Rina Foygel Barber, Emmanuel J. Candès.

Source: The Annals of Statistics, Volume 47, Number 5, 2504--2537.

Abstract:
This paper develops a framework for testing for associations in a possibly high-dimensional linear model where the number of features/variables may far exceed the number of observational units. In this framework, the observations are split into two groups, where the first group is used to screen for a set of potentially relevant variables, whereas the second is used for inference over this reduced set of variables; we also develop strategies for leveraging information from the first part of the data at the inference step for greater power. In our work, the inferential step is carried out by applying the recently introduced knockoff filter, which creates a knockoff copy—a fake variable serving as a control—for each screened variable. We prove that this procedure controls the directional false discovery rate (FDR) in the reduced model controlling for all screened variables; this says that our high-dimensional knockoff procedure “discovers” important variables as well as the directions (signs) of their effects, in such a way that the expected proportion of wrongly chosen signs is below the user-specified level (thereby controlling a notion of Type S error averaged over the selected set). This result is nonasymptotic, and holds for any distribution of the original features and any values of the unknown regression coefficients, so that inference is not calibrated under hypothesized values of the effect sizes. We demonstrate the performance of our general and flexible approach through numerical studies, showing more power than existing alternatives. Finally, we apply our method to a genome-wide association study to find locations on the genome that are possibly associated with a continuous phenotype.

Joshua A. Gordon
May 15, 1996; 16:3274-3286
Articles

EL Bienenstock
Jan 1, 1982; 2:32-48
Articles

Despite their opposing actions on food intake, POMC and NPY/AgRP neurons in the arcuate nucleus of the hypothalamus (ARH) are derived from the same progenitors that give rise to ARH neurons. However, the mechanism whereby common neuronal precursors subsequently adopt either the anorexigenic (POMC) or the orexigenic (NPY/AgRP) identity remains elusive. We hypothesize that POMC and NPY/AgRP cell fates are specified and maintained by distinct intrinsic factors. In search of them, we profiled the transcriptomes of developing POMC and NPY/AgRP neurons in mice. Moreover, cell-type-specific transcriptomic analyses revealed transcription regulators that are selectively enriched in either population, but whose developmental functions are unknown in these neurons. Among them, we found the expression of the PR domain-containing factor 12 (Prdm12) was enriched in POMC neurons but absent in NPY/AgRP neurons. To study the role of Prdm12 in vivo, we developed and characterized a floxed Prdm12 allele. Selective ablation of Prdm12 in embryonic POMC neurons led to significantly reduced Pomc expression as well as early-onset obesity in mice of either sex that recapitulates symptoms of human POMC deficiency. Interestingly, however, specific deletion of Prdm12 in adult POMC neurons showed that it is no longer required for Pomc expression or energy balance. Collectively, these findings establish a critical role for Prdm12 in the anorexigenic neuron identity and suggest that it acts developmentally to program body weight homeostasis. Finally, the combination of cell-type-specific genomic and genetic analyses provides a means to dissect cellular and functional diversity in the hypothalamus whose neurodevelopment remains poorly studied.

SIGNIFICANCE STATEMENT POMC and NPY/AgRP neurons are derived from the same hypothalamic progenitors but have opposing effects on food intake. We profiled the transcriptomes of genetically labeled POMC and NPY/AgRP neurons in the developing mouse hypothalamus to decipher the transcriptional codes behind the versus orexigenic neuron identity. Our analyses revealed 29 transcription regulators that are selectively enriched in one of the two populations. We generated new mouse genetic models to selective ablate one of POMC-neuron enriched transcription factors Prdm12 in developing and adult POMC neurons. Our studies establish a previously unrecognized role for Prdm12 in the anorexigenic neuron identity and suggest that it acts developmentally to program body weight homeostasis.

