Pneumatic tube systems (PTSs) provide an efficient means of rapidly transporting samples within hospitals. The feasibility of using hospital PTSs to transport therapeutic monoclonal antibody (mAb) dosing solutions in intravenous infusion containers has been evaluated in recent years. In this study, we evaluated the impact of transporting therapeutic mAb drug products in vials-both lyophilized and reconstituted-via a laboratory-scale PTS that simulates typical hospital transport stresses, and assessed the resulting effects on product quality for two test mAbs. Samples were subjected to increasing cumulative PTS stress (transport distances of 586 m, 1,172 m, or 2,344 m) to represent typical hospital transportation scenarios and were assessed using multiple analytical methods. Mechanical shock and vibration caused breakage of lyophilized cakes and increased subvisible particle counts after reconstitution. For both mAbs, higher cumulative PTS stress (longer distances) was associated with higher subvisible particle counts, with mAb2 exhibiting greater sensitivity. PTS transport of the same mAb drug products after reconstitution had a negligible effect on subvisible particle counts. PTS transport had minimal impact on other quality attributes tested. Overall, these results contribute to the growing body of knowledge on PTS transport and indicate that PTS transport poses a significantly elevated risk to quality of lyophilized mAb products but may pose a low risk to quality of the reconstituted solution derived from the same lyophilized product. The decision to allow PTS transport for a given mAb product should be made on a case-by-case basis based on relative risk toward product quality impact for that particular mAb, which may be assessed using representative PTS stress conditions.

Liver fibrosis is a pathological consequence of chronic liver injury that is reversible but can potentially progress to serious complications. Currently, due to the incomplete elucidation of its key molecular driving mechanisms, effective targeted therapeutic approaches remain lacking in clinical practice. The mitochondrial inner membrane transporter Timm13 has been implicated in cellular stress and disease, yet its role in liver fibrosis remains unclear. This study found that Timm13 is significantly upregulated in human fibrotic liver tissue. Functional studies demonstrated that Timm13 overexpression promotes the activation, proliferation, migration, and extracellular matrix production of hepatic stellate cells (HSCs), whereas Timm13 knockdown inhibits these processes. In a carbon tetrachloride (CCl4)-induced mouse model of liver fibrosis, Timm13 overexpression exacerbated collagen deposition and histopathological damage, whereas its knockdown exerted a protective effect. Mechanistically, Co-immunoprecipitation and proteomic analyses identified heat shock protein 90α family member 1 (Hsp90aa1) as a direct binding partner of Timm13. We further demonstrate that this interaction is critical for activating the TGFβ signaling pathway, as Hsp90aa1 silencing reverses the Timm13-driven upregulation of fibrotic markers and TGFβ1 protein levels. This indicates that Timm13 promotes fibrogenic activity by regulating TGFβ1 expression through Hsp90aa1. Furthermore, studies indicate that Timm13 significantly induces mitochondrial dysfunction in hepatic stellate cells, manifested by reduced mitochondrial membrane potential, elevated reactive oxygen species levels, and impaired ATP synthesis. Our results defined a previously unrecognized Timm13/Hsp90aa1/TGFβ signaling axis, revealing how Timm13, through its interaction with Hsp90aa1, concurrently regulates TGFβ1 expression and integrates mitochondrial stress signals to drive the fibrogenic activity of hepatic stellate cells. This study establishes Timm13 as a potential diagnostic biomarker and therapeutic target for liver fibrosis.

Frequent extreme high-temperature events have severely restricted the development of the cold-water rainbow trout (Oncorhynchus mykiss) aquaculture industry, yet the systemic tolerance mechanisms behind the selection of heat-tolerant strains remain elusive. In this study, a 28-day thermal stress challenge (14-28 °C) was performed on common rainbow trout (N trout) and family-selected heat-tolerant rainbow trout (T trout, F2 generation). Integrated biochemical, histopathological, and transcriptomic-metabolomic analyses were conducted. The results indicated that T trout effectively delayed the collapse of the antioxidant defense system under high temperatures (with significantly higher SOD, CAT, and GSH activities and significantly lower lipid peroxidation product MDA, P < 0.01). Gill cell apoptosis rates were significantly reduced (P < 0.05). At the extreme temperature of 28 °C, T trout exhibited prominent compensatory structural remodeling of the gills (P < 0.01) and sustained blood glucose mobilization capacity, thereby avoiding the metabolic failure seen in N trout. Multi-omics analysis further revealed that, distinct from the endoplasmic reticulum stress and cell membrane damage observed in N trout, T trout maintained protein homeostasis, likely through the upregulation of heat shock proteins (e.g., hspa1s, hsp90). Moreover, T trout achieved efficient thermal adaptation through metabolic reprogramming (restructuring carbohydrate metabolic to support the glutathione antioxidant system, upregulating gpx4a, and accumulating CHAC) and adaptive regulation of membrane lipids (e.g., arachidonic acid and sphingolipid pathways). This study systematically clarifies the key mechanisms of high-temperature tolerance in selectively bred rainbow trout, providing a vital theoretical basis and candidate markers (e.g., hspa1s and gpx4a) for molecular marker-assisted breeding of stress-resistant cold-water fish.

