breeders' Cup Classic future wagering pool, is coming off a big win as the 3-10 favorite in the Awesome Again Stakes at Saratoga. But should you bet him?

The Democratic presidential candidate criticized the president for saying there were “people on both sides” of the exoneration of the men wrongly accused of raping a jogger.

The "Father of the Bride" star spent two days meeting with lawmakers to press for foreign assistance funding.

The musician pointed out that she hails from Texas and is familiar with hunting and gun culture, but she said the recent tragedies deserve a different response.

Review of 'The Case for Nationalism: How It Made Us Powerful, United, and Free' by Rich Lowry

What is a Cron Job? Cron Jobs are basically scheduled tasks that are created automatically by WordPress or a WordPress Plugin that you have activated on your website. They are similar to an alarm clock that goes off regularly to trigger an automative response. Before we delve into explaining how you can view and control […]

The post How to View and Control WordPress Cron Jobs appeared first on Tips and Tricks HQ.

Octoshape announced a partnership with INVISO to deliver Internet TV contribution services throughout Latin America.

"At INVISO, we seek the most innovative and high quality products to serve our customers through the brands we represent,” said Jose Luis Reyes, Vice President for Sales and Operations, INVISO. “In the case of Octoshape, we found a company and a product that bring these qualities to our supply chain, sales and service.”

Octoshape offers an innovative cloud-based solution that provides instant infrastructure for the distribution of both linear and video on demand content. The Octoshape Infinite Uplink service provides point-to-point distribution of TV signals over the Internet for source signal acquisition to traditional IPTV and cable headends as an alternative to traditional methods like satellite and video fiber.

Andreas Georgiou offered an honest accounting of Greece’s financial situation. He’s still paying for it.

oscon: RT @mmmpork: If you're going to #oscon and you're into #perl check out my talk on contributing http://t.co/HEJaPF5JbY

Starting a business is not an easy decision to make. Building a company requires work and money as well, and time. The resources of time, work, and money mean you will have to make sacrifices to start your business. It takes someone who has a lot of passion and confidence to establish a startup. These […]

The post These Teen AI Entrepreneurs Will Amaze You appeared first on ReadWrite.

The US leads the world in terms of number of positive coronavirus cases, with a total of more than 1.2 million.

velocityconf: @da3mon Sorry! We were just doing some followup with past Velocity attendees. Sorry to intrude. No harm intended. :-(

Stay up to date with the latest news, views and analysis as the number of coronavirus cases in SA increases.

The author made sense in both the argument put forth as well as in accurately describing the actual policing festive season operations on the ground in the bustling and busy city centre of Durban.

Make no mistake, the ratings agencies are scratching their heads on whether to downgrade us to junk status or to skip directly to leaking styrofoam cup status.

News24 asked young people to send to us their reasons for leaving and what would have to be different for them to return to South Africa, or not leave at all. The response was overwhelming.

The Amateur Traveler talks to Friedel from TravellingTwo.com about their continuing bike tour around the world. Friedel was on previously talking about travel to Iran and has come back to talk about the Central Asian countries of Turkmenistan, Uzbekistan, Kazakhstan, and Kyrgyzstan. This region of the world offers adventure travelers deserts, tea houses, camels, wild horses, yurts, tea houses, spectacular mountains, friendly locals and fermented mares milk.

The Amateur Traveler talks to Carole Terwilliger Meyers the author of Weekend Adventures in San Francisco and Northern California. Carole comes on the show to talk about California’s traditional wine country of Napa and Sonoma Valleys. This episode focuses primarily on the Napa Valley and the many wineries that can be found there. We explore recreated castles, wine tasting , tours as well as other none wine related sites like mud baths, petrified forests, and the CIA (The Culinary Institute). We talk about restaurants and hotels, picnics and the wine train. Whether you are a connoisseur, a foodie or just a tourist, the Napa Valley has much to offer.

Hear about a road trip in Central Mexico as the Amateur Traveler talks to Amie from thetravelingtogetherjournal.com about a trip they took near Mexico City.

Hear about travel to Columbus and central Ohio as the Amateur Traveler talks to Matthew Caracciolo from matthewcaracciolo.com about what to do in the Buckeye State.

Hear about travel to Central Europe (Prague, Krakow, Budapest) as the Amateur Traveler talks to 4 people who joined me on this year's Amateur Traveler trip: Darrell, Derrick, Loraine and Holly.

