Observational Behavioral Coding in the Pediatric Emergency Department: Development of the Emergency Department Child Behavior Coding System.

BACKGROUND: Despite improvements over the past decade, children continue to experience significant pain and distress surrounding invasive procedures in the emergency department (ED). To assess the impact of newly developed interventions, we must create more reliable and valid behavioral assessment tools that have been validated for the unique settings of pediatric EDs. OBJECTIVE: This study aimed to create and test the Emergency Department Child Behavior Coding System (ED-CBCS) for the assessment of child distress and nondistress behaviors surrounding pediatric ED procedures. METHODS: Via an iterative process, a multidisciplinary expert panel developed the ED-CBCS, an advanced time-based behavioral coding measure. Inter-rater reliability and concurrent validity were examined using 38 videos of children aged from 2 to 12 years undergoing laceration procedures. Face, Legs, Activity, Cry, Consolability (FLACC) scale scores were used to examine concurrent validity. RESULTS: The final ED-CBCS included 27 child distress and nondistress behaviors. Time-unit κ values from 0.64 to 0.98 and event alignment κ values from 0.62 to 1.00 indicated good to excellent inter-rater reliability for all but one of the individual codes. ED-CBCS distress (B = 1.26; p < 0.001) and nondistress behaviors (B = -0.69, p = 0.025) were independently significantly associated with FLACC scores, indicating concurrent validity. CONCLUSIONS: We developed a psychometrically sound tool tailored for pediatric ED procedures. Future work could use this measure to better identify behavioral targets and test the effects of interventions to relieve pediatric ED pain and distress.

Chain End-Functionalized Dense Polymer Brushes from an Inimer Coating by SI-RAFT.

Polymer brushes with controllable grafting density are grown on an inimer coating bearing Reversible Addition-Fragmentation Chain Transfer polymerization (RAFT) chain transfer agents (CTAs). The inimer coating is cross-linked on the substrate to provide an initiator layer that is stable during exposure to organic solvents at high temperatures. Surface-initiated RAFT is conducted to grow poly(2-vinylpyridine) (P2VP) brushes on the coating at grafting densities approaching the theoretical limits. This methodology allows facile end-group functionalization using an efficient thiol-ene click chemistry. Chain ends were functionalized with low surface energy groups to modulate the location of the untethered chain ends by thermal annealing. At lower grafting densities, the low surface energy groups segregate to the surface upon annealing. This effect is less pronounced at higher grafting densities. Detailed characterization of the brushes at varying grafting densities using X-ray photoelectron spectroscopy (XPS) is presented. In tandem with experiments, Monte Carlo simulations examine the effect of the chain-end group size and selectivity on the conformation of the polymer brush, providing numerical evidence of laterally non-uniform distributions of functional groups at different locations in the brush. Simulations further predict the existence of morphologies with an interlayer formed by spherical micelles rich in functional end groups, demonstrating the possibility of end-group functionalization for synthetic modulation of both brush conformation and chain-end location.

Pro-Apoptotic Activity and Cell Cycle Arrest of Caulerpa sertularioides against SKLU-1 Cancer Cell in 2D and 3D Cultures.

Cancer is a disease with the highest mortality and morbidity rate worldwide. First-line drugs induce several side effects that drastically reduce the quality of life of people with this disease. Finding molecules to prevent it or generate less aggressiveness or no side effects is significant to counteract this problem. Therefore, this work searched for bioactive compounds of marine macroalgae as an alternative treatment. An 80% ethanol extract of dried Caulerpa sertularioides (CSE) was analyzed by HPLS-MS to identify the chemical components. CSE was utilized through a comparative 2D versus 3D culture model. Cisplatin (Cis) was used as a standard drug. The effects on cell viability, apoptosis, cell cycle, and tumor invasion were evaluated. The IC50 of CSE for the 2D model was 80.28 µg/mL versus 530 µg/mL for the 3D model after 24 h of treatment exposure. These results confirmed that the 3D model is more resistant to treatments and complex than the 2D model. CSE generated a loss of mitochondrial membrane potential, induced apoptosis by extrinsic and intrinsic pathways, upregulated caspases-3 and -7, and significantly decreased tumor invasion of a 3D SKLU-1 lung adenocarcinoma cell line. CSE generates biochemical and morphological changes in the plasma membrane and causes cell cycle arrest at the S and G2/M phases. These findings conclude that C. sertularioides is a potential candidate for alternative treatment against lung cancer. This work reinforced the use of complex models for drug screening and suggested using CSE's primary component, caulerpin, to determine its effect and mechanism of action on SKLU-1 in the future. A multi-approach with molecular and histological analysis and combination with first-line drugs must be included.