Gamma-band oscillations (GBOs) elicited by transient nociceptive stimuli are one of the most promising biomarkers of pain across species. Still, whether these GBOs reflect stimulus encoding in the primary somatosensory cortex (S1) or nocifensive behavior in the primary motor cortex (M1) is debated. Here we recorded neural activity simultaneously from the brain surface as well as at different depths of the bilateral S1/M1 in freely-moving male rats receiving nociceptive stimulation. GBOs measured from superficial layers of S1 contralateral to the stimulated paw not only had the largest magnitude, but also showed the strongest temporal and phase coupling with epidural GBOs. Also, spiking of superficial S1 interneurons had the strongest phase coherence with epidural GBOs. These results provide the first direct demonstration that scalp GBOs, one of the most promising pain biomarkers, reflect neural activity strongly coupled with the fast spiking of interneurons in the superficial layers of the S1 contralateral to the stimulated side.

SIGNIFICANCE STATEMENT Nociceptive-induced gamma-band oscillations (GBOs) measured at population level are one of the most promising biomarkers of pain perception. Our results provide the direct demonstration that these GBOs reflect neural activity coupled with the spike firing of interneurons in the superficial layers of the primary somatosensory cortex (S1) contralateral to the side of nociceptive stimulation. These results address the ongoing debate about whether nociceptive-induced GBOs recorded with scalp EEG or epidurally reflect stimulus encoding in the S1 or nocifensive behavior in the primary motor cortex (M1), and will therefore influence how experiments in pain neuroscience will be designed and interpreted.

One of the first signs of viral infection is body-wide aches and pain. Although this type of pain usually subsides, at the extreme, viral infections can induce painful neuropathies that can last for decades. Neither of these types of pain sensitization is well understood. A key part of the response to viral infection is production of interferons (IFNs), which then activate their specific receptors (IFNRs) resulting in downstream activation of cellular signaling and a variety of physiological responses. We sought to understand how type I IFNs (IFN-α and IFN-β) might act directly on nociceptors in the dorsal root ganglion (DRG) to cause pain sensitization. We demonstrate that type I IFNRs are expressed in small/medium DRG neurons and that their activation produces neuronal hyper-excitability and mechanical pain in mice. Type I IFNs stimulate JAK/STAT signaling in DRG neurons but this does not apparently result in PKR-eIF2α activation that normally induces an anti-viral response by limiting mRNA translation. Rather, type I IFNs stimulate MNK-mediated eIF4E phosphorylation in DRG neurons to promote pain hypersensitivity. Endogenous release of type I IFNs with the double-stranded RNA mimetic poly(I:C) likewise produces pain hypersensitivity that is blunted in mice lacking MNK-eIF4E signaling. Our findings reveal mechanisms through which type I IFNs cause nociceptor sensitization with implications for understanding how viral infections promote pain and can lead to neuropathies.

SIGNIFICANCE STATEMENT It is increasingly understood that pathogens interact with nociceptors to alert organisms to infection as well as to mount early host defenses. Although specific mechanisms have been discovered for diverse bacterial and fungal pathogens, mechanisms engaged by viruses have remained elusive. Here we show that type I interferons, one of the first mediators produced by viral infection, act directly on nociceptors to produce pain sensitization. Type I interferons act via a specific signaling pathway (MNK-eIF4E signaling), which is known to produce nociceptor sensitization in inflammatory and neuropathic pain conditions. Our work reveals a mechanism through which viral infections cause heightened pain sensitivity

Her face is full of joy, and her excitement for her job is clear. But how well do you know TeenStreet translator Agnes' story?

History’s census enumerators came back with the numbers and some very tall tales

1. Bharat Biotech to lead human monoclonal antibodies project  The Hindu
2. Bharat Biotech leads CSIR project to develop antibodies against Covid-19  Times of India
3. Bharat Biotech To Lead CSIR’s Project To Develop Human Monoclonal Antibodies For Covid-19 Therapy  Swarajya
4. Bharat Biotech to develop human antibodies for COVID-19 therapy  Telangana Today
5. Bharat Biotech vows human monoclonal antibodies to neutralise COVID-19 in 6 months  Mumbai Mirror
6. View Full coverage on Google News

Nearly one-third of all US children are overweight or obese, with even higher prevalence among Mexican American children. Overweight and obesity increase systemic inflammation, contributing to increased risk for chronic diseases, such as type 2 diabetes mellitus and cardiovascular disease.

