Standard Error Of The Mean Research Articles

Pest characteristics of Trigonorhinus sp. (Coleoptera: Anthribidae) and the developmental process of its galls in Caragana liouana Zhao Y. Chang and Yakovlev

Abstract In recent years, a Trigonorhinus sp. (Coleoptera) has caused serious damage to Caragana liouana Zhao Y. Chang and Yakovlev, a major ecological restoration shrub in China. Here, we survey the occurrence and damage pattern of Trigonorhinus sp. and its galls and compare the growth of affected and unaffected plants. Trigonorhinus sp. larvae usually infest the main trunk and lateral branches of the plant, causing the affected branches to become partially swollen and verrucose. Galls develop in stages depending on whether eggs are laid in May or July and proceed in sequence over three years from egg-laying to formation, expansion, dormancy, maturity, dormancy, maturity, and death. Galls inhibit plant development to some extent. On average, six (at least one, and no more than 18; standard error of the mean = 3) larvae occupy each gall, and the number of larvae within a gall did not significantly affect gall size. Gall size significantly affected branch dieback, and large-diameter infested branches had larger galls. This study clarifies the growth dynamics of Trigonorhinus sp. galls and provides a basis for further research into the growth mechanism of the species’ galls.

Effects of a single transdermal administration of flunixin meglumine in early postpartum Holstein Friesian dairy cows: Part 1. Inflammatory and metabolic markers, uterine health, and indicators of pain.

The objectives of this study were to assess the effects of a single transdermal administration of flunixin meglumine (FM) in early postpartum Holstein Friesian dairy cows on serum concentrations of inflammatory and metabolic markers, uterine health, and indicators of pain. The hypothesis was that the anti-inflammatory, antipyretic, and analgetic effects of the pharmaceutic agent would reduce systemic inflammation, resulting in improved metabolic and inflammatory profile, diminished incidence of metritis, and reduced expression of pain. A total of 500 cows (153 primiparous, 347 multiparous) from 3 different commercial dairy farms in the northeast of Germany were included in a randomized controlled clinical trial. Farms were preselected based on high haptoglobin concentrations in their fresh lactating cows. Cows were excluded if they had experienced dystocia, stillbirth, or twin birth, or if they showed any signs of milk fever, retained fetal membranes, or fever (>40°C). The cows were treated once with either FM (3.33 mg/kg) or a placebo as control (CON) through transdermal administration between 24 to 36 h postpartum (d 2). General health examinations were performed (daily from d 2-8 and additionally on d 15 postpartum), vaginal discharge was assessed using the Metricheck device (d 8 and 15 postpartum) and serum samples were analyzed for inflammatory and metabolic markers (d 2, 4, and 6 postpartum). Effects of treatment, parity, sampling day, and their interactions were evaluated using mixed effects models. Primiparous cows treated with FM showed lower serum haptoglobin concentrations (0.90 ± 0.08 vs. 1.17 ± 0.07 g/L; ± standard error of the mean) and higher serum albumin concentrations (35.5 ± 0.31 vs. 34.8 ± 0.31 g/L) on d 6 postpartum. They also had a lower risk for purulent vaginal discharge with or without a fever compared with CON cows on d 15 postpartum (odds ratio for CON vs. FM: 1.63, 95% CI: 1.26-2.00), and body temperature was lower throughout the first 15 d in milk (39.1 ± 0.11 vs. 39.2 ± 0.11°C). Multiparous cows treated with FM had lower serum β-hydroxybutyrate concentrations on d 4 postpartum (0.71 ± 0.05 vs. 0.78 ± 0.05 mmol/L) and d 6 postpartum (0.74 ± 0.05 vs. 0.80 ± 0.05 mmol/L). Regardless of parity, FM-treated cows were significantly less likely to abduct their tail from their body (14.3 vs. 23.6%) and show an arched back (27.9 vs. 39.7%) on the day after treatment compared with CON cows. It can be concluded that FM treatment slightly reduced inflammation and diminished the risk for metritis in primiparous cows, improved metabolic profile in multiparous cows, and reduced expressions of pain in all cows.

Somatostatin Receptor PET/MR Imaging of Inflammation in Patients With Large Vessel Vasculitis and Atherosclerosis

BackgroundAssessing inflammatory disease activity in large vessel vasculitis (LVV) can be challenging by conventional measures. ObjectivesWe aimed to investigate somatostatin receptor 2 (SST2) as a novel inflammation-specific molecular imaging target in LVV. MethodsIn a prospective, observational cohort study, in vivo arterial SST2 expression was assessed by positron emission tomography/magnetic resonance imaging (PET/MRI) using 68Ga-DOTATATE and 18F-FET-βAG-TOCA. Ex vivo mapping of the imaging target was performed using immunofluorescence microscopy; imaging mass cytometry; and bulk, single-cell, and single-nucleus RNA sequencing. ResultsSixty-one participants (LVV: n = 27; recent atherosclerotic myocardial infarction of ≤2 weeks: n = 25; control subjects with an oncologic indication for imaging: n = 9) were included. Index vessel SST2 maximum tissue-to-blood ratio was 61.8% (P < 0.0001) higher in active/grumbling LVV than inactive LVV and 34.6% (P = 0.0002) higher than myocardial infarction, with good diagnostic accuracy (area under the curve: ≥0.86; P < 0.001 for both). Arterial SST2 signal was not elevated in any of the control subjects. SST2 PET/MRI was generally consistent with 18F-fluorodeoxyglucose PET/computed tomography imaging in LVV patients with contemporaneous clinical scans but with very low background signal in the brain and heart, allowing for unimpeded assessment of nearby coronary, myocardial, and intracranial artery involvement. Clinically effective treatment for LVV was associated with a 0.49 ± 0.24 (standard error of the mean [SEM]) (P = 0.04; 22.3%) reduction in the SST2 maximum tissue-to-blood ratio after 9.3 ± 3.2 months. SST2 expression was localized to macrophages, pericytes, and perivascular adipocytes in vasculitis specimens, with specific receptor binding confirmed by autoradiography. SSTR2-expressing macrophages coexpressed proinflammatory markers. ConclusionsSST2 PET/MRI holds major promise for diagnosis and therapeutic monitoring in LVV. (PET Imaging of Giant Cell and Takayasu Arteritis [PITA], NCT04071691; Residual Inflammation and Plaque Progression Long-Term Evaluation [RIPPLE], NCT04073810)

Evaluation of right ventricle echocardiographic variables in apparently healthy domestic shorthair cats.

Right ventricular structural and functional changes result from many cardiovascular disorders in cats. Accordingly, echocardiographic evaluation of the size and function of the right ventricle (RV) provides important prognostic information in many conditions affecting the right heart. Therefore, detection of these changes is clinically important and needs reference values to ease the diagnosis. The present study was designed to calculate structural and functional right ventricular variables in 10 apparently healthy cats (six males and four females, average age 1 year old and body weight 2.70 - 4.80 kg) with no sedation. For this purpose, The minimum, maximum, mean ± standard error of the mean of right ventricle internal dimension in systole and diastole (cm), right ventricle free wall thickness in systole and diastole (cm), fractional shortening (%), right ventricle volume in systole and diastole (mL), right ventricle area in systole and diastole (cm2), ejection fraction (%) and fractional area change (%) were measured and reported. Descriptive statistics were provided for all calculated variables. Statistical correlation of the collected variables with body weight, gender and heart rate were analyzed. Results showed that there is a significant correlation between heart rate with body weight and between systolic and diastolic RV volumes with gender. The results of the present study would help clinicians in the diagnosis of the right heart changes in DSH cats.

Evaluation and estimation of diuretic activity of the linalyl acetate in the rats.

This study aimed to explore the diuretic activity of linalyl acetate (LA). LA is an essential oil, it is an integral phyto-constituent of various plants. In this study, acute and chronic diuretic activities were explored by measuring the levels of different electrolytes and pH in the urine of experimental rats. Rats were divided into five groups. The control group was given 10 mg/kg normal saline, the treated group was given 10 mg/kg furosemide, and the remaining 3 groups received different doses of LA including 25, 50, and 75 mg/kg through intraperitoneal route, to determine its diuretic potential. Urine volume for acute diuretic activity was measured for 6 hours however for chronic diuretic activity was measured for 6 days. For a comparative study of LA with a control group and treated group with reference drug, diuretic index was used. Moreover, the underlying mechanism of the diuretic activity was also explored by comparing atropine, L-NAME, and indomethacin. The results of each group with 6 rats in each group were obtained by ± standard error of the mean of every group. Analysis of Variance (ANOVA) was used for statistical analysis. Results revealed that the LA 75 mg/kg dose showed comparable results as of furosemide. Moreover, this study revealed the involvement of muscarinic receptors to produce diuresis in comparison with atropine with very little involvement of prostanoids and no effect on NO pathway induced by indomethacin and L-NAME respectively. It is concluded that LA possess anti-diuretic potential. Muscarinic receptors might be involved in producing diuretic effects.

Energy-entropy multiscale cell correlation method to predict toluene-water log P in the SAMPL9 challenge.

