In vitro, human fetal lung type II pneumocytes and their response to cocaine by phospholipid analysis

Previously, it was found that lung explants from mid-trimester human abortuses differentiate spontaneously in organ culture in serum-free defined medium in an atmosphere of 95% air-5% CO2. Dibutyryl adenosine 3',5'-cyclic monophosphate (DBcAMP) treatment of human fetal lung in culture increases the rate of morphological differentiation and enhances expression of the surfactant protein A (SP-A) gene. To begin to define the factors responsible for this accelerated in vitro differentiation, we analyzed the effects of atmospheric oxygen on the morphological and biochemical development of human fetal lung in culture and on responsiveness of the cultured tissue to DBcAMP. We found that when lung explants were maintained in an atmosphere containing 1% oxygen they failed to differentiate spontaneously and no induction of SP-A gene expression was apparent. Furthermore, at 1% oxygen, DBcAMP had no effect to stimulate morphological differentiation or SP-A gene expression. When lung tissues that had been maintained for 5 days in 1% oxygen were transferred to an environment containing 20% oxygen, there was rapid morphological development and induction of SP-A gene expression. The effects on morphological development were manifest within 24 h of transfer to the 20% oxygen environment; within 72 h, a marked stimulatory effect of DBcAMP on SP-A gene expression also was observed. Our findings further suggest that the effects of oxygen on the levels of SP-A and SP-A mRNA are concentration dependent. Interestingly, the inductive effects of DBcAMP on SP-A gene expression were apparent only at oxygen concentrations > or = 10%. Morphological differentiation of the cultured human fetal lung tissue also was influenced by oxygen in a concentration-dependent manner. These findings suggest that oxygen plays an important permissive role in the spontaneous differentiation of human fetal lung in vitro.

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Differentiation of Type II Cells during Explant Culture of Human Fetal Lung Is Accelerated by Endogenous Prostanoids and Adenosine 3′,5′-Monophosphate*

Endocrinology ◽

10.1210/endo-128-6-2916 ◽

1991 ◽

Vol 128 (6) ◽

pp. 2916-2924 ◽

Cited By ~ 35

Author(s):

PHILIP L. BALLARD ◽

LINDA W. GONZALES ◽

MARY C. WILLIAMS ◽

JAMES M. ROBERTS ◽

MARK M. JACOBS

Keyword(s):

Fetal Lung ◽

Explant Culture ◽

Type Ii Cells ◽

Type Ii ◽

Human Fetal Lung

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Acetaminophen decreases intracellular glutathione levels and modulates cytokine production in human alveolar macrophages and type II pneumocytes in vitro

The International Journal of Biochemistry & Cell Biology ◽

10.1016/j.biocel.2005.03.005 ◽

2005 ◽

Vol 37 (8) ◽

pp. 1727-1737 ◽

Cited By ~ 39

Author(s):

Svetlana Dimova ◽

Peter H.M. Hoet ◽

David Dinsdale ◽

Benoit Nemery

Keyword(s):

Alveolar Macrophages ◽

Cytokine Production ◽

Type Ii ◽

Type Ii Pneumocytes ◽

Human Alveolar Macrophages ◽

Intracellular Glutathione

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Effect of tunicamycin on maturation of fetal mouse lung

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.1993.265.3.l250 ◽

1993 ◽

Vol 265 (3) ◽

pp. L250-L259

Author(s):

E. H. Webster ◽

S. R. Hilfer ◽

R. L. Searls ◽

J. Kornilow

Keyword(s):

Extracellular Matrix ◽

Fetal Lung ◽

Mouse Lung ◽

Type I ◽

Type Ii ◽

Fetal Mouse ◽

Type Ii Pneumocytes ◽

Separate Control ◽

Normal Controls

The mesodermal capsule of the fetal lung plays a role in differentiation of the respiratory region. It has been proposed for other epithelial organs that the mesodermal capsule influences development by modifying the basal lamina or the extended extracellular matrix. The effect could be on deposition or turnover of collagens, proteoglycans, and/or glycoproteins. This study tests the role of glycoproteins in differentiation of respiratory endings by inhibiting their synthesis with the antibiotic tunicamycin (TM). Lungs at 16 and 18 days gestation and 3 days after birth were cultured with TM and examined for morphological and biochemical differences from normal controls. With TM, alveolar regions did not expand properly and formed fewer type I pneumocytes, although type II pneumocytes were unaffected. The epithelium of untreated respiratory regions showed greater incorporation of radioactive mannose than the airways region or mesenchyme. This incorporation was diminished in TM, but the pattern persisted. Comparison with the results obtained with beta-xyloside suggested that differentiation of type I and type II pneumocytes is under separate control.

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TMPRSS2 Is the Major Activating Protease of Influenza A Virus in Primary Human Airway Cells and Influenza B Virus in Human Type II Pneumocytes

Journal of Virology ◽

10.1128/jvi.00649-19 ◽

2019 ◽

Vol 93 (21) ◽

Cited By ~ 29

Author(s):

Hannah Limburg ◽

Anne Harbig ◽

Dorothea Bestle ◽

David A. Stein ◽

Hong M. Moulton ◽

...

