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American Journal of Physiology. Lung... Nov 2008Hyaluronan (HA), a large glycosaminoglycan found in the ECM, has major roles in lung and vascular biology and disease. However, its role in idiopathic pulmonary arterial...
Hyaluronan (HA), a large glycosaminoglycan found in the ECM, has major roles in lung and vascular biology and disease. However, its role in idiopathic pulmonary arterial hypertension (IPAH) is unknown. We hypothesized that HA metabolism is abnormal in IPAH. We measured the plasma levels of HA in IPAH and healthy individuals. We also evaluated HA synthesis and the expression of HA synthases and hyaluronidases in pulmonary artery smooth muscle cells (PASMCs) from explanted lungs. Plasma HA levels were markedly elevated in IPAH compared with controls [HA (ng/ml, mean +/- SD): IPAH 325 +/- 80, control 28 +/- 9; P = 0.02]. In vitro, unstimulated IPAH PASMCs produced high levels of HA compared with control cells [HA in supernatant (microg/ml, mean +/- SD): IPAH 12 +/- 2, controls 6 +/- 0.9; P = 0.04]. HA levels were also higher in IPAH PASMC lysates. The increased HA was biologically relevant as shown by tissue staining and increased HA-specific binding of mononuclear cells to IPAH compared with control PASMCs [number of bound cells x 10(4) (mean +/- SD): IPAH 9.5 +/- 3, control 3.0 +/- 1; P = 0.01]. This binding was abrogated by the addition of hyaluronidase. HA synthase-2 and hyaluronidase-2 were predominant in control and IPAH PASMCs. Interestingly, the expressions of HA synthase-2 and hyaluronidase-2 were approximately 2-fold lower in IPAH compared with controls [HA synthase-2 (relative expression mean +/- SE): IPAH 4.3 +/- 0.02, control 7.8 +/- 0.1; P = 0.0004; hyaluronidase-2 (relative expression mean +/- SE): IPAH 4.2 +/- 0.06, control 7.6 +/- 0.07; P = 0.008]. Thus patients with IPAH have higher circulating levels of HA, and PASMCs derived from IPAH lungs produce more HA compared with controls. This is associated with increased tissue levels and increased binding of inflammatory cells suggesting a role for HA in remodeling and inflammation in IPAH.
Topics: Adult; Case-Control Studies; Cell Adhesion; Endoplasmic Reticulum; Female; Gene Expression Regulation, Enzymologic; Glucuronosyltransferase; Humans; Hyaluronic Acid; Hyaluronoglucosaminidase; Hypertension, Pulmonary; Inflammation; Isoenzymes; Lung; Male; Middle Aged; Models, Biological; Molecular Weight; Myocytes, Smooth Muscle; Pulmonary Artery; U937 Cells
PubMed: 18776053
DOI: 10.1152/ajplung.90306.2008 -
International Journal of Molecular... Sep 2022We conducted intratracheal instillations of different molecular weights of polyacrylic acid (PAA) into rats in order to examine what kinds of physicochemical...
BACKGROUND
We conducted intratracheal instillations of different molecular weights of polyacrylic acid (PAA) into rats in order to examine what kinds of physicochemical characteristics of acrylic acid-based polymer affect responses in the lung.
METHODS
F344 rats were intratracheally exposed to a high molecular weight (HMW) of 598 thousand g/mol or a low molecular weight (LMW) of 30.9 thousand g/mol PAA at low and high doses. Rats were sacrificed at 3 days, 1 week, 1 month, 3 months and 6 months post exposure.
RESULTS
HMW PAA caused persistent increases in neutrophil influx, cytokine-induced neutrophil chemoattractants (CINC) in the bronchoalveolar lavage fluid (BALF), and heme oxygenase-1 (HO-1) in the lung tissue from 3 days to 3 months and 6 months following instillation. On the other hand, LMW PAA caused only transient increases in neutrophil influx, CINC in BALF, and HO-1 in the lung tissue from 3 days to up to 1 week or 1 month following instillation. Histopathological findings of the lungs demonstrated that the extensive inflammation and fibrotic changes caused by the HMW PAA was greater than that in exposure to the LMW PAA during the observation period.
CONCLUSION
HMW PAA induced persistence of lung disorder, suggesting that molecular weight is a physicochemical characteristic of PAA-induced lung disorder.
Topics: Acrylic Resins; Animals; Bronchoalveolar Lavage Fluid; Chemotactic Factