Norovirus Detection and Genotyping for Children with Gastroenteritis, Brazil

During 1998–2005, we analyzed stool samples from 289 children in Rio de Janeiro to detect and genotype norovirus strains. Previous tests showed all samples to be negative for rotavirus and adenovirus. Of 42 (14.5%) norovirus-positive specimens, 20 (47.6%) were identified as genogroup GI and 22 (52.3%) as GII

TOI-2015b: A Warm Neptune with Transit Timing Variations Orbiting an Active mid M Dwarf

We report the discovery of a close-in ($P_{\mathrm{orb}} = 3.349\:\mathrm{days}$) warm Neptune with clear transit timing variations (TTVs) orbiting the nearby ($d=47.3\:\mathrm{pc}$) active M4 star, TOI-2015. We characterize the planet's properties using TESS photometry, precise near-infrared radial velocities (RV) with the Habitable-zone Planet Finder (HP) Spectrograph, ground-based photometry, and high-contrast imaging. A joint photometry and RV fit yields a radius $R_p~=~3.37_{-0.20}^{+0.15} \:\mathrm{R_\oplus}$, mass $m_p~=~16.4_{-4.1}^{+4.1}\:\mathrm{M_\oplus}$, and density $\rho_p~=~2.32_{-0.37}^{+0.38} \:\mathrm{g cm^{-3}}$ for TOI-2015b, suggesting a likely volatile-rich planet. The young, active host star has a rotation period of $P_{\mathrm{rot}}~=~8.7 \pm~0.9~\mathrm{days}$ and associated rotation-based age estimate of $1.1~\pm~0.1\:\mathrm{Gyr}$. Though no other transiting planets are seen in the TESS data, the system shows clear TTVs of super period $P_{\mathrm{sup}}~\approx~430\:\mathrm{days}$ and amplitude $\sim$$100\:\mathrm{minutes}$. After considering multiple likely period ratio models, we show an outer planet candidate near a 2:1 resonance can explain the observed TTVs while offering a dynamically stable solution. However, other possible two-planet solutions -- including 3:2 and 4:3 resonance -- cannot be conclusively excluded without further observations. Assuming a 2:1 resonance in the joint TTV-RV modeling suggests a mass of $m_b~=~13.3_{-4.5}^{+4.7}\:\mathrm{M_\oplus}$ for TOI-2015b and $m_c~=~6.8_{-2.3}^{+3.5}\:\mathrm{M_\oplus}$ for the outer candidate. Additional transit and RV observations will be beneficial to explicitly identify the resonance and further characterize the properties of the system.Comment: 28 pages, 15 figures, 6 tables. As submitted to AAS Journal

Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead

Adding 6 months of androgen deprivation therapy to postoperative radiotherapy for prostate cancer: a comparison of short-course versus no androgen deprivation therapy in the RADICALS-HD randomised controlled trial

Background Previous evidence indicates that adjuvant, short-course androgen deprivation therapy (ADT) improves metastasis-free survival when given with primary radiotherapy for intermediate-risk and high-risk localised prostate cancer. However, the value of ADT with postoperative radiotherapy after radical prostatectomy is unclear. Methods RADICALS-HD was an international randomised controlled trial to test the efficacy of ADT used in combination with postoperative radiotherapy for prostate cancer. Key eligibility criteria were indication for radiotherapy after radical prostatectomy for prostate cancer, prostate-specific antigen less than 5 ng/mL, absence of metastatic disease, and written consent. Participants were randomly assigned (1:1) to radiotherapy alone (no ADT) or radiotherapy with 6 months of ADT (short-course ADT), using monthly subcutaneous gonadotropin-releasing hormone analogue injections, daily oral bicalutamide monotherapy 150 mg, or monthly subcutaneous degarelix. Randomisation was done centrally through minimisation with a random element, stratified by Gleason score, positive margins, radiotherapy timing, planned radiotherapy schedule, and planned type of ADT, in a computerised system. The allocated treatment was not masked. The primary outcome measure was metastasis-free survival, defined as distant metastasis arising from prostate cancer or death from any cause. Standard survival analysis methods were used, accounting for randomisation stratification factors. The trial had 80% power with two-sided α of 5% to detect an absolute increase in 10-year metastasis-free survival from 80% to 86% (hazard ratio [HR] 0·67). Analyses followed the intention-to-treat principle. The trial is registered with the ISRCTN registry, ISRCTN40814031, and ClinicalTrials.gov, NCT00541047. Findings Between Nov 22, 2007, and June 29, 2015, 1480 patients (median age 66 years [IQR 61–69]) were randomly assigned to receive no ADT (n=737) or short-course ADT (n=743) in addition to postoperative radiotherapy at 121 centres in Canada, Denmark, Ireland, and the UK. With a median follow-up of 9·0 years (IQR 7·1–10·1), metastasis-free survival events were reported for 268 participants (142 in the no ADT group and 126 in the short-course ADT group; HR 0·886 [95% CI 0·688–1·140], p=0·35). 10-year metastasis-free survival was 79·2% (95% CI 75·4–82·5) in the no ADT group and 80·4% (76·6–83·6) in the short-course ADT group. Toxicity of grade 3 or higher was reported for 121 (17%) of 737 participants in the no ADT group and 100 (14%) of 743 in the short-course ADT group (p=0·15), with no treatment-related deaths. Interpretation Metastatic disease is uncommon following postoperative bed radiotherapy after radical prostatectomy. Adding 6 months of ADT to this radiotherapy did not improve metastasis-free survival compared with no ADT. These findings do not support the use of short-course ADT with postoperative radiotherapy in this patient population

