Search

The experiment tested the effect of non-contact boxing training on the frequency and timing of anticipatory postural adjustments (APAs) resulting from self-induced postural perturbations in healthy adults.The 8-week non-contact boxing intervention study involved 33 healthy adults between 18 and 27 years of age who had no boxing experience (control group = 17 participants, boxing group = 16 participants). Pretests and post-tests utilized rapid bilateral arm raising as a focal movement to elicit APAs. EMG in the anterior deltoid, thoracic and lumbar erector spinae, semitendinosus, and soleus muscles was recorded. The boxing group completed twenty 90-min non-contact boxing training sessions over 8 weeks, whereas the control group kept physical activity consistent during the intervention period.Non-contact boxing training caused APAs to become more frequent during the focal movement, in comparison to the control group, in the soleus and in the semitendinosus after an outlier was removed. Non-contact boxing training caused earlier APA onset during the focal movement, in comparison to the control group, in the lumbar erector spinae after an outlier was removed.Non-contact boxing training had a modest positive effect on the frequency and timing of APAs resulting from self-induced postural perturbations in healthy adults.

Background: Many people with aphasia (PWA) want to read books. Text-to-speech (TTS) technology sometimes provides comprehension and processing time benefits when PWA read short, multisentence passages. Currently, no research examines the effect of TTS support when PWA read books. Aims: This study's primary purpose was to examine comprehension accuracy and total processing time of PWA and neurotypical healthy adults (NHAs) when reading book sections in read-only versus TTS-supported conditions. A secondary aim was to examine condition preference and perceived degree of understanding by people in both participant groups. Method and Procedure: Ten PWA and 10 NHAs alternated between read-only and TTS-supported conditions to read a book. Participants answered comprehension questions and provided feedback about their reading experience, condition preference, and desire to use TTS technology for future book reading. Outcomes and Result: Overall, PWA exhibited less accurate comprehension and slower processing times compared to NHAs in both conditions. No significant comprehension accuracy difference occurred between conditions for either group. However, four PWA exhibited a 10% or greater increase in comprehension accuracy when receiving TTS support. A significant processing time difference occurred with PWA processing text faster with TTS support, whereas NHAs did not demonstrate processing time differences. Most PWA preferred the TTS condition and expressed a desire to use TTS technology in the future. Most NHAs expressed the opposite preference. Conclusions: TTS support during book reading promotes faster processing without compromising comprehension for PWA. Clinicians should discuss with PWA the relative importance of comprehension accuracy, processing time, and comfort with technology when determining whether using TTS support during book reading is desirable.

Alcohol use and high-risk alcohol drinking behaviours among women are rapidly rising. In rodent models, females typically consume more ethanol (EtOH) than males. Here, we used the four core genotypes (FCG) mouse model to investigate the influence of gonadal hormones and sex chromosome complement on EtOH drinking behaviours. FCG mice were given access to escalating concentrations of EtOH in a two-bottle, 24-h continuous access drinking paradigm to assess consumption and preference. Relapse-like behaviour was measured by assessing escalated intake following repeated cycles of deprivation and re-exposure. Twenty-four-hour EtOH consumption was greater in mice with ovaries (Sry-), relative to those with testes, and in mice with the XX chromosome complement, relative to those with XY sex chromosomes. EtOH preference was higher in XX versus XY mice. For both consumption and preference, the influences of the Sry gene and sex chromosomes were concentration dependent. Escalated intake following repeated cycles of deprivation and re-exposure emerged only in XX mice (vs. XY). Mice with ovaries (Sry- FCG mice and C57BL/6J females) were also found to consume more water than mice with testes. These results demonstrate that aspects of EtOH drinking behaviour may be independently regulated by sex hormones and chromosomes and inform our understanding of the neurobiological mechanisms which contribute to EtOH dependence in male and female mice. Future investigation of the contribution of sex chromosomes to EtOH drinking behaviours is warranted. We used the FCG mouse model to investigate the influence of gonadal hormones and sex chromosome complement on EtOH drinking behaviours, including the alcohol deprivation effect. Escalated intake following repeated cycles of deprivation and re-exposure emerged only in XX mice (vs. XY). These results demonstrate that aspects of EtOH drinking behaviour may be independently regulated by sex hormones and chromosomes.

The CD34 cell surface antigen is widely expressed in tissues on cells with progenitor-like properties and on mature vascular endothelia. In adult human bone marrow, CD34 marks hematopoietic stem and progenitor cells (HSPCs) starting from the bulk of hematopoietic stem cells with long-term repopulating potential (LT-HSCs) throughout expansion and differentiation of oligopotent and unipotent progenitors. CD34 protein surface expression is typically lost as cells mature into terminal effectors. Because of this expression pattern of HSPCs, CD34 has had a central role in the evaluation or selection of donor graft tissue in HSC transplant (HSCT). Given its clinical importance, it is surprising that the biological functions of CD34 are still poorly understood. This enigma is due, in part, to CD34's context-specific role as both a pro-adhesive and anti-adhesive molecule and its potential functional redundancy with other sialomucins. Moreover, there are also critical differences in the regulation of CD34 expression on HSPCs in humans and experimental mice. In this review, we highlight some of the more well-defined functions of CD34 in HSPCs with a focus on proposed functions most relevant to HSCT biology.