DNA aptamer selection for SARS-CoV-2 spike glycoprotein detection.

The rapid spread of SARS-CoV-2 infection throughout the world led to a global public health and economic crisis triggering an urgent need for the development of low-cost vaccines, therapies and high-throughput detection assays. In this work, we used a combination of Ideal-Filter Capillary Electrophoresis SELEX (IFCE-SELEX), Next Generation Sequencing (NGS) and binding assays to isolate and validate single-stranded DNA aptamers that can specifically recognize the SARS-CoV-2 Spike glycoprotein. Two selected non-competing DNA aptamers, C7 and C9 were successfully used as sensitive and specific biological recognition elements for the development of electrochemical and fluorescent aptasensors for the SARS-CoV-2 Spike glycoprotein with detection limits of 0.07 fM and 41.87 nM, respectively.

Risk of acquiring Ascaris lumbricoides infection in an endemically infected rural community in Venezuela.

Soil-transmitted helminths, such as Ascaris lumbricoides, are the most prevalent parasites globally. Optimal anthelmintic treatment for A. lumbricoides in endemically infected communities is challenged by several host-related and environmental factors influencing infection acquisition. We assessed the risk of A. lumbricoides (re)infection after treatment in a Venezuelan rural community. Individual merthiolate-iodine-formaldehyde-fixed faecal samples were collected from 224 persons before a single-dose pyrantel treatment and at 1, 3, 6, 9 and 15 months after treatment. Effects of age, sex and socioeconomic status (SES) on A. lumbricoides prevalence, eggs/gram faeces (EPG) and infection (re)acquisition were assessed using both generalised linear mixed-effects models and survival analysis. Pre-treatment A. lumbricoides prevalence was 39.7%. Higher prevalence was associated with younger age and lower SES. Higher EPG values were observed among young children. Median time to A. lumbricoides infection was six months after treatment: at 1, 3, 6, 9 and 15 months post-treatment, cumulative incidence was 6.7%, 18.9%, 34.6%, 42.2%, and 52.6%, respectively. Younger age, lower SES, and pre-treatment A. lumbricoides infection status showed significantly elevated hazard ratios. Mass drug administration protocols would benefit from considering these factors in selective treatment strategies and possibly more than just annual or biannual treatments in the target population.

High-resolution phase-shifting Ronchi test.

A method adding phase-shifting capacity in two mutually perpendicular axes to the Ronchi test is presented in this work. The phase of the object with the position of the reflected ray on the grating was identified and used to solve the equation of reflection in two orthogonal directions. In this manner, the test-surface figure was obtained. The procedure was demonstrated with an inverse qualitative test and a direct, quantitative test. Both tests give results comparable to Fizeau interferometry, with the precision of the order of 25 nm peak to valley. This technique is a good alternative to interferometry because, in addition to its inherent high-resolution, it is possible to obtain the radius of curvature and conic constant, which interferometers, requiring auxiliary optics, cannot provide. This method also has a high dynamic range and is not as susceptible to vibrations or turbulence. The setup can be built with low-cost, readily available components, is easily aligned, uses a white light source, and can be made very lightweight and compact, which makes it ideal for mounting onto existing polishing machines in any optical fabrication workshop, to perform in situ surface metrology.

Non-Functionalized Gold Nanoparticles Inhibit Human Papillomavirus (HPV) Infection.

The spontaneous interaction between human papillomavirus type 16 (HPV16) L1 virus-like particles (VLPs) and non-functionalized gold nanoparticles (nfGNPs) interferes with the nfGNPs' salt-induced aggregation, inhibiting the red-blue color shift in the presence of NaCl. Electron microscopy and competition studies showed that color-shift inhibition is a consequence of direct nfGNP-VLP interaction and, thus, may produce a negative impact on the virus entry cell process. Here, an in vitro infection system based on the HPV16 pseudovirus (PsV) was used to stimulate the natural infection process in vitro. PsVs carry a pseudogenome with a reporter gene, resulting in a fluorescent signal when PsVs infect a cell, allowing quantification of the viral infection process. Aggregation assays showed that nfGNP-treated PsVs also inhibit color shift in the presence of NaCl. High-resolution microscopy confirmed nfGNP-PsV complex formation. In addition, PsVs can interact with silver nanoparticles, suggesting a generalized interaction of metallic nanoparticles with HPV16 capsids. The treatment of PsVs with nfGNPs produced viral infection inhibition at a higher level than heparin, the canonical inhibitor of HPV infection. Thus, nfGNPs can efficiently interfere with the HPV16 cell entry process and may represent a potential active component in prophylactic formulations to reduce the risk of HPV infection.

