Viral DNAemia and DNA virus seropositivity and mortality in pediatric sepsis

IMPORTANCE: Sepsis is a leading cause of pediatric mortality. Little attention has been paid to the association between viral DNA and mortality in children and adolescents with sepsis. OBJECTIVE: To assess the association of the presence of viral DNA with sepsis-related mortality in a large multicenter study. DESIGN, SETTING, AND PARTICIPANTS: This cohort study compares pediatric patients with and without plasma cytomegalovirus (CMV), Epstein-Barr virus (EBV), herpes simplex virus 1 (HSV-1), human herpesvirus 6 (HHV-6), parvovirus B19 (B19V), BK polyomavirus (BKPyV), human adenovirus (HAdV), and torque teno virus (TTV) DNAemia detected by quantitative real-time polymerase chain reaction or plasma IgG antibodies to CMV, EBV, HSV-1, or HHV-6. A total of 401 patients younger than 18 years with severe sepsis were enrolled from 9 pediatric intensive care units (PICUs) in the Eunice Kennedy Shriver National Institute of Child Health and Human Development Collaborative Pediatric Critical Care Research Network. Data were collected from 2015 to 2018. Samples were assayed from 2019 to 2022. Data were analyzed from 2022 to 2023. MAIN OUTCOMES AND MEASURES: Death while in the PICU. RESULTS: Among the 401 patients included in the analysis, the median age was 6 (IQR, 1-12) years, and 222 (55.4%) were male. One hundred fifty-four patients (38.4%) were previously healthy, 108 (26.9%) were immunocompromised, and 225 (56.1%) had documented infection(s) at enrollment. Forty-four patients (11.0%) died in the PICU. Viral DNAemia with at least 1 virus (excluding TTV) was detected in 191 patients (47.6%) overall, 63 of 108 patients (58.3%) who were immunocompromised, and 128 of 293 (43.7%) who were not immunocompromised at sepsis onset. After adjustment for age, Pediatric Risk of Mortality score, previously healthy status, and immunocompromised status at sepsis onset, CMV (adjusted odds ratio [AOR], 3.01 [95% CI, 1.36-6.45]; P = .007), HAdV (AOR, 3.50 [95% CI, 1.46-8.09]; P = .006), BKPyV (AOR. 3.02 [95% CI, 1.17-7.34]; P = .02), and HHV-6 (AOR, 2.62 [95% CI, 1.31-5.20]; P = .007) DNAemia were each associated with increased mortality. Two or more viruses were detected in 78 patients (19.5%), with mortality among 12 of 32 (37.5%) who were immunocompromised and 9 of 46 (19.6%) who were not immunocompromised at sepsis onset. Herpesvirus seropositivity was common (HSV-1, 82 of 246 [33.3%]; CMV, 107 of 254 [42.1%]; EBV, 152 of 251 [60.6%]; HHV-6, 253 if 257 [98.4%]). After additional adjustment for receipt of blood products in the PICU, EBV seropositivity was associated with increased mortality (AOR, 6.10 [95% CI, 1.00-118.61]; P = .049). CONCLUSIONS AND RELEVANCE: The findings of this cohort study suggest that DNAemia for CMV, HAdV, BKPyV, and HHV-6 and EBV seropositivity were independently associated with increased sepsis mortality. Further investigation of the underlying biology of these viral DNA infections in children with sepsis is warranted to determine whether they only reflect mortality risk or contribute to mortality

Interleukin-7 restores lymphocytes in septic shock: The IRIS-7 randomized clinical trial