Duration of androgen deprivation therapy with postoperative radiotherapy for prostate cancer: a comparison of long-course versus short-course androgen deprivation therapy in the RADICALS-HD randomised trial

Background Previous evidence supports androgen deprivation therapy (ADT) with primary radiotherapy as initial treatment for intermediate-risk and high-risk localised prostate cancer. However, the use and optimal duration of ADT with postoperative radiotherapy after radical prostatectomy remains uncertain. Methods RADICALS-HD was a randomised controlled trial of ADT duration within the RADICALS protocol. Here, we report on the comparison of short-course versus long-course ADT. Key eligibility criteria were indication for radiotherapy after previous radical prostatectomy for prostate cancer, prostate-specific antigen less than 5 ng/mL, absence of metastatic disease, and written consent. Participants were randomly assigned (1:1) to add 6 months of ADT (short-course ADT) or 24 months of ADT (long-course ADT) to radiotherapy, using subcutaneous gonadotrophin-releasing hormone analogue (monthly in the short-course ADT group and 3-monthly in the long-course ADT group), daily oral bicalutamide monotherapy 150 mg, or monthly subcutaneous degarelix. Randomisation was done centrally through minimisation with a random element, stratified by Gleason score, positive margins, radiotherapy timing, planned radiotherapy schedule, and planned type of ADT, in a computerised system. The allocated treatment was not masked. The primary outcome measure was metastasis-free survival, defined as metastasis arising from prostate cancer or death from any cause. The comparison had more than 80% power with two-sided α of 5% to detect an absolute increase in 10-year metastasis-free survival from 75% to 81% (hazard ratio [HR] 0·72). Standard time-to-event analyses were used. Analyses followed intention-to-treat principle. The trial is registered with the ISRCTN registry, ISRCTN40814031, and ClinicalTrials.gov , NCT00541047 . Findings Between Jan 30, 2008, and July 7, 2015, 1523 patients (median age 65 years, IQR 60–69) were randomly assigned to receive short-course ADT (n=761) or long-course ADT (n=762) in addition to postoperative radiotherapy at 138 centres in Canada, Denmark, Ireland, and the UK. With a median follow-up of 8·9 years (7·0–10·0), 313 metastasis-free survival events were reported overall (174 in the short-course ADT group and 139 in the long-course ADT group; HR 0·773 [95% CI 0·612–0·975]; p=0·029). 10-year metastasis-free survival was 71·9% (95% CI 67·6–75·7) in the short-course ADT group and 78·1% (74·2–81·5) in the long-course ADT group. Toxicity of grade 3 or higher was reported for 105 (14%) of 753 participants in the short-course ADT group and 142 (19%) of 757 participants in the long-course ADT group (p=0·025), with no treatment-related deaths. Interpretation Compared with adding 6 months of ADT, adding 24 months of ADT improved metastasis-free survival in people receiving postoperative radiotherapy. For individuals who can accept the additional duration of adverse effects, long-course ADT should be offered with postoperative radiotherapy. Funding Cancer Research UK, UK Research and Innovation (formerly Medical Research Council), and Canadian Cancer Society

Effect of angiotensin-converting enzyme inhibitor and angiotensin receptor blocker initiation on organ support-free days in patients hospitalized with COVID-19