The energy-entropy multiscale cell correlation (EE-MCC) method is used to calculate toluene-water log P values of 16 drug molecules in the SAMPL9 physical properties challenge. EE-MCC calculates the free energy, energy and entropy from molecular dynamics (MD) simulations of the water and toluene solutions. Specifically, MCC evaluates entropy by partitioning the system into cells of correlated atoms at multiple length scales and further partitioning the local coordinates into energy wells, yielding vibrational and topographical terms from the energy-well sizes and probabilities. The log P values calculated by EE-MCC using three 200 ns MD simulations have a mean average error of 0.82 and standard error of the mean of 0.97 versus experiment, which is comparable with the best methods entered in SAMPL9. The main contribution to log P is from energy. Less polar drugs have more favourable energies of transfer. The entropy of transfer consists of increased solute vibrational and conformational terms in toluene due to weaker interactions, fewer solute positions in the larger-molecule solvent, reduced water vibrational entropy, negligible change in toluene vibrational entropy, and gains in solvent orientational entropy. The solvent entropy contributions here may be slightly underestimated because software limitations and statistical fluctuations meant that only the first shell could be included while averaged over the whole solution. Nonetheless, such issues will be addressed in future software to offer a general method to calculate entropy directly from MD simulation and to provide molecular understanding or guide system design.

Manufacture and Characterization of Good Manufacturing Practice-Compliant SARS-COV-2 Cytotoxic T Lymphocytes.

Coronavirus disease 2019 (COVID-19) is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). SARS-CoV-2 virus-specific cytotoxic T-cell lymphocytes (vCTLs) could provide a promising modality in COVID-19 treatment. We aimed to screen, manufacture, and characterize SARS-CoV-2-vCTLs generated from convalescent COVID-19 donors using the CliniMACS Cytokine Capture System (CCS). Donor screening was done by stimulation of convalescent COVID-19 donor peripheral blood mononuclear cells with viral peptides and identification of interferonγ (IFN-γ)+ CD4 and CD8 T cells using flow cytometry. Clinical-grade SARS-CoV-2-vCTLs were manufactured using the CliniMACS CCS. The enriched SARS-CoV-2-vCTLs were characterized by T-cell receptor sequencing, mass cytometry, and transcriptome analysis. Of the convalescent donor blood samples, 93% passed the screening criteria for clinical manufacture. Three validation runs resulted in enriched T cells that were 79% (standard error of the mean 21%) IFN-γ+ T cells. SARS-CoV-2-vCTLs displayed a highly diverse T-cell receptor repertoire with enhancement of both memory CD8 and CD4 T cells, especially in CD8 TEM, CD4 TCM, and CD4 TEMRA cell subsets. SARS-CoV-2-vCTLs were polyfunctional with increased gene expression in T-cell function, interleukin, pathogen defense, and tumor necrosis factor superfamily pathways. Highly functional SARS-CoV-2-vCTLs can be rapidly generated by direct cytokine enrichment (12 hours) from convalescent donors. NCT04896606.

Effect of potassium tellurite concentration in a chromogenic agar medium on isolation of tellurite-resistant “Top Seven” Shiga toxin-producing Escherichia coli from ground beef

The effect of potassium tellurite concentration in a chromogenic agar medium on the detection of tellurite-resistant “top seven” Shiga toxin-producing Escherichia coli (STEC) in beef was evaluated. Samples of ground beef were inoculated with tellurite-resistant STEC O26, O45, O103, O111, O121, O145, or O157 strains at geometric mean (±standard error of the mean) levels of 0, 49 (±1), 490 (±1), or 4900 (±1) CFU/10 g and enriched 1:10 (90 mL) in EC broth (40°C for 6 h). Following enrichment, aliquots of broth culture were treated by immunomagnetic separation with one of three pools of beads against STEC serogroups; pool I: O26, O45, and O121; pool II: O103, O111, and O145; and pool III: O157. After immunomagnetic separation, 50 μL of washed bead suspensions in buffered peptone water was spiral plated onto a modified Possé medium containing 0.5, 1.0, or 1.5 mg/L potassium tellurite, and incubated at 37°C for 18 h. Up to four isolated colonies were picked from each spiral plate based on expected colony phenotypes for STEC, and isolate identity was confirmed with an 11-plex PCR assay targeting the O serogroups and virulence genes. Overall, across all inoculum levels and strains, modified Possé media containing 0.5, 1.0, or 1.5 mg/L potassium tellurite each had a positive predictive value of 100%, and medium containing 0.5 mg/L potassium tellurite had numerically the highest sensitivity (100%) and negative predictive value (100%), which was significantly different from 1.5 mg/L (92.9% and 40.0%, respectively; P < 0.05). Similarly, there was an inverse relationship between potassium tellurite concentration and analytical specificity (number of colonies tested that were STEC-positive): 0.5 (1463 of 1482; 98.7%), 1.0 (1356 of 1411; 96.1%), and 1.5 mg/L (1187 of 1278; 92.9%; P < 0.05). These results suggest that 0.5 mg/L gives better performance than 1.0 or 1.5 mg/L of potassium tellurite in Possé medium for isolation of tellurite-resistant “top seven” STEC from ground beef.