Dominant and recessive mutations in podocalyxin (PODXL) are associated with human kidney disease. Interestingly, some PODXL mutations manifest as anuria while others are associated with proteinuric kidney disease. PODXL heterozygosity is associated with adult-onset kidney disease and podocalyxin shedding into the urine is a common biomarker of a variety nephrotic syndromes. It is unknown, however, how various lesions in PODXL contribute to these disparate disease pathologies. Here we generated two mouse stains: one that deletes Podxl in developmentally mature podocytes (Podxl∆Pod) and a second that is heterozygous for podocalyxin in all tissues (Podxl+/−). We used histologic and ultrastructural analyses, as well as clinical chemistry assays to evaluate kidney development and function in these strains. In contrast to null knockout mice (Podxl−/−), which die shortly after birth from anuria and hypertension, Podxl∆Pod mice develop an acute congenital nephrotic syndrome characterized by focal segmental glomerulosclerosis (FSGS) and proteinuria. Podxl+/− mice, in contrast, have a normal lifespan, and fail to develop kidney disease under normal conditions. Intriguingly, although wild-type C57Bl/6 mice are resistant to puromycin aminonucleoside (PA)-induced nephrosis (PAN), Podxl+/− mice are highly sensitive and PA induces severe proteinuria and collapsing FSGS. In summary, we find that the developmental timepoint at which podocalyxin is ablated (immature vs. mature podocytes) has a profound effect on the urinary phenotype due to its critical roles in both the formation and the maintenance of podocyte ultrastructure. In addition, Podxl∆Pod and Podxl+/− mice offer powerful new mouse models to evaluate early biomarkers of proteinuric kidney disease and to test novel therapeutics.

The COVID-19 pandemic continues to challenge the capacities of hospital ICUs which currently lack the ability to identify prospectively those patients who may require extended management. In this study of 90 ICU COVID-19 patients, we evaluated serum levels of four cytokines (IL-1β, IL-6, IL-10 and TNFα) as well as standard clinical and laboratory measurements. On 42 of these patients (binned into Initial and Replication Cohorts), we further performed CyTOF-based deep immunophenotyping of peripheral blood mononuclear cells with a panel of 38 antibodies. All measurements and patient samples were taken at time of ICU admission and retrospectively linked to patient clinical outcomes through statistical approaches. These analyses resulted in the definition of a new measure of patient clinical outcome: patients who will recover after short ICU stays (< 6 days) and those who will subsequently die or recover after long ICU stays (> 6 days). Based on these clinical outcome categories, we identified blood prognostic biomarkers that, at time of ICU admission, prospectively distinguish, with 91% sensitivity and 91% specificity (positive likelihood ratio 10.1), patients in the two clinical outcome groups. This is achieved through a tiered evaluation of serum IL-10 and targeted immunophenotyping of monocyte subsets, specifically, CD11clow classical monocytes. Immunophenotyping revealed clear predictors of clinical outcome in COVID-19 providing a highly sensitive and specific prognostic test that could prove useful in guiding clinical resource allocation.Graphical Abstract

Metastatic breast cancer is challenging to effectively treat, highlighting the need for an improved understanding of host factors that influence metastatic tumor cell colonization and growth in distant tissues. The lungs are a common site of breast cancer metastasis and are host to a population of tissue-resident eosinophils. Eosinophils are granulocytic innate immune cells known for their prominent roles in allergy and Th2 immunity. Though their presence in solid tumors and metastases have been reported for decades, the influence of eosinophils on metastatic tumor growth in the lungs is unclear. We used transgenic mouse models characterized by elevated pulmonary eosinophils (IL5Tg mice) and eosinophil-deficiency (ΔdblGATA mice), as well as antibody-mediated depletion of eosinophils, to study the role of eosinophils in EO771 mammary tumor growth in the lungs. We found that IL5Tg mice exhibit reduced pulmonary metastatic colonization and decreased metastatic tumor burden compared to wild-type (WT) mice or eosinophil-deficient mice. Eosinophils co-cultured with tumor cellsex vivoproduced peroxidase activity and induced tumor cell death, indicating that eosinophils are capable of releasing eosinophil peroxidase (EPX) and killing EO771 tumor cells. We found that lung eosinophils expressed phenotypic markers of activation during EO771 tumor growth in the lungs, and that metastatic growth was accelerated in eosinophil-deficient mice and in WT mice after immunological depletion of eosinophils. Our results highlight an important role for eosinophils in restricting mammary tumor cell growth in the lungs and support further work to determine whether strategies to trigger local eosinophil degranulation may decrease pulmonary metastatic growth.

Forward-viewing endoscopic optical coherence tomography (OCT) provides 3D imaging

Forward-viewing endoscopic optical coherence tomography (OCT) provides 3D imaging in vivo, and can be combined with widefield fluorescence imaging by use of a double-clad fiber. However, it is technically challenging to build a high-performance miniaturized 2D scanning system with a large field-of-view. In this paper we demonstrate how a 1D scanning probe, which produces cross-sectional OCT images (B-scans) and 1D fluorescence T-scans, can be transformed into a 2D scanning probe by manual scanning along the second axis. OCT volumes are assembled from the B-scans using speckle decorrelation measurements to estimate the out-of-plane motion along the manual scan direction. Motion within the plane of the B-scans is corrected using image registration by normalized cross correlation. En-face OCT slices and fluorescence images, corrected for probe motion in 3D, can be displayed in real-time during the scan. For a B-scan frame rate of 250 Hz, and an OCT lateral resolution of approximately 20 micrometers, the approach can handle out-of-plane motion at speeds of up to 4 mm/s., 16 pages

Autonomic cardiac function can be indirectly detected non-invasively by measuring the variation in microtiming of heart beats by a method known as heart rate variability (HRV). Aerobic training for sport is associated with reduced risk for some factors associated with cardiovascular diseases (CVD), but effects on autonomic function in different athlete types are less known. To compare cardiac autonomic modulation using a standard protocol and established CVD risk factors in highly trained intercollegiate athletes competing in aerobic, explosive, and cross-trained sports. A total of 176 college athletes were categorized in distinct sports as explosive (EA), aerobic (AA), or cross-trained (mixed) athletes. Eight different HRV measures obtained at rest were compared across training type and five health factors: systolic (SBP), diastolic blood pressure (DBP), body weight (BW), sex, and race. All athletic types shared favorable HRV measures that correlated with low CVD risk factors and indicated normal sympathovagal balance. A significant correlation was reported between DBP and pNN50 (% RR intervals &gt, 50 ms) (β = −0.214, p = 0.011) and between BW and low-frequency (LF) power (β = 0.205, p = 0.006). Caucasian and African American athletes differed significantly (p &lt, 0.05) with respect to four HRV variables: pNN50, HF power, LF power, and LF/HF ratios. Explosive, aerobic and mixed athletes had similar cardiovascular and autonomic HRV results in all eight HRV parameters measured. All athletes reported LF and pNN50 values that were significantly correlated with two CVD risk factors: DBP and BW. Compared with Caucasian teammates, African American athletes demonstrated lower LF/HF and higher pNN50, indicating an even more favorable resting sympathovagal activity and healthy CV function.

Podocalyxin (Podxl) is broadly expressed on the luminal face of most blood vessels in adult vertebrates, yet its function on these cells is poorly defined. In the present study, we identified specific functions for Podxl in maintaining endothelial barrier function. Using electrical cell substrate impedance sensing and live imaging, we found that, in the absence of Podxl, human umbilical vein endothelial cells fail to form an efficient barrier when plated on several extracellular matrix substrates. In addition, these monolayers lack adherens junctions and focal adhesions and display a disorganized cortical actin cytoskeleton. Thus, Podxl has a key role in promoting the appropriate endothelial morphogenesis required to form functional barriers. This conclusion is further supported by analyses of mutant mice in which we conditionally deleted a floxed allele of Podxl in vascular endothelial cells (vECs) using Tie2Cre mice (Podxl(ΔTie2Cre)). Although we did not detect substantially altered permeability in naïve mice, systemic priming with lipopolysaccharide (LPS) selectively disrupted the blood–brain barrier (BBB) in Podxl(ΔTie2Cre) mice. To study the potential consequence of this BBB breach, we used a selective agonist (TFLLR-NH(2)) of the protease-activated receptor-1 (PAR-1), a thrombin receptor expressed by vECs, neuronal cells, and glial cells. In response to systemic administration of TFLLR-NH(2), LPS-primed Podxl(ΔTie2Cre) mice become completely immobilized for a 5-min period, coinciding with severely dampened neuroelectric activity. We conclude that Podxl expression by CNS tissue vECs is essential for BBB maintenance under inflammatory conditions.

In the last two decades, our understanding of the genetic underpinnings of inherited podocytopathies has advanced immensely. By sequencing the genomes of a large pool of families affected by focal segmental glomerulosclerosis (FSGS), researchers have identified a common theme: familial podocytopathies are frequently caused by genes selectively expressed in podocytes. Podocalyxin is a podocyte-specific surface sialomucin that has long been known to play important roles in podocyte morphogenesis and function. Few studies, however, have shown a conclusive link between mutations in the gene and FSGS complemented by functional evidence. In a fascinating new paper published in Clinical Science, Lin et al. identify two unrelated pedigrees in which dominant loss-of-function mutations in PODXL lead to adult-onset FSGS. Nonsense-mediated decay of the mutated PODXL transcripts leads to protein insufficiency, which in turn cause podocyte dysfunction through defects in motility and cytoskeletal organization. This is the first study to date that demonstrates, mechanistically, how autosomal dominant mutations in podocalyxin can lead to FSGS and renal insufficiency. Here, we summarize the experimental findings of this manuscript and propose, perhaps, a more controversial hypothesis: down-regulation of podocalyxin protein expression from podocytes is a critical turning point in the progression of most podocytopathies and may be mechanistically relevant to glomerulopathies in which podocyte damage is not necessarily induced by genetic lesions.

Objectives The level of detail included when describing nailfold videocapillaroscopy (NVC) methods varies among research studies, making interpretation and comparison of results challenging. The overarching objective of the present study was to seek consensus on the reporting standards in NVC methodology for clinical research in rheumatic diseases and to propose a pragmatic reporting checklist. Methods Based on the items derived from a systematic review focused on this topic, a three-step web-based Delphi consensus on minimum reporting standards in NVC was performed among members of the European League against Rheumatism (EULAR) Study Group on Microcirculation in Rheumatic Diseases and the Scleroderma Clinical Trials Consortium. Results A total of 319 articles were selected by the systematic review, and 46 items were proposed in the Delphi process. This Delphi exercise was completed by 80 participants from 31 countries, including Australia and countries within Asia, Europe, North America and South America. Agreement was reached on items covering three main areas: patient preparation before NVC (15 items), device description (5 items) and examination details (13 items). Conclusion Based on the available evidence, the description of NVC methods was highly heterogeneous in the identified studies and differed markedly on several items. A reporting checklist of 33 items, based on practical suggestions made (using a Delphi process) by international participants, has been developed to provide guidance to improve and standardize the NVC methodology to be applied in future clinical research studies.