Obese Mexican American children had concurrent alterations in both inflammatory markers and traditional disease risk markers, relative to healthy weight children. Our results provide evidence partially explaining the health disparity for disease in Mexican American children who are overweight/obese. (Read the full article)

Honey is recommended as a cough medication by the World Health Organization. To date, the efficacy of this treatment has been shown in 2 studies: one tested only buckwheat honey and the other study was not blinded.

In a randomized controlled trial, we compared 3 types of honey versus placebo as a treatment of upper respiratory tract infection–associated cough. These types of honey were superior to placebo in alleviating cough. (Read the full article)

Three years after the launch of the M10, Leica is bringing a black-and-white Monochrom version to market, with an all-new 40MP imager and the classic M rangefinder design.

In vitro and in vivo interactions of minocycline and azoles including itraconazole, voriconazole, and posaconazole against filamentous pathogenic fungi were investigated. A total of 56 clinical isolates were studied in vitro via broth microdilution checkerboard technique, including 20 strains of Aspergillus fumigatus, 7 strains of A. flavus, 16 strains of Exophiala dermatitidis, 10 strains of Fusarium solani and 3 strain s of F. oxysporum. The results revealed that minocycline individually did not exhibit any significant antifungal activity against all tested strains. However, favorable synergy of minocycline with itraconazole, voriconazole, or posaconazole were observed against 34 (61%), 28 (50%), and 38 (69%) isolates, respectively, including azole resistant A. fumigatus and Fusarium spp. with inherently high MICs of azoles. Synergistic combinations resulted in 4 fold to 16-fold reduction of effective MICs of minocycline and azoles. No antagonism was observed. In vivo effect of minocycline-azole combinations were evaluated by survival assay in Galleria mellonella model infected with E. dermatitidis strain BMU00034, F. solani strain FS9, A. fumigatus strain AF293, AFR1 and AFR2 . Minocycline acted synergistically with azoles and significantly increased larvae survival in all isolates (P<0.001), including azole resistant A. fumigatus and azole-inactive Fusarium spp.. In conclusion, the results suggested that minocycline combined with azoles may help to enhance the antifungal susceptibilities of azoles against pathogenic fungi and had the potential to overcome azole resistance issues.

The IMpact-RSV Study Group
Sep 1, 1998; 102:531-537
ARTICLES

The eight group stage winners and eight play-off victors discovered their UEFA Youth League round of 16 opponents when the draw was made on Friday.

From bicycle culture to electric vehicles, the automotive industry is changing and Covid-19 may be accelerating the pace of change.

UPDATE: This event will take place at Kent County’s Brecknock Park – all other details are the same. For directions: click here   “Forestry for the future” – Students to test forestry skills at Brecknock Park on June 25. Contact: Ashley Peebles, 302-698-4551 ashley.peebles@delaware.gov Who:     – Delaware school chapters of the Future Farmers of America (FFA) and […]

This project brings together National Centre for Cell Science (NCCS), Pune; Indian Institute of Technology, Indore, and Gurgaon-based PredOmix Technologies in a collaborative mode for a public health emergency.

1. Bharat Biotech to lead human monoclonal antibodies venture  Microbioz India
2. Bharat Biotech to lead monoclonal antibodies project for Covid-19 therapy  The Financial Express
3. Bharat Biotech leads CSIR project to develop antibodies against Covid-19  Times of India
4. CSIR approves project to develop human monoclonal antibodies that can neutralize COVID-19 in patients  Times Now
5. Bharat Biotech to lead project on monoclonal antibodies therapy for COVID-19  The New Indian Express
6. View Full coverage on Google News

France defender Samuel Umtiti has picked up a calf injury in just the second session since Barcelona returned to training from the coronavirus quarantine, the Spanish champions announced on Saturday. Barca said that Umiti picked up an injury to his right calf during Saturday’s training session at the club’s Joan Gamper facility, but did not
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The post Barca’s Samuel Umtiti suffers calf knock in second training session appeared first on Vanguard News.

La enseñanza bíblica en profundidad de John MacArthur lleva la verdad transformadora de la Palabra de Dios a millones de personas cada día.

La enseñanza bíblica en profundidad de John MacArthur lleva la verdad transformadora de la Palabra de Dios a millones de personas cada día.