IMPORTANCE Overactivation of the renin-angiotensin system (RAS) may contribute to poor clinical outcomes in patients with COVID-19. Objective To determine whether angiotensin-converting enzyme (ACE) inhibitor or angiotensin receptor blocker (ARB) initiation improves outcomes in patients hospitalized for COVID-19. DESIGN, SETTING, AND PARTICIPANTS In an ongoing, adaptive platform randomized clinical trial, 721 critically ill and 58 non–critically ill hospitalized adults were randomized to receive an RAS inhibitor or control between March 16, 2021, and February 25, 2022, at 69 sites in 7 countries (final follow-up on June 1, 2022). INTERVENTIONS Patients were randomized to receive open-label initiation of an ACE inhibitor (n = 257), ARB (n = 248), ARB in combination with DMX-200 (a chemokine receptor-2 inhibitor; n = 10), or no RAS inhibitor (control; n = 264) for up to 10 days. MAIN OUTCOMES AND MEASURES The primary outcome was organ support–free days, a composite of hospital survival and days alive without cardiovascular or respiratory organ support through 21 days. The primary analysis was a bayesian cumulative logistic model. Odds ratios (ORs) greater than 1 represent improved outcomes. RESULTS On February 25, 2022, enrollment was discontinued due to safety concerns. Among 679 critically ill patients with available primary outcome data, the median age was 56 years and 239 participants (35.2%) were women. Median (IQR) organ support–free days among critically ill patients was 10 (–1 to 16) in the ACE inhibitor group (n = 231), 8 (–1 to 17) in the ARB group (n = 217), and 12 (0 to 17) in the control group (n = 231) (median adjusted odds ratios of 0.77 [95% bayesian credible interval, 0.58-1.06] for improvement for ACE inhibitor and 0.76 [95% credible interval, 0.56-1.05] for ARB compared with control). The posterior probabilities that ACE inhibitors and ARBs worsened organ support–free days compared with control were 94.9% and 95.4%, respectively. Hospital survival occurred in 166 of 231 critically ill participants (71.9%) in the ACE inhibitor group, 152 of 217 (70.0%) in the ARB group, and 182 of 231 (78.8%) in the control group (posterior probabilities that ACE inhibitor and ARB worsened hospital survival compared with control were 95.3% and 98.1%, respectively). CONCLUSIONS AND RELEVANCE In this trial, among critically ill adults with COVID-19, initiation of an ACE inhibitor or ARB did not improve, and likely worsened, clinical outcomes. TRIAL REGISTRATION ClinicalTrials.gov Identifier: NCT0273570

Regulation of viral reactivation and cell death by inflammatory cytokine signaling during murine gammaherpesvirus infection

Gammaherpesviruses (γHV) cause multiple cancers in immunocompromised individuals. Due to the species\u27 specificity of human γHV, which include Epstein-Barr and Kaposi\u27s sarcoma herpesviruses, we use murine gammaherpesvirus 68 (MHV68) as an animal model to study these viruses in each stage of the viral life cycle. The goal of this research is to define the role of cytokines involved in the innate immune response in control of lytic infection and reactivation from latency. One of these cytokines is TNFα, which is involved in death receptor signaling. To test the hypothesis that MHV68 regulates TNFα-induced cell death, we treated infected 3T3 cells with TNFα either alone or in combination with cycloheximide (CHX). In uninfected cells, TNFα simultaneously activates pro-survival and apoptotic signaling pathways, resulting in cell activation but not cell death. CHX blocks NF-κB-dependent synthesis of pro-survival proteins, causing apoptosis when administered with TNFα. We found that cells infected with MHV68 for 8 hours underwent rapid TNFα-induced cell death even in the absence of CHX. We also found that a replication-competent virus is necessary for the cell death, suggesting that pro-survival pathways are blocked by early steps in MHV68 infection. We show that anti-apoptotic pathways are intact by demonstrating that the NF-κB subunit p65 translocates to the nucleus and functions as a transcription factor. We have also identified a modification of the anti-apoptotic protein cellular FLICE-like inhibitory protein Raji (cFLIPR) during MHV68 infection, which, we hypothesize, predisposes the infected cells to death upon TNFα treatment in a novel way. To date, no other viruses have been shown to modify cFLIP to induce cell death in this manner. Another innate immune response cytokine known to slow the spread of infection is IFNαβ. We found that mice lacking receptors for IFNαβ (Ifnar1-/-) were able to clear MHV68 infection but experienced a significant increase in reactivation of latent virus ex vivo. To determine which specific cell types respond to IFNαβ to control reactivation in vivo, we used tissue-specific Cre-recombinase-transgenic mice to delete Ifnar1 only in macrophages. Macrophages are a known latency reservoir for MHV68, and this technique allowed us to target the role of macrophages and only in macrophages. Macrophages are a known latency reservoir for MHV68, and this technique allowed us to target the role of macrophages and IFNαβ signaling during latency. Macrophage-specific Ifnar1-/- mice were infected with MHV68 and viral reactivation assessed at 16 days post-infection. We found that removal of IFNAR1 function from macrophages causes a 10-fold increase in reactivation over wild-type in peritoneal cells, as well as a 2.5-fold increase in splenocytes. These data indicates an essential role for IFNαβ signaling in macrophages to control MHV68 reactivation. Determining the role of IFNαβ signaling and the specific cell types that are important for controlling γHV reactivation from latency, as well as understanding how MHV68 modulates cell death pathways, will help guide future research into therapeutics for these diseases