Do you have the equivalent of a will for all your digital possessions—photos, email, online accounts, and all the rest? If not, you should, and we have a book that shows you what to do. Plus, Interesting Thing of the Day readers can save 30% on it.

Digital cameras make it easy to take way too many pictures. Need help sorting, organizing, storing, and managing them? We've got a great book for you—and Interesting Thing of the Day readers can save 30% on it.

By easing distancing measures, the Greater Montreal area could experience a substantial increase in the number of deaths per day due to the coronavirus, according to a new document published by the Institut national de santé publique du Québec (INSPQ) in collaboration with experts from Laval University.

I’m excited to introduce something I’ve wanted to do for quite a long time — private one-on-one calls between me and my readers! This new product is a way for us to have private one-on-one conversations tailored to whatever you want. I got into this business is because I wanted to help women achieve their …

INTRODUCING One-on-Ones with Adventurous Kate! Read More »

The post INTRODUCING One-on-Ones with Adventurous Kate! appeared first on Adventurous Kate.

Since starting this blog, I’ve shared hundreds of books with you. Books have been the second topic on this site as long as I can remember, and sharing books with you has been one of my great pleasures. And it goes both ways — you guys have introduced me to some truly wonderful reads as …

Introducing Adventurous Kate’s Book Club! Read More »

The post Introducing Adventurous Kate’s Book Club! appeared first on Adventurous Kate.

The DML News App offers the best in news reporting.

The post REPORT: US issues new visa restrictions for country’s journalists appeared first on Dennis Michael Lynch.

The DML News App offers the best in news reporting.

The post REPORT: US slams new restrictions on journalists from this country appeared first on Dennis Michael Lynch.

The Ontario government gave garden centres and nurseries the green light to open their doors to the public on Friday.

The following article, BREAKING: Sen Marsha Blackburn Introduces Stop COVID Act…Allowing US Citizens To Sue Communist China For Damage They’ve Inflicted On Our Nation, was first published on 100PercentFedUp.com.

Yesterday, Senator Marsha Blackburn (R-TN), along with Senator Martha McSally (R-AZ) introduced the Stop COVID Act, giving Americans the ability to sue Communist China for the damage they’ve inflicted on our nation. Senator Blackburn appeared on Fox News with host Judge Jeanine where she explained the act to Jeanine Pirro. Blackburn told the Fox News […]

Continue reading: BREAKING: Sen Marsha Blackburn Introduces Stop COVID Act…Allowing US Citizens To Sue Communist China For Damage They’ve Inflicted On Our Nation ...

Health officials with the Manitoba government announced on Friday that a COVID-19 outbreak at the Health Sciences Centre has officially been declared over.

The empty space next to Winnipeg City Hall that once housed restaurants could be turned into a place to help exploited women.

Type 2 diabetes mellitus (T2DM) is characterized by impaired glucose-stimulated insulin secretion and increased peripheral insulin resistance. Unremitting endoplasmic reticulum (ER) stress can lead to beta-cell apoptosis and has been linked to type 2 diabetes. Although many studies have attempted to link ER stress and T2DM, the specific effects of ER stress on beta-cell function remain incompletely understood. To determine the interrelationship between ER stress and beta-cell function, here we treated insulin-secreting INS-1(832/13) cells or isolated mouse islets with the ER stress–inducer tunicamycin (TM). TM induced ER stress as expected, as evidenced by activation of the unfolded protein response. Beta cells treated with TM also exhibited concomitant alterations in their electrical activity and cytosolic free Ca2+ oscillations. As ER stress is known to reduce ER Ca2+ levels, we tested the hypothesis that the observed increase in Ca2+ oscillations occurred because of reduced ER Ca2+ levels and, in turn, increased store-operated Ca2+ entry. TM-induced cytosolic Ca2+ and membrane electrical oscillations were acutely inhibited by YM58483, which blocks store-operated Ca2+ channels. Significantly, TM-treated cells secreted increased insulin under conditions normally associated with only minimal release, e.g. 5 mm glucose, and YM58483 blocked this secretion. Taken together, these results support a critical role for ER Ca2+ depletion–activated Ca2+ current in mediating Ca2+-induced insulin secretion in response to ER stress.

