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Alzheimer's Disease (AD) is characterized by cognitive impairment and the presence of amyloid-β (Aβ) plaques and tau tangles. This study was conducted to assess the effects of white button mushroom (WBM) supplementation on spatial memory and plaque formation in mice with mutations in amyloid (Aβ). Mice with amyloid precursor protein (hAPP) mutations and their wildtype (WT) littermates were fed a 10% white button mushroom (WBM) feed

Acute graft-versus-host disease (aGVHD) is a significant complication of allogeneic hematopoietic stem cell transplant (HSCT) and negatively affects T cell reconstitution. Extracorporeal photopheresis (ECP) reduces aGVHD, but the mechanisms remain incompletely understood. Our objective was to examine the impact of ECP on thymopoiesis in pediatric aGVHD and the mechanisms at a cellular and transcriptional level. Sixteen pediatric HSCT patients were recruited: 6 with ECP-treated aGVHD, 5 without aGVHD, and 5 with aGVHD treated with corticosteroids only. Thymopoiesis was evaluated by measuring naive T cells, TRECs, IL-7, and T cell receptor repertoire diversity. Regulatory T cell (Treg) enumeration and function and dendritic cell (DC) enumeration and phenotype were analyzed using flow cytometry. T cell transcriptome analysis was performed on ECP patients after treatment and responders pre- and post-treatment. Four ECP responders demonstrated thymic-dependent T cell recovery, and superior median naïve T cell numbers at 8 and 12 months post-HSCT compared to the aGVHD corticosteroid group. Increased Tregs and Treg suppressive function, reduced cDC/pDC and DC co-stimulatory marker expression in ECP responders suggest upregulated peripheral tolerance; these findings were not observed in partial responders. Responder post-ECP CD3+ T cell transcriptional profile demonstrated 3333 downregulated and 364 upregulated genes, with significant downregulation of ERRα and GαS pathways, and reduced expression of pro-inflammatory and adhesion proteins. Thymic function improves with successful ECP treatment. ECP reduces T cell activation and impacts peripheral tolerance via DCs and Tregs. Differences in thymic recovery, DC, and Treg cellular patterns and the T cell transcriptome were observed between ECP responders and partial responders and require further validation and investigation in additional patients. Supplementary Information The online version contains supplementary material available at 10.1007/s10875-021-00991-y.

The thymus is critical for central tolerance and diverse T-lymphocyte repertoire development, to provide lifelong defense against pathogens while maintaining self-tolerance. Peak thymic output occurs in utero, during infancy, and in early childhood, diminishing throughout life. Infants with congenital heart disease requiring sternotomy often undergo thymectomy to clear the surgical field. The long-term effects of early thymectomy are just being appreciated. Many patients remain asymptomatic despite immunologic findings mirroring those of immunosenescence. Few develop increased infection or lymphoreticular malignancy risk. When considering the effects of infant thymectomy, patients with partial DiGeorge syndrome or hypomorphic recombination-activating gene (RAG) mutations may be instructive. These patients are lymphocytopenic, with increased early-onset infection and autoimmunity risk that is not seen in most patients who underwent thymectomy during infancy. The thymic structure of patients with partial DiGeorge syndrome or hypomorphic RAG is abnormal, with disrupted architecture inclining to perturbation of central tolerance. Similar findings may be seen in patients with myasthenia gravis, although disrupted peripheral tolerance may play a greater role in autoimmunity development. In conclusion, thymectomy during infancy may increase future risk of infection or autoimmunity, with premature immunosenescence mediated through disruption of central and peripheral tolerance mechanisms initiated by early cessation or diminution of thymic output. Ideally, some thymic tissue should be preserved at the time of surgery.

RNAi efficiency in insects is different from species to species; some species in Coleoptera are relatively more amenable to RNA interference (RNAi) than other species. One of the major factors is the presence of dsRNA-degrading enzymes, called dsRNases, in saliva, gut, or hemolymph in insects, which degrade the double-stranded RNA (dsRNA) introduced, resulting in the low efficacy of RNAi. In this study, we report a dsRNA-degrading activity in the gut homogenates from the spotted-wing drosophila, Drosophila suzukii, by ex vivo assay. Then, we identified two Drosophila suzukii dsRNase genes, named DrosudsRNase1 and DrosudsRNase2. In silico analysis shows that the gene structures are similar to dsRNases found in other insects. When dsRNases expressed in Sf9 cells were compared for their dsRNA degrading activities, dsRNase1 was more vital than dsRNase2. Both dsRNases were expressed highly and exclusively in the gut compared to the rest of body. Also, they were highly expressed during larval and adult stages but not in embryonic and pupal stages, suggesting the dsRNases protect foreign RNA molecules received during the feeding periods. DsRNase1 was expressed at a higher level in adults, whereas dsRNase2 showed more expression in early larvae. Our study on the tissue and development-specific patterns of dsRNases provides an improved understanding of the RNAi application for the management of D. suzukii.