La enseñanza bíblica en profundidad de John MacArthur lleva la verdad transformadora de la Palabra de Dios a millones de personas cada día.

La enseñanza bíblica en profundidad de John MacArthur lleva la verdad transformadora de la Palabra de Dios a millones de personas cada día.

A recent decision concerning ownership of copyright has served as a stark reminder to businesses to ensure they consider and clearly capture ownership of intellectual property at the outset when commissioning works. The case concerned rights in the ...

In the lead-up to the Truth Matters conference in October, we will be focusing our attention on the sufficiency, authority, and clarity of Scripture. Of our previous blog series, none better embodies that emphasis than Frequently Abused Verses. The following entry from that series originally appeared on October 5, 2015. -ed.

Is it really “abuse” if a verse is used inaccurately to make an important point?

The short answer is, “Yes.” We should not be so careless and cavalier with Scripture, or think so highly of ourselves, that we can impose new meaning—even if it is valid—on the inerrant, sufficient Word of God. If the point is worth making, it’s worth making from the appropriate text.

Which brings us to the verse before us today: Revelation 3:20 is certainly one of the most familiar and frequently-quoted verses in the church. It’s a particular favorite for evangelists, camp preachers, and anyone else who wants to lend some urgency to the call of God on a sinner’s life

“Behold, I stand at the door and knock; if anyone hears My voice and opens the door, I will come in to him and will dine with him, and he with Me” (Revelation 3:20). In the hands of many preachers and evangelists, the verse paints an attractive, compelling picture of Christ’s pursuit of the sinner, and highlights the need for an immediate response.

But is that an accurate interpretation of the verse—is Christ truly at the doorstep of each sinner’s heart, pleading to come in? And if not, on whose door is the Lord knocking? Let’s tackle those issues one at a time.

Is Christ Knocking?

We use a lot of clichés as shorthand in the church, and not all of them are helpful or even accurate. For example, many Christians talk about “asking Jesus into your heart.” And while that phrase might have some vaguely biblical underpinnings, it doesn’t shed any light on what it truly means to repent and believe. If anything, it muddles the sinner’s responsibility in salvation; it dulls some of the sharp edges of the gospel.

In the same way, the common misapplication of Revelation 3:20 has done more harm than good. Yes, the mental image of Christ knocking on the door of a sinner’s heart is moving. But it’s not accurate—it’s a caricature at best, and it comes at a high theological cost.

Put simply, Christ isn’t pleading on every sinner’s spiritual doorstep. Jesus doesn’t need to beg or badger anyone into the kingdom of heaven (John 10:27-28). Salvation isn’t merely a matter of the Lord getting a foot inside the door of your heart—it’s a work of total transformation (Ezekiel 36:26). And most important of all, salvation is not triggered by an act of the sinner’s will—it is God’s intervening work that rescues us from the just penalty of our sin (Ephesians 2:4-9).

In fact, the abuse of Revelation 3:20 often goes hand-in-hand with talk of “asking Jesus into your heart” and other man-centered versions of the gospel message. One way to protect yourself and your evangelism from such skewed perspectives is to closely adhere to biblical language when you’re explaining the gospel.

And you were dead in your trespasses and sins, in which you formerly walked according to the course of this world, according to the prince of the power of the air, of the spirit that is now working in the sons of disobedience. Among them we too all formerly lived in the lusts of our flesh, indulging the desires of the flesh and of the mind, and were by nature children of wrath, even as the rest. But God, being rich in mercy, because of His great love with which He loved us, even when we were dead in our transgressions, made us alive together with Christ (by grace you have been saved), and raised us up with Him, and seated us with Him in the heavenly places in Christ Jesus, so that in the ages to come He might show the surpassing riches of His grace in kindness toward us in Christ Jesus. For by grace you have been saved through faith; and that not of yourselves, it is the gift of God; not as a result of works, so that no one may boast. (Ephesians 2:1-9, emphasis added)

Train yourself to think about the gospel in those terms, and you’ll insulate yourself from the influence of man-centered theology, and the temptation to reinterpret God’s Word.

Whose Door?