During infection with highly pathogenic Avian Influenza virus (HPAIV), heat shock proteins (HSPs) play roles in host immune responses by interacting with various regulators of cell signaling pathways and in mediating cellular homeostasis. However, the tissue-specific regulation of these chaperones, particularly their potential association with the NF-κB pathway, remains poorly defined in avian species. Chickens were infected with HPAIV (A/chicken/Vietnam/NA01/2019 (H5N1), and the expression patterns of a comprehensive range of HSPs (small HSPs to canonical classes) were analyzed in lung and spleen at 1 and 3 days post-infection (dpi). As a result, HPAIV infection induced significant temporal up-regulation of mRNA of small sHSPs (sHSPs; HSPB7, HSPB9), HSPE1, and a collagen-specific molecular chaperone, SERPINH1, in both tissues. To investigate transcriptional regulation, DF-1 cells were stimulated with Poly(I: C) in the presence or absence of NF-κB inhibitors. Notably, NF-κB inhibition was associated with an up-regulation of HSPB9 and a depression of SERPINH1 in PIC-treated DF-1. These results suggest that specific HSPs may be influenced by pro-inflammatory signaling during viral infection, with NF-κB signalling potentially contributing to their negative regulation during viral stress response. Collectively, these findings provide preliminary insights into the complex molecular dynamics of HPAIV pathogenesis and highlight the importance of host-mediated signaling pathways in modulating the host cellular stress response in poultry.

Epigenetic modifications to DNA are proposed to underpin plastic responses to environmental change, and the manner in which DNA methylation contributes to plasticity likely differs among tissues. However, few studies have investigated tissue-specific DNA methylation responses to ecologically relevant environmental stressors in natural settings. Here, we used reduced representation bisulfite sequencing to examine the influence of in situ microhabitats on DNA methylation in gill and foot of juvenile California mussels (Mytilus californianus), a foundation species with widespread dispersal and little evidence of genetic population structure. We examined mussels from a one-month reciprocal transplant experiment between a cool, wave-exposed and a warm, wave-protected microhabitat. These manipulations-which were previously shown to alter juvenile mussels' heat tolerance-led to significant and tissue-specific changes in CpG methylation, including within a number of genes with roles in stress response pathways. Differentially methylated genes were involved in processes including heat shock response, proteolysis, and temperature sensing. In gills, differentially methylated CpGs were more likely to occur in introns relative to other inter- and intragenic features. This study expands on previous research that examined environmentally driven shifts in DNA methylation by documenting plastic and tissue-specific changes in DNA methylation between microhabitats in a natural setting.

Glucocorticoids (GCs) are used as anti-inflammatory and immunosuppressive drugs in many immune mediated diseases, but their use in sepsis and shock is controversial. This is caused in part by a lack of information regarding the responding cell types and GC-regulated genes in vivo. We used large collections of public blood transcriptomic datasets and different GC-induced query genes to obtain 2 robust gene expression correlation signatures of GC induction, either in the absence or in the presence of severe inflammation. GC signature 1 originated from circadian cortisol with biases for gene expression in NK cells and neutrophils. GC signature 2 originated from GC in severe inflammation, mainly with biases for gene expression in monocytes and neutrophils. Many genes upregulated by GC treatment in septic shock and burn shock were also present as high-ranking genes in GC signatures, which pointed to their direct regulation by GC. Robust GC signatures were also obtained from dataset collections of different tissues for comparison, and of monocytes and neutrophils, separately. Gene induction by GC was put into a wider framework of gene expression in circulating monocytes and neutrophils in health and during systemic inflammation, which included macrophage-like cells and a hypoxia-related gene expression signature in monocytes in severe inflammation. We present and interpret a large number of GC-regulated genes in different blood cells and tissues in vivo, and select 2 whole blood transcriptomic biomarker gene sets, GC-1 and GC-2, for monitoring cortisol action in health, and in severe inflammation, respectively.

Aquaporins (AQPs) are a family of water channel proteins with a significant role in immune cell function. Although aquaporins have been studied in sepsis, their expression in critical illness remains underexplored. Our group aimed to examine the mRNA expression profile of AQP isoforms in polymorphonuclear (PMN) and peripheral blood mononuclear cells (PBMCs) of critically ill patients admitted to the intensive care unit (ICU). To this end, we designed a prospective, longitudinal, observational study performed in the ICU of "Evangelismos" General Hospital between February 2024 and October 2024. In total, 40 critically ill patients and 25 healthy controls were enrolled in the study. Blood samples were collected at four time points: upon ICU admission (24-48h), and at days 4 (D4), 8 (D8), and 14 (D14). Total RNA was extracted from PMNs and PBMCs, and RT-qPCR was performed to examine mRNA levels of AQPs 1, 2, 3, 4, 5, 7, and 9. Our results revealed downregulation of AQP isoforms, except AQP1, in PMNs, whereas PBMCs exhibited sustained upregulation of all AQPs except AQP3, which was downregulated. Our findings suggest that differential AQP expression in immune cell subsets may reflect adaptive immune responses during critical illness, providing insights into leukocyte function and potential therapeutic targets.