Advanced glycated end-products inhibit dilation through constitutive endothelial RAGE and Nox1/4 in rat isolated skeletal muscle arteries.

OBJECTIVE: This study investigated the actions of advanced glycated end-products (AGE), their receptors (RAGE), and NAD(P)H oxidase (Nox) subtypes 1, 2, and 4 on mechanisms of endothelium-dependent dilation of the rat cremaster muscle artery (CMA). METHODS: Immunofluorescence studies were used to examine expression of RAGE in rat arteries. ROS accumulation was measured using luminescence and fluorescence assays. Functional studies were performed using pressure myography. RESULTS: High levels of RAGE expression were shown in the endothelial cells of the CMA, compared with low endothelial expression in middle cerebral and mesenteric arteries and the aorta. Exogenous AGE (in vitro glycated bovine serum albumin) stimulated H2O2 accumulation in CMA, which was prevented by the RAGE antagonist FPS-ZM1, the NAD(P)H oxidase (Nox) inhibitor apocynin and inhibited by the Nox1/4 inhibitor setanaxib, but not the Nox2 inhibitor GSK2795039. In functional studies, AGE inhibited vasodilation of CMA stimulated by acetylcholine, sodium nitroprusside, and the BKCa activator NS1619, but not adenosine-induced dilation. FPS-ZM1, apocynin, and setanaxib prevented the inhibitory effects of AGE on responses to acetylcholine and NS-1619. CONCLUSION: These observations suggest RAGE are constitutively expressed in the endothelium of the rat CMA and may be activated by AGE to stimulate Nox1/4 and ROS formation with resulting inhibition of NO and BKCa-mediated endothelium-dependent dilation.

Improved Durability to SARS-CoV-2 Vaccine Immunity following Coimmunization with Molecular Adjuvant Adenosine Deaminase-1.

Although severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccines have demonstrated strong immunogenicity and protection against severe disease, concerns about the duration and breadth of these responses remain. In this study, we show that codelivery of plasmid-encoded adenosine deaminase-1 (pADA) with SARS-CoV-2 spike glycoprotein DNA enhances immune memory and durability in vivo. Coimmunized mice displayed increased spike-specific IgG of higher affinity and neutralizing capacity as compared with plasmid-encoded spike-only-immunized animals. Importantly, pADA significantly improved the longevity of these enhanced responses in vivo. This coincided with durable increases in frequencies of plasmablasts, receptor-binding domain-specific memory B cells, and SARS-CoV-2-specific T follicular helper cells. Increased spike-specific T cell polyfunctionality was also observed. Notably, animals coimmunized with pADA had significantly reduced viral loads compared with their nonadjuvanted counterparts in a SARS-CoV-2 infection model. These data suggest that pADA enhances immune memory and durability and supports further translational studies.

Population Based Time Trends in the Epidemiology and Mortality of Gastroesophageal Junction and Esophageal Adenocarcinoma.

BACKGROUND: Limited data are available on the epidemiology of gastroesophageal junction adenocarcinoma (GEJAC), particularly in comparison to esophageal adenocarcinoma (EAC). With the advent of molecular non-endoscopic Barrett's esophagus (BE) detection tests which sample the esophagus and gastroesophageal junction, early detection of EAC and GEJAC has become a possibility and their epidemiology has gained importance. AIMS: We sought to evaluate time trends in the epidemiology and survival of patients with EAC and GEJAC in a population-based cohort. METHODS: EAC and GEJAC patients from 1976 to 2019 were identified using ICD 9 and 10 diagnostic codes from the Rochester Epidemiology Project (REP). Clinical data and survival status were abstracted. Poisson regression was used to calculate incidence rate ratios (IRR). Survival analysis and Cox proportional models were used to assess predictors of survival. RESULTS: We included 443 patients (287 EAC,156 GEJAC). The incidence of EAC and GEJAC during 1976-2019 was 1.40 (CI 1.1-1.74) and 0.83 (CI 0.61-1.11) per 100,000 people, respectively. There was an increase in the incidence of EAC (IRR = 2.45, p = 0.011) and GEJAC (IRR = 3.17, p = 0.08) from 2000 to 2004 compared to 1995-1999, plateauing in later time periods. Most patients had associated BE and presented at advanced stages, leading to high 5-year mortality rates (66% in EAC and 59% in GEJAC). Age and stage at diagnosis were predictors of mortality. CONCLUSION: The rising incidence of EAC/GEJAC appears to have plateaued somewhat in the last decade. However, both cancers present at advanced stages with persistently poor survival, underscoring the need for early detection.