The door in Revelation 3:20 was not a vague spiritual metaphor—it was a specific door. And while Christ wasn’t physically knocking, His words were directed to a specific group of people, and should not be watered down or applied carelessly to just anyone.

The context of Revelation 3:20 is Christ’s letter to the church at Laodicea—also known as the lukewarm church. In Revelation 3:14-22, the Lord condemns them for their spiritual self-deception and apathy. Christ says, “I know your deeds, that you are neither cold nor hot; I wish that you were cold or hot” (v. 15). They did not openly reject Christ, but neither did they exhibit any spiritual zeal or authentic love for God or His Word. They professed to know Christ, but He had no place in their assembly.

And lost in their self-deception, they risked being spat out of God’s mouth altogether (v. 16). Their only hope was to truly repent (v. 19).

In the context of Revelation 3, then, Christ was standing at the door of the Laodicean church, eager to re-enter the congregation through the genuine repentance and salvation of its members. In his commentary on this passage, John MacArthur explains the imagery of verse 20:

Though this verse has been used in countless tracts and evangelistic messages to depict Christ’s knocking on the door of the sinner’s heart, it is broader than that. The door on which Christ is knocking is not the door to a single human heart, but to the Laodicean church. Christ was outside this apostate church and wanted to come in—something that could only happen if the people repented.

The invitation is, first of all, a personal one, since salvation is individual. But He is knocking on the door of the church, calling the many to saving faith, so that He may enter the church. If one person (anyone) opened the door by repentance and faith, Christ would enter that church through that individual. The picture of Christ outside the Laodicean church seeking entrance strongly implies that, unlike Sardis, there were no believers there at all.

Christ’s offer to dine with the repentant church speaks of fellowship, communion, and intimacy. Sharing a meal in ancient times symbolized the union of people in loving fellowship. Believers will dine with Christ at the marriage supper of the Lamb (Revelation 19:9), and in the millennial kingdom (Luke 22:16, 29-30). Dine is from deipneō, which refers to the evening meal, the last meal of the day. The Lord Jesus Christ urged them to repent and have fellowship with Him before the night of judgment fell and it was too late forever. ^[1] John MacArthur, The MacArthur New Testament Commentary: Revelation 1-11 (Chicago: Moody Press, 1999) 140.

What does repentance look like? Far from merely opening the door of your heart to Christ, true repentance reflects the conviction of your sin and the deep desire for righteousness. Here’s how D. Martyn Lloyd-Jones defined this important doctrine:

Repentance means that you realize that you are a guilty, vile sinner in the presence of God, that you deserve the wrath and punishment of God, that you are hell-bound. It means that you begin to realize that this thing called sin is in you, that you long to get rid of it, and that you turn your back on it in every shape and form. You renounce the world whatever the cost, the world in its mind and outlook as well as its practice, and you deny yourself, and take up the cross and go after Christ. ^[2] D. Martyn Lloyd-Jones, Studies in the Sermon on the Mount (Grand Rapids: Eerdmans, 1974) 2:248.

The Urgent Call of the Gospel

When it comes to applying and interpreting Scripture, the details matter; good intentions are not enough. We bring the authority of Scripture to bear in sinners’ lives only inasmuch as we handle it accurately. We have a responsibility to the Lord, to each other, and to the unsaved world to proclaim the excellence, inerrancy, and sufficiency of the Bible. And we can’t fulfill that responsibility if we’re assigning our own meaning to God’s immutable truth.

With that in mind, you may still want to inject some urgency into the call to repent the next time you share the gospel with friends or family. Rather than falling back on a misappropriation of Christ’s words in Revelation, why not make a biblically sound argument? Here are a couple passages that convey the sinner’s urgent spiritual needs.

