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Passive siRNA transfection method for gene knockdown in air-liquid interface airway epithelial cell cultures
Am J Physiol Lung Cell Mol Physiol. 2021 May 26. doi: 10.1152/ajplung.00122.2021. Online ahead of print.ABSTRACTDifferentiation of human bronchial epithelial cells (HBEs) in air-liquid interface (ALI) cultures recapitulates organotypic modeling of the in vivo environment. Although ALI cultures are invaluable for studying the respiratory epithelial barrier, loss-of-function studies are limited by potentially cytotoxic reagents in classical transfection methods, the length of the differentiation protocol, and the number of primary epithelial cell passages. Here, we present the efficacy and utility of a simple method for siRN...
Source: American Journal of Physiology. Lung Cellular and Molecular Physiology - May 26, 2021 Category: Cytology Authors: Colleen M Bartman Kimberly E Stelzig David R Linden Y S Prakash Sergio E Chiarella Source Type: research

A Primate-Specific RNA-Binding Protein (RBMXL3) Is Involved in Glucocorticoid Regulation of Human Pulmonary Surfactant Protein B (SP-B) mRNA Stability
Am J Physiol Lung Cell Mol Physiol. 2021 Mar 10. doi: 10.1152/ajplung.00022.2020. Online ahead of print.ABSTRACTThe ability of pulmonary surfactant to reduce alveolar surface tension requires adequate levels of surfactant protein B (SP-B). Dexamethasone (DEX) increases human SP-B expression, in part, through increased SP-B mRNA stability. A 30 nt-long hairpin element (RBE) in the 3'-untranslated region of human SP-B mRNA mediates both DEX-induced and intrinsic mRNA stability, but the mechanism is unknown. Proteomic analysis of RBE-interacting proteins identified a primate-specific protein; RNA binding motif X-linked-like-3...
Source: American Journal of Physiology. Lung Cellular and Molecular Physiology - March 15, 2021 Category: Cytology Authors: Lidan Liu Xiangli Liu Weizhen Bi Joseph L Alcorn Source Type: research

Sodium coupled neutral amino acid transporter SNAT2 counteracts cardiogenic pulmonary edema by driving alveolar fluid clearance.
Abstract The constant transport of ions across the alveolar epithelial barrier regulates alveolar fluid homeostasis. Dysregulation or inhibition of Na+ transport causes fluid accumulation in the distal airspaces resulting in impaired gas exchange and respiratory failure. Previous studies have primarily focused on the critical role of amiloride-sensitive epithelial sodium channel (ENaC) in alveolar fluid clearance (AFC), yet activation of ENaC failed to attenuate pulmonary edema in clinical trials. Since 40% of AFC is amiloride-insensitive, Na+ channels/transporters other than ENaC such as Na+-coupled neutral amino...
Source: American Journal of Physiology. Lung Cellular and Molecular Physiology - January 13, 2021 Category: Cytology Authors: Weidenfeld S, Chupin C, Langner DI, Zetoun T, Rozowsky S, Kuebler WM Tags: Am J Physiol Lung Cell Mol Physiol Source Type: research

UCHL1, a Deubiquitinating Enzyme, Regulates Lung Endothelial Cell Permeability In Vitro and In Vivo.
Abstract Increasing evidence suggests an important role for deubiquitinating enzymes (DUBs) in modulating a variety of biological functions and diseases. We previously identified the upregulation of the DUB, ubiquitin carboxyl terminal hydrolase 1 (UCHL1) in murine ventilator-induced lung injury (VILI). However, the role of UCHL1 in modulating vascular permeability, a cardinal feature of acute lung injury (ALI) in general, remains unclear. We investigated the role of UCHL1 in pulmonary endothelial cell (EC) barrier function in vitro and in vivo and examined effects of UCHL1 on VE-cadherin and claudin-5 regulation,...
Source: American Journal of Physiology. Lung Cellular and Molecular Physiology - January 13, 2021 Category: Cytology Authors: Mitra S, Epshtein Y, Sammani S, Quijada H, Chen W, Bandela M, Desai AA, Garcia JGN, Jacobson JR Tags: Am J Physiol Lung Cell Mol Physiol Source Type: research

Cellular Clocks in Hyperoxia Effects on Ca2+i Regulation in Developing Human Airway Smooth Muscle.
We report that human fASM express core clock machinery (PER1, PER2, CRY1, ARNTL/BMAL1, CLOCK) that is responsive to dexamethasone and altered by O2. Disruption of the clock via siRNA-mediated PER1 or ARNTL knockdown alters store-operated calcium entry (SOCE) and [Ca2+]i response to histamine in hyperoxia. Effects of O2 on [Ca2+]i are rescued by driving expression of clock proteins, via effects on the Ca2+ channels IP3R and Orai1. These data reveal a functional fASM clock that modulates [Ca2+]i regulation, particularly in hyperoxia. Harnessing clock biology may be a novel therapeutic consideration for neonatal airway diseas...
Source: American Journal of Physiology. Lung Cellular and Molecular Physiology - January 6, 2021 Category: Cytology Authors: Bartman CM, Matveyenko A, Pabelick CM, Prakash YS Tags: Am J Physiol Lung Cell Mol Physiol Source Type: research

TNF α Induces Mitochondrial Fragmentation and Biogenesis in Human Airway Smooth Muscle.
TNFα Induces Mitochondrial Fragmentation and Biogenesis in Human Airway Smooth Muscle. Am J Physiol Lung Cell Mol Physiol. 2020 Nov 04;: Authors: Delmotte PF, Marin Mathieu N, Sieck GC Abstract In human airway smooth muscle (hASM), mitochondrial volume density is greater in asthmatic patients compared to normal controls. There is also an increase in mitochondrial fragmentation in hASM of moderate asthmatics associated with an increase in Drp1 and a decrease in Mfn2 expression, mitochondrial fission and fusion proteins, respectively. Pro-inflammatory cytokines such TNFα contribute to hASM hyperreacti...
Source: American Journal of Physiology. Lung Cellular and Molecular Physiology - November 4, 2020 Category: Cytology Authors: Delmotte PF, Marin Mathieu N, Sieck GC Tags: Am J Physiol Lung Cell Mol Physiol Source Type: research

Mitochondrial 8-Oxoguanine DNA Glycosylase Mitigates Alveolar Epithelial Cell PINK1 Deficiency, Mitochondrial DNA Damage, Apoptosis and Lung Fibrosis.
Abstract Alveolar epithelial cell (AEC) apoptosis arising from mitochondrial dysfunction and mitophagy defects are important in mediating idiopathic pulmonary fibrosis (IPF). Our group established a role for the mitochondrial (mt) DNA base excision repair enzyme, 8-oxoguanine-DNA glycosylase 1 (mtOGG1), in preventing oxidant-induced AEC mtDNA damage and apoptosis and showed that OGG1-deficient mice have increased lung fibrosis. Herein, we determined whether mice over-expressing mtOGG1 transgene (MtOgg1tg) are protected against lung fibrosis and whether AEC mtOGG1 preservation of mtDNA integrity mitigates PINK1 def...
Source: American Journal of Physiology. Lung Cellular and Molecular Physiology - March 24, 2020 Category: Cytology Authors: Kim SJ, Cheresh P, Jablonski RP, Rachek L, Yeldandi A, Piseaux-Aillon R, Ciesielski MJ, Ridge KM, Gottardi CJ, Lam AP, Pardo A, Selman M, Natarajan V, Kamp DW Tags: Am J Physiol Lung Cell Mol Physiol Source Type: research

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