STING agonist 8803 reprograms the immune microenvironment and increases survival in preclinical models of glioblastoma.

STING agonists can reprogram the tumor microenvironment to induce immunological clearance within the central nervous system. Using multiplexed sequential immunofluorescence (SeqIF) and the Ivy Glioblastoma Atlas, STING expression was found in myeloid populations and in the perivascular space. The STING agonist 8803 increased median survival in multiple preclinical models of glioblastoma, including QPP8, an immune checkpoint blockade-resistant model, where 100% of mice were cured. Ex vivo flow cytometry profiling during the therapeutic window demonstrated increases in myeloid tumor trafficking and activation, alongside enhancement of CD8+ T cell and NK effector responses. Treatment with 8803 reprogrammed microglia to express costimulatory CD80/CD86 and iNOS, while decreasing immunosuppressive CD206 and arginase. In humanized mice, where tumor cell STING is epigenetically silenced, 8803 therapeutic activity was maintained, further attesting to myeloid dependency and reprogramming. Although the combination with a STAT3 inhibitor did not further enhance STING agonist activity, the addition of anti-PD-1 antibodies to 8803 treatment enhanced survival in an immune checkpoint blockade-responsive glioma model. In summary, 8803 as a monotherapy demonstrates marked in vivo therapeutic activity, meriting consideration for clinical translation.

Protocol to quantify immune cell distribution from the vasculature to the glioma microenvironment on sequential immunofluorescence multiplex images.

Although myeloid-derived immune cells can be dispersed throughout the tumor microenvironment (TME), anti-tumor effector cells are confined to the perivascular space. Here, we present a protocol to quantify immune cell distribution from tumor vasculature to its glioma microenvironment on sequential immunofluorescence multiplex images. We describe steps for sequential immunofluorescence multiplex staining, image generation, and storage. We then detail the procedures for tissue, vessel, and nuclei segmentation; cell phenotyping; data extraction; and training using RStudio and Spyder.

Intestinal Transcriptome Analysis Reveals Enrichment of Genes Associated with Immune and Lipid Mechanisms, Favoring Soybean Meal Tolerance in High-Growth Zebrafish (Danio Rerio).

The molecular mechanisms underlying fish tolerance to soybean meal (SBM) remain unclear. Identifying these mechanisms would be beneficial, as this trait favors growth. Two fish replicates from 19 experimental families were fed fishmeal-(100FM) or SBM-based diets supplemented with saponin (50SBM + 2SPN) from juvenile to adult stages. Individuals were selected from families with a genotype-by-environment interaction higher (HG-50SBM + 2SPN, 170 ± 18 mg) or lower (LG-50SBM + 2SPN, 76 ± 10 mg) weight gain on 50SBM + 2SPN for intestinal transcriptomic analysis. A histological evaluation confirmed middle intestinal inflammation in the LG- vs. HG-50SBM + 2SPN group. Enrichment analysis of 665 differentially expressed genes (DEGs) identified pathways associated with immunity and lipid metabolism. Genes linked to intestinal immunity were downregulated in HG fish (mpx, cxcr3.2, cftr, irg1l, itln2, sgk1, nup61l, il22), likely dampening inflammatory responses. Conversely, genes involved in retinol signaling were upregulated (rbp4, stra6, nr2f5), potentially favoring growth by suppressing insulin responses. Genes associated with lipid metabolism were upregulated, including key components of the SREBP (mbtps1, elov5l, elov6l) and cholesterol catabolism (cyp46a1), as well as the downregulation of cyp7a1. These results strongly suggest that transcriptomic changes in lipid metabolism mediate SBM tolerance. Genotypic variations in DEGs may become biomarkers for improving early selection of fish tolerant to SMB or others plant-based diets.

Effect of Non-Conventional Drying Methods on In Vitro Starch Digestibility Assessment of Cooked Potato Genotypes.

Potatoes (Solanum tuberosum L.) are a good dietary source of carbohydrates in the form of digestible starch (DS) and resistant starch (RS). As increased RS content consumption can be associated with decreased chronic disease risk, breeding efforts have focused on identifying potato varieties with higher RS content, which requires high-throughput analysis of starch profiles. For this purpose, freeze drying of potatoes has been used but this approach leads to inaccurate RS values. The present study objective was to assess the starch content (RS, DS and total starch (TS)) of three cooked potato genotypes that were dried using freeze drying and innovative drying techniques (microwave vacuum drying, instant controlled pressure drop drying and conductive hydro-drying) relative to freshly cooked potato samples. Depending on the genotype, all drying methods showed one or more starch measures that were significantly different from freshly cooked values. The combination of ultrasound and infrared assisted conductive hydro-drying was the only method identified to be associated with accurate assessment of DS and TS content relative to fresh samples. The drying treatments were all generally associated with highly variable RS content relative to fresh controls. We conclude that freshly cooked samples must be used for selecting varieties with a high proportion of RS starch as drying of cooked potatoes leads to unreliable RS measurements.

Frecuencia y factores asociados al acoso escolar en colegios públicos / Frequency and factors associated with bullying in public schools

Resumen El presente estudio tuvo como objetivo identificar la frecuencia del acoso y ciberacoso escolar y los factores asociados en una muestra representativa de 475 estudiantes de secundaria en tres instituciones públicas del municipio de Chinchiná, en el departamento de Caldas, Colombia, en el año 2014. La población fue seleccionada mediante un muestreo probabilístico estratificado por grado y género. Aplicamos las escalas: APGAR familiar, CAGE (Cut, Annoyed, Guilty, Eye), Escala Hospitalaria de Ansiedad y Depresión (HAD), la SCOFF (Scik Control On Fast Food), el acoso escolar por medio de la encuesta sobre convivencia escolar para alumnos, y el ciberacoso por medio de Escala de Patchin. De acuerdo al cuestionario empleado se encontró una proporción del más alto nivel de acoso del 1.9%, 0.4% acosadores, 1.3% ciberacoso, 1.9% ciberacosadores, en rango medio, 14% acoso, 8.5% acosador, 5.5% ciberacoso, 4.9% ciberagresores. Se encontraron relaciones significativas entre nivel de acoso y funcionalidad familiar, consumo de alcohol, ansiedad, depresión, consumo de sustancias, nivel de ciberacoso también con trastornos de la conducta alimentaria. Los factores asociados con acoso y ciberacoso afectan por igual a acosados y acosadores, resultado también encontrado en otros estudios. En esta población se presentan principalmente niveles de acoso medio y bajo.

Abstract This study aimed to identify the frequency of bullying and cyberbullying and associated factors in a representative sample of 475 high school students in three public institutions of Chinchiná (Colombia) in 2014. This is a cross sectional study. Demographic variables were taken; family functioning was assessed by the family APGAR scale instrument, taken with the CAGE (Cut, Annoyed, Guilty, Eye) scale, anxiety and depression, with hospital scale of anxiety and depression (HAD), eating disorders SCOFF (Scik Control On Fast Food), scale bullying staff through the survey about coexistence school for students and cyberbullying through Patchin scale. According to the questionnaire employed a high ranking proportion of 1.9% was found, 4% bullies, cyber bullying 1.3%, 1.9% cyberbulliesin the middle bullying 14%, 8.5% stalker 5.5% cibera-sew, 4.9% cyber attackers. Significant relationships between the level of harassment and family functioning, alcohol consumption, anxiety, depression, substance abuse levels were found, level of cyberbullying also disordered eating behavior. We found that factors associated Aphek-as both bullies and bullied, result similar to that found in other studies. In this population, high levels of harassment have low proportions, not harassment levels low and medium.