Isaiah preached to the apostate nation of Israel pleading with them to return to the Lord:

Seek the Lord while He may be found; call upon Him while He is near. Let the wicked forsake his way and the unrighteous man his thoughts; and let him return to the Lord, and He will have compassion on him, and to our God, for He will abundantly pardon. (Isaiah 55:6-7)

And in Acts 17 Paul ended his gospel appeal to a crowd of philosophers with these words:

Therefore having overlooked the times of ignorance, God is now declaring to men that all people everywhere should repent, because He has fixed a day in which He will judge the world in righteousness through a Man whom He has appointed, having furnished proof to all men by raising Him from the dead. (Acts 17:30-31)

These and other passages (cf. Acts 2:37-40; Hebrews 4:6-7) can be rightly used to urge unbelievers or those lost in self-deception to respond to the gospel by repenting and turning to Christ. What good is our evangelistic zeal if we aren’t biblically sound?

The 26-year-old joined Barca from Lyon in 2016.

President Donald Trump needled his Democratic rival Joe Biden Friday for limiting his campaign appearances to virtual events from the basement of his home in Delaware.

Title: Health Tip: Knocked Out Teeth
Category: Health News
Created: 1/2/2020 12:00:00 AM
Last Editorial Review: 1/2/2020 12:00:00 AM

ABSTRACT

Arthritogenic alphaviruses such as Ross River and Chikungunya viruses cause debilitating muscle and joint pain and pose significant challenges in the light of recent outbreaks. How host immune responses are orchestrated after alphaviral infections and lead to musculoskeletal inflammation remains poorly understood. Here, we show that myositis induced by Ross River virus (RRV) infection is driven by CD11b^hi Ly6C^hi inflammatory monocytes and followed by the establishment of a CD11b^hi Ly6C^lo CX₃CR1⁺ macrophage population in the muscle upon recovery. Selective modulation of CD11b^hi Ly6C^hi monocyte migration to infected muscle using immune-modifying microparticles (IMP) reduced disease score, tissue damage, and inflammation and promoted the accumulation of CX₃CR1⁺ macrophages, enhancing recovery and resolution. Here, we detail the role of immune pathology, describing a poorly characterized muscle macrophage subset as part of the dynamics of alphavirus-induced myositis and tissue recovery and identify IMP as an effective immunomodulatory approach. Given the lack of specific treatments available for alphavirus-induced pathologies, this study highlights a therapeutic potential for simple immune modulation by IMP in infected individuals in the event of large alphavirus outbreaks.

IMPORTANCE Arthritogenic alphaviruses cause debilitating inflammatory disease, and current therapies are restricted to palliative approaches. Here, we show that following monocyte-driven muscle inflammation, tissue recovery is associated with the accumulation of CX₃CR1⁺ macrophages in the muscle. Modulating inflammatory monocyte infiltration using immune-modifying microparticles (IMP) reduced tissue damage and inflammation and enhanced the formation of tissue repair-associated CX₃CR1⁺ macrophages in the muscle. This shows that modulating key effectors of viral inflammation using microparticles can alter the outcome of disease by facilitating the accumulation of macrophage subsets associated with tissue repair.

ABSTRACT

In the absence of a vaccine, multidrug-resistant Neisseria gonorrhoeae has emerged as a major human health threat, and new approaches to treat gonorrhea are urgently needed. N. gonorrhoeae pili are posttranslationally modified by a glycan that terminates in a galactose. The terminal galactose is critical for initial contact with the human cervical mucosa via an interaction with the I-domain of complement receptor 3 (CR3). We have now identified the I-domain galactose-binding epitope and characterized its galactose-specific lectin activity. Using surface plasmon resonance and cellular infection assays, we found that a peptide mimic of this galactose-binding region competitively inhibited the N. gonorrhoeae-CR3 interaction. A compound library was screened for potential drugs that could similarly prohibit the N. gonorrhoeae-CR3 interaction and be repurposed as novel host-targeted therapeutics for multidrug-resistant gonococcal infections in women. Two drugs, methyldopa and carbamazepine, prevented and cured cervical cell infection by multidrug-resistant gonococci by blocking the gonococcal-CR3 I-domain interaction.