Background/Aims The range of approved immunosuppressive and immunomodulatory (IM) agents has grown considerably with an increasing list of indications across paediatric specialties. At present, there is limited evidence supporting best practice for prescribing and monitoring of IM agents in children and young people (CYP). We present a staged service development project exploring cross-specialty prescribing and monitoring of IM agents at a tertiary children’s hospital (Great North Children’s Hospital, GNCH) and data sharing with local hospitals across northeast England. Methods In Phase 1, we searched pharmacy databases and surveyed specialty teams in GNCH to identify clinicians regularly prescribing IM agents to CYP over a twelve-month period. Phase 2 was a cross-specialty retrospective case-notes review of prescribing, monitoring and infection surveillance in a representative sample of CYP on IM agents. Phase 3 explored information sharing with six other hospitals in the region and acute presentations to these sites involving CYP on IM agents. Results Phase 1 identified 9 paediatric and 2 adult specialties prescribing IM agents to 416 CYP. 32 discrete IM therapies were prescribed with significant between-specialty overlap in drugs prescribed but a wide range of prescribing and monitoring practices. Phase 2 assessed 77 CYP on IM agents in detail - 57% were prescribed >1 IM agent, 100% had FBC measured at least once (range once only to weekly), 18% developed lymphopenia at least once and 40% were prescribed prophylactic antibiotics. Previous varicella exposure had been assessed in 70%. Phase 3 data are summarised in Table 1. P011 Table 1:Information sharing and acute presentations to regional hospitals local to immunosuppressed patientsTotal number of patients141Mean age in years (range)11 (2 - 17)NDiagnosisJIA without uveitis108JIA with uveitis9Uveitis alone8Systemic JIA4Period fever4Behçet’s disease2Juvenile dermatomyositis2Scleroderma1Juvenile systemic lupus erythematosus1Mixed connective tissue disease1Granulomatosis with polyangiitis1Immunosuppressive or immunomodulatory agent usedAdalimumab65Methotrexate42Tocilizumab22Mycophenolate mofetil10Etanercept10Infliximab5Sulfasalazine5Prednisolone4Abatacept4Leflunomide4Canakinumab2Colchicine2Anakinra2Rituximab2Cyclophosphamide1Number of immunosuppressive or immunomodulatory agents per patient3 agents52 agents441 agent92Number of acute presentations by diagnosis or presenting complaint (n = 19)Fever4Chickenpox4Viral upper respiratory tract infection2Joint pain2Abdominal pain2Rash2Eye infection1Tonsilitis1Wheeze1Yes (%)No (%)Named local consultant (n = 129)3763Correct diagnosis recorded locally (n = 130)8020Correct immunosuppressive or immunomodulatory agent recorded locally (n = 130)5050Open access for febrile illness (n = 116)4159Reviewed in past 2 years for acute illness (n = 109)1783Note: presented numbers for immunosuppressive or immunomodulatory agents are not mutually exclusive. JIA, juvenile idiopathic arthritis Conclusion IM agents are central to modern paediatric clinical care across a wide range of diseases. This staged project identified significant variation in IM prescribing and monitoring practice between specialties at GNCH. Communication between specialty and local teams is inadequate. Particular areas of concern include limited diagnostic, blood monitoring and medication information sharing and limited local information governing management of intercurrent illness and vaccination. Although different disease processes can necessitate different advice and prescribing practices, sharing examples of good practice will minimise unnecessary variation. We propose the development of a regional immunosuppression working group to improve quality and safety across our region. Disclosure J.L. Bennett: None. C. Tsilifis: None. A. Flinn: None. T. Altmann: None. N. Jansen: None. H. Tumelty: None. K. Aitken: None. S. Bhopal: None. E. Harrison: None. S. Ravenscroft: None. E. Sen: None. E. Williams: None. T. Flood: None. S. Sampath: None. A. Battersby: None. F. McErlane: None.

Chronic traumatic encephalopathy (CTE) is a progressive neurodegenerative condition caused by repetitive mild traumatic brain injury (TBI) that leads to impaired executive functioning, emotional disturbances, and disordered memory, warranting both basic and translational research of potential therapeutic targets. One area of research concerns prophylactic zinc (Zn) supplementation; however, Zn supplementation remains poorly understood. This study explored the effects of Zn supplementation in a mouse model of repetitive mild TBI. Four-week-old male (n = 50) and female (n = 50) C57BL/6J mice consumed tap water or 10 parts per million Zn-supplemented water for eight weeks prior to injury. At 12 weeks of age, mice underwent either five sham procedures or five closed-head injuries spaced apart by 48 h after which they completed behavioral tests. Zinc-supplemented injured mice righted themselves and regained normal ambulatory function as fast as non-injured mice across four out of the five injury days. In contrast, non-supplemented injured mice exhibited impairment in normal ambulation by days 4 and 5. Injury also reduced free, ionic Zn in the dentate gyrus and CA3 region of the hippocampus and Zn supplementation partially remediated this reduction but not to the levels of non-injured mice. There were no structural differences in cortex, hippocampus, striatum, and corpus callosum, suggesting that Zn reduction was not due to macroscopic abnormalities. Overall, these results suggest that Zn may improve short-term and physical neurological recovery, but it may not be sufficient as a single pre-treatment for consequences of repetitive mild TBI such as cognitive impairment. These results further demonstrate the need for additional research documenting the underlying mechanisms of Zn in TBI-related neuropathology.

Aims Immunosuppression has become integral to the management of a wide range of childhood illness. Although total numbers of children and young people (CYP) on immunosuppression is unknown, they are thought to be increasing. Multiple paediatric sub-specialities initiate and monitor different immunosuppressive therapies with anecdotal variation in prescribing and monitoring practices. To understand existing variations in care, we undertook a retrospective review, focussing on: Number of CYP on immunosuppression; Drug monitoring and antimicrobial prophylaxis; Numbers of serious infections. Methods CYP attending our hospital on immunosuppressive medication were identified from speciality teams (excluding immunology/oncology patients). Immunosuppressive agents were defined as: Long–term/high–dose steroids Cytotoxic drugs Biologic agents/monoclonal antibodies Data were collected on a representative sample of patients under each speciality: Diagnosis; Current/past immunosuppression; Current/past antimicrobial prophylaxis; Consideration of risk for specific infections; Serious infective episodes. Existing speciality/departmental guidance on prescribing, monitoring and surveillance was collected. Results 416 CYP on immunosuppression identified; data were collected on 77 (18.5%), with cross-speciality representation. 47/77 children are currently prescribed ≥2 agents with 8/77 presently off immunosuppression. 46/77 were not prescribed prophylaxis at any point. In those who were, cotrimoxazole (n=28) was commonest. 54/77 patients had past VZV exposure documented or tested. 7/77 patients attended after chickenpox exposure; 2 required admission for treatment. 10/77 patients were hospitalised for possible bacterial infection; none had proven bacteraemia but 2 developed cryptosporidiosis. All patients had FBC checked with varying frequency (weekly–once only). 14/77 developed lymphopaenia; of these, 8 had subsets checked. There was no clear relationship between speciality and monitoring frequency. Two specialities were able to provide departmental guidance for the management of intercurrent infection. Conclusions There is a large population of CYP on a wide range of immunosuppression. There is inter-speciality variation in the agents prescribed, monitoring schedules, infection risk stratification and antimicrobial prophylaxis. Departmental protocols are uncommon and not readily accessible outside of speciality or hospital, making out-of-hours decisions unnecessarily challenging. This wide variation in practice and lack of evidence-based guidance is unacceptable. Regional immunosuppression guidance may improve the quality of care offered to immunosuppressed children in our region.

