Incidence and outcomes of neonatal acute kidney injury (AWAKEN): a multicentre, multinational, observational cohort study

Background: Single-center studies suggest that neonatal acute kidney injury (AKI) is associated with poor outcomes. However, inferences regarding the association between AKI, mortality, and hospital length of stay are limited due to the small sample size of those studies. In order to determine whether neonatal AKI is independently associated with increased mortality and longer hospital stay, we analyzed the Assessment of Worldwide Acute Kidney Epidemiology in Neonates (AWAKEN) database. Methods: All neonates admitted to 24 participating neonatal intensive care units from four countries (Australia, Canada, India, United States) between January 1 and March 31, 2014, were screened. Of 4273 neonates screened, 2022 (47·3%) met study criteria. Exclusion criteria included: no intravenous fluids ≥48 hours, admission ≥14 days of life, congenital heart disease requiring surgical repair at <7 days of life, lethal chromosomal anomaly, death within 48 hours, inability to determine AKI status or severe congenital kidney abnormalities. AKI was defined using a standardized definition -i.e., serum creatinine rise of ≥0.3 mg/dL (26.5 mcmol/L) or ≥50% from previous lowest value, and/or if urine output was <1 mL/kg/h on postnatal days 2 to 7. Findings: Incidence of AKI was 605/2022 (29·9%). Rates varied by gestational age groups (i.e., ≥22 to <29 weeks =47·9%; ≥29 to <36 weeks =18·3%; and ≥36 weeks =36·7%). Even after adjusting for multiple potential confounding factors, infants with AKI had higher mortality compared to those without AKI [(59/605 (9·7%) vs. 20/1417 (1·4%); p< 0.001; adjusted OR=4·6 (95% CI=2·5-8·3); p=<0·0001], and longer hospital stay [adjusted parameter estimate 8·8 days (95% CI=6·1-11·5); p<0·0001]. Interpretation: Neonatal AKI is a common and independent risk factor for mortality and longer hospital stay. These data suggest that neonates may be impacted by AKI in a manner similar to pediatric and adult patients

Rapid Cloning of Novel Rhesus Adenoviral Vaccine Vectors

ABSTRACT Human and chimpanzee adenovirus vectors are being developed to circumvent preexisting antibodies against common adenovirus vectors such as Ad5. However, baseline immunity to these vectors still exists in human populations. Traditional cloning of new adenovirus vaccine vectors is a long and cumbersome process that takes 2 months or more and that requires rare unique restriction enzyme sites. Here we describe a novel, restriction enzyme-independent method for rapid cloning of new adenovirus vaccine vectors that reduces the total cloning procedure to 1 week. We developed 14 novel adenovirus vectors from rhesus monkeys that can be grown to high titers and that are immunogenic in mice. All vectors grouped with the unusual adenovirus species G and show extremely low seroprevalence in humans. Rapid cloning of novel adenovirus vectors is a promising approach for the development of new vector platforms. Rhesus adenovirus vectors may prove useful for clinical development. IMPORTANCE: To overcome baseline immunity to human and chimpanzee adenovirus vectors, we developed 14 novel adenovirus vectors from rhesus monkeys. These vectors are immunogenic in mice and show extremely low seroprevalence in humans. Rhesus adenovirus vectors may prove useful for clinical development

Vaccine Protection Against Zika Virus from Brazil

Zika virus (ZIKV) is a flavivirus that is responsible for an unprecedented current epidemic in Brazil and the Americas1,2. ZIKV has been causally associated with fetal microcephaly, intrauterine growth restriction, and other birth defects in both humans3–8 and mice9–11. The rapid development of a safe and effective ZIKV vaccine is a global health priority1,2, but very little is currently known about ZIKV immunology and mechanisms of immune protection. Here we show that a single immunization of a plasmid DNA vaccine or a purified inactivated virus vaccine provides complete protection in susceptible mice against challenge with a ZIKV outbreak strain from northeast Brazil. This ZIKV strain has recently been shown to cross the placenta and to induce fetal microcephaly and other congenital malformations in mice11. We produced DNA vaccines expressing full-length ZIKV pre-membrane and envelope (prM-Env) as well as a series of deletion mutants. The full-length prM-Env DNA vaccine, but not the deletion mutants, afforded complete protection against ZIKV as measured by absence of detectable viremia following challenge, and protective efficacy correlated with Env-specific antibody titers. Adoptive transfer of purified IgG from vaccinated mice conferred passive protection, and CD4 and CD8 T lymphocyte depletion in vaccinated mice did not abrogate protective efficacy. These data demonstrate that protection against ZIKV challenge can be achieved by single-shot subunit and inactivated virus vaccines in mice and that Env-specific antibody titers represent key immunologic correlates of protection. Our findings suggest that the development of a ZIKV vaccine for humans will likely be readily achievable