Choriodecidual leukocytes display a unique gene expression signature in spontaneous labor at term.

Prior to and during the process of human labor, maternal circulating leukocytes infiltrate the maternal-fetal interface (choriodecidua) and become activated resembling choriodecidual leukocytes. Since, there is no evidence comparing maternal circulating and choriodecidual leukocytes, herein, we characterized their transcriptome and explored the biological processes enriched in choriodecidual leukocytes. From women undergoing spontaneous term labor we isolated circulating and choriodecidual leukocytes, performed microarray analysis (n = 5) and qRT-PCR validation (n = 9) and interaction network analysis with up-regulated genes. We found 270 genes up-regulated and only 17 genes down-regulated in choriodecidual leukocytes compared to maternal circulating leukocytes. The most up-regulated genes were CCL18, GPNMB, SEPP1, FN1, RNASE1, SPP1, C1QC, and PLTP. The biological processes enriched in choriodecidual leukocytes were cell migration and regulation of immune response, chemotaxis, and humoral immune responses. Our results show striking differences between the transcriptome of choriodecidual and maternal circulating leukocytes. Choriodecidual leukocytes are enriched in immune mediators implicated in the spontaneous process of labor at term.

MYPT1 is targeted by miR-145 inhibiting viability, migration and invasion in 2D and 3D HeLa cultures.

The miR-143/145 cluster is down-regulated in cervical tumor cells suggesting a role in tumorigenesis including cytoskeleton remodeling, a key event for tumor progression. The aim of the present work was to determine the role of miR-143/145 in the modulation of the myosin regulator phospho-myosin light chain (pMLC). HeLa monolayer and tridimensional cultures were transfected with miR-143 or miR-145 mimics inhibiting cell viability, proliferation, migration and invasion, mainly through miR-145. MiR-145 transfection increased pMLC levels by targeting the MYPT1 subunit of the regulatory myosin phosphatase. MYPT1 knockdown by siRNAs reproduced miR-145 effects suggesting miR-145 as a tumor suppressor through MYPT1 targeting, leading to a subsequent increase of pMLC levels with implications for cervical cell viability, migration and invasion.

Ultraviolet solar radiation in the tropical central Andes (12.0°S).

Ultraviolet (UV) solar irradiance measurements performed in the central Andes, Huancayo, Peru (12.0°S, 75.3°W, 3313 m asl) at 1 min intervals between January 2003 and December 2006 were used to analyse daily, monthly, and annual cycles of UV solar irradiance. The measurements were performed using a GUV-511 multi-channel filter radiometer at four wavelengths: 305, 320, 340, and 380 nm. UV irradiance data under clear sky and all sky conditions were separated using a procedure based on calculation of normalized irradiance. In February, the highest hourly mean value at noon for the UV Index reached 18.8 for clear sky conditions and 15.5 for all sky conditions, with outlier peaks close to UVI = 28. In addition, the highest mean value for the daily erythemal dose was found also in February, reaching 7.5 kJ m-2 d-1 with a maximum outlier value close to 10.1 kJ m-2 d-1. Comparisons between the clear sky GUV measurements and TUV model estimations were evaluated with statistical quantities showing values of R2 close to 0.98. The total ozone column and trace gases were obtained from OMI. The aerosol parameters were obtained from MODIS. The enhancements due to clouds of spectral irradiance at 340 nm as compared to a cloudless sky reach maxima of 20%. These results indicate that tropical central Andes has among the highest incident ultraviolet solar radiation in the world.

Common-pull, multiple-push, vacuum-activated telescope mirror cell.