Acute graft-versus-host disease (aGVHD) complicates allogeneic hematopoietic stem cell transplantation (HSCT), and is treated with topical and/or systemic corticosteroids. Systemic corticosteroids and aGVHD damage thymic tissue. We compared thymopoietic effect of topical steroid therapy, corticosteroids and extracorporeal photopheresis (ECP) in 102 pediatric allogeneic HSCT patients. We categorized patients into 4 groups: - no aGVHD, aGVHD treated with topical or systemic steroid, or ECP. Naive CD4+CD45RA+CD27+ T-lymphocyte values at 3, 6, 9, 12months post-HSCT were recorded: for ECP patients, values were recorded at 3, 6, 9, 12months during ECP. Differences were compared using the Kruskal-Wallis test. 41 patients had no aGVHD, 23 had aGVHD treated topically or systemically (25), 13 received ECP. Rate of thymopoiesis was significantly different between all groups at all time-points post-transplant (p=0.002, p

Hyperphosphorylated tau protein is a key pathology in Alzheimer's disease (AD), frontotemporal dementia, chronic traumatic encephalopathy, and Parkinson's disease. The essential trace element zinc exacerbates tauopathy in vitro as well as in a Drosophila model of AD. However, the interaction has never been assessed behaviorally or biochemically in mammals. Zinc supplementation is prevalent in society, finding use as a treatment for macular degeneration and cataracts, and is also taken as an immune system booster with high levels appearing in multivitamins marketed toward the elderly. Using a transgenic mouse model that contains the human gene for tau protein (P301L), we assessed the effects of excess chronic zinc supplementation on tau pathology. Behavioral tests included nest building, circadian rhythm, Morris Water Maze, fear conditioning, and open field. Biochemically, total tau and Ser396 phosphorylation were assessed using western blot. Number of tangles were assessed by Thioflavin-S and free zinc levels were assessed by Zinpyr-1. Tau mice demonstrated behavioral deficits compared to control mice. Zinc supplementation exacerbated tauopathic deficits in circadian rhythm, nesting behavior, and Morris Water Maze. Biochemically, zinc-supplemented tau mice showed increased phosphorylation at pSer396. Zinc supplementation in tau mice also increased tangle numbers in the hippocampus while decreasing free-zinc levels, demonstrating that tangles were sequestering zinc. These results show that zinc intensified the deficits in behavior and biochemistry caused by tau.

The European and Asian earthworms introduced to eastern North American forests have great potential to facilitate plant invasions, in part through selective seed predation and dispersal. The invasive plant Alliaria petiolata (Garlic Mustard) contains secondary metabolites that may deter earthworms from eating its seeds. In 2 growth-chamber experiments, I determined whether the invasive earthworms Lumbricus terrestris (Nightcrawler) and Eisenia fetida (Red Wiggler) could aid the spread of Garlic Mustard by ingesting its seeds at lower rates than the similar-sized seeds of the native forest herb Geranium maculatum (Wild Geranium). Earthworms had similar rates of seed ingestion regardless of earthworm or plant species and digested the majority of seeds they ate (67–73%). There was no interaction between earthworm and plant species. Given a choice between Garlic Mustard and Wild Geranium, seed selectivity cannot explain positive associations between earthworm abundance and Garlic Mustard invasion.

Circadian rhythms are altered in several diseases associated with aging, one of which is Alzheimer's disease (AD). One example of a circadian rhythm is the rest-activity cycle, which can be measured in mice by monitoring their wheel-running. The present study sought to investigate differences in light phase/dark phase activity between a mouse model of late onset AD (APP/E4) and control (C57Bl6J) mice, in both the pre-plaque and post-plaques stages of the disease. To assess activity level, 24-h wheel running behavior was monitored at six months (pre-plaque) and twelve months (post-plaque) for a period of nine days. The following measures were analyzed: counts (wheel rotations) during the dark phase, counts during the light phase, hour of activity onset, and hour of activity offset. Key findings indicate that activity onset is delayed in APP/E4 mice at six and twelve months, and activity profiles for APP/E4 and C57Bl6J mice differ during the light and dark phase in such a way that APP/E4 mice run less in the early hours of the dark phase and more in the later hours of the dark phase compared to C57Bl6J mice. These findings imply that rest-activity cycle is altered in the pre-plaque stages of AD in APP/E4 mice, as they show impairments as early as six months of age.

Serpentine barrens are globally rare, savanna habitats that support distinctive vegetation. In eastern North America, two major threats to serpentine barrens are the spread of invasive species and the encroachment of trees into open habitats. We assessed these threats by comparing serpentine savannas and woodlands in the State Line serpentine barrens, southeast Pennsylvania. We asked which invasive species were most frequent and abundant in each habitat and how they may affect plant diversity and community composition and how succession of savannas to woodlands may affect plant diversity and community composition. We compared species richness, evenness, diversity, and beta diversity between savannas and adjacent woodlands and quantified differences in community composition. Invasive species comprised 16% of total plant cover, and the most frequent invasive species were Microstegium vimineum (Trin.) A. Camus, Lonicera japonica Thunb., and Elaeagnus umbellata Thunb. Savannas had substantially great...

Alzheimer’s disease (AD) is a neurocognitive disorder that impacts both the brain and behavior. Metal ions, including zinc (Zn), have been seen to play an important role in AD-related pathology. In this study, we show alterations in wheel-running behavior both early and late in disease progression in a novel dual Tg mouse model of AD. This mouse includes both amyloid and tau pathology through its cross with the J20 (hAPP) and P301L (Tau) parentage. Animals were given either lab water or water that had been supplemented with 10 ppm Zn. Wheel running was assessed through individually housing mice and measuring wheel-running activity in both the light and dark cycles. Dual Tg mice showed significantly less activity in the first part of the dark cycle than WT mice at both 3.5 and 7 months of age (p < 0.05). Dual Tg mice given Zn water showed less activity compared to dual Tg mice on lab water, tau mice on Zn water, or WT mice given either lab or Zn water (p < 0.05) at 7 months. Female mice in this study consistently showed higher activity compared to male mice in all groups whereas Zn led to reduced activity. Daily activity rhythm was altered in both the tau and dual Tg mice, and Zn impacted this alteration through effects on amyloid, tau, and through circadian pathways.