Prominins (proms) are transmembrane glycoproteins conserved throughout the animal kingdom. They are associated with plasma membrane protrusions, such as primary cilia, as well as extracellular vesicles derived thereof. Primary cilia host numerous signaling pathways affected in diseases known as ciliopathies. Human PROM1 (CD133) is detected in both somatic and cancer stem cells and is also expressed in terminally differentiated epithelial and photoreceptor cells. Genetic mutations in the PROM1 gene result in retinal degeneration by impairing the proper formation of the outer segment of photoreceptors, a modified cilium. Here, we investigated the impact of proms on two distinct examples of ciliogenesis. First, we demonstrate that the overexpression of a dominant-negative mutant variant of human PROM1 (i.e. mutation Y819F/Y828F) significantly decreases ciliary length in Madin–Darby canine kidney cells. These results contrast strongly to the previously observed enhancing effect of WT PROM1 on ciliary length. Mechanistically, the mutation impeded the interaction of PROM1 with ADP-ribosylation factor–like protein 13B, a key regulator of ciliary length. Second, we observed that in vivo knockdown of prom3 in zebrafish alters the number and length of monocilia in the Kupffer's vesicle, resulting in molecular and anatomical defects in the left-right asymmetry. These distinct loss-of-function approaches in two biological systems reveal that prom proteins are critical for the integrity and function of cilia. Our data provide new insights into ciliogenesis and might be of particular interest for investigations of the etiologies of ciliopathies.

Interferon-regulated myxovirus resistance protein B (MxB) is an interferon-induced GTPase belonging to the dynamin superfamily. It inhibits infection with a wide range of different viruses, including HIV-1, by impairing viral DNA entry into the nucleus. Unlike the related antiviral GTPase MxA, MxB possesses an N-terminal region that contains a nuclear localization signal and is crucial for inhibiting HIV-1. Because MxB previously has been shown to reside in both the nuclear envelope and the cytoplasm, here we used bioinformatics and biochemical approaches to identify a nuclear export signal (NES) responsible for MxB's cytoplasmic location. Using the online computational tool LocNES (Locating Nuclear Export Signals or NESs), we identified five putative NES candidates in MxB and investigated whether their deletion caused nuclear localization of MxB. Our results revealed that none of the five deletion variants relocates to the nucleus, suggesting that these five predicted NES sequences do not confer NES activity. Interestingly, deletion of one sequence, encompassing amino acids 505–527, abrogated the anti-HIV-1 activity of MxB. Further mutation experiments disclosed that amino acids 515–519, and Pro-515 in particular, regulate MxB oligomerization and its binding to HIV-1 capsid, thereby playing an important role in MxB-mediated restriction of HIV-1 infection. In summary, our results indicate that none of the five predicted NES sequences in MxB appears to be required for its nuclear export. Our findings also reveal several residues in MxB, including Pro-515, critical for its oligomerization and anti-HIV-1 function.

Research Event

Event participants

Osamah Al Rawhani, Deputy Director, Sana’a Center for Strategic Studies
Moderator: Nadim Houry, Executive Director, Arab Reform Initiative

NAD+ is a central metabolite participating in core metabolic redox reactions. The prokaryotic NAD synthetase enzyme NadE catalyzes the last step of NAD+ biosynthesis, converting nicotinic acid adenine dinucleotide (NaAD) to NAD+. Some members of the NadE family use l-glutamine as a nitrogen donor and are named NadEGln. Previous gene neighborhood analysis has indicated that the bacterial nadE gene is frequently clustered with the gene encoding the regulatory signal transduction protein PII, suggesting a functional relationship between these proteins in response to the nutritional status and the carbon/nitrogen ratio of the bacterial cell. Here, using affinity chromatography, bioinformatics analyses, NAD synthetase activity, and biolayer interferometry assays, we show that PII and NadEGln physically interact in vitro, that this complex relieves NadEGln negative feedback inhibition by NAD+. This mechanism is conserved in distantly related bacteria. Of note, the PII protein allosteric effector and cellular nitrogen level indicator 2-oxoglutarate (2-OG) inhibited the formation of the PII-NadEGln complex within a physiological range. These results indicate an interplay between the levels of ATP, ADP, 2-OG, PII-sensed glutamine, and NAD+, representing a metabolic hub that may balance the levels of core nitrogen and carbon metabolites. Our findings support the notion that PII proteins act as a dissociable regulatory subunit of NadEGln, thereby enabling the control of NAD+ biosynthesis according to the nutritional status of the bacterial cell.