Background Asthma was identified as the most common comorbidity in hospitalized patients during the 2009 H1N1 influenza pandemic. We determined using a murine model of allergic asthma whether these mice experienced increased morbidity from pandemic H1N1 (pH1N1) viral infection and whether blockade of interleukin-4 receptor α (IL-4Rα), a critical mediator of Th2 signalling, improved their outcomes. Methods Male BALB/c mice were intranasally sensitized with house dust mite antigen (Der p 1) for 2 weeks; the mice were then inoculated intranasally with a single dose of pandemic H1N1 (pH1N1). The mice were administered intraperitoneally anti-IL-4Rα through either a prophylactic or a therapeutic treatment strategy. Results Infection with pH1N1 of mice sensitized to house dust mite (HDM) led to a 24% loss in weight by day 7 of infection (versus 14% in non-sensitized mice; p Conclusion Together, these data implicate allergic asthma as a significant risk factor for H1N1-related morbidity and reveal a potential therapeutic role for IL-4Rα signalling blockade in reducing the severity of influenza infection in those with allergic airway disease.

High-grade serous ovarian carcinoma (HGSOC) is an aggressive tumor with a 5-year disease-free survival of roughly 15%, partly because it is usually diagnosed at an advanced stage. Podocalyxin (Podxl) is a highly glycosylated sialomucin normally expressed by vascular endothelia and kidney podocytes. Strikingly, Podxl expression is frequently upregulated by a variety of tumors (including HGSOC) and is consistently associated with poor prognosis. We capitalized on the fact that glycosylation pathways are frequently dysregulated in cancer to develop an antibody, PODO447, that recognizes a tumor-restricted glycoform of Podxl not expressed on normal tissue. While the exact epitope remains to be identified, our results suggest that PODO447 binds an epitope comprising a peptide domain of Podxl in combination with the core 1 O-GalNAc glycan (T-antigen). When coupled to a cytotoxin, a PODO447-antibody-drug conjugate (ADC) effectively kills human tumor cells in vitro and in xenografted mice. While the vast majority of ovarian tumors highly express the Podxl core protein, only a subset of these express the PODO447 epitope. Strikingly, tumors that express a high level of PODO447 epitope tend to be those that lack infiltrating CD8+ T cells and CD20+ B cells: a phenotype that has previously been linked to immune evasion and poorest disease-free survival. Furthermore, we find that PODO447 is a more consistent marker of these immunologically “cold” tumors than a number of other markers, including CA125, mesothelin and folate receptor α. These results highlight the PODO447-epitope as a highly selective diagnostic marker of poor outcome tumors and the PODO447-ADC as a novel strategy for therapeutic intervention. This research was supported by the Canadian Institutes of Health Research (Grant Number: PJT-166180), the School of Biomedical Engineering (The University of British Columbia) postdoctoral fellowship and the Michael Smith Foundation for Health Research (MSFHR) research trainee award.

Text mining is widely used within the life sciences as an evidence stream for inferring relationships between biological entities. In most cases, conventional string matching is used to identify cooccurrences of given entities within sentences. This limits the utility of text mining results, as they tend to contain significant noise due to weak inclusion criteria. We show that, in the indicative case of protein-protein interactions (PPIs), the majority of sentences containing cooccurrences (∽75%) do not describe any causal relationship. We further demonstrate the feasibility of fine tuning a strong domain-specific language model, BioBERT, to analyse sentences containing cooccurrences and accurately (F1 score: 88.95%) identify functional links between proteins. These strong results come in spite of the deep complexity of the language involved, which limits the accuracy even of expert curators. We establish guidelines for best practices in data creation to this end, including an examination of inter-annotator agreement, of semisupervision, and of rules based alternatives to manual curation, and explore the potential for downstream use of the model to accelerate curation of interactions in the SIGNOR database of causal protein interactions and the IntAct database of experimental evidence for physical protein interactions.

Innate lymphoid cells (ILCs) are a recently identified subset of leukocytes that play a central role in pathogen surveillance and resistance, modulation of immune response, and tissue repair. They are remarkably similar to CD4+ T-helper subsets in terms of function and transcription factors required for their development but are distinguished by their lack of antigen-specific receptors. Despite their similarities, the absence of a surface T-cell receptor (TCR) and presence of ILCs and precursors in adult bone marrow has led to speculation that ILCs and T cells develop separately from lineages that branch at the point of precursors within the bone marrow. Considering the common lineage markers and effector cytokine profiles shared between ILCs and T cells, it is surprising that the status of the TCR loci in ILCs was not fully explored at the time of their discovery. Here, we demonstrate that a high proportion of peripheral tissue ILC2s have TCRγ chain gene rearrangements and TCRδ locus deletions. Detailed analyses of these loci show abundant frameshifts and premature stop codons that would encode nonfunctional TCR proteins. Collectively, these data argue that ILC2 can develop from T cells that fail to appropriately rearrange TCR genes, potentially within the thymus.