Nest building is an innate behavior in male and female rodents, even when raised in laboratory settings. As such, many researchers provide rodents synthetic and/or natural materials (such as twine, tissue, cotton, paper, and hay) as a gauge of their overall well-being and as an ancillary assessment to predict the possible decline in cognition. Typically, changes in nesting behaviors, such as failure to create a nest, indicate a change in health or welfare. In addition, nesting behavior is sensitive to many environmental and physiological challenges, as well as many genetic mutations underlying pathological disease states. The following protocol describes a nesting behavior paradigm that explores the usage of four types of nesting material. In addition, the protocol utilizes intraclass correlations to demonstrate that inter-rater reliability is higher when nests are constructed out of shredded paper compared to other common nesting materials such as cotton squares, paper twists, and soft cob bedding. The chosen methodology and statistical considerations (i.e., intraclass correlation) for this assay may be of interest for those conducting experiments assessing the quality of living of mice.

MHC class II deficiency is a rare, but life-threatening, primary combined immunodeficiency. Hematopoietic cell transplantation (HCT) remains the only curative treatment for this condition, but transplant survival in the previously published result was poor. We analyzed the outcome of 25 patients with MHC class II deficiency undergoing first HCT at Great North Children's Hospital between 1995 and 2018. Median age at diagnosis was 6.5 months (birth to 7.5 years). Median age at transplant was 21.4 months (0.1-7.8 years). Donors were matched family donors (MFDs; n = 6), unrelated donors (UDs; n = 12), and haploidentical donors (HIDs; n = 7). Peripheral blood stem cells were the stem cell source in 68% of patients. Conditioning was treosulfanbased in 84% of patients; 84% received alemtuzumab (n = 14) or anti-thymocyte globulin (n = 8) as serotherapy. With a 2.9-year median follow-up, OS improved from 33% (46-68%) for HCT before 2008 (n = 6) to 94% (66-99%) for HCT after 2008 (n = 19; P = .003). For HCT after 2008, OS according to donor was 100% for MFDs and UDs and 85% for HIDs (P = .40). None had grade III-IV acute or chronic graft-versus-host disease. Latest median donor myeloid and lymphocyte chimerism were 100% (range, 0-100) and 100% (range, 64-100), respectively. Latest CD4+ T-lymphocyte number was significantly lower in transplant survivors (n = 14) compared with posttransplant disease controls (P = .01). All survivors were off immunoglobulin replacement and had protective vaccine responses to tetanus and Haemophilus influenzae. None had any significant infection or autoimmunity. Changing transplant strategy in Great North Children's Hospital has significantly improved outcomes for MHC class II deficiency.

Background: Fixation of brain tissue is a common practice which allows preservation of tissue and aids in preventing structural and chemical abnormalities. However, fixation procedures may disrupt the levels of biometals such as zinc when compared to tissue that is fresh-frozen. Thus, we sought to determine if any differences in free-zinc levels exist between perfused and fresh-frozen tissue. Zinc is an essential biometal critical for cellular communication and memory and exists in both bound and free forms; the latter playing critical roles in synaptic communication. New method: C57BL/6 J mice were divided into two water types: those given lab water and those given water supplemented with 10 ppm zinc carbonate. Perfusion was carried out with 4% paraformaldehyde on half of the animals in each water group to assess the impact on levels of free Zn as measured through Zinpyr-1 fluorescence. Results: There were significant differences in Zn fluorescence values between Zn-supplemented and lab water groups as well as between perfused and fresh-frozen tissues in the dentate gyrus and CA3 regions of the hippocampus, regions critical in learning & memory. Comparison with existing methods: These results show that when determining a method for euthanasia, any future histological techniques involving assessment of metal content should first be considered. Conclusions: Researchers must be cautious with the way in which tissue is collected and treated since this can lead to misleading conclusions when linking changes in behavior and relative levels of trace metals.

Diagnosis of Kawasaki Disease (KD) can be challenging due to lack of a diagnostic test, and some children present with 'incomplete' KD when not all diagnostic criteria are met. Treatment with intravenous immunoglobulin (IVIG) and aspirin reduces the risk of coronary artery complications. There is sub-group of patients who are resistant to IVIG/aspirin therapy and are at increased risk of complications. Recent evidence suggests that additional treatment of this high-risk group with corticosteroids is beneficial in reducing this risk. We examine the treatment and coronary artery outcomes, by retrospective review of medical records, of a cohort of 32 paediatric patients with KD admitted to a single Irish tertiary centre from January 2010-December 2014. Twenty-eight percent of patients (9/32) had an incomplete diagnosis of KD; these patients received IVIG later compared to those with a complete KD diagnosis. 15/32 (47%) had abnormal echocardiogram findings in the acute phase, 8/32 (25%) had echocardiogram abnormalities at 6-week follow-up, and 4/32 (12.5%) had persisting abnormalities. This study highlights the potential for adverse outcome in KD, the difficulty in diagnosis in 'incomplete' cases, and the need to identify children at higher risk for adverse outcome where adjunctive therapies would be most beneficial.

Environmental factors play a significant role in well-being of laboratory animals. Regulations and guidelines recommend, if not require, that stressors such as bright lighting, smells, and noises are eliminated or reduced to maximize animal well-being. A factor that is often overlooked is handling and how researchers interact with their animals. Researchers, lab assistants, and husbandry staff in animal facilities may use inconsistent handling methods when interacting with rodents, but humans should be considered a part of the animal’s social environment. This study examined the effects of different handling techniques on depressive-like behavior, measured by the Porsolt forced swim test, in adult C57BL/6J male mice. The same two researchers handled the mice in a gentle, aggressive, or minimal (control) fashion over approximately two weeks prior to testing. The results demonstrated a beneficial effect of gentle handling: gentle handling reduced swimming immobility in the forced swim test compared to mice that were aggressively or minimally handled. We argue that gentle handling, rather than methodical handling, can foster a better relationship between the handlers and rodents. Although handling is not standardized across labs, consistent gentle handling allows for less challenging behavioral testing, better data collection, and overall improved animal welfare.