A new concept for push-pull active optics is presented, where the push-force is provided by means of individual airbag type actuators and a common force in the form of a vacuum is applied to the entire back of the mirror. The vacuum provides the pull-component of the system, in addition to gravity. Vacuum is controlled as a function of the zenithal angle, providing correction for the axial component of the mirror's weight. In this way, the push actuators are only responsible for correcting mirror deformations, as well as for supporting the axial mirror weight at the zenith, allowing for a uniform, full dynamic-range behavior of the system along the telescope's pointing range. This can result in the ability to perform corrections of up to a few microns for low-order aberrations. This mirror support concept was simulated using a finite element model and was tested experimentally at the 2.12 m San Pedro Mártir telescope. Advantages such as stress-free attachments, lighter weight, large actuator area, lower system complexity, and lower required mirror-cell stiffness could make this a method to consider for future large telescopes.

Low-cost, phase-shifting mechanisms for Newton-type interferometers.

Using standard optical shop equipment, it is possible to implement simple, low-cost, phase-shifting Newton interferometers sufficiently accurate for surface evaluation. The simplification of the phase-shifting mechanism is compensated with image-processing algorithms that can deal with vibrations and uneven, nonsequential steps. The results are cross-compared with a Fizeau phase-shifting interferometer to verify the effectiveness of the method.

HyDRa: polishing with a vortex.

We present a hydrodynamic, deterministic polishing tool (HyDRa) based on the fluid-jet polishing (FJP) principle. In contrast to other FJP methods, the polishing flux is accelerated with pressurized air and then expelled at high velocity, forming a radial, grazing abrasive pattern that exerts no net force of the tool on the surface to be polished, since the vacuum and thrust forces that are created at the tool's output balance each other out. The grazing effect minimizes microroughness, making it appropriate for finishing high-quality surfaces. The principle of operation as well as polishing results of a series of small etalon plates are presented.

HyDRa: polishing process convergence rate optimization.

In an effort to optimize the hydrodynamic radial (HyDRa) polishing process for applications where the amount of material that has to be removed implies long polishing times, we have developed a method to determine the optimum correction fraction that has to be made for a given error map, in terms of the level of determinism of the process, the number of iterations, and their associated polishing runs as well as run times.

Inflammation- and cancer-related microRNAs in rat renal cortex after subchronic exposure to fluoride.

The proximal tubule is a target of subchronic exposure to fluoride (F) in the kidney. Early markers are used to classify kidney damage, stage, and prognosis. MicroRNAs (miRNAs) are small sequences of non-coding single-stranded RNA that regulate gene expression and play an essential role in developing many pathologies, including renal diseases. This study aimed to evaluate the expression of Cytokine-Chemokine molecules (IL-1α/1ß/4/6/10, INF-γ, MIP-1α, MCP-1, RANTES, and TGF ß1/2/3) and inflammation-related miRNAs to evidence the possible renal mechanisms involved in subchronic exposure to F. Total protein and miRNAs were obtained from the renal cortex of male Wistar rats exposed to 0, 15 and 50 mg NaF/L through drinking water during 40 and 80 days. In addition, cytokines-chemokines were analyzed by multiplexing assay, and a panel of 77 sequences of inflammatory-related miRNAs was analyzed by qPCR. The results show that cytokines-chemokines expression was concentration- and time-dependent with F, where the 50 mg NaF/L were the main altered groups. The miRNAs expression resulted in statistically significant differences in thirty-four miRNAs in the 50 mg NaF/L groups at 40 and 80 days. Furthermore, a molecular interaction network analysis was performed. The relevant pathways modified by subchronic exposure to fluoride were related to extracellular matrix-receptor interaction, Mucin type O-glycan biosynthesis, Gap junction, and miRNAs involved with renal cell carcinoma. Thus, F-induced cytokines-chemokines suggest subchronic inflammation; detecting miRNAs related to cancer and proliferation indicates a transition from renal epithelium to pathologic tissue after fluoride exposure.

Coarse-Grained Simulation of Bottlebrush: From Single-Chain Properties to Self-Assembly.

Bottlebrush polymers consist of a linear backbone with densely grafted side chains. They are known to have a range of properties of interest, such as enhanced mechanical strength and rapid self-assembly into large domains, and have attracted attention as promising candidates for applications in photonics, lithography, energy storage, organic optoelectronics, and drug delivery. Here, we present a coarse-grained model of bottlebrush polymers that is able to reproduce their experimentally observed persistence lengths and chain conformations in the melt. The model is then used to investigate the morphologies of this class of materials for various chain architectures and grafting densities.