Large-scale, pragmatic randomized trials in the era of big data, precision medicine and machine learning. Valid and necessary, or outdated and a waste of resources?

During the past decade, more and more large-scale pragmatic clinical trials have been carried out in orthopedic trauma surgery. This trend is fueled by the common belief that the larger the numbers in a trial, the broader the eligibility criteria, and the less strict the regimentation of local treatment standards by protocol, the more trustworthy the findings would be. However, it must also be taken into account that the precision of an outcome measurement does not depend on the sample size alone, but the homogeneity of the studied population. Consequently, a small trial with stringent entry and assessment criteria may offer similarly precise answers as a large trial with less strict entry and assessment criteria because of the basic mathematical correlation between standard deviation and standard error of the mean. There is now a lively and controversial debate about the role of randomized controlled trials (RCT) in an era of stratified medicine driven by the ever increasing understanding and clinical measurability of molecular pathways, making a certain intervention more effective in patients who show a distinct genetic variant. Cluster and pattern recognition by artificial intelligence (AI) and its methodological variety applied to huge datasets and population-based cohorts further propel the spiral of knowledge. Advanced adaptive RCT concepts like enrichment designs, basket and bucket trials, master protocols etc. were developed to combine classic principles of the scientific method with big data, the latter of which have not arrived yet in trauma care.In spite of all biomedical and methodological achievements made, surprisingly such key questions remain unanswered as a) is a certain treatment causally responsible for making a difference in patient-centered outcomes compared to placebo, a control treatment, or the standard of care, b) do the results of a controlled experiment are relevant enough to change clinical practice, and c) under which conditions and assumptions shall we conduct large-scale pragmatic RCTs, focused confirmatory RCTs, or personalized analyses with or without AI support.

Translation, cross-cultural adaptation, validity and reliability of the Brazilian Portuguese version of Activity Scale for Kids questionnaire.

The Activity Scale for Kids (ASK) assesses the physical disability of children (5 to 15 years old) with neurological, orthopaedic or rheumatic diseases. The objective of this study was to translate and cross-culturally adapt the ASK for Brazilian Portuguese and assess the validity and reliability of the instrument. A total of 67 children and adolescents with musculoskeletal, neurological or rheumatic diseases participated in the study. We evaluated the comprehension of the pre-final version of the questionnaire in 24 participants and reliability and validity in the other 43 participants. The translation and adaptation of ASK to Brazilian Portuguese followed guidelines from previous studies. The validity of the Brazilian Portuguese version of the ASK was verified through Spearman's correlation with the Pediatric Quality of Life Inventory™ Version 4.0 (PedQL). Intraclass correlation coefficient verified inter- and intra-evaluator reliability, while internal consistency was assessed using Cronbach's alpha. Scores were used to assess the standard error of the mean and minimal detectable change. The Brazilian Portuguese version of the ASK presented excellent reliability, internal consistency, agreement and moderate correlation with PedsQL (0.522, P< 0.001) between ASK performance and PedsQL; 0.537, P< 0.001 between ASK capacity and PedsQL. The Brazilian Portuguese version of the ASK has good validity and reliability and may be used by professionals and researchers to assess the functioning of children and adolescents with disabilities.

Heat treatment and homogenization of bovine milk loosened gastric curd structure and increased gastric emptying in growing pigs

