Rapid and Efficient Enrichment of Mouse Spinal Cord Microglia.

The vertebral column defines a vertebrate and shapes the spinal canal, a cavity that encloses and safeguards the spinal cord. Proper development and function of the mammalian central nervous system rely significantly on the activity of resident macrophages known as microglia. Microglia display heterogeneity and multifunctionality, enabling distinct gene expression and behavior within the spinal cord and brain. Numerous studies have explored cerebral microglia function, detailing purification methods extensively. However, the purification of microglia from the spinal cord in mice lacks a comprehensive description. In contrast, the utilization of a highly purified collagenase, as opposed to an unrefined extract, lacks reporting within central nervous system tissues. In this study, the vertebral column and spinal cord were excised from 8-10 week-old C57BL/6 mice. Subsequent digestion employed a highly purified collagenase, and microglia purification utilized a density gradient. Cells underwent staining for flow cytometry, assessing viability and purity through CD11b and CD45 staining. Results yielded an average viability of 80% and a mean purity of 95%. In conclusion, manipulation of mouse microglia involved digestion with a highly purified collagenase, followed by a density gradient. This approach effectively produced substantial spinal cord microglia populations.

IL-36γ is secreted through an unconventional pathway using the Gasdermin D and P2X7R membrane pores.

Mucosal innate immunity functions as the first line of defense against invading pathogens. Members of the IL-1 family are key cytokines upregulated in the inflamed mucosa. Inflammatory cytokines are regulated by limiting their function and availability through their activation and secretion mechanisms. IL-1 cytokines secretion is affected by the lack of a signal peptide on their sequence, which prevents them from accessing the conventional protein secretion pathway; thus, they use unconventional protein secretion pathways. Here we show in mouse macrophages that LPS/ATP stimulation induces cytokine relocalization to the plasma membrane, and conventional secretion blockade using monensin or Brefeldin A triggers no IL-36γ accumulation within the cell. In silico modeling indicates IL-36γ can pass through both the P2X7R and Gasdermin D pores, and both IL-36γ, P2X7R and Gasdermin D mRNA are upregulated in inflammation; further, experimental blockade of these receptors' limits IL-36γ release. Our results demonstrate that IL-36γ is secreted mainly by an unconventional pathway through membrane pores formed by P2X7R and Gasdermin D.

Eficacia terapéutica de la buprenorfina para tratamiento de dolor agudo postoperatorio en cirugía torácica por toracoscopía / Therapeutic efficacy of buprenorphine for treatment of acute postoperative pain in thoracic surgery by thoracoscopy

Resumen: Introducción: El dolor postoperatorio en toracoscopía tiene una incidencia del 80%. Estos procedimientos se realizan bajo anestesia general o sedoanalgesia. La buprenorfina en dosis de 4 μg/kg o más es un fármaco pilar para el tratamiento de dolor postoperatorio; sin embargo, dosis menores pueden ser suficientes. El objetivo de este estudio fue demostrar que dosis de 1-3 μg/kg de buprenorfina aporta una adecuada analgesia con menores efectos adversos durante el postoperatorio en pacientes sometidos a cirugía toracoscópica. Material y métodos: Se realizó el estudio prospectivo, transversal y doble ciego en 48 pacientes sometidos a toracoscopía, divididos en 3 grupos: A (1 μg/kg), B (2 μg/kg) y C (3 μg/kg) tratados con buprenorfina, se evaluaron efectos adversos, intensidad de dolor por escala verbal numérica (EVN) y sedación con escala de agitación-sedación de Richmond (RASS) de los 0 a los 120 minutos postanestesia. Resultados: Las dosis de 1 a 3 μg/kg de buprenorfina no ocasionaron efectos adversos. El mantenimiento de analgesia es mayor con 2 μg/kg (p = 0.019) y 3 μg/kg (p = 0.045). Las dosis de 1 y 2 μg/kg presentan un nivel de sedación menor. Conclusiones: La analgesia multimodal disminuyó un 200% el dolor postquirúrgico inmediato. Las dosis de 2-3 μg/kg de buprenorfina son efectivas para el control del dolor con menor incidencia de efectos adversos y un nivel de sedación superficial durante el postoperatorio.

Abstract: Introduction: Posterior pain in thoracoscopy has an 80% incidence. These procedures are done under general anesthesia or sedoanalgesia. The buprenorphin in dosage of 4 μg/kg it's the foundation for post-operatory pain treatment, nevertheless, smaller dosage can be sufficient. The objective of this study is to demonstrate that a dosage of buprenorphin from 1-3 μg/kg provides suitable analgesia with less side effects during post-operatory in patients who underwent thoracoscopic surgery. Material and methods: We conducted a double-blind, prospective, transversal study, in 48 patients which underwent a thoracoscopy divided in 3 groups: A (1 μg/kg), B (2 μg/kg), and C (3 μg/kg) treated with buprenorphin. We studied side effects, pain intensity (NVE), and sedative effect (RASS) from 0 to 120 minutes postanesthesia. Results: Dosage from 1 to 3 μg/kg did not generate side effects. The analgesia effect is higher with 2 μg/kg (p = 0.019) and 3 μg/kg (p = 0.045). Dosage of 1 and 2 μg/kg have lower sedative effect. Conclusion: The multimodal analgesia was diminished by 200% inmidiate postsurgical pain. Dosage from 2-3 μg/kg of buprenorphine are effective for pain control with less side effects and a superficial sedative effect during post-operatory.