In this classroom activity, students build a phylogeny for woody plant species based on the morphology of their twigs. Using any available twigs, students can practice the process of cladistics to test evolutionary hypotheses for real organisms. They identify homologous characters, determine polarity through outgroup comparison, and construct a parsimonious tree based on synapomorphies (shared derived characters). This activity efficiently demonstrates many systematics concepts, including homology, homoplasy (convergence and reversal), polarity, synapomorphy, symplesiomorphy, autapomorphy, polytomy, and parsimony. It also engages students in inquiry, promotes student collaboration, raises awareness of plant structure, and exposes students to the diversity of common local trees.

Background: When intubating newborns, clinicians aim to place the tip of the endotracheal tube (ETT) in the mid-trachea. Clinicians usually estimate the ETT insertion depth based on weight. ETT tips are often incorrectly positioned in newborns. Estimating the insertion depth based on gestation may be more accurate. Objective: To determine whether estimating the ETT insertion depth using gestation, compared to weight, results in more correctly placed ETTs. Methods: Newborn infants without congenital anomalies who were intubated orally were randomised to having their ETT insertion depth estimated using weight [insertion depth (cm) = weight (kg) + 6] or gestation [value determined from a table]. The primary outcome was correct ETT position, defined as an ETT tip between the upper border of the first thoracic vertebra (T1) and the lower border of the second thoracic vertebra (T2) on a chest X-ray. The primary outcome was determined by one paediatric radiologist who was masked to group assignment. Results: Ninety infants were enrolled and the groups were well matched. The proportion of correctly placed ETTs was not significantly different between the groups [weight, 25/49 (51%), vs. gestation, 16/41 (39%), p = 0.293]. We found no significant differences in the secondary outcomes measured. Conclusion: Estimating the ETT insertion depth in newborns using gestation compared to weight did not result in more correctly placed ETTs.

Tubulin Polymerization Promoting Protein (TPPP/p25) modulates the dynamics and stability of the microtubule network by its bundling and acetylation enhancing activities that can be modulated by the binding of zinc to TPPP/p25. Its expression is essential for the differentiation of oligodendrocytes, the major constituents of the myelin sheath, and has been associated with neuronal inclusions. In this paper, evidence is provided for the expression and localization of TPPP/p25 in the zinc-rich retina and in the oligodendrocytes in the optic nerve. Localization of TPPP/p25 was established by confocal microscopy using calbindin and synaptophysin as markers of specific striations in the inner plexiform layer (IPL) and presynaptic terminals, respectively. Postsynaptic nerve terminals in striations S1, S3 and S5 in the IPL and a subset of amacrine cells show immunopositivity against TPPP/p25 both in mice and human eyes. The co-localization of TPPP/p25 with acetylated tubulin was detected in amacrine cells, oligodendrocyte cell bodies and in synapses in the IPL. Quantitative Western blot revealed that the TPPP/p25 level in the retina was 0.05–0.13 ng/μg protein, comparable to that in the brain. There was a central (from optic nerve head) to peripheral retinal gradient in TPPP/p25 protein levels. Our in vivo studies revealed that the oral zinc supplementation of mice significantly increased TPPP/p25 as well as acetylated tubulin levels in the IPL. These results suggest that TPPP/p25, a microtubule stabilizer can play a role in the organization and reorganization of synaptic connections and visual integration in the eye.

As exotic species invasions continue to transform patterns of biological diversity, it becomes increasingly urgent to understand invasions' full consequences. Woody exotics that colonize closed-canopy forests have the potential for strong effects on plant communities. Berberis thunbergii DC, one such species, has invaded forests throughout eastern North America. Here we quantified the impacts of B. thunbergii invasions on the diversity and composition of native plant communities, both directly and through possible modifications of nutrient, moisture, and light availability. Plots with and without B. thunbergii had similar species richness, evenness, and diversity. Only two species were less likely to occur in plots with B. thunbergii. We also found no effect of B. thunbergii on soil moisture or other key soil properties such as pH and organic matter content. Plots with B. thunbergii had lower light levels at 10 cm from the ground, as under any shrub. It is possible that the invasion we observed may develop into a dense thicket with more severe impacts; however, the effects of B. thunbergii were modest at this time.

Old-growth forests dominated by shade-tolerant tree species are among the plant communities most likely to be in equilibrium. However, the assumption that these forests show long-term stability has rarely been tested. In this study, we resurveyed plots established 86 yr before in an old-growth beech-maple-hemlock forest in northeast Ohio to examine changes in tree community composition, diversity, and structure. Species richness and diversity remained constant from 1932 to 2018, but we found dramatic shifts in the relative abundances of species and the size structure of populations. Fagus grandifolia remained the most frequent and abundant tree species in the forest, while changing dramatically in size structure; larger Fagus grandifolia trees died and were replaced by large numbers of root suckers, increasing the overall stem density of the forest by 50%. Tsuga canadensis declined in frequency and stem density, shifted away from smaller size classes, and had almost no saplings. Plots dominated by Tsuga canadensis in 1932 showed the greatest magnitude of compositional change over time, shifting toward dominance by Acer saccharum, which along with Acer rubrum, increased greatly in frequency, stem density, and basal area. Eight species including Castanea dentata and Sassafras albidum were lost from the plots. In sum, Fagus grandifolia and Acer saccharum are displacing Tsuga canadensis in this stand. Tree diseases, pests, and deer herbivory likely contributed to these compositional shifts. Given the time scale of the directional changes we observed, even relatively undisturbed communities such as old-growth forests might be best viewed as not in equilibrium.