IMPORTANCE Novel therapies that avert the problem of Neisseria gonorrhoeae with acquired antibiotic resistance are urgently needed. Gonococcal infection of the human cervix is initiated by an interaction between a galactose modification made to its surface appendages, pili, and the I-domain region of (host) complement receptor 3 (CR3). By targeting this crucial gonococcal–I-domain interaction, it may be possible to prevent cervical infection in females. To this end, we identified the I-domain galactose-binding epitope of CR3 and characterized its galactose lectin activity. Moreover, we identified two drugs, carbamazepine and methyldopa, as effective host-targeted therapies for gonorrhea treatment. At doses below those currently used for their respective existing indications, both carbamazepine and methyldopa were more effective than ceftriaxone in curing cervical infection ex vivo. This host-targeted approach would not be subject to N. gonorrhoeae drug resistance mechanisms. Thus, our data suggest a long-term solution to the growing problem of multidrug-resistant N. gonorrhoeae infections.

ABSTRACT

Hantaviruses are the etiological agent of hemorrhagic fever with renal syndrome (HFRS) and hantavirus cardiopulmonary syndrome (HCPS). The latter is associated with case fatality rates ranging from 30% to 50%. HCPS cases are rare, with approximately 300 recorded annually in the Americas. Recently, an HCPS outbreak of unprecedented size has been occurring in and around Epuyén, in the southwestern Argentinian state of Chubut. Since November of 2018, at least 29 cases have been laboratory confirmed, and human-to-human transmission is suspected. Despite posing a significant threat to public health, no treatment or vaccine is available for hantaviral disease. Here, we describe an effort to identify, characterize, and develop neutralizing and protective antibodies against the glycoprotein complex (Gn and Gc) of Andes virus (ANDV), the causative agent of the Epuyén outbreak. Using murine hybridoma technology, we generated 19 distinct monoclonal antibodies (MAbs) against ANDV GnGc. When tested for neutralization against a recombinant vesicular stomatitis virus expressing the Andes glycoprotein (GP) (VSV-ANDV), 12 MAbs showed potent neutralization and 8 showed activity in an antibody-dependent cellular cytotoxicity reporter assay. Escape mutant analysis revealed that neutralizing MAbs targeted both the Gn and the Gc. Four MAbs that bound different epitopes were selected for preclinical studies and were found to be 100% protective against lethality in a Syrian hamster model of ANDV infection. These data suggest the existence of a wide array of neutralizing antibody epitopes on hantavirus GnGc with unique properties and mechanisms of action.

IMPORTANCE Infections with New World hantaviruses are associated with high case fatality rates, and no specific vaccine or treatment options exist. Furthermore, the biology of the hantaviral GnGc complex, its antigenicity, and its fusion machinery are poorly understood. Protective monoclonal antibodies against GnGc have the potential to be developed into therapeutics against hantaviral disease and are also great tools to elucidate the biology of the glycoprotein complex.

Objective

To investigate the pathogenicity of the TGM6 variant for spinocerebellar ataxia 35 (SCA35), which was previously reported to be caused by pathogenic mutations in the gene TGM6.

The lignin biosynthetic pathway is highly conserved in angiosperms, yet pathway manipulations give rise to a variety of taxon-specific outcomes. Knockout of lignin-associated 4-coumarate:CoA ligases (4CLs) in herbaceous species mainly reduces guaiacyl (G) lignin and enhances cell wall saccharification. Here we show that CRISPR-knockout of 4CL1 in poplar (Populus tremula x alba) preferentially reduced syringyl (S) lignin, with negligible effects on biomass recalcitrance. Concordant with reduced S-lignin was downregulation of ferulate 5-hydroxylases (F5Hs). Lignification was largely sustained by 4CL5, a low-affinity paralog of 4CL1 typically with only minor xylem expression or activity. Levels of caffeate, the preferred substrate of 4CL5, increased in line with significant upregulation of caffeoyl shikimate esterase1. Upregulation of caffeoyl-CoA O-methyltransferase1 and downregulation of F5Hs are consistent with preferential funneling of 4CL5 products toward G-lignin biosynthesis at the expense of S-lignin. Thus, transcriptional and metabolic adaptations to 4CL1-knockout appear to have enabled 4CL5 catalysis at a level sufficient to sustain lignification. Finally, genes involved in sulfur assimilation, the glutathione-ascorbate cycle, and various antioxidant systems were upregulated in the mutants, suggesting cascading responses to perturbed thioesterification in lignin biosynthesis.