Summary Background/Objectives Vocal changes in the male singing voice associated with puberty are variable and often unpredictable resulting in challenges for the singer and the choral director. Limited knowledge regarding the physiologic changes in the vocal mechanism as they correlate to perceptual variations observed in the male adolescent singer exists in the literature. The purpose of this study was to examine pitch breaks and perceptual characteristics of vocal quality during singing tasks for boys in various stages of the male changing voice. Study Design Prospective Study. Methods Twenty-eight boys were initially evaluated at Cooksey Stage 0 (Pubertal Unchanged; n = 15) or Cooksey Stage 1 (Mid-Voice; n = 13). Range of age was 8–13 years old. Participants performed vocal slide intervals (1-3-1, 1-5-1, 1-8-1) with discrete starting frequencies on G3, C4, F4, and A4 and sang the “Star-Spangled Banner” in the key of Ab. Pitch breaks and perceptual qualities were evaluated on the recorded tasks by expert raters. Seven boys were evaluated again when they progressed to Cooksey Stage 4 (Baritone) performing the same singing tasks. Results For the participants evaluated at Cooksey Stage 0/1, pitch breaks were observed more in the higher frequencies and increased interval spacing regardless of starting frequency. Participants at Cooksey Stage 0 had more pitch breaks than Stage 1. At Cooksey Stage 4, an increase in the number of pitch breaks was observed in comparison to their tasks performed at Stage 0/1 and the perceptual quality of breathiness was significantly greater. Conclusions Pitch breaks are a characteristic perceptual change that indicates a young man may be transitioning through puberty. Findings from the present study demonstrate that in addition to perceived pitch breaks, breathiness was noted to significantly increase as the male progressed through puberty. Breathiness was noted to be more significant than vocal timbre and overall vocal quality. This research provides acoustic evidence to enhance the perceptual characteristics of voice change for those who teach and train male voices through puberty.

Objectives: This study examined the association between pure tone hearing sensitivity and music listening behaviors among traditional college-aged students and sought to determine factors that mediate hearing sensitivity, including health and fitness levels, gender, and personal listening device (PLD) use. Methods: A convenience sample of college students (N = 182; 133 females, 49 males, mean age = 19.8 ± 1.4 year, average PLD use = 1.52 ± 7.1 hours•day−1) completed hearing assessments, music listening behavior questionnaires, and health and fitness tests. Results: Most students listened to music at safe intensity levels (Conclusion: The majority of college students had healthy music listening behavior and fitness, contributing to normal hearing sensitivity in most. In cases where greater hearing threshold levels at one or more frequencies was detected, TC:HDL and TC:TG were statistically related and at 2 kHz, males were more likely to demonstrate higher listening levels compared with females of similar health and fitness level.

Humans employ anticipatory muscle activation when catching under conditions of load uncertainty. Questions addressed were (a) on what information referent do catchers base their anticipatory neuromotor control when catching balls of unknown weight?, and (b) how do catchers use this functional referent? Thirty-six participants caught visually identical balls dropped from 0.75 m. Participants performed 40 trials, half with knowledge of ball weight and half without. Group L caught balls with a large weight range, while group S caught balls with a smaller range of weights. EMG integrals were computed for the ball flight period in five muscles. Anticipatory EMG integrals in the unknown weight condition were normalized to anticipatory EMG integrals for the maximum, minimum and average ball weights in the known ball weight condition. We assumed participants would base anticipatory control in the unknown weight condition on similar information, regardless of group. Therefore, differences in normalized EMG integrals between groups L and S would suggest that the specific referent tested (e.g., minimum possible ball weight) was not used to scale anticipatory muscle activation under load uncertainty. Independent sample t tests ascertained differences in normalized EMG integrals between groups L and S. The results suggested that the information referent participants used to catch balls of an unknown weight was knowledge of the maximum ball weight. Participants used this referent to generate a submaximal level of anticipatory muscle activation, i.e., about 93.2% of that used to catch the heaviest ball when ball weight was known in advance.

A critical question in invasion biology involves the relative importance of propagule rain and community invasibility. For plant invasions, invasibility is often related to disturbance, but few studies of forest invaders have simultaneously investigated both canopy and ground-level disturbance. We investigated the relative importance of seed rain, canopy disturbance, and soil disturbance in a mature forest in Maryland on the recruitment of four invasive species: wine raspberry (Rubus phoenicolasiusMaxim.), Japanese barberry (Berberis thunbergiiDC), multiflora rose (Rosa multifloraThunb.), and Japanese stiltgrass [Microstegium vimineum(Trin.) A. Camus]. Using complete censuses of a 9-ha plot at two points in time (2011–12 and 2014), we mapped new recruits, and related their locations to canopy and soil disturbance, as well as to a seed rain index based on locations of reproducing plants and seed-dispersal kernels.We found that propagule rain, as measured by the seed rain index, was a significant predictor of recruitment forB. thunbergii,R. phoenicolasius, andM. vimineum. ForR. multiflora, seed sources were not located, precluding assessment of propagule rain, but recruitment was linked to canopy disturbance, as was recruitment ofM. vimineum. However, because reproduction ofR. phoenicolasiusand, in some years, ofB. thunbergiiis higher in treefall gaps, these gaps experience higher propagule rain, with the result that recruitment is indirectly associated with these gaps. Ground-layer disturbance was an important predictor of recruitment only forB. thunbergii. Our findings reveal that the importance of propagule rain is the most consistent driver of recruitment, but canopy or ground-layer disturbance promotes recruitment of some invasive plant species.