Zinc (Zn) is abundantly enriched in sub-retinal pigment epithelial (RPE) deposits, the hallmarks of age-related macular degeneration (AMD), and is thought to play a role in the formation of these deposits. However, it is not known whether Zn is the only metal relevant for sub-RPE deposit formation. Because of their involvement in the pathogenesis of AMD, we determined the concentration and distribution of calcium (Ca), iron (Fe) and copper (Cu) and compared these with Zn in isolated and sectioned macular (MSD), equatorial (PHD) and far peripheral (FPD) sub-RPE deposits from an 86 year old donor eye with post mortem diagnosis of early AMD. The sections were mounted on Zn free microscopy slides and analyzed by microprobe synchrotron X-ray fluorescence (μSXRF). Metal concentrations were determined using spiked sectioned sheep brain matrix standards, prepared the same way as the samples. The heterogeneity of metal distributions was examined using pixel by pixel comparison. The orders of metal concentrations were Ca ⋙ Zn > Fe in all three types of deposits but Cu levels were not distinguishable from background values. Zinc and Ca were consistently present in all deposits but reached highest concentration in MSD. Iron was present in some but not all deposits and was especially enriched in FPD. Correlation analysis indicated considerable variation in metal distribution within and between sub-RPE deposits. The results suggest that Zn and Ca are the most likely contributors to deposit formation especially in MSD, the characteristic risk factor for the development of AMD in the human eye.

This study investigated why some homeless individuals seem unable to transition towards self-reliance, following traditional supportive services. It was hypothesized that this may be due to some cognitive dysfunction. Chronically homeless adults were compared to controls on three tests of prefrontal competency: the Iowa Gambling Task, Word Fluency (FAS), and the Burglar's Story; they performed significantly worse than controls on all three tests. These results indicate a relationship between chronic homelessness and possible pre-frontal deficits. This may explain why some long-term homeless fail to learn from the consequences of unproductive behavior and to develop more constructive behaviors needed to attain stability.

Allogeneic hematopoietic hematopoietic stem cell transplant (HSCT) is used to treat increasing numbers of malignant and non-malignant disorders. Despite significant advances in improved HLA-typing techniques, less toxic conditioning regimens and better supportive care, resulting in improved clinical outcomes, acute graft-versus-host disease (aGvHD) continues to be a major obstacle and, although it principally involves the skin, gastrointestinal tract and liver, the thymus is also a primary target. An important aim following HSCT is to achieve complete and durable immunoreconstitution with a diverse T-cell receptor (TCR) repertoire to recognise a broad range of pathogens providing adequate long-term adaptive T-lymphocyte immunity, essential to reduce the risk of infection, disease relapse and secondary malignancies. Reconstitution of adaptive T-lymphocyte immunity is a lengthy and complex process which requires a functioning and structurally intact thymus responsible for the production of new naive T-lymphocytes with a broad TCR repertoire. Damage to the thymic microenvironment, secondary to aGvHD and the effect of corticosteroid treatment, disturbs normal signalling required for thymocyte development, resulting in impaired T-lymphopoiesis and reduced thymic export. Primary immunodeficiencies, in which failure of central or peripheral tolerance is a major feature, because of intrinsic defects in hematopoietic stem cells leading to abnormal T-lymphocyte development, or defects in thymic stroma, can give insights into critical processes important for recovery from aGvHD. Extracorporeal photopheresis (ECP) is a potential alternative therapy for aGvHD, which acts in an immunomodulatory fashion, through the generation of regulatory T-lymphocytes (Tregs), alteration of cytokine patterns and modulation of dendritic cells (DCs). Promoting normal central and peripheral immune tolerance, with selective down-regulation of immune stimulation, could reduce aGvHD, and enable a reduction in other immunosuppression, facilitating thymic recovery, restoration of normal T-lymphocyte ontogeny, and complete immunoreconstitution with improved clinical outcome as the ability to fight infections improves and risk of secondary malignancy or relapse diminishes.

Acute graft-versus-host disease (aGvHD) continues to be a major obstacle to allogeneic haematopoietic stem cell transplantation. Thymic damage secondary to aGvHD along with corticosteroids and other non-selective T lymphocyte-suppressive agents used in the treatment of aGvHD concurrently impair thymopoiesis and negatively impact on immunoreconstitution of the adaptive immune compartment and ultimately adversely affect clinical outcome. Extracorporeal photopheresis (ECP) is an alternative therapeutic strategy that appears to act in an immunomodulatory fashion, potentially involving regulatory T lymphocytes and dendritic cells. By promoting immune tolerance and simultaneously avoiding systemic immunosuppression, ECP could reduce aGvHD and enable a reduction in other immunosuppression, allowing thymic recovery, restoration of normal T lymphopoiesis, and complete immunoreconstitution with improved clinical outcome. Although the safety and efficacy of ECP has been demonstrated, further randomised controlled studies are needed as well as elucidation of the underlying mechanisms responsible and the effect of ECP on thymic recovery.

The amyloid-β protein (Aβ) is a metalloprotein with affinity for the metal ions zinc (Zn), copper (Cu), and iron (Fe), which are found in high concentrations in the plaques of Alzheimer's disease (AD). Increasing attention is focused on the role of these metals in AD, and much of the evidence suggests a dyshomeostasis between these metal ions may significantly affect Aβ aggregation and deposition in the brain. While the effect of these metals on Aβ has been shown in vitro, there is less behavioral data supporting a direct role in cognitive impairment. In order to investigate the cognitive consequences of metal dyshomeostasis, we sought to directly increase metal levels in the brain by dietary means in a transgenic mouse model (Tg2576). We have now examined the effect of increased Zn (10 ppm) and Fe (10 ppm) levels in the drinking water in the Tg2576 mouse. Since increased dietary Zn can lead to Cu deficiency, a Zn group supplemented with copper was also examined (Zn (10 ppm)+Cu (0.025 ppm)). Significant increases in latency and fewer platform crossings on probe trials, which are considered measures of spatial memory impairment, were seen in both Fe and Zn supplemented transgenic mice, compared to those raised on lab water. No significant differences were seen between the Zn + Cu group and in transgenic mice raised on lab water. These data suggest that the negative consequences of Zn may be due to a reduction in copper levels and, therefore, an imbalance between these metal ions rather than a direct effect of increased Zn.