During gastric digestion, bovine milk forms a curd, which consists largely of proteins and lipids. However, it is unknown how processing-induced changes to curd structure affects the gastric emptying of milk proteins and lipids. This study aimed to determine the impact of heat treatment and homogenization on gastric curd formation, and gastric emptying of dry matter (DM), proteins and lipids from bovine milk fed to pigs as a human model. Growing pigs (n = 180, mean ± standard error of the mean (SEM) bodyweight 22.4 ± 0.13 kg) consumed raw, or pasteurized non-homogenized (PNH), or pasteurized homogenized (PH), or ultra-high temperature treated homogenized (UHT) milk diets. A protein-lipid-free lactose (PLFL) solution was also fed as a test diet. At 0, 20, 60, 120, 180 and 300 min postprandially the entire gastrointestinal tract was dissected out. The gastric chyme (curd and liquid) fractions were collected after separation using a mesh screen. The DM, protein, and lipid contents of these fractions were quantified. Confocal, transmission electron microscopy, cryo-scanning electron microscopy and rheological analyses were conducted to determine the micro- and macrostructure of the curd. Overall, both heat treatment and homogenization influenced the in vivo gastric curd structure formed of bovine milk, although to different extents. The gastric emptying of DM, proteins, and lipids increased with the extent of processing. Gastric emptying rates of DM and proteins followed the pattern UHT > PH > PNH = raw, while emptying rates of lipid also differed between PNH and raw milk. Curd structure was the main gastric parameter affected in PNH milk.

Increasing levels of calcium salts of palm fatty acids affect production responses during the immediate postpartum and carryover periods in dairy cows.

Our objective was to determine the dose-response effects of calcium salts of palm fatty acids (CSPF) on nutrient digestibility and production responses of early-lactation dairy cows grazing on tropical pastures and to evaluate carryover effects throughout mid and late lactation. Forty multiparous dairy cows (Jersey × Holstein) with (mean ± standard error of the mean) 20 ± 1.69 kg of milk/d and 20 ± 5.0 d in milk were used in a randomized complete block design. During the treatment period, all cows were kept in a grazing system. The treatments were offered for 90 d (treatment period) and consisted of 4 increasing levels of CSPF: 0 (0 kg/d), 0.2 (0.2 kg/d), 0.4 (0.4 kg/d), and 0.6 (0.6 kg/d). Each treatment had 10 animals. Increasing CSPF from 0 to 0.6 kg/d replaced an equivalent amount of a corn-based concentrate supplement offered at 10 kg/d on an as-fed basis (8.96 kg/d as a dry matter basis). All cows were housed and received a diet without fat inclusion fed as total mixed ration once a day from 91 to 258 d of the experiment (carryover period). During the treatment period, increasing CSPF linearly decreased dry matter intake (1.20 kg/d), linearly increased neutral detergent fiber digestibility (3.90 percentage units), and quadratically increased total fat digestibility (6.30 percentage units at 0.4 kg/d CSPF). Increasing CSPF linearly increased the yields of milk (4.10 kg/d), milk fat (0.11 kg/d), milk lactose (0.19 kg/d), energy-corrected milk (ECM; 3.30 kg/d), and feed efficiency (ECM/dry matter intake, 0.34 kg/kg), and linearly decreased milk protein content (0.38 g/100 g), body weight change (0.05 kg/d), and body condition score (0.37). We observed interactions between CSPF and time during the carryover period. Overall, CSPF supplementation linearly increased or tended to increase milk yield until 202 d of the experiment with a similar pattern observed for all the other yield variables. In conclusion, supplementing CSPF from 0 to 0.6 kg/d during 90 d increased neutral detergent fiber and total fat digestibility and the yields of milk, milk fat, and ECM in early-lactation dairy cows grazing on tropical pastures. Most production measurements linearly increased during the treatment period, indicating that 0.6 kg/d CSPF was the best dose. Also, supplementing CSPF from 0 to 0.6 kg/d for 90 d during early lactation had positive carryover effects across mid and late lactation.

Factors Associated With Unplanned Reoperation After 4-Corner Arthrodesis: A Study of 478 Wrists

The 4-corner arthrodesis (FCA) is a reliable, motion-sparing technique used to treat scapholunate advanced collapse and scaphoid nonunion advanced collapse arthritis, particularly in stage III wrists in which the capitolunate articulation is compromised. Surgical technique and patient-level variables may influence complications following FCA. We sought to evaluate the rate of complications in a large, combined database and manual chart review study. Current Procedural Terminology codes were used to search the United States Veteran's Health Administration corporate database to identify wrists treated with FCA over a 24-year period. A retrospective chart review was completed to collect data regarding scapholunate advanced collapse/scaphoid nonunion advanced collapse stage, implant used, the use of a bone graft, smoking status, and comorbidities for all patients undergoing an FCA. A multivariable cox proportional hazards regression was used to assess hazard ratios for reoperation. Incidence rates and the standard error of the mean for reoperation and conversion to total wrist fusion were calculated after grouping patients by 10-year age categories. A total of 478 wrists underwent FCA during the study period, with a mean follow-up of 63 months. Seventy-three (16%) wrists required reoperation. The most frequent secondary procedures included unplanned implant removal (8.2%), total wrist arthrodesis (4.6%), and revision FCA (1.7%). Positive smoking history increased the risk of reoperation, whereas posterior interosseous nerve neurectomy, arthritis stage, and fixation type did not have a statistically significant association with reoperation. Younger age demonstrated an increased incidence of overall reoperation and wrist fusion. The most common reason for reoperation after FCA was implant removal. Smoking history is associated with increased rates of reoperation and wrist arthrodesis. Knowledge of these factors may assist with accurately counseling and indicating patients for FCA. Therapeutic III.