The mammalian gastrointestinal tract harbors a microbial community with metabolic activity critical for host health, including metabolites that can modulate effector functions of immune cells. Mice treated with vancomycin have an altered microbiome and metabolite profile, exhibit exacerbated T helper type 2 cell (Th2) responses, and are more susceptible to allergic lung inflammation. Here we show that dietary supplementation with short-chain fatty acids (SCFAs) ameliorates this enhanced asthma susceptibility by modulating the activity of T cells and dendritic cells (DCs). Dysbiotic mice treated with SCFAs have fewer interleukin-4 (IL4)-producing CD4+ T cells and decreased levels of circulating immunoglobulin E (IgE). In addition, DCs exposed to SCFAs activate T cells less robustly, are less motile in response to CCL19 in vitro, and exhibit a dampened ability to transport inhaled allergens to lung draining nodes. Our data thus demonstrate that gut dysbiosis can exacerbate allergic lung inflammation through both T cell- and DC-dependent mechanisms that are inhibited by SCFAs.

The use of antioxidants in tissue regeneration has been studied, but their mechanism of action is not well understood. Here, we analyze the role of the antioxidant N-acetylcysteine (NAC) in retina regeneration. Embryonic chicks are able to regenerate their retina after its complete removal from retinal stem/progenitor cells present in the ciliary margin (CM) of the eye only if a source of exogenous factors, such as FGF2, is present. This study shows that NAC modifies the redox status of the CM, initiates self-renewal of the stem/progenitor cells, and induces regeneration in the absence of FGF2. NAC works as an antioxidant by scavenging free radicals either independently or through the synthesis of glutathione (GSH), and/or by reducing oxidized proteins through a thiol disulfide exchange activity. We dissected the mechanism used by NAC to induce regeneration through the use of inhibitors of GSH synthesis and the use of other antioxidants with different biochemical structures and modes of action, and found that NAC induces regeneration through its thiol disulfide exchange activity. Thus, our results provide, for the first time, a biochemical basis for induction of retina regeneration. Furthermore, NAC induction was independent of FGF receptor signaling, but dependent on the MAPK (pErk1/2) pathway.

Cold stress is an important abiotic factor that can impact insect physiology, behavior, and overall fitness. Upon exposure to cold temperature, many insects enter a reversible state of immobility called chill coma. If the cold stress is brief and mild enough, insects can recover and regain full mobility upon return to warmer temperatures. However, the long-term impact of sublethal cold stress on insect behavior has been understudied. Here, sexually naive adult male Acheta domesticus crickets were exposed to a single 0 °C cold stress for 6 h. One week later, the ability of these males to mate with a female was examined. For mating trials, a cold stressed male cricket was paired with a non-cold stressed, control female. Control pairs were comprised of a non-cold stressed control male and control female. Cold exposed males were less successful at mating than control males because most did not carry a spermatophore at the time of their mating trials. However, when these cold stressed males were allowed 1 h of chemosensory contact with a female, most produced a spermatophore. Males that produced spermatophores were given the opportunity to mate once with a female, and stressed males that successfully mated sired as many offspring as did control males. However, our results support that a single cold stress exposure can negatively impact the reproductive fitness of male crickets since it reduced their capacity to carry spermatophores and, as a consequence, to attract females.

A critical event during kidney organogenesis is the differentiation of podocytes, specialized epithelial cells that filter blood plasma to form urine. Podocytes derived from human pluripotent stem cells (hPSC-podocytes) have recently been generated in nephron-like kidney organoids, but the developmental stage of these cells and their capacity to reveal disease mechanisms remains unclear. Here, we show that hPSC-podocytes phenocopy mammalian podocytes at the capillary loop stage (CLS), recapitulating key features of ultrastructure, gene expression, and mutant phenotype. hPSC-podocytes in vitro progressively establish junction-rich basal membranes (nephrin+podocin+ZO-1+) and microvillus-rich apical membranes (podocalyxin+), similar to CLS podocytes in vivo. Ultrastructural, biophysical, and transcriptomic analysis of podocalyxin-knockout hPSCs and derived podocytes, generated using CRISPR/Cas9, reveals defects in the assembly of microvilli and lateral spaces between developing podocytes, resulting in failed junctional migration. These defects are phenocopied in CLS glomeruli of podocalyxin-deficient mice, which cannot produce urine, thereby demonstrating that podocalyxin has a conserved and essential role in mammalian podocyte maturation. Defining the maturity of hPSC-podocytes and their capacity to reveal and recapitulate pathophysiological mechanisms establishes a powerful framework for studying human kidney disease and regeneration.

Mutations and ectopic FAT1 cadherin expression are implicated in a broad spectrum of diseases ranging from developmental disorders to cancer. The regulation of FAT1 and its downstream signalling pathways remain incompletely understood. We hypothesized that identification of additional proteins interacting with the FAT1 cytoplasmic tail would further delineate its regulation and function. A yeast two-hybrid library screen carried out against the juxtamembrane region of the cytoplasmic tail of FAT1 identified the E3 ubiquitin-protein ligase SH3RF1 as the most frequently recovered protein-binding partner. Ablating SH3RF1 using siRNA increased cellular FAT1 protein levels and stabilized expression at the cell surface, while overexpression of SH3RF1 reduced FAT1 levels. We conclude that SH3RF1 acts as a negative post-translational regulator of FAT1 levels.