Fluorescence-Linked Aptamer Assay for SARS-CoV-2 Spike-Protein: A Step-by-Step Performance Analysis in Clinical Samples.

The COVID-19 pandemic has been a main concern over the last two years and has become one of the most important crises in the history of human health. Today, there is still a need for affordable and reliable diagnostic tests for massive disease monitoring. Previously, a set of highly specific DNA-aptamers (C7/C9) binding to the SARS-CoV-2 Spike (S) protein were isolated but its performance in clinical samples remained to be tested. Here, 242 samples were collected through three different methods and subjected to florescence-linked aptamer assays (FLAA) based on C7/C9 aptamers through two readout protocols. Then, a step-by-step statistical approach which included agreement tests, proportion comparisons and binomial and multinomial logistic regressions was used to predict optimal conditions for the novel C7/C9 FLAA test. RTqPCR threshold cycles, symptoms onset and processing time were influential factors on FLAA test results. Naturally occurring mutations on S were also detected and analyzed. Aminoacidic substitutions D614G and T732A appeared relevant for aptamer recognition although further studies are necessary. The methodology presented here is the first step to determine the performance and diagnosis across a range of clinical contexts and it might serve as a base for a complete analysis applicable to other designs of new diagnostic tests.

ZO-2 favors Hippo signaling, and its re-expression in the steatotic liver by AMPK restores junctional sealing.

ZO-2 is a peripheral tight junction (TJ) protein whose silencing in renal epithelia induces cell hypertrophy. Here, we found that in ZO-2 KD MDCK cells, in compensatory renal hypertrophy triggered in rats by a unilateral nephrectomy and in liver steatosis of obese Zucker (OZ) rats, ZO-2 silencing is accompanied by the diminished activity of LATS, a kinase of the Hippo pathway, and the nuclear concentration of YAP, the final effector of this signaling route. ZO-2 appears to function as a scaffold for the Hippo pathway as it associates to LATS1. ZO-2 silencing in hypertrophic tissue is due to a diminished abundance of ZO-2 mRNA, and the Sp1 transcription factor is critical for ZO-2 transcription in renal cells. Treatment of OZ rats with metformin, an activator of AMPK that blocks JNK activity, augments ZO-2 and claudin-1 expression in the liver, reduces the paracellular permeability of hepatocytes, and serum bile acid content. Our results suggest that ZO-2 silencing is a common feature of hypertrophy, and that ZO-2 is a positive regulator of the Hippo pathway that regulates cell size. Moreover, our observations highlight the importance of AMPK, JNK, and ZO-2 as therapeutic targets for blood-bile barrier dysfunction.

Prevalence of filaggrin loss-of-function variants in Chilean population with and without atopic dermatitis.

BACKGROUND: Filaggrin (FLG) loss-of-function variants are major genetic risk factors for atopic dermatitis (AD), but these have not been studied in Latin American populations with and without AD. METHODS: FLG variants R501X and 2282del4 were genotyped in 275 Chilean adults with and without AD from the "Early origins of allergy and asthma" (ARIES) cohort and in 227 patients from an AD cohort based in Santiago, Chile. RESULTS: Among adults in the ARIES cohort, 3.3% were carriers of R501X and 2.9% of 2282del4 variants, all heterozygotes. In this cohort, 6.2% were FLG variant carriers: 11.1% of subjects reporting AD were carriers of FLG variants vs. 5.2% in those without AD (P = 0.13). In this first cohort, FLG variants were not significantly associated with asthma, allergic rhinitis, or food allergy. In the AD cohort, the prevalence of FLG variants was 7% for R501X, 2.2% for the 2282del4 variant, and 9.3% for the combined genotype. In this cohort, FLG variants were present in 15.5% of severe AD vs. 7.1% of mild-to-moderate AD subjects (P = 0.056). Evaluation of Chilean population from both cohorts combined (n = 502) revealed that FLG variants were not significantly associated with AD (OR = 1.92 [95% CI 0.95-3.9], P = 0.067) but were associated with asthma (OR = 2.16 [95% CI 1.02-4.56], P = 0.039). CONCLUSIONS: This is the first study to evaluate FLG loss-of-function variants R501X and 2282del4 in Latin American population, revealing a similar prevalence of these FLG variant carriers to that of European populations. Among Chileans, FLG variants were significantly associated with asthma but not AD.