BACKGROUND: A defining pathophysiologic feature of sepsis is profound apoptosis-induced death and depletion of CD4+ and CD8+ T cells. Interleukin-7 (IL-7) is an antiapoptotic common γ-chain cytokine that is essential for lymphocyte proliferation and survival. Clinical trials of IL-7 in over 390 oncologic and lymphopenic patients showed that IL-7 was safe, invariably increased CD4+ and CD8+ lymphocyte counts, and improved immunity. METHODS: We conducted a prospective, randomized, double-blind, placebo-controlled trial of recombinant human IL-7 (CYT107) in patients with septic shock and severe lymphopenia. Twenty-seven patients at academic sites in France and the United States received CYT107 or placebo for 4 weeks. Primary aims were to determine the safety of CYT107 in sepsis and its ability to reverse lymphopenia. RESULTS: CYT107 was well tolerated without evidence of inducing cytokine storm or worsening inflammation or organ dysfunction. CYT107 caused a 3- to 4-fold increase in absolute lymphocyte counts and in circulating CD4+ and CD8+ T cells that persisted for weeks after drug administration. CYT107 also increased T cell proliferation and activation. CONCLUSIONS: This is the first trial of an immunoadjuvant therapy targeting defects in adaptive immunity in patients with sepsis. CYT107 reversed the marked loss of CD4+ and CD8+ immune effector cells, a hallmark of sepsis and a likely key mechanism in its morbidity and mortality. CYT107 represents a potential new way forward in the treatment of patients with sepsis by restoring adaptive immunity. Such immune-based therapy should be broadly protective against diverse pathogens including multidrug resistant bacteria that preferentially target patients with impaired immunity. TRIAL REGISTRATION: Trials registered at clinicaltrials.gov: NCT02640807 and NCT02797431. FUNDING: Revimmune, NIH National Institute of General Medical Sciences GM44118

Comparison of monocyte human leukocyte antigen-DR expression and stimulated tumor necrosis factor alpha production as outcome predictors in severe sepsis: A prospective observational study

BACKGROUND: Identifying patients in the immunosuppressive phase of sepsis is essential for development of immunomodulatory therapies. Little data exists comparing the ability of the two most well-studied markers of sepsis-induced immunosuppression, human leukocyte antigen (HLA)-DR expression and lipopolysaccharide (LPS)-induced tumor necrosis factor alpha (TNF-ɑ) production, to predict mortality and morbidity. The purpose of this study was to compare HLA-DR expression and LPS-induced TNF-ɑ production as predictors of 28-day mortality and acquisition of secondary infections in adult septic patients. METHODS: A single-center, prospective observational study of 83 adult septic patients admitted to a medical or surgical intensive care unit. Blood samples were collected at three time points during the septic course (days 1–2, days 3–4, and days 6–8 after sepsis diagnosis) and assayed for HLA-DR expression and LPS-induced TNF-ɑ production. A repeated measures mixed model analysis was used to compare values of these immunological markers among survivors and non-survivors and among those who did and did not develop a secondary infection. RESULTS: Twenty-five patients (30.1 %) died within 28 days of sepsis diagnosis. HLA-DR expression was significantly lower in non-survivors as compared to survivors on days 3–4 (p = 0.04) and days 6–8 (p = 0.002). The change in HLA-DR from days 1–2 to days 6–8 was also lower in non-survivors (p = 0.04). Median HLA-DR expression decreased from days 1–2 to days 3–4 in patients who developed secondary infections while it increased in those without secondary infections (p = 0.054). TNF-ɑ production did not differ between survivors and non-survivors or between patients who did and did not develop a secondary infection. CONCLUSIONS: Monocyte HLA-DR expression may be a more accurate predictor of mortality and acquisition of secondary infections than LPS-stimulated TNF-ɑ production in adult medical and surgical critically ill patients. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13054-016-1505-0) contains supplementary material, which is available to authorized users

Modification and characterisation of material hydrophobicity for surface acoustic wave driven microfluidics

Surface acoustic waves (SAW) generated in a piezoelectric substrate may be used to manipulate micro-scale droplets of liquid in a digital microfluidic system for lab-on-a-chip applications. The wettability of the surface over which a droplet is driven determines the ease and speed with which the droplet is propelled. This provides the opportunity to achieve fine control of SAW driven droplets simply by patterning of the surface into areas with different levels of wettability. This paper evaluates a number of different materials and surface preparation techniques and assesses their manufacturability and efficacy for this application. Test structures have been designed and developed to help optimise a fabrication process using the biocompatible polymer Parylene. Early results obtained using airflow as a driving force show that it is possible to manipulate droplets through direction changes of up to 60°. Additional work has been done using surface acoustic waves as the driving force to determine the extent to which droplets can be guided to desired locations

Adverse outcomes and an immunosuppressed endotype in septic patients with reduced IFN-γ ELISpot