Muscle activation was measured using EMG in 28 males (n = 28) while participants caught visually identical balls of known and unknown weights (50, 1.32, 2.18, and 2.99 kg) under variable (1–10s) and constant (3s) foreperiods. EMG integrals were computed for three time intervals before the catch (anticipatory), and one after (compensatory). Load uncertainty caused the CNS to use an anticipatory strategy characterized by preparation to catch balls of an unknown weight by utilizing about 92% of the muscle activation used to catch the heaviest possible ball under the known weight condition. The CNS appeared to scale anticipatory muscle activation to afford an opportunity to catch a ball of an unknown weight between .50 and 2.99 kg. The constant 3s foreperiod, which permitted temporal anticipation, did not influence the anticipatory neuromotor strategy adopted by the CNS to cope with load uncertainty. Load uncertainty also altered compensatory neuromotor control in catching.

Innate lymphoid cells (ILCs) are critical for host defense and tissue repair but can also contribute to chronic inflammatory diseases. The transcription factor RORα is required for ILC2 development but is also highly expressed by other ILC subsets where its function remains poorly defined. We previously reported that Rorasg/sg bone marrow chimeric mice (C57BL/6J) were protected from Salmonella-induced intestinal fibrosis due to defective ILC3 responses. In this study, single-cell RNA analysis of ILCs isolated from inflamed tissues indicates that RORα perturbation led to a reduction in ILC3 lineages. Furthermore, residual Rorasg/sg ILC3s have decreased expression of key signature genes, including Rorc and activating cytokine receptors. Collectively, our data suggest that RORα plays a key role in preserving functional ILC3s by modulating their ability to integrate environmental cues to efficiently produce cytokines.

To reveal how the CNS copes with load uncertainty in catching, electromyography (EMG) was recorded in 15 females and 14 males while catching visually identical balls of known and unknown weights under varied (1-10 s) and constant (1 s) foreperiods (warning time). EMG integrals, which represented total muscle activity, were computed for three time intervals prior to the catch (anticipatory), and one interval after (compensatory). Load uncertainty caused the CNS to utilize an anticipatory strategy in several muscles, primarily during the ball-flight interval, characterized by preparation to catch balls of unknown weight by utilizing an average of 99.7% of the muscle activation used to catch the heaviest ball under the known weight condition. The constant 1 s foreperiod, which permitted precise temporal anticipation of ball release, did not influence the anticipatory strategy adopted by the CNS to cope with load uncertainty. There were no observed differences in the neuromotor control used by men and women to manage load uncertainty in catching, although there was an interesting difference in the way men and women employed the triceps to prepare to catch balls of a known weight.

Celebrity suicides that are reported heavily in the media may increase risk for others' suicidal behavior. This study examined whether Internet search volumes for suicide-related terms changed after three celebrity suicide deaths (Robin Williams, Chester Bennington, and Alexander McQueen) and three celebrities who died by means other than suicide (David Bowie, Azzedine Alaia, and Paul Walker). Suicide search terms included suicide, how to suicide, commit suicide, depression, hanging, and suicide prevention. Observed suicide search volumes in the United States were collected from Google Trends for the 10 weeks before and the 2 weeks following each celebrity's death. Predicted search volumes for the 14 days postdeath were forecasted from the predeath search volumes and predicted search volumes were then compared to the true, observed search volumes. Search volumes for suicide terms significantly increased following Robin Williams' suicide death. Some of the terms increased in search volume following Chester Bennington's and Alexander McQueen's suicide deaths, but not significantly. Most search volumes for nonsuicide celebrity deaths did not change following their deaths. Celebrity suicide deaths can lead to significant, national increases in Internet search volumes for suicide-related terms for celebrities of high prominence. Results highlight the critical importance of reporting suicide deaths in the media responsibly.

Genome-wide association studies have linked gene variants of the receptor patched homolog 1 (PTCH1) with chronic obstructive pulmonary disease (COPD). However, its biological role in the disease is unclear. Our objective was to determine the expression pattern and biological role of PTCH1 in the lungs of patients with COPD. Airway epithelial-specific PTCH1 protein expression and epithelial morphology were assessed in lung tissues of control and COPD patients. PTCH1 mRNA expression was measured in bronchial epithelial cells obtained from individuals with and without COPD. The effects of PTCH1 siRNA knockdown on epithelial repair and mucous expression were evaluated using human epithelial cell lines. Ptch1+/− mice were used to assess the effect of decreased PTCH1 on mucous expression and airway epithelial phenotypes. Airway epithelial-specific PTCH1 protein expression was significantly increased in subjects with COPD compared to controls, and its expression was associated with total airway epithelial cell count and thickness. PTCH1 knockdown attenuated wound closure and mucous expression in airway epithelial cell lines. Ptch1+/− mice had reduced mucous expression compared to wildtype mice following mucous induction. PTCH1 protein is up-regulated in COPD airway epithelium and may upregulate mucous expression. PTCH1 provides a novel target to reduce chronic bronchitis in COPD patients.