Blood Glucose Levels Regulate Pancreatic β-Cell Proliferation during Experimentally-Induced and Spontaneous Autoimmune Diabetes in Mice

Type 1 diabetes mellitus is caused by immune-mediated destruction of pancreatic beta-cells leading to insulin deficiency, impaired intermediary metabolism, and elevated blood glucose concentrations. While at autoimmune diabetes onset a limited number of beta-cells persist, the cells' regenerative potential and its regulation have remained largely unexplored. Using two mouse autoimmune diabetes models, this study examined the proliferation of pancreatic islet ss-cells and other endocrine and non-endocrine subsets, and the factors regulating that proliferation.We adapted multi-parameter flow cytometry techniques (including DNA-content measurements and 5'-bromo-2'-deoxyuridine [BrdU] incorporation) to study pancreatic islet single cell suspensions. These studies demonstrate that beta-cell proliferation rapidly increases at diabetes onset, and that this proliferation is closely correlated with the diabetic animals' elevated blood glucose levels. For instance, we show that when normoglycemia is restored by exogenous insulin or islet transplantation, the beta-cell proliferation rate returns towards low levels found in control animals, yet surges when hyperglycemia recurs. In contrast, other-than-ss endocrine islet cells did not exhibit the same glucose-dependent proliferative responses. Rather, disease-associated alterations of BrdU-incorporation rates of delta-cells (minor decrease), and non-endocrine islet cells (slight increase) were not affected by blood glucose levels, or were inversely related to glycemia control after diabetes onset (alpha-cells).We conclude that murine beta-cells' ability to proliferate in response to metabolic need (i.e. rising blood glucose concentrations) is remarkably well preserved during severe, chronic beta-cell autoimmunity. These data suggest that timely control of the destructive immune response after disease manifestation could allow spontaneous regeneration of sufficient beta-cell mass to restore normal glucose homeostasis

Sesquiterpene Induction by the Balsam Woolly Adelgid (Adelges piceae) in Putatively Resistant Fraser Fir (Abies fraseri)

Fraser fir, Abies fraseri (Pursh) Poir., is a tree endemic to the Southern Appalachians and is found only in a few isolated populations at high elevations. Fraser firs are also cultivated on a commercial scale as Christmas trees. The species is imperiled by an introduced insect, the balsam woolly adelgid, Adelges piceae Ratzeburg (BWA). The insect severely damages Christmas tree crops and has caused substantial Fraser fir mortality in natural stands. Foliar terpenoids are one mechanism of host plant defense against invading insects and may be one focus of future Christmas tree breeding efforts. This study examines the correlation of foliar terpenoids with Fraser fir performance when infested with BWA. GC-MS and GC-FID analysis of artificially infested Fraser fir foliage reveals that increased concentrations of four terpenoid compounds are associated with BWA infestations. Foliar concentrations of two sesquiterpenes, camphene and humulene, are significantly higher in putatively resistant Fraser fir clones than in more susceptible clones after sustained adelgid feeding for a period of 20 weeks. Although it is unclear if the induction of these sesquiterpenes in the host fir is directly contributing to adelgid resistance, these compounds could serve as effective indicators while screening for BWA resistance in future Christmas tree breeding programs

PHYSIOLOGICAL RESPONSES OF EASTERN HEMLOCK (TSUGA CANADENSIS) TO SILVICULTURAL RELEASE AND VARIABLE SITE HISTORY: IMPLICATIONS FOR HEMLOCK RESTORATION