The Epigenetic Reader Protein SP140 Regulates Dendritic Cell Activation, Maturation and Tolerogenic Potential.

SP140 is an epigenetic reader protein expressed predominantly in immune cells. GWAS studies have shown an association between SP140 single nucleotide polymorphisms (SNPs) and diverse autoimmune and inflammatory diseases, suggesting a possible pathogenic role for SP140 in immune-mediated diseases. We previously demonstrated that treatment of human macrophages with the novel selective inhibitor of the SP140 protein (GSK761) reduced the expression of endotoxin-induced cytokines, implicating a role of SP140 in the function of inflammatory macrophages. In this study, we investigated the effects of GSK761 on in vitro human dendritic cell (DC) differentiation and maturation, assessing the expression of cytokines and co-stimulatory molecules and their capacity to stimulate T-cell activation and induce phenotypic changes. In DCs, lipopolysaccharide (LPS) stimulation induced an increase in SP140 expression and its recruitment to transcription start sites (TSS) of pro-inflammatory cytokine genes. Moreover, LPS-induced cytokines such as TNF, IL-6, and IL-1ß were reduced in GSK761- or SP140 siRNA- treated DCs. Although GSK761 did not significantly affect the expression of surface markers that define the differentiation of CD14+ monocytes into immature DCs (iDCs), subsequent maturation of iDCs to mature DCs was significantly inhibited. GSK761 strongly reduced expression of the maturation marker CD83, the co-stimulatory molecules CD80 and CD86, and the lipid-antigen presentation molecule CD1b. Finally, when the ability of DCs to stimulate recall T-cell responses by vaccine-specific T cells was assessed, T cells stimulated by GSK761-treated DCs showed reduced TBX21 and RORA expression and increased FOXP3 expression, indicating a preferential generation of regulatory T cells. Overall, this study suggests that SP140 inhibition enhances the tolerogenic properties of DCs, supporting the rationale of targeting SP140 in autoimmune and inflammatory diseases where DC-mediated inflammatory responses contribute to disease pathogenesis.

Typical m. triceps surae morphology and architecture measurement from 0 to 18 years: A narrative review.

The aim of this review was to report on the imaging modalities used to assess morphological and architectural properties of the m. triceps surae muscle in typically developing children, and the available reliability analyses. Scopus and MEDLINE (Pubmed) were searched systematically for all original articles published up to September 2020 measuring morphological and architectural properties of the m. triceps surae in typically developing children (18 years or under). Thirty eligible studies were included in this analysis, measuring fibre bundle length (FBL) (n = 11), pennation angle (PA) (n = 10), muscle volume (MV) (n = 16) and physiological cross-sectional area (PCSA) (n = 4). Three primary imaging modalities were utilised to assess these architectural parameters in vivo: two-dimensional ultrasound (2DUS; n = 12), three-dimensional ultrasound (3DUS; n = 9) and magnetic resonance imaging (MRI; n = 6). The mean age of participants ranged from 1.4 years to 18 years old. There was an apparent increase in m. gastrocnemius medialis MV and pCSA with age; however, no trend was evident with FBL or PA. Analysis of correlations of muscle variables with age was limited by a lack of longitudinal data and methodological variations between studies affecting outcomes. Only five studies evaluated the reliability of the methods. Imaging methodologies such as MRI and US may provide valuable insight into the development of skeletal muscle from childhood to adulthood; however, variations in methodological approaches can significantly influence outcomes. Researchers wishing to develop a model of typical muscle development should carry out longitudinal architectural assessment of all muscles comprising the m. triceps surae utilising a consistent approach that minimises confounding errors.

The reliability and validity of triceps surae muscle volume assessment using freehand three-dimensional ultrasound in typically developing infants.

This study assessed the intra-acquirer, intra- and inter-processor reliability, and validity of the in vivo assessment of the medial gastrocnemius (MG), lateral gastrocnemius (LG) and soleus (SOL) muscle volumes using freehand 3D ultrasound (3DUS) in typically developing infants. Reliability assessments of freehand 3DUS were undertaken in infants across three ages groups: three, six and twelve months of age, with validity testing completed against magnetic resonance imaging (MRI) in infants at 3 months of age. Freehand 3DUS scanning was carried out by a single acquirer, with two independent processors manually segmenting images to render volumes. MRI images were segmented independently by a separate processor, with the volumes compared to those obtained via freehand 3DUS. Reliability was assessed using intraclass correlation (ICC), coefficient of variance (CV) and minimal detectable change (MDC) across each assessment time point. Validity was assessed using the limits of agreement. ICCs for intra-acquirer reliability of the acquisition process for freehand 3DUS ranged from 0.91 to 0.99 across all muscles. ICCs for intra-processor and inter-processor reliability for the segmentation process of freehand 3DUS ranged from 0.80 to 0.98 across all muscles. Acceptable levels of agreement between muscle volume obtained by freehand 3DUS and MRI were found for all muscles; however, freehand 3DUS overestimated muscle volume of MG and LG and underestimate the SOL compared with MRI, with average absolute differences of MG = 0.3 ml, LG = 0.3 ml and Sol = 1.2 ml. Freehand 3DUS is a reliable method for measuring in vivo triceps surae muscle volume in typically developing infants. We conclude that freehand 3DUS is a useful tool to assess changes in muscle volume in response to growth and interventions in infants.

Innovations in Screening Tools for Barrett's Esophagus and Esophageal Adenocarcinoma.

PURPOSE OF REVIEW: Esophageal adenocarcinoma (EAC) is a lethal disease with rapidly rising incidence. Screening for EAC and its metaplastic precursor, Barrett's esophagus (BE), followed by endoscopic surveillance and endoscopic treatment of dysplasia or early EAC are promising approaches to decreasing EAC incidence and EAC mortality. Historically, screening for EAC has been completed with a traditional per-oral esophagogastroduodenoscopy (EGD); however, this method has limitations including cost, tolerability, and accessibility. For this reason, much effort has been put forward to develop more effective, minimally invasive, and accessible BE and EAC screening tools. The purpose of this review is to describe recent developments of these novel tools. RECENT FINDINGS: While endoscopic alternatives such as transnasal endoscopy are cheaper and well tolerated, they have not gained acceptance. Non-endoscopic modalities namely, swallowable cell collection devices coupled with biomarker analysis have been found to have excellent performance characteristics, tolerability, and cost effectiveness. In this article, we provide an update on innovative developments in EAC/BE screening modalities including transnasal endoscopy, capsule endomicroscopy, swallowable cell collection devices, and exhaled volatile organic compound analyses.

Improving robustness and accuracy of predicted daily methane emissions of dairy cows using milk mid-infrared spectra.

BACKGROUND: A robust proxy for estimating methane (CH4 ) emissions of individual dairy cows would be valuable especially for selective breeding. This study aimed to improve the robustness and accuracy of prediction models that estimate daily CH4 emissions from milk Fourier transform mid-infrared (FT-MIR) spectra by (i) increasing the reference dataset and (ii) adjusting for routinely recorded phenotypic information. Prediction equations for CH4 were developed using a combined dataset including daily CH4 measurements (n = 1089; g d-1 ) collected using the SF6 tracer technique (n = 513) and measurements using respiration chambers (RC, n = 576). Furthermore, in addition to the milk FT-MIR spectra, the variables of milk yield (MY) on the test day, parity (P) and breed (B) of cows were included in the regression analysis as explanatory variables. RESULTS: Models developed based on a combined RC and SF6 dataset predicted the expected pattern in CH4 values (in g d-1 ) during a lactation cycle, namely an increase during the first weeks after calving followed by a gradual decrease until the end of lactation. The model including MY, P and B information provided the best prediction results (cross-validation statistics: R2 = 0.68 and standard error = 57 g CH4 d-1 ). CONCLUSIONS: The models developed accounted for more of the observed variability in CH4 emissions than previously developed models and thus were considered more robust. This approach is suitable for large-scale studies (e.g. animal genetic evaluation) where robustness is paramount for accurate predictions across a range of animal conditions. © 2020 Society of Chemical Industry.

Explaining negative kin discrimination in a cooperative mammal society.

Kin selection theory predicts that, where kin discrimination is possible, animals should typically act more favorably toward closer genetic relatives and direct aggression toward less closely related individuals. Contrary to this prediction, we present data from an 18-y study of wild banded mongooses, Mungos mungo, showing that females that are more closely related to dominant individuals are specifically targeted for forcible eviction from the group, often suffering severe injury, and sometimes death, as a result. This pattern cannot be explained by inbreeding avoidance or as a response to more intense local competition among kin. Instead, we use game theory to show that such negative kin discrimination can be explained by selection for unrelated targets to invest more effort in resisting eviction. Consistent with our model, negative kin discrimination is restricted to eviction attempts of older females capable of resistance; dominants exhibit no kin discrimination when attempting to evict younger females, nor do they discriminate between more closely or less closely related young when carrying out infanticidal attacks on vulnerable infants who cannot defend themselves. We suggest that in contexts where recipients of selfish acts are capable of resistance, the usual prediction of positive kin discrimination can be reversed. Kin selection theory, as an explanation for social behavior, can benefit from much greater exploration of sequential social interactions.

Aspirin in Primary and Secondary Prevention of Cardiovascular Disease.

Aspirin is one of the most widely used drugs for the treatment of cardiovascular disease. While its use in patients with known cardiovascular disease has been supported with trials which have included mortality benefit, the utility of aspirin therapy in patients without established cardiovascular disease has been less clear. Early trials appeared to demonstrate benefit with the use of aspirin, but trials after 2000 did not consistently substantiate using aspirin for primary prevention of cardiovascular disease. Despite the lack of robust supportive evidence, aspirin was recommended and used extensively for primary prevention in patients at higher risk for cardiovascular events. More recently in 2018, results of three randomized, controlled trials: ARRIVE, ASCEND, ASPREE demonstrated modest to no benefit in preventing cardiovascular events and mortality with aspirin use for primary prevention. These trials also demonstrated an increased risk of bleeding in these patients who were on aspirin for primary prevention.

Biopsychosocial Pain Management and its Effect on Postoperative Opioid Use.

INTRODUCTION: Medical use and overuse of opioids have become an increasing problem over the past several decades. Postoperative pain control is the strongest indication for the use of opioid analgesics. Previous studies have demonstrated benefit from complementary and alternative therapy (CAT) for postoperative pain relief. A prior study conducted by Riswold et al. found that a unit staff training session on CAT improved patient experiences postoperatively following total joint replacement. The study was limited in that it did not examine if there were any changes in opioid usage following this intervention. METHODS: This study is a continuation of the Riswold et al. study on CAT training intervention. In July 2017, a four-hour staff training session on alternative comfort measures and pain medication administration took place. Opioid administration data was extracted from the PYXIS software for all patients who had received more than three opioid administrations across their hospital stay in the three months prior to CAT training and the three months post-training. Opioid administrations were converted to total oral morphine equivalents. The pre- and post-intervention groups were compared using independent sample t-tests using SPSS software. RESULTS: Statistically significant reduction of total oral morphine equivalents occurred following CAT training intervention (p=.034, CI 2.76, 69.81). Average oral morphine equivalents per day (p=0.023, CI 1.26, 16.57) and per administration (p=0.00048, CI 0.64, 2.25) also were significantly reduced following the CAT training intervention. CONCLUSION: This study strengthens the findings of prior studies, showing that CAT can improve patient satisfaction while also reducing overall opioid burden for post-surgical patients.

Pif1 helicase unfolding of G-quadruplex DNA is highly dependent on sequence and reaction conditions.

In addition to unwinding double-stranded nucleic acids, helicase activity can also unfold noncanonical structures such as G-quadruplexes. We previously characterized Pif1 helicase catalyzed unfolding of parallel G-quadruplex DNA. Here we characterized unfolding of the telomeric G-quadruplex, which can fold into antiparallel and mixed hybrid structures and found significant differences. Telomeric DNA sequences are unfolded more readily than the parallel quadruplex formed by the c-MYC promoter in K+ Furthermore, we found that under conditions in which the telomeric quadruplex is less stable, such as in Na+, Pif1 traps thermally melted quadruplexes in the absence of ATP, leading to the appearance of increased product formation under conditions in which the enzyme is preincubated with the substrate. Stable telomeric G-quadruplex structures were unfolded in a stepwise manner at a rate slower than that of duplex DNA unwinding; however, the slower dissociation from G-quadruplexes compared with duplexes allowed the helicase to traverse more nucleotides than on duplexes. Consistent with this, the rate of ATP hydrolysis on the telomeric quadruplex DNA was reduced relative to that on single-stranded DNA (ssDNA), but less quadruplex DNA was needed to saturate ATPase activity. Under single-cycle conditions, telomeric quadruplex was unfolded by Pif1, but for the c-MYC quadruplex, unfolding required multiple helicase molecules loaded onto the adjacent ssDNA. Our findings illustrate that Pif1-catalyzed unfolding of G-quadruplex DNA is highly dependent on the specific sequence and the conditions of the reaction, including both the monovalent cation and the order of addition.

Impact of food allergy on food insecurity and health literacy in a tertiary care pediatric allergy population.

BACKGROUND: Food insecurity (FI), limited availability of or access to nutritional foods, is linked to poor child/caregiver health. We examined FI in food-allergic and non-food-allergic children to determine whether dietary limitations associated with food allergy increases risk of FI. METHODS: Food-allergic and non-food-allergic children (1-17 years) were recruited from Arkansas Children's Hospital allergy/asthma clinics. The USDA Food Security Survey, the Newest Vital Sign Health Literacy (HL) questionnaire, and the Food Allergy Impact Scale QOL survey were administered. Logistic regression and analysis of covariance models were utilized for data analysis. RESULTS: Subjects (n = 650) included 325 food-allergic and 325 non-food-allergic children. Overall rate of FI was 21.5% (food allergic 22.2% and non-food allergic 20.9%) with no significant difference in the prevalence of FI between groups (OR = 1.30; 95% CI 0.86-1.96; P = 0.21). FI was increased in households of children with both milk and egg allergy when compared to those without food allergy and those with single food allergy (OR = 2.5; 95% CI 1.4-4.6; P = 0.003). Mean HL rates were higher in the food-secure vs food-insecure groups (mean diff = 0.31; 95% CI 0.03-0.59; P = 0.03). Among food-allergic children, QOL was better in the food-secure vs food-insecure group (mean diff = 0.61; 95% CI 0.002-1.23; P = 0.049). CONCLUSION: Food allergy to milk and egg was associated with increased risk of household FI. Food-insecure participants had lower HL than their food-secure counterparts. Further work is needed to define risks associated with FI among food-allergic children to improve screening and management strategies.

Infrared ion spectroscopy: an analytical tool for the study of metabolites.

Vibrational ion spectroscopy techniques coupled with mass spectrometry are applied to standard metabolites as a proof-of-principle demonstration for the structural identification of unknown metabolites. The traditional room temperature infrared multiple photon dissociation (IRMPD) spectroscopy technique is shown to differentiate chemical moieties in isobaric and isomeric variants. These results are compared to infrared spectra of cryogenically cooled analyte ions, showing enhanced spectral resolution, and thus also improved differentiation between closely related molecules, such as isomers. The cryogenic spectroscopy is effected in a recently developed mass-selective cryogenic linear ion trap, which is capable of high sensitivity and the ability to measure the IR spectra of multiple analytes simultaneously.

Probing the Structures of Solvent-Complexed Ions Formed in Electrospray Ionization Using Cryogenic Infrared Photodissociation Spectroscopy.

The gas-phase infrared photodissociation (IRPD) spectra of solvent-tagged small biomolecules are studied in a cryogenic ion trap at 17 K. In this study para-aminobenzoic acid (PABA) and tyramine molecules are noncovalently tagged with water or acetonitrile in the electrospray ionization (ESI) source. The complexes are then cooled in the cryogenic trap prior to spectroscopic measurements. These molecules provide two putative sites for solvent attachment: the protonated amine (NH3+) and the OH groups. Comparisons of the experimental IR spectra to theoretical spectra obtained with density functional theory show that the NH3+ site is mainly favored. Evidence for the formation of both NH3-bound and OH-bound conformers is found only in tyramine, despite having similar solution- and gas-phase energetics to that of PABA. Since the structures cannot interconvert in the gas phase, this suggests an isomerization during the electrospray process.