Proinflammatory cytokines produced during immune responses to infectious stimuli are well-characterized to have secondary effects on the function of hematopoietic progenitor cells in the BM. However, these effects on the BM are poorly characterized during chronic infection with intestinal helminth parasites. In this study, we use the Trichuris muris model of infection and show that Th1 cell-associated, but not acute Th2 cell-associated, responses to chronic T. muris infection cause a major, transient expansion of CD48- CD150- multipotent progenitor cells in the BM that is dependent on the presence of adaptive immune cells and IFN-γ signaling. Chronic T. muris infection also broadly stimulated proliferation of BM progenitor cells including CD48- CD150+ hematopoietic stem cells. This shift in progenitor activity during chronic T. muris infection correlated with a functional increase in myeloid colony formation in vitro as well as neutrophilia in the BM and peripheral blood. In parallel, we observed an accumulation of CD4+ , CD8+ , and CD4- CD8- (double negative) T cells that expressed IFN-γ, displaying activated and central memory-type phenotypes in the bone marrow during chronic infection. Thus, these results demonstrate that Th1 cell-driven responses in the intestine during chronic helminth infection potently influence upstream hematopoietic processes in the BM via IFN-γ.

Antignano, Zaph, and collaborators show that the lysine methyltransferase G9a plays a critical role in determining the developmental programs of group 2 and 3 innate lymphoid cells., Innate lymphoid cells (ILCs) are emerging as important regulators of homeostatic and disease-associated immune processes. Despite recent advances in defining the molecular pathways that control development and function of ILCs, the epigenetic mechanisms that regulate ILC biology are unknown. Here, we identify a role for the lysine methyltransferase G9a in regulating ILC2 development and function. Mice with a hematopoietic cell–specific deletion of G9a (Vav.G9a−/− mice) have a severe reduction in ILC2s in peripheral sites, associated with impaired development of immature ILC2s in the bone marrow. Accordingly, Vav.G9a−/− mice are resistant to the development of allergic lung inflammation. G9a-dependent dimethylation of histone 3 lysine 9 (H3K9me2) is a repressive histone mark that is associated with gene silencing. Genome-wide expression analysis demonstrated that the absence of G9a led to increased expression of ILC3-associated genes in developing ILC2 populations. Further, we found high levels of G9a-dependent H3K9me2 at ILC3-specific genetic loci, demonstrating that G9a-mediated repression of ILC3-associated genes is critical for the optimal development of ILC2s. Together, these results provide the first identification of an epigenetic regulatory mechanism in ILC development and function.

BackgroundThe success of new targeted cancer therapies has been dependent on the identification of tumor-specific antigens. Podocalyxin (Podxl) is upregulated on tumors with high metastatic index and its presence is associated with poor outcome, thus emerging as an important prognostic and theragnostic marker in several human cancers. Moreover, in human tumor xenograft models, Podxl expression promotes tumor growth and metastasis. Although a promising target for immunotherapy, the expression of Podxl on normal vascular endothelia and kidney podocytes could hamper efforts to therapeutically target this molecule. Since pathways regulating post-translational modifications are frequently perturbed in cancer cells, we sought to produce novel anti-Podxl antibodies (Abs) that selectively recognize tumor-restricted glycoepitopes on the extracellular mucin domain of Podxl.MethodsSplenic B cells were isolated from rabbits immunized with a Podxl-expressing human tumor cell line. Abs from these B cells were screened for potent reactivity to Podxl+ neoplastic cell lines but not Podxl+ primary endothelial cells. Transcripts encoding heavy and light chain variable regions from promising B cells were cloned and expressed as recombinant proteins. Tumor specificity was assessed using primary normal tissue and an ovarian cancer tissue microarray (TMA). Mapping of the tumor-restricted epitope was performed using enzyme-treated human tumor cell lines and a glycan array.ResultsOne mAb (PODO447) showed strong reactivity with a variety of Podxl+ tumor cell lines but not with normal primary human tissue including Podxl+ kidney podocytes and most vascular endothelia. Screening of an ovarian carcinoma TMA (219 cases) revealed PODO447 reactivity with the majority of tumors, including 65% of the high-grade serous histotype. Subsequent biochemical analyses determined that PODO447 reacts with a highly unusual terminal N-acetylgalactosamine beta-1 (GalNAcβ1) motif predominantly found on the Podxl protein core. Finally, Ab–drug conjugates showed specific efficacy in killing tumor cells in vitro.ConclusionsWe have generated a novel and exquisitely tumor-restricted mAb, PODO447, that recognizes a glycoepitope on Podxl expressed at high levels by a variety of tumors including the majority of life-threatening high-grade serous ovarian tumors. Thus, tumor-restricted PODO447 exhibits the appropriate specificity for further development as a targeted immunotherapy.

Gut dysbiosis alters the development and severity of atopic disease. We previously demonstrated that nursing dams and newborn mice treated with low-dose vancomycin alters gut microbial diversity with a marked loss of bacteria that produce short-chain fatty acids (including butyrate). Vancomycin-induced gut dysbiosis enhances the TH2 response to lung allergens due to altered dendritic cell trafficking and activation in addition to modifying the behavior of other mature leukocyte lineages. Butyrate supplementation reverses the vancomycin-induced TH2 pro-inflammatory phenotype. Butyrate is known to exert some of its effects on target cells by inhibiting histone deacetylases (HDACs) with consequent effects on gene expression. Consistent with a role for epigenetic skewing of the hematopoietic compartment, we found that engraftment of total bone marrow from dysbiotic mice transferred enhanced TH2 proclivity in normobiotic recipients. Strikingly, we found unique regulatory states (H3K27ac marks) in purified hematopoietic stem and progenitor cells (HSPC) of TH2-skewed recipient mice. Single cell RNA sequence analyses identified a distinct transcriptomic signature in HSPC of dysbiotic mice that was reversed by butyrate supplementation. Together, these data suggest that the gut microbiome alters gene expression in blood progenitor cells with long term consequences on the immune response to peripheral allergens.