The development of post-agricultural forests provides a unique opportunity to understand the mechanisms controlling plant community assembly. In particular, identifying traits associated with colonization can point to the causes of community development. The trait of association with the soil pH gradient has not yet been explored in this context. I test whether acidophilic species are more frequent in post-agricultural forests and discuss several possible explanations for this pattern. Species with low pH optima clearly tended to be more frequent in post-agricultural forests, yet the mean pH of post-agricultural forest soils did not differ from adjacent forests that had never been cleared. Calciphilic species nevertheless could remain absent from post-agricultural forests if they depended on high-pH microsites that agriculture had destroyed, or they could be less able to tolerate the lower nutrient status of post-agricultural forest soils. Alternatively, acidophilic species may have more effectiv...

Summary 1. Dispersal ability can influence the importance of dispersal relative to other processes organizing metacommunities,suchasspeciessortingamonghabitatsalongenvironmental gradients. 2. We compare plants with different dispersal modes and habitat affinities, evaluating the roles of environmental and spatial controls on plant community composition in 128 wetlands within 10 km 2 of old-growthmaple-beechforestinsouthern Quebec,Canada. 3. We address two hypotheses. First, we ask whether species with short-distance dispersal mecha- nisms are more dispersal-limited than species with adaptations for long-distance dispersal. Second, becausewetland habitatsaremorefragmented thanupland habitats inthisforestedarea, wetestthe hypothesis that wetland species are more dispersal-limited than upland species growing in the same wetlands(e.g. onhummockswithinaswamp). 4. Variation partitioning based on constrained ordinations showed that the distributions of species with short-distance dispersal mechanisms related more strongly to spatial factors than the distribu- tions of long-distance dispersers,supporting the interpretationthatthesespecies are moredispersal- limited. The distributions of short-distance dispersers also showed finer-scale spatial patterns than thedistributions of long-distancedispersers. 5. Distributions of wetland species related more strongly to environmental conditions than the dis- tributions of upland species growing in the same wetlands, suggesting that wetland species are actu- ally less dispersal-limited than upland species. Wetland and upland species had similar patterns of spatial variationincommunitycomposition. 6. Synthesis. The processes of dispersal limitation and species sorting along environmental gradi- ents have differential importance to plants that grow within the same communities,but differ in dis- persal ability and habitat affinity. This result emphasizes the impact of dispersal ability on the organizationofmetacommunities.

There is considerable evidence suggesting that metals play a central role in the pathogenesis of Alzheimer's disease. Reports suggest that elevated dietary metals may both precipitate and potentiate an Alzheimer's disease phenotype. Despite this, there remain few studies that have examined the behavioral consequences of elevated dietary metals in wild type and Alzheimer's disease animals. To further investigate this in the current study, two separate transgenic models of AD (Tg2576 and TgCRND8), together with wild type littermates were administered 10 ppm (0.153 mM) Zn. Tg2576 animals were maintained on a zinc-enriched diet both pre- and postnatally until 11 months of age, while TgCRND8 animals were treated for five months following weaning. Behavioral testing, consisting of "Atlantis" and "moving" platform versions of the Morris water maze, were conducted at the end of the study, and tissues were collected for immunohistochemical analysis of amyloid-beta burden. Our data demonstrate that the provision of a zinc-enriched diet potentiated Alzheimer-like spatial memory impairments in the transgenic animals and was associated with reduced hippocampal amyloid-beta plaque deposits. Zinc-related behavioral deficits were also demonstrated in wild type mice, which were sometimes as great as those present in the transgenic animals. However, zinc-related cognitive impairments in transgenic mice were greater than the summation of zinc effects in the wild type mice and the transgene effects.

Though often overlooked, small wetlands in an upland matrix can support diverse plant communities that increase both local and regional species richness. Here we characterize the full range of wetland vegetation within an upland forest landscape and compare the diversity and composition of different wetland plant communities. In an old-growth forest reserve in southern Quebec, Canada, we sampled wet habitats including lakeshores, permanent and seasonal ponds, swamps, glades, and streamsides. We used clustering, indicator species analysis, and nonmetric multidimensional scaling ordination to identify and compare vegetation types. The wetlands contained 280 species of vascular plants, 45% of the reserve ’ s fl ora, in only 1.1% of its area. Local diversity averaged 24 ± 0.7 species per 7 m 2 , much higher than in the surrounding upland forests. Plant communities sorted into fi ve types, whose strongest indicator species were Osmunda regalis , Glyceria striata , O. cinnamomea , Deparia acrostichoides , and Matteuccia struthiopteris , respectively. Both local species richness and compositional variation among sites differed among the vegetation types. By combining species representative of the region ’ s major wetlands with species from the upland forest matrix, the plant assemblages of these wetlands make disproportionately important contributions to landscape-level diversity.