The rapid loss of eastern hemlock (Tsuga canadensis) in the southern Appalachian Mountains due to hemlock woolly adelgid (Adelges tsugae) infestation has resulted in substantial changes to ecosystem structure and function. Several restoration strategies have been proposed, including silvicultural treatments that increase incident light in forest understories. We conducted a four-year manipulative field experiment on surviving midstory hemlock trees to investigate the effects of release from light limitation on hemlock woolly adelgid infestation and physiological parameters, expecting that higher light levels would improve tree carbon balance. Mixed hardwood forest sites were either previously uninfested with hemlock woolly adelgid, infested with hemlock woolly adelgid, or infested with hemlock woolly adelgid and had a history of predatory beetle releases for biological control. At each site, we identified ten eastern hemlock trees in the mid-story and cut ~15 m radius canopy gaps around half of them while leaving the canopy intact over the other half. We compared short- and long-term indices of carbon gain and stress: leaf net photosynthesis; leaf fluorescence; leaf total nonstructural carbohydrate concentration; new shoot growth; hemlock woolly adelgid density; and basal area growth. We found that trees experienced greater leaf-level stress in gaps and when hemlock woolly adelgid was actively feeding. Despite being more stressed, trees in gaps fixed two times more carbon than those in reference conditions. High net leaf photosynthesis in the spring translated into high leaf total nonstructural carbohydrate concentration in the spring, coinciding with when hemlock woolly adelgid was actively feeding. Although infested and uninfested trees had similar leaf total nonstructural carbohydrate concentration maxima, infestation prevented trees from allocating this carbon to shoot and basal area growth; this was particularly true for reference trees. Greater shoot growth in gap trees translated to greater annual basal area growth—by the end of the study, trees in gaps were growing nine times more than trees in reference conditions, and this was generally regardless of infestation status. In terms of growth and carbon balance, eastern hemlock consistently benefitted from the increased light and soil moisture found in gaps; there was inconsistent and rather weak evidence that predator beetles conferred an additional advantage. Our results indicate that silvicultural treatments may improve long-term health and survival of infested trees and that integration of such treatments with existing strategies is worthy of continued exploration

Assessment of Worldwide Acute Kidney Injury Epidemiology in Neonates: Design of a Retrospective Cohort Study.

INTRODUCTION: Acute kidney injury (AKI) affects ~30% of hospitalized neonates. Critical to advancing our understanding of neonatal AKI is collaborative research among neonatologists and nephrologists. The Neonatal Kidney Collaborative (NKC) is an international, multidisciplinary group dedicated to investigating neonatal AKI. The AWAKEN study (Assessment of Worldwide Acute Kidney injury Epidemiology in Neonates) was designed to describe the epidemiology of neonatal AKI, validate the definition of neonatal AKI, identify primary risk factors for neonatal AKI, and investigate the contribution of fluid management to AKI events and short-term outcomes. METHODS AND ANALYSIS: The NKC was established with at least one pediatric nephrologist and neonatologist from 24 institutions in 4 countries (USA, Canada, Australia, and India). A Steering Committee and four subcommittees were created. The database subcommittee oversaw the development of the web-based database (MediData Rave™) that captured all NICU admissions from 1/1/14 to 3/31/14. Inclusion and exclusion criteria were applied to eliminate neonates with a low likelihood of AKI. Data collection included: (1) baseline demographic information; (2) daily physiologic parameters and care received during the first week of life; (3) weekly snapshots ; (4) discharge information including growth parameters, final diagnoses, discharge medications, and need for renal replacement therapy; and (5) all serum creatinine values. ETHICS AND DISSEMINATION: AWAKEN was proposed as human subjects research. The study design allowed for a waiver of informed consent/parental permission. NKC investigators will disseminate data through peer-reviewed publications and educational conferences. DISCUSSION: The purpose of this publication is to describe the formation of the NKC, the establishment of the AWAKEN cohort and database, future directions, and a few lessons learned. The AWAKEN database includes ~325 unique variables and \u3e4 million discrete data points. AWAKEN will be the largest, most inclusive neonatal AKI study to date. In addition to validating the neonatal AKI definition and identifying risk factors for AKI, this study will uncover variations in practice patterns related to fluid provision, renal function monitoring, and involvement of pediatric nephrologists during hospitalization. The AWAKEN study will position the NKC to achieve the long-term goal of improving the lives, health, and well-being of newborns at risk for kidney disease