Evidence That G-quadruplex DNA Accumulates in the Cytoplasm and Participates in Stress Granule Assembly in Response to Oxidative Stress.

Cells engage numerous signaling pathways in response to oxidative stress that together repair macromolecular damage or direct the cell toward apoptosis. As a result of DNA damage, mitochondrial DNA or nuclear DNA has been shown to enter the cytoplasm where it binds to "DNA sensors," which in turn initiate signaling cascades. Here we report data that support a novel signaling pathway in response to oxidative stress mediated by specific guanine-rich sequences that can fold into G-quadruplex DNA (G4DNA). In response to oxidative stress, we demonstrate that sequences capable of forming G4DNA appear at increasing levels in the cytoplasm and participate in assembly of stress granules. Identified proteins that bind to endogenous G4DNA in the cytoplasm are known to modulate mRNA translation and participate in stress granule formation. Consistent with these findings, stress granule formation is known to regulate mRNA translation during oxidative stress. We propose a signaling pathway whereby cells can rapidly respond to DNA damage caused by oxidative stress. Guanine-rich sequences that are excised from damaged genomic DNA are proposed to enter the cytoplasm where they can regulate translation through stress granule formation. This newly proposed role for G4DNA provides an additional molecular explanation for why such sequences are prevalent in the human genome.

Biased escorts: offspring sex, not relatedness explains alloparental care patterns in a cooperative breeder.

Kin selection theory predicts that animals should direct costly care where inclusive fitness gains are highest. Individuals may achieve this by directing care at closer relatives, yet evidence for such discrimination in vertebrates is equivocal. We investigated patterns of cooperative care in banded mongooses, where communal litters are raised by adult 'escorts' who form exclusive caring relationships with individual pups. We found no evidence that escorts and pups assort by parentage or relatedness. However, the time males spent escorting increased with increasing relatedness to the other group members, and to the pup they had paired with. Thus, we found no effect of relatedness in partner choice, but (in males) increasing helping effort with relatedness once partner choices had been made. Unexpectedly, the results showed clear assortment by sex, with female carers being more likely to tend to female pups, and male carers to male pups. This sex-specific assortment in helping behaviour has potential lifelong impacts on individual development and may impact the future size and composition of natal groups and dispersing cohorts. Where relatedness between helpers and recipients is already high, individuals may be better off choosing partners using other predictors of the costs and benefits of cooperation, without the need for possibly costly within-group kin discrimination.

Chronic inorganic arsenic exposure in vitro induces a cancer cell phenotype in human peripheral lung epithelial cells.

Inorganic arsenic is a human lung carcinogen. We studied the ability of chronic inorganic arsenic (2 µM; as sodium arsenite) exposure to induce a cancer phenotype in the immortalized, non-tumorigenic human lung peripheral epithelial cell line, HPL-1D. After 38 weeks of continuous arsenic exposure, secreted matrix metalloproteinase-2 (MMP2) activity increased to over 200% of control, levels linked to arsenic-induced cancer phenotypes in other cell lines. The invasive capacity of these chronic arsenic-treated lung epithelial (CATLE) cells increased to 320% of control and colony formation increased to 280% of control. CATLE cells showed enhanced proliferation in serum-free media indicative of autonomous growth. Compared to control cells, CATLE cells showed reduced protein expression of the tumor suppressor gene PTEN (decreased to 26% of control) and the putative tumor suppressor gene SLC38A3 (14% of control). Morphological evidence of epithelial-to-mesenchymal transition (EMT) occurred in CATLE cells together with appropriate changes in expression of the EMT markers vimentin (VIM; increased to 300% of control) and e-cadherin (CDH1; decreased to 16% of control). EMT is common in carcinogenic transformation of epithelial cells. CATLE cells showed increased KRAS (291%), ERK1/2 (274%), phosphorylated ERK (p-ERK; 152%), and phosphorylated AKT1 (p-AKT1; 170%) protein expression. Increased transcript expression of metallothioneins, MT1A and MT2A and the stress response genes HMOX1 (690%) and HIF1A (247%) occurred in CATLE cells possibly in adaptation to chronic arsenic exposure. Thus, arsenic induced multiple cancer cell characteristics in human peripheral lung epithelial cells. This model may be useful to assess mechanisms of arsenic-induced lung cancer.