According to press reports in December of 2015, terrorist Tashfeen Malik posted her allegiance to ISIS on her Facebook account before killing 14 innocent civilians at the County Health Department in San Bernardino, California. Though Facebook had removed her account as violative of internal company rules, the government was not immediately alerted by the company to the existence of the post -- or the possibility of an attack. In a more recent example, gunman Omar Mateen checked his own Facebook posts and other social media accounts during the five-hour standoff in the Orlando bar where he killed 49 people on June 19, 2016, to verify that his pledge to Abu Bakr al-Baghdadi, the leader of the Islamic State, had been properly publicized. We suggest in this article that social media companies, like other corporate entities, should be legally required to institute compliance programs that ferret out and report terrorist activity at the earliest possible opportunity. Most of these companies, such as Facebook, Twitter, YouTube, and Instagram, already have internal rules against messages that might violate the federal prohibition against material support to terrorists or to a Foreign Terrorist Organization. Lesser known and new sites, such as Tumblr and Soundcloud, and even nonprofits such as Internet Archive in San Francisco, should be forced to follow suite. Additionally, many of these companies already have both a method of internal reporting by other users against rule-breakers, as well as computer programs that seek out key words to alert company monitors that a breach of internal rules might be occurring. We suggest two supplementary federal proposals.The first would create a new substantive offense; it would criminalize the failure by social media companies to discover terrorism-related posts by its users and to immediately release the posts to the government. A social media company would be guilty of this new crime if it knowingly, recklessly, or even negligently failed to institute a government-approved compliance program and report any suspicious results it discovered through its program to federal authorities. We realize that this proposal is strong medicine. However, we believe that the danger of online terror activity warrants such a vigorous federal response. This proposal does not replicate the Online Terrorism Activity Act recently proposed by Senator Feinstein, though we agree that her bill ought to be enacted. We are not suggesting merely that the social media companies be required to report known terrorist activity to federal law enforcement agents. Rather, we would require such companies to monitor customers for compliance with 18 U.S.C. sections 2339 to 2339D and other terrorism offenses on pain of criminal liability. And rather than automatically shutting such accounts down when they are discovered, which may have adverse unintended consequences, we would shift those decisions to the FBI experts best suited to make them. In some cases, it might serve intelligence needs to allow the postings to continue. Moving the loci of such decision-making from a private company to the government might also allow innocent and aggrieved users to pursue avenues of redress.The second proposal is to simply grant these companies leniency at sentencing should they be found liable under the federal doctrine of respondeat superior for the material support crimes of their agents. The federal government does this already with corporations, primarily in the white collar crime arena, to prevent the violation of federal criminal statutes or, failing that, to assist the government in discovering whom within the corporation committed the offense, and to prevent its recurrence. This strategy will likely not be nearly as effective as a tool against terrorism as our first proposal, as federal prosecutors have not yet attempted to charge social media companies for the crimes committed or assisted by their agents, and such agents may not have the required mens rea regarding postings by the users. Such a strategy works best when the corporation is facing criminal liability, with its attendant high-dollar fines, for its own violations. It is doubtful that social media companies will consider themselves sufficiently exposed under current substantive terrorism law to bother with the expense of such programs. However, because it will be less effective at criminalizing corporate behavior, and because it does not directly impinge on the privacy rights of social media users, this proposal might be politically palatable; it effects only those entities already found to have committed a serious federal felony, and it does not require offending companies to reveal offending posts to the government.In Part I of this Article, we review the development of terror activity in today's globalized environment, including the high rate of reliance on the Internet and mobile applications. In describing the very well-known danger of terrorism, we focus on "lone-wolf" terrorists and the enormous difficulty of finding such individuals and stopping them before they attack. The internet has made this problem all but impossible to solve, and therefore companies that make their fortunes utilizing the internet must become part of the solution. A Brookings Institute Report estimates 46,000 to 70,000 Twitter accounts used by ISIS supporters from September to December of 2014, and a George Washington University Study counted 900 active FBI investigations against ISIS sympathizers, 250 Americans who have traveled or attempted to travel to Syria/Iraq to join the Islamic State, and 300 Americans and/or U.S.-based ISIS sympathizers active on social media. In Part II, we will respond to perceived insufficiencies in existing legislation and in recent legislative proposals. We will also set forth proposals to address the liabilities of companies to enable the governmental review and discretion of potential terror activity online. In addition to both of our proposals, we offer precedents for such governmental action, including the Federal Guidelines pertaining to organizations, criminal and civil regulations regarding the Foreign Corrupt Practices Act, the Bank Secrecy Act, and international bodies in the enforcement of copyright law. Once compared to these other criminal and regulatory measures, the proposals are not as shocking as they might first appear.In Part III, we respond to both historical and anticipated opposition, grounded in constitutional arguments, to the proposed legislative framework in Part II. We believe that neither proposal would violate the Fourth Amendment protection against unreasonable searches and seizures, nor the First Amendment's protection of speech and association. A long line of precedent confirms that the Fourth Amendment offers no reasonable expectation of privacy in communications voluntarily revealed to third parties. And the Court's relatively recent 6-3 opinion in Holder v. Humanitarian Law Project, upholding the material support statute against a First Amendment freedom of speech and freedom of association and Fifth Amendment Due Process vagueness challenge, lends significant support to the validity of our proposals.

Objective To characterize the morphology of flash visual evoked potentials (fVEPs) obtained from infants hospitalized with severe, chronic malnutrition (marasmus). Methods A covariance-based principal component analysis with Promax factor rotation was applied to fVEPs obtained from malnourished infants and age-matched control subjects. Results The analysis suggests the presence of a late positive complex in the fVEP, with at least one of its components being significantly diminished in marasmic infants. The N3 component was also diminished in marasmic infants. Following remediation, the marasmic group no longer differed with respect to these components. However, an abnormally large late, positive deflection was evident at discharge. Conclusions The fVEP morphology of infants hospitalized with severe malnutrition was found to be significantly different from age-matched controls. Moreover, although there was evidence of recovery following remediation, fVEPs continued to show abnormality at discharge, suggesting the possibility that nutritional rehabilitation did not fully eliminate the physiological deficit. Significance Malnourishment during early infancy results in altered neurophysiological functioning, possibly in cortical areas responsible for higher order visual processing.

Temperate deciduous forests across much of Europe and eastern North America reflect legacies of past land use, particularly in the diversity and composition of plant communities. Intense disturbances, such as clearing forests for agriculture, may cause persistent environmental changes that continue to shape vegetation patterns as landscapes recover. We assessed the long-term consequences of agriculture for environmental conditions in central New York forests, including tree community structure and composition, soil physical and chemical properties, and light availability. To isolate the effects of agriculture, we compared 20 adjacent pairs of forests that were never cleared for agriculture (primary forests) and forests that established 85-100 years ago on plowed fields (secondary forests). Tree communities in primary and secondary forests had similar stem density, though secondary forests had 14% greater basal area. Species composition differed dramatically between the two forest types, with primary forests dominated by Acer saccharum and Fagus grandifolia and secondary forests by Acer rubrum and Pinus strobus. Primary and secondary forests showed no consistent differences in soil physical properties or in the principal gradient of soil fertility associated with soil pH. Within stands, however, soil water content and pH were more variable in primary forests. Secondary forest soils had 15% less organic matter, 16% less total carbon, and 29% less extractable phosphorus in the top 10 cm than adjacent primary stands, though the ranges of the forest types mostly overlapped. Understory light availability in primary and secondary forests was similar. These results suggest that, within 100 years, post-agricultural stands have recovered conditions comparable to less disturbed forests in many attributes, including tree size and number, soil physical properties, soil chemical properties associated with pH, and understory light availability. The principal legacies of agriculture that remain in these forests are the reduced levels of soil organic matter, carbon, and phosphorus; the spatial homogenization of soil properties; and the altered species composition of the vegetation.