Reduction in Endothelial Adhesion of Red Blood Cells Under Crizanlizumab: Assessment of Standardized Microfluidic Platform in Sickle Cell Disease Population

Introduction: Monoclonal P-selectin blocking antibody (Crizanlizumab, ADAKVEO®, Novartis, Switzerland) targets reduction of vaso-occlusive crises (VOCs) in adult and pediatric patients (≥16 years) with sickle cell disease (SCD). Patients with SCD have been reported with endothelial p-selectin to be initiatory for endothelial adhesion of sickle red blood cells (RBCs). Herein, we report the novel endothelium-on-a-chip (microfluidic platform - MFP) that enables assessment of Crizanlizumab on the adhesion of RBCs to perfusion-cultured human endothelial cells (ECs) in acute and chronic activation setup. Methods: Whole blood samples collected from SCD subjects (n=14), HbSS and in EDTA and sodium citrate. RBCs were isolated via centrifugation from whole blood and resuspended in basal cell culture medium (EBM; Lonza, Morristown, NJ, USA) at a hematocrit of 20% with 10 mM of HEPES. Human umbilical vein endothelial cells (HUVECs; Lonza, Morristown, NJ, USA) were cultured within the microfluidic platform channels at 15 dyne/cm2 for at least 48 hours prior to experiments. To mimic the acute EC activation, blood samples were supplemented with 40 µM heme +/- 100 µg/ml Crizanlizumab and injected through the microfluidic channels for 15 minutes. Instead to mimic chronic pre-activation in SCD HUVECs were pretreated for 4 hours with TNFα (20 ng/ml) or 50% plasma of SCD patients in basal media before, +/- 100 µg/ml Crizanlizumab followed by injection of blood samples through the microfluidic channels. Non-adherent RBCs were removed via washing solution with or without Crizanlizumab and the remaining RBCs were imaged using an inverted microscope and the manual quantification was completed based on characteristics previous published. Paired t-test to evaluate the effects of the Crizanlizumab on RBCs adherent to activated EC has been used. Results: We observed a statistically significant reduction of RBC adhesion due to Crizanlizumab treatment in 15-minutes acutely heme activated ECs (135±40 vs 1513±617, p≤0.05). The effect of Crizanlizumab on decreasing RBC adhesion was also statistically significant when ECs were chronically pre-activated by TNF-α followed by a 15-minute heme activation (4404±1393 vs 2016±609, p≤0.05). To further elaborate on the effects of Crizanlizumab in the atherogenic SCD population, we activated the EC with subjects' autologous plasma for 4 hours followed by a 15-minute heme activation, (5876±2579 vs 2397±1381p≤0.05). Discussion and Conclusions: Per label use of Crizanlizumab to reduce VOC, application of novel MFP associated with Crizanlizumab use displayed a reduced RBC adhesion to acutely heme-activated EC. Effects were preserved in presence or absence of chronic TNFα or autologous plasma pre-activation. Endothelium-on-a-chip MFP has been shown as reliable in monitoring patient response to anti-adhesive therapies in SCD population. Fig 1. Endothelium-on-a-chip MFP set up (A). The phase-contrast image of HUVECs demonstrate a confluent layer of cells aligned with flow. (C) RBCs adhesion to activated HUVECs. Effects of Crizanlizumab on RBC adhesion levels to HUVECs activated for 15-minutes with heme (D); 4-hours with TNF-α and 15-minutes with heme (E); 4-hours with plasma and 15-minutes with heme(F). Microscope images at 10X. Scale bare 100um. Error bars represent the standard error of the mean (SEM). Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Novel Application of 3D Scaffolds of Poly(E-Caprolactone)/Graphene as Osteoinductive Properties in Bone Defect.