BACKGROUNDSepsis remains a major clinical challenge for which successful treatment requires greater precision in identifying patients at increased risk of adverse outcomes requiring different therapeutic approaches. Predicting clinical outcomes and immunological endotyping of septic patients generally relies on using blood protein or mRNA biomarkers, or static cell phenotyping. Here, we sought to determine whether functional immune responsiveness would yield improved precision.METHODSAn ex vivo whole-blood enzyme-linked immunosorbent spot (ELISpot) assay for cellular production of interferon γ (IFN-γ) was evaluated in 107 septic and 68 nonseptic patients from 5 academic health centers using blood samples collected on days 1, 4, and 7 following ICU admission.RESULTSCompared with 46 healthy participants, unstimulated and stimulated whole-blood IFN-γ expression was either increased or unchanged, respectively, in septic and nonseptic ICU patients. However, in septic patients who did not survive 180 days, stimulated whole-blood IFN-γ expression was significantly reduced on ICU days 1, 4, and 7 (all P \u3c 0.05), due to both significant reductions in total number of IFN-γ-producing cells and amount of IFN-γ produced per cell (all P \u3c 0.05). Importantly, IFN-γ total expression on days 1 and 4 after admission could discriminate 180-day mortality better than absolute lymphocyte count (ALC), IL-6, and procalcitonin. Septic patients with low IFN-γ expression were older and had lower ALCs and higher soluble PD-L1 and IL-10 concentrations, consistent with an immunosuppressed endotype.CONCLUSIONSA whole-blood IFN-γ ELISpot assay can both identify septic patients at increased risk of late mortality and identify immunosuppressed septic patients.TRIAL REGISTRYN/A.FUNDINGThis prospective, observational, multicenter clinical study was directly supported by National Institute of General Medical Sciences grant R01 GM-139046, including a supplement (R01 GM-139046-03S1) from 2022 to 2024

Burst mitofusin activation reverses neuromuscular dysfunction in murine CMT2A

Charcot-Marie-Tooth disease type 2A (CMT2A) is an untreatable childhood peripheral neuropathy caused by mutations of the mitochondrial fusion protein, mitofusin (MFN) 2. Here, pharmacological activation of endogenous normal mitofusins overcame dominant inhibitory effects of CMT2A mutants in reprogrammed human patient motor neurons, reversing hallmark mitochondrial stasis and fragmentation independent of causa

A DNA nanoswitch incorporating the fluorescent base analogue 2-aminopurine detects single nucleotide mismatches in unlabelled targets

DNA nanoswitches can be designed to detect unlabelled nucleic acid targets and have been shown to discriminate between targets which differ in the identity of only one base. This paper demonstrates that the fluorescent base analogue 2-aminopurine (AP) can be used to discriminate between nanoswitches with and without targets and to discriminate between matched and mismatched targets. In particular, we have used both steady-state and time-resolved fluorescence spectroscopy to determine differences in AP environment at the branchpoint of nanoswitches assembled using complementary targets and targets which incorporate single base mismatches

Epigenome-wide change and variation in DNA methylation in childhood:Trajectories from birth to late adolescence

DNA methylation (DNAm) is known to play a pivotal role in childhood health and development, but a comprehensive characterization of genome-wide DNAm trajectories across this age period is currently lacking. We have therefore performed a series of epigenome-wide association studies in 5019 blood samples collected at multiple time-points from birth to late adolescence from 2348 participants of two large independent cohorts. DNAm profiles of autosomal CpG sites (CpGs) were generated using the Illumina Infinium HumanMethylation450 BeadChip. Change over time was widespread, observed at over one-half (53%) of CpGs. In most cases, DNAm was decreasing (36% of CpGs). Inter-individual variation in linear trajectories was similarly widespread (27% of CpGs). Evidence for non-linear change and inter-individual variation in non-linear trajectories was somewhat less common (11 and 8% of CpGs, respectively). Very little inter-individual variation in change was explained by sex differences (0.4% of CpGs) even though sex-specific DNAm was observed at 5% of CpGs. DNAm trajectories were distributed non-randomly across the genome. For example, CpGs with decreasing DNAm were enriched in gene bodies and enhancers and were annotated to genes enriched in immune-developmental functions. In contrast, CpGs with increasing DNAm were enriched in promoter regions and annotated to genes enriched in neurodevelopmental functions. These findings depict a methylome undergoing widespread and often non-linear change throughout childhood. They support a developmental role for DNA methylation that extends beyond birth into late adolescence and has implications for understanding life-long health and disease. DNAm trajectories can be visualized at http://epidelta.mrcieu.ac.uk

Reply to: New Meta- and Mega-analyses of Magnetic Resonance Imaging Findings in Schizophrenia: Do They Really Increase Our Knowledge About the Nature of the Disease Process?

This work was supported by National Institute of Biomedical Imaging and Bioengineering Grant No. U54EB020403 (to the ENIGMA consortium)

Crop Updates 1999 - Oilseeds

This article contains eighteen papers INTRODUCTION, Paul Carmody, Agriculture Western Australia PLENARY SESSION Transgenic canola in Western Australia: Outlook and challenges, Phil Salisbury, University of Melbourne Farming system issues for herbicide tolerant canola, Rick Madin, Rick Madin and Associates, David Bowran, Agriculture Western Australia Beating blackleg in 1999, Martin Barbetti, Ravjit Khangura, Paul Carmody, Graham Walton, Agriculture Western Australia The Mustard Industry in Australia – Opportunities for a new oilseed, Phil Parker, NSW Agriculture Management of blackleg with fungicides, Ravjit Khangura and Martin Barbetti, Agriculture Western Australia Effect of aphid feeding damage on canola yields in 1998, Francoise Berlandier and Linnet Cartwright, Agriculture Western Australia Post-anthesis duration and rainfall affect oil content of canola, Ping Si, University of West Australia, Graham Walton, Agriculture Western Australia, Nick Galwey and David Turner, University of West Australia Canola responded to high rates of fertiliser in 1998, Wayne Pluske, CSBP Impact of agronomic practices on canola quality, Graham Walton, Agriculture Western Australia Survey reveals widespread infection with two virus diseases in Western Australian canola crops, Roger Jones and Brenda Coutts, Agriculture Western Australia Calculating canola yields and oil contents as a function of soil and fertiliser nitrogen supply, Bill Bowden and Isabel Arevalo-Vigne, Agriculture Western Australia Canola benchmarks 1997/98 – Central Eastern District, Jeff Russell, Agriculture Western Australia Seeding rate affects the yield and some architectural features of canola, Syed H. Zaheer, Nick W. Galwey and David Turner, University of Western Australia Foliar applied fungicides for blackleg, Andrew Simon and Art Diggle, Agriculture Western Australia Farm based demonstration 1998 canola N – Wheel evaluation, Jeff Russell, Agriculture Western Australia Effect of sowing date on seed yield of canola, Dave Eksteen, Agriculture Western Australia Investigating water use of summer crops on the South Coast of Western Australia, Arjen Ryder, Agriculture Western Australia, Bill Crabtree, Western Australia No Till Farming Association, Serena Wyatt, Catchment Landcare Coordinator, Wellstead, Jim Baily, Subasio Downs, Wellstead INTRODUCTION, Paul Carmody, Agriculture Western Australia PLENARY SESSION Transgenic canola in Western Australia: Outlook and challenges, Phil Salisbury, University of Melbourne Farming system issues for herbicide tolerant canola, Rick Madin, Rick Madin and Associates, David Bowran, Agriculture Western Australia Beating blackleg in 1999, Martin Barbetti, Ravjit Khangura, Paul Carmody, Graham Walton, Agriculture Western Australia The Mustard Industry in Australia – Opportunities for a new oilseed, Phil Parker, NSW Agriculture Management of blackleg with fungicides, Ravjit Khangura and Martin Barbetti, Agriculture Western Australia Effect of aphid feeding damage on canola yields in 1998, Francoise Berlandier and Linnet Cartwright, Agriculture Western Australia Post-anthesis duration and rainfall affect oil content of canola, Ping Si, University of West Australia, Graham Walton, Agriculture Western Australia, Nick Galwey and David Turner, University of West Australia Canola responded to high rates of fertiliser in 1998, Wayne Pluske, CSBP Impact of agronomic practices on canola quality, Graham Walton, Agriculture Western Australia Survey reveals widespread infection with two virus diseases in Western Australian canola crops, Roger Jones and Brenda Coutts, Agriculture Western Australia Calculating canola yields and oil contents as a function of soil and fertiliser nitrogen supply, Bill Bowden and Isabel Arevalo-Vigne, Agriculture Western Australia Canola benchmarks 1997/98 – Central Eastern District, Jeff Russell, Agriculture Western Australi