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Sheep primary epithelial cells are short-lived in cell culture systems. For long-term in vitro studies, primary cells need to be immortalized. This study aims to establish and characterize T immortalized sheep embryo kidney cells (TISEKC). In this study, we used fetal lamb kidneys to derive primary cultures of epithelial cells. We subsequently immortalized these cells using the large T SV40 antigen to generate crude TISEKC and isolate TISEKC clones. Among numerous clones of immortalized cells, the selected TISEKC-5 maintained active division and cell growth over 20 passages but lacked expression of the oncogenic large T SV40 antigen. Morphologically, TISEKC-5 maintained their epithelial aspect similar to the parental primary epithelial cells. However, their growth properties showed quite different patterns. Crude TISEKC, as well as the clones of TISEKC proliferated highly in culture compared to the parental primary cells. In the early passages, immortalized cells showed heterogeneous polyploidy but in the late passages the karyotype of immortalized cells became progressively stable, identical to that of the primary cells, because the TISEKC-5 cell line has lost the large SV40 T antigen expression, this cell line is a valuable tool for veterinary sciences and biotechnological productions.The origin, migratory pathways and adult derivatives of neural crest cells (NCCs) are well known. However, less is known about how these cells migrate. In this study, in a laboratory based in a low-resource setting, a hanging drop culture assay was utilised to study the movement of individual avian trunk neural crest cells. Mode of migration by means of lamellipodia and filopodia was studied in live cell cultures with a laser scanning confocal microscope and Airyscan module. Both distance migrated and speed of migration were calculated. NCCs migrated in a chain soon after emerging from the explanted neural tube, but were more dispersed and had random movements when they reached the periphery of the culture. While the distances travelled by these NCCs were less and the cells were slower on gelatine than on other extracellular matrices reported in the literature, the assay afforded detailed observation of actin filament distribution and cytoplasmic protrusions. The study has provided unique evidence of individual NCC movements in vitro, in a simple hanging drop assay optimized for the study of NCCs. The assay could be used for further analysis of the behaviour of NCCs on different extracellular matrices or with targeted action.The bioinsecticidal Cry1Ac proteins (protoxin and toxin) are potent immunogens that can activate macrophages by inducing upregulation of costimulatory molecules, pro-inflammatory cytokines, and mitogen-activated protein kinase (MAPK) signaling pathways. Besides, by the oral route, Cry1Ac toxin is mildly allergenic and induces intestinal lymphoid hyperplasia in mice. Given the potential utility of Cry1Ac protoxin as an adjuvant, as well as the human consumption of Cry1Ac toxin in transgenic crops, it is necessary to more deeply evaluate the toxicological potential of these proteins in mammalian immune cells. Here, were used in vitro evaluations in leukocyte and macrophage cell lines to test the potential toxicity of various doses of Cry1Ac proteins, by means of Alamar Blue, MTT, Annexin V, and JC1 assays. Our results indicated that neither Cry1Ac protoxin nor toxin elicited acute toxic effects, after monitoring the cell activity for 4, 8, 10, and 24 h of exposure. By flow cytometry and confocal microscopy analysis, it was observed that neither Cry1Ac toxin nor protoxin generated mitochondrial damage or depolarization or induced apoptosis or necrosis. In conclusion, despite their immunostimulatory effects, it was demonstrated that Cry1Ac proteins did not have cytotoxic effects, even at high concentrations, in primary leukocytes or macrophages or cell lines.Elasmobranchs are exposed to mercury (Hg) through a variety of pathways in the environment. This study assessed maternal offloading and diet-based Hg exposure for neonatal and juvenile blacktip sharks (Carcharhinus limbatus) from Charlotte Harbor located along southwest Florida's coast, a recognized Hg hotspot. Neonates (n = 57) had highest total Hg (THg) concentrations in the kidney (0.56 ± 0.26 mg kg-1; n = 38) and muscle (0.53 ± 0.17 mg kg-1; n = 57), followed by liver (0.31 ± 0.11 mg kg-1; n = 38), and blood (0.05 ± 0.033 mg kg-1; n = 57). Juveniles (n = 13) exhibited a different distribution with highest THg in the liver (0.868 ± 0.54 mg kg-1; n = 6), followed by the muscle (0.84 ± 0.28 mg kg-1; n = 13), kidney (0.55 ± 0.22 mg kg-1; n = 6), and blood (0.11 ± 0.04 mg kg-1; n = 11). The distribution of THg among tissues and liver-to-muscle ratios indicated that Hg originated primarily from maternal offloading in neonates, whereas juveniles continued to accumulate Hg through dietary exposure post-parturition. Additionally, comparisons between results of the present study and previous Florida blacktip shark surveys suggested that Hg levels have not declined in southwest Florida estuaries for over two decades.In most situations, we are able to tell those outcomes we cause from those we do not. By now, research has provided us with a reasonably good understanding of the cognitive processes that underlie this sense of agency - it is thought to be produced by a comparison between a prediction of the outcome and the actual outcome that occurs. LOXO-292 molecular weight What is less clear is whether having a sense of agency can, itself, influence cognition. In the current study, we examined the possibility that sense of agency can affect memory, and we report evidence that stimuli that one feels a sense of agency over are, in fact, better remembered than counterparts without this. This self-agency effect can be distinguished from previously described control-related memory enhancements and adds to what we know of the cognitive consequences of having a sense of agency.
Immune checkpoint inhibitors have become standard of care for many patients with non-small cell lung cancer (NSCLC). These agents often cause immune-related adverse events (IRAEs), which have been associated with increased overall survival (OS). Intracranial disease control and OS for patients experiencing IRAEs with metastatic NSCLC and brain metastases have not yet been described.
We performed a single-institution, retrospective review of patients with NSCLC and existing diagnosis of brain metastasis, who underwent pembrolizumab treatment and developed any grade IRAE. The primary outcome of the study was intracranial time to treatment failure (TTF), defined from time of pembrolizumab initiation to new intracranial disease progression or death. Kaplan-Meier and Cox proportional hazard analyses were performed.
A total of 63 patients with NSCLC brain metastasis were identified, and 24 developed IRAEs. Patients with any grade IRAEs had longer OS (21 vs. 10months, p = 0.004), systemic TTF (15 vs. 4months, p < 0.