Scaffolds provided a surface on which cells could attach, proliferate, and differentiate. Nowadays, bone tissue engineering offers hope for treating bone cancer. Poly(e-caprolactone) (PCL)/graphene have capability as an osteogenic and regenerative therapy. It could be used to produce bone tissue engineering scaffolds. The purpose of this study was to investigate the ability of PCL/graphene to enhance the osteoinductive mechanism. The PCL/graphene scaffold was developed utilizing a particulate-leaching process and cultured with osteoblast-like cells MG63 at 0.5, 1.5, and 2.5 wt% of graphene. We evaluated the porosity, pore size, migratory cells, and cell attachment of the scaffold. Data was expressed as the mean ± standard error of the mean and statistical analyses were performed using one-way analysis of variance and Tukey's post hoc at a level of p-value < 0.05. Porosity of scaffold with various percentage of graphene was nonsignificant (p > 0.05). There were differences in the acceleration of cell migration following wound closure between groups at 24 hours (p < 0.01) and 48 hours (p < 0.00). Adding the graphene on the scaffolds enhanced migration of osteoblast cells culture and possibility to attach. Graphene on 2.5 wt% exhibited good characteristics over other concentrations. This finding suggests that PCL/graphene composites may have potential applications in bone tissue engineering.

Migration patterns and habitat use by molt migrant temperate‐breeding Canada geese in James Bay, Canada

Numbers of temperate‐breeding Atlantic and Mississippi Flyway Canada geese have greatly increased since the 1980s. Consequently, numbers of yearlings, sub‐adults and failed breeders undertaking pre‐molt migration to northern latitudes has also increased, potentially providing additional hunting opportunities for Cree hunters living near James Bay, Canada. We described movement patterns and habitat use of molt migrant Canada geese Brenta canadensis maxima along the east coast of James Bay based on nine geese fitted with GSM–GPS devices during 11 northward and eight southward migrations between 2015 and 2019. Geese staged for 2.8 ± 0.6 days (mean ± standard error of the mean) at 3.2 ± 0.6 staging sites (mostly tidal flats and salt marshes) from the first week of June in spring and 3.8 ± 1.8 days at 2.0 ± 0.5 staging sites (mostly inland freshwater wetlands, peatlands and tidal flats) from the first week of September when returning south. Shallow and deep water habitats were used as resting sites during both migrations. In spring, molt migrants were mostly harvested in June well after the migration of sub‐arctic breeding geese whereas in autumn, both subspecies were harvested in September. Molt migrant temperate‐breeding geese can increase harvest opportunities and represent supplementary wildlife food for Cree communities. However, the current number of molt migrant geese harvested by Cree hunters is not sufficient to significantly impact populations of temperate‐breeding geese.

A monitoring and data analysis method for microplastics in marine sediments

In Europe, policy frameworks demand the monitoring of microplastics in marine sediments. Here we provide a monitoring and data analysis method for microplastic particles designed to be used in the context of Marine Strategy Framework Directive (MSFD) and OSPAR policy frameworks. Microplastics were analysed in marine sediments at four different locations in Dutch coastal and transitional waters using replicate sampling to investigate micro-spatial variation. Particle size distribution followed a power law with slope 3.76. Thirteen polymers were identified, with their composition varying between sediments near densely populated West coast areas versus the more rural Wadden Sea area. We quantify differences in the micro-spatial variation of microplastic concentrations between locations using the relative standard error of the mean (RSEM). This metric provides an opportunity to optimize the sensitivity of trend detection in microplastic monitoring networks by selecting locations with relatively low micro-spatial variation. We provide a method to optimize the number of replicate samples for a given location using its relationship with the RSEM. Two replicate samples appear to be cost-effective for relatively homogenous locations, whereas more heterogenous locations require four replicates.

An inducible expression system for the manipulation of autophagic flux in vivo

ABSTRACT Much of our understanding of the intracellular regulation of macroautophagy/autophagy comes from in vitro studies. However, there remains a paucity of knowledge about how this process is regulated within different tissues during development, aging and disease in vivo. Because upregulation of autophagy is considered a promising therapeutic strategy for the treatment of diverse disorders, it is vital that we understand how this pathway functions in different tissues and this is best done by in vivo analysis. Similarly, to understand the role of autophagy in the pathogenesis of disease, it is important to study this process in the whole animal to investigate how tissue-specific changes in flux and cell-autonomous versus non-cell-autonomous effects alter disease progression. To this end, we have developed an inducible expression system to up- or downregulate autophagy in vivo, in zebrafish. We have used a modified version of the Gal4-UAS expression system to allow inducible expression of autophagy up- or downregulating transgenes by addition of tamoxifen. Using this inducible expression system, we have tested which transgenes robustly up- or downregulate autophagy and have validated these tools using Lc3-II blots and puncta analysis and disease rescue in a zebrafish model of neurodegeneration. These tools allow the temporal control of autophagy via the administration of tamoxifen and spatial control via tissue or cell-specific ERT2-Gal4 driver lines and will enable the investigation of how cell- or tissue-specific changes in autophagic flux affect processes such as aging, inflammation and neurodegeneration in vivo. Abbreviations: ANOVA: analysis of variance; Atg: autophagy related; Bcl2l11/Bim: BCL2 like 11; d.p.f.: days post-fertilization; Cryaa: crystallin, alpha a: DMSO: dimethyl sulfoxide; Elavl3: ELAV like neuron-specific RNA binding protein 3; ER: estrogen receptor; ERT2: modified ligand-binding domain of human ESR1/estrogen receptor α; Gal4: galactose-responsive transcription factor 4; GFP: green fluorescent protein; h.p.f.: hours post-fertilization; HSP: heat-shock protein; Map1lc3/Lc3: microtubule-associated protein 1 light chain 3; RFP: red fluorescent protein; SD: standard deviation; SEM: standard error of the mean; UAS: upstream activating sequence; Ubb: ubiquitin b