Interferon-regulated myxovirus resistance protein B (MxB) is an interferon-induced GTPase belonging to the dynamin superfamily. It inhibits infection with a wide range of different viruses, including HIV-1, by impairing viral DNA entry into the nucleus. Unlike the related antiviral GTPase MxA, MxB possesses an N-terminal region that contains a nuclear localization signal and is crucial for inhibiting HIV-1. Because MxB previously has been shown to reside in both the nuclear envelope and the cytoplasm, here we used bioinformatics and biochemical approaches to identify a nuclear export signal (NES) responsible for MxB's cytoplasmic location. Using the online computational tool LocNES (Locating Nuclear Export Signals or NESs), we identified five putative NES candidates in MxB and investigated whether their deletion caused nuclear localization of MxB. Our results revealed that none of the five deletion variants relocates to the nucleus, suggesting that these five predicted NES sequences do not confer NES activity. Interestingly, deletion of one sequence, encompassing amino acids 505–527, abrogated the anti-HIV-1 activity of MxB. Further mutation experiments disclosed that amino acids 515–519, and Pro-515 in particular, regulate MxB oligomerization and its binding to HIV-1 capsid, thereby playing an important role in MxB-mediated restriction of HIV-1 infection. In summary, our results indicate that none of the five predicted NES sequences in MxB appears to be required for its nuclear export. Our findings also reveal several residues in MxB, including Pro-515, critical for its oligomerization and anti-HIV-1 function.

Support from the Bill & Melinda Gates Foundation has enabled Chatham House to develop a global health security track at the Munich Security Conference (MSC).

Chatham House is pleased to announce that Rob Yates has been appointed as head of the Centre on Global Health Security.

12 March 2015 , Volume 91, Number 2

6 February 2020

Viviane Baladi and Mark F. Demers
J. Amer. Math. Soc. 33 (2020), 381-449.
Abstract, references and article information

Purinergic signaling by extracellular ATP regulates a variety of cellular events and is implicated in both normal physiology and pathophysiology. Several molecules have been associated with the release of ATP and other small molecules, but their precise contributions have been difficult to assess because of their complexity and heterogeneity. Here, we report on the results of a gain-of-function screen for modulators of hypotonicity-induced ATP release using HEK-293 cells and murine cerebellar granule neurons, along with bioluminescence, calcium FLIPR, and short hairpin RNA–based gene-silencing assays. This screen utilized the most extensive genome-wide ORF collection to date, covering 90% of human, nonredundant, protein-encoding genes. We identified two ABCG1 (ABC subfamily G member 1) variants, which regulate cellular cholesterol, as modulators of hypotonicity-induced ATP release. We found that cholesterol levels control volume-regulated anion channel–dependent ATP release. These findings reveal novel mechanisms for the regulation of ATP release and volume-regulated anion channel activity and provide critical links among cellular status, cholesterol, and purinergic signaling.

Kindlins are focal adhesion proteins that regulate integrin activation and outside-in signaling. The kindlin family consists of three members, kindlin-1, -2, and -3. Kindlin-2 is widely expressed in multiple cell types, except those from the hematopoietic lineage. A previous study has reported that the Drosophila Fit1 protein (an ortholog of kindlin-2) prevents abnormal spindle assembly; however, the mechanism remains unknown. Here, we show that kindlin-2 maintains spindle integrity in mitotic human cells. The human neuroblastoma SH-SY5Y cell line expresses only kindlin-2, and we found that when SH-SY5Y cells are depleted of kindlin-2, they exhibit pronounced spindle abnormalities and delayed mitosis. Of note, acetylation of α-tubulin, which maintains microtubule flexibility and stability, was diminished in the kindlin-2–depleted cells. Mechanistically, we found that kindlin-2 maintains α-tubulin acetylation by inhibiting the microtubule-associated deacetylase histone deacetylase 6 (HDAC6) via a signaling pathway involving AKT Ser/Thr kinase (AKT)/glycogen synthase kinase 3β (GSK3β) or paxillin. We also provide evidence that prolonged hypoxia down-regulates kindlin-2 expression, leading to spindle abnormalities not only in the SH-SY5Y cell line, but also cell lines derived from colon and breast tissues. The findings of our study highlight that kindlin-2 regulates mitotic spindle assembly and that this process is perturbed in cancer cells in a hypoxic environment.