Keyword(s):

Influenza A ◽

Influenza B Virus ◽

Influenza B ◽

Virus Infectivity ◽

Type Ii ◽

Proteolytic Activation ◽

Human Airway ◽

Type Ii Pneumocytes ◽

Airway Cells

ABSTRACT Cleavage of influenza virus hemagglutinin (HA) by host cell proteases is essential for virus infectivity and spread. We previously demonstrated in vitro that the transmembrane protease TMPRSS2 cleaves influenza A virus (IAV) and influenza B virus (IBV) HA possessing a monobasic cleavage site. Subsequent studies revealed that TMPRSS2 is crucial for the activation and pathogenesis of H1N1pdm and H7N9 IAV in mice. In contrast, activation of H3N2 IAV and IBV was found to be independent of TMPRSS2 expression and supported by an as-yet-undetermined protease(s). Here, we investigated the role of TMPRSS2 in proteolytic activation of IAV and IBV in three human airway cell culture systems: primary human bronchial epithelial cells (HBEC), primary type II alveolar epithelial cells (AECII), and Calu-3 cells. Knockdown of TMPRSS2 expression was performed using a previously described antisense peptide-conjugated phosphorodiamidate morpholino oligomer, T-ex5, that interferes with splicing of TMPRSS2 pre-mRNA, resulting in the expression of enzymatically inactive TMPRSS2. T-ex5 treatment produced efficient knockdown of active TMPRSS2 in all three airway cell culture models and prevented proteolytic activation and multiplication of H7N9 IAV in Calu-3 cells and H1N1pdm, H7N9, and H3N2 IAV in HBEC and AECII. T-ex5 treatment also inhibited the activation and spread of IBV in AECII but did not affect IBV activation in HBEC and Calu-3 cells. This study identifies TMPRSS2 as the major HA-activating protease of IAV in human airway cells and IBV in type II pneumocytes and as a potential target for the development of novel drugs to treat influenza infections. IMPORTANCE Influenza A viruses (IAV) and influenza B viruses (IBV) cause significant morbidity and mortality during seasonal outbreaks. Cleavage of the viral surface glycoprotein hemagglutinin (HA) by host proteases is a prerequisite for membrane fusion and essential for virus infectivity. Inhibition of relevant proteases provides a promising therapeutic approach that may avoid the development of drug resistance. HA of most influenza viruses is cleaved at a monobasic cleavage site, and a number of proteases have been shown to cleave HA in vitro. This study demonstrates that the transmembrane protease TMPRSS2 is the major HA-activating protease of IAV in primary human bronchial cells and of both IAV and IBV in primary human type II pneumocytes. It further reveals that human and murine airway cells can differ in their HA-cleaving protease repertoires. Our data will help drive the development of potent and selective protease inhibitors as novel drugs for influenza treatment.

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Bismuth(III) bromide-thioamide complexes: synthesis, characterization and cytotoxic properties

Main Group Metal Chemistry ◽

10.1515/mgmc-2018-0035 ◽

2018 ◽

Vol 41 (5-6) ◽

pp. 143-154 ◽

Cited By ~ 2

Author(s):

M. Cakmak ◽

I.I. Ozturk ◽

C.N. Banti ◽

M. Manoli ◽

E. Moushi ◽

...

Keyword(s):

Fourier Transform ◽

Fetal Lung ◽

Molecular Structures ◽

X Ray Diffraction ◽

Vis Spectroscopy ◽

Normal Human ◽

Ft Ir ◽

Human Fetal Lung ◽

Heterocyclic Thioamides

Abstract New bismuth(III) bromine compounds of the heterocyclic thioamides were prepared and structurally characterized. The reaction of heterocyclic thioamides with bismuth(III) bromide resulted in the formation of the {[BiBr2(μ2-Br)(MMI)2]2·CH3COCH3·H2O} (1), {[BiBr2(MBZIM)4]·Br·2H2O} (2), {[BiBr2(μ2-Br)(tHPMT)2]2·CH3CN} (3), {[BiBr2(μ2-Br)(PYT)2]2·CH3CN} (4) and {[BiBr2(μ2-Br)(MBZT)2]2 2CH3OH} (5) complexes (MMI: 2-mercapto-1-methylimidazole, MBZIM: 2-mercaptobenzimidazole, tHPMT: 2-mercapto-3,4,5,6-tetrahydro-pyrimidine, PYT: 2-mercaptopyridine and MBZT: 2-mercaptobenzothiazole). The complexes 1–5 were characterized by melting point (m.p.), elemental analysis (e.a.), molar conductivity, Fourier-transform infrared (FT-IR), Fourier-transform Raman (FT-Raman), nuclear magnetic resonance (1H and 13CNMR) spectroscopy, UV-Vis spectroscopy and thermogravimetric-differential thermal analysis (TG-DTA). The molecular structures of 1–5 were determined by single-crystal X-ray diffraction. Complex 2 is a first ionic monomuclear octahedral bismuth(III) bromide, while the complexes 1, 3–5 are the first examples of dinuclear bismuth(III) bromide derivatives. Complexes 1–5 were evaluated in terms of their in vitro cytotoxic activity against human adenocarcinoma breast (MCF-7) and cervix (HeLa) cells. The toxicity on normal human fetal lung fibroblast cells (MRC-5) was also evaluated. Moreover, the complexes 1–5 and free heterocyclic thioamide ligands were studied upon the catalytic peroxidation of the linoleic acid by the enzyme lipoxygenase (LOX).

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