Single-incision cochlear implantation and hearing evaluation in piglets and minipigs

ObjectivesVarious animal models have been established and applied in hearing research. In the exploration of novel cochlear implant developments, mainly rodents have been used. Despite their important contribution to the understanding of auditory function, translation of experimental observations from rodents to humans is limited due to the size differences and genetic variability. Large animal models with better representation of the human cochlea are sparse. For this reason, we evaluated domestic piglets and Aachen minipigs for the suitability as a cochlear implantation animal model with commercially available cochlear implants. MethodsFour domestic piglets (two male and two female) and six Aachen minipigs were implanted with either MED-EL Flex24 or Flex20 cochlear implants respectively, after a step-by-step surgical approach was trained with pig cadavers. Electrophysiological measurements were performed before, during and after implantation for as long as 56 days after surgery. Auditory brainstem responses, electrocochleography as well as electrically and acoustically evoked compound action potentials were recorded. Selected cochleae were further analyzed histologically or with micro-CT imaging. ResultsA surgical approach was established using a retroauricular single incision. Baseline auditory thresholds were 27 ± 3 dB sound pressure level (SPL; auditory brainstem click responses, mean ± standard error of the mean) and ranged between 30 and 80 dB SPL in frequency-specific responses (0.5 – 32 kHz). Follow-up measurements revealed deafness within the first two weeks after surgery, but some animals partially recovered to a hearing threshold of 80 dB SPL in certain frequencies as well as in click responses. Electrically evoked compound action potential thresholds increased within the first week after surgery, which led to lower stimulation responses or increase of necessary charge input. Immune reactions and consecutive scalar fibrosis following implantation were confirmed with histological analysis of implanted cochleae and may result in increased impedances. A three-dimensional minipig micro-CT segmentation revealed cochlear volumetric data similar to human inner ear dimensions. ConclusionsThis study underlines the feasibility of cochlear implantation with clinically used cochlear implants in a large animal model with representative inner ear dimensions comparable to humans. To bridge the gap between small animal models and humans in translational research and to account for the structural and size differences, we recommend the minipig as a valuable animal model for hearing research. First insights into the induced trauma in minipigs after cochlear implant surgery and a partial hearing recovery present important data of the cochlear health changes in large animal cochleae.

The reliability of the measurement of muscle volume using magnetic resonance imaging in typically developing infants by two raters

To assess intra-rater and inter-rater reliability of the manual segmentation of Magnetic Resonance Imaging (MRI) for the in vivo measurement of infant muscle volume of the knee extensor and flexor muscles by two raters. Muscles of the knee extensor and flexor muscle of ten typically developing infants (86 days ± 7 days) were scanned with MRI (Proton density sequence). Scans were then segmented using Slicer software, and volumes rendered by two raters. Intra-rater and inter-rater reliability were assessed using intra-class correlation (ICC), with mean difference (MD), standard error of the mean (SEM), and minimal detectable change (MDC) for each muscle calculated. ICCs for Intra-rater reliability of the segmentation process for the muscle volume of the muscles of the knee extensors and flexor muscles were 0.901–0.972, and 0.776–0.945 respectively, with inter-rater reliabilities between 0.914–0.954 and 0.848–0.978, for the knee extensor and flexors muscles respectively. For intra-rater reliability, MD ≤ − 0.47 cm3, MDCs for were < 1.09 cm3 and for inter-rater MD ≤ − 1.40 cm3, MDCs for were < 1.63 cm3 for all muscles. MRI segmentation for muscle volumes showed good to excellent reliability, though given the small volumes of the muscles themselves, variations between raters are amplified. Care should be taken in the reporting and interpretation of infant muscle volume.