2 March 2020

Heme-regulatory motifs (HRMs) are present in many proteins that are involved in diverse biological functions. The C-terminal tail region of human heme oxygenase-2 (HO2) contains two HRMs whose cysteine residues form a disulfide bond; when reduced, these cysteines are available to bind Fe3+-heme. Heme binding to the HRMs occurs independently of the HO2 catalytic active site in the core of the protein, where heme binds with high affinity and is degraded to biliverdin. Here, we describe the reversible, protein-mediated transfer of heme between the HRMs and the HO2 core. Using hydrogen-deuterium exchange (HDX)-MS to monitor the dynamics of HO2 with and without Fe3+-heme bound to the HRMs and to the core, we detected conformational changes in the catalytic core only in one state of the catalytic cycle—when Fe3+-heme is bound to the HRMs and the core is in the apo state. These conformational changes were consistent with transfer of heme between binding sites. Indeed, we observed that HRM-bound Fe3+-heme is transferred to the apo-core either upon independent expression of the core and of a construct spanning the HRM-containing tail or after a single turnover of heme at the core. Moreover, we observed transfer of heme from the core to the HRMs and equilibration of heme between the core and HRMs. We therefore propose an Fe3+-heme transfer model in which HRM-bound heme is readily transferred to the catalytic site for degradation to facilitate turnover but can also equilibrate between the sites to maintain heme homeostasis.

NAD+ is a central metabolite participating in core metabolic redox reactions. The prokaryotic NAD synthetase enzyme NadE catalyzes the last step of NAD+ biosynthesis, converting nicotinic acid adenine dinucleotide (NaAD) to NAD+. Some members of the NadE family use l-glutamine as a nitrogen donor and are named NadEGln. Previous gene neighborhood analysis has indicated that the bacterial nadE gene is frequently clustered with the gene encoding the regulatory signal transduction protein PII, suggesting a functional relationship between these proteins in response to the nutritional status and the carbon/nitrogen ratio of the bacterial cell. Here, using affinity chromatography, bioinformatics analyses, NAD synthetase activity, and biolayer interferometry assays, we show that PII and NadEGln physically interact in vitro, that this complex relieves NadEGln negative feedback inhibition by NAD+. This mechanism is conserved in distantly related bacteria. Of note, the PII protein allosteric effector and cellular nitrogen level indicator 2-oxoglutarate (2-OG) inhibited the formation of the PII-NadEGln complex within a physiological range. These results indicate an interplay between the levels of ATP, ADP, 2-OG, PII-sensed glutamine, and NAD+, representing a metabolic hub that may balance the levels of core nitrogen and carbon metabolites. Our findings support the notion that PII proteins act as a dissociable regulatory subunit of NadEGln, thereby enabling the control of NAD+ biosynthesis according to the nutritional status of the bacterial cell.

3-Mercaptopyruvate sulfur transferase (MPST) catalyzes the desulfuration of 3-mercaptopyruvate (3-MP) and transfers sulfane sulfur from an enzyme-bound persulfide intermediate to thiophilic acceptors such as thioredoxin and cysteine. Hydrogen sulfide (H2S), a signaling molecule implicated in many physiological processes, can be released from the persulfide product of the MPST reaction. Two splice variants of MPST, differing by 20 amino acids at the N terminus, give rise to the cytosolic MPST1 and mitochondrial MPST2 isoforms. Here, we characterized the poorly-studied MPST1 variant and demonstrated that substitutions in its Ser–His–Asp triad, proposed to serve a general acid–base role, minimally affect catalytic activity. We estimated the 3-MP concentration in murine liver, kidney, and brain tissues, finding that it ranges from 0.4 μmol·kg−1 in brain to 1.4 μmol·kg−1 in kidney. We also show that N-acetylcysteine, a widely-used antioxidant, is a poor substrate for MPST and is unlikely to function as a thiophilic acceptor. Thioredoxin exhibits substrate inhibition, increasing the KM for 3-MP ∼15-fold compared with other sulfur acceptors. Kinetic simulations at physiologically-relevant substrate concentrations predicted that the proportion of sulfur transfer to thioredoxin increases ∼3.5-fold as its concentration decreases from 10 to 1 μm, whereas the total MPST reaction rate increases ∼7-fold. The simulations also predicted that cysteine is a quantitatively-significant sulfane sulfur acceptor, revealing MPST's potential to generate low-molecular-weight persulfides. We conclude that the MPST1 and MPST2 isoforms are kinetically indistinguishable and that thioredoxin modulates the MPST-catalyzed reaction in a physiologically-relevant concentration range.