TOI-2015 b: A Warm Neptune with Transit Timing Variations Orbiting an Active Mid-type M Dwarf

We report the discovery of a close-in ( P _orb = 3.349 days) warm Neptune with clear transit timing variations (TTVs) orbiting the nearby ( d = 47.3 pc) active M4 star, TOI-2015. We characterize the planet's properties using Transiting Exoplanet Survey Satellite (TESS) photometry, precise near-infrared radial velocities (RVs) with the Habitable-zone Planet Finder Spectrograph, ground-based photometry, and high-contrast imaging. A joint photometry and RV fit yields a radius ${R}_{p}={3.37}_{-0.20}^{+0.15}\ {R}_{\oplus }$ , mass ${m}_{p}={16.4}_{-4.1}^{+4.1}\ {M}_{\oplus }$ , and density ${\rho }_{p}\,={2.32}_{-0.37}^{+0.38}\ {\rm{g}}\ {\mathrm{cm}}^{-3}$ for TOI-2015 b, suggesting a likely volatile-rich planet. The young, active host star has a rotation period of P _rot = 8.7 ± 0.9 days and associated rotation-based age estimate of 1.1 ± 0.1 Gyr. Though no other transiting planets are seen in the TESS data, the system shows clear TTVs of super-period ${P}_{\sup }\approx 430\ \mathrm{days}$ and amplitude ∼100 minutes. After considering multiple likely period-ratio models, we show an outer planet candidate near a 2:1 resonance can explain the observed TTVs while offering a dynamically stable solution. However, other possible two-planet solutions—including 3:2 and 4:3 resonances—cannot be conclusively excluded without further observations. Assuming a 2:1 resonance in the joint TTV-RV modeling suggests a mass of ${m}_{b}={13.3}_{-4.5}^{+4.7}\ {M}_{\oplus }$ for TOI-2015 b and ${m}_{c}={6.8}_{-2.3}^{+3.5}\ {M}_{\oplus }$ for the outer candidate. Additional transit and RV observations will be beneficial to explicitly identify the resonance and further characterize the properties of the system

Extreme drought impacts have been underestimated in grasslands and shrublands globally.

Climate change is increasing the frequency and severity of short-term (~1 y) drought events-the most common duration of drought-globally. Yet the impact of this intensification of drought on ecosystem functioning remains poorly resolved. This is due in part to the widely disparate approaches ecologists have employed to study drought, variation in the severity and duration of drought studied, and differences among ecosystems in vegetation, edaphic and climatic attributes that can mediate drought impacts. To overcome these problems and better identify the factors that modulate drought responses, we used a coordinated distributed experiment to quantify the impact of short-term drought on grassland and shrubland ecosystems. With a standardized approach, we imposed ~a single year of drought at 100 sites on six continents. Here we show that loss of a foundational ecosystem function-aboveground net primary production (ANPP)-was 60% greater at sites that experienced statistically extreme drought (1-in-100-y event) vs. those sites where drought was nominal (historically more common) in magnitude (35% vs. 21%, respectively). This reduction in a key carbon cycle process with a single year of extreme drought greatly exceeds previously reported losses for grasslands and shrublands. Our global experiment also revealed high variability in drought response but that relative reductions in ANPP were greater in drier ecosystems and those with fewer plant species. Overall, our results demonstrate with unprecedented rigor that the global impacts of projected increases in drought severity have been significantly underestimated and that drier and less diverse sites are likely to be most vulnerable to extreme drought

Extreme drought impacts have been underestimated in grasslands and shrublands globally

Climate change is increasing the frequency and severity of short-term (~1 y) drought events—the most common duration of drought—globally. Yet the impact of this intensification of drought on ecosystem functioning remains poorly resolved. This is due in part to the widely disparate approaches ecologists have employed to study drought, variation in the severity and duration of drought studied, and differences among ecosystems in vegetation, edaphic and climatic attributes that can mediate drought impacts. To overcome these problems and better identify the factors that modulate drought responses, we used a coordinated distributed experiment to quantify the impact of short-term drought on grassland and shrubland ecosystems. With a standardized approach, we imposed ~a single year of drought at 100 sites on six continents. Here we show that loss of a foundational ecosystem function—aboveground net primary production (ANPP)—was 60% greater at sites that experienced statistically extreme drought (1-in-100-y event) vs. those sites where drought was nominal (historically more common) in magnitude (35% vs. 21%, respectively). This reduction in a key carbon cycle process with a single year of extreme drought greatly exceeds previously reported losses for grasslands and shrublands. Our global experiment also revealed high variability in drought response but that relative reductions in ANPP were greater in drier ecosystems and those with fewer plant species. Overall, our results demonstrate with unprecedented rigor that the global impacts of projected increases in drought severity have been significantly underestimated and that drier and less diverse sites are likely to be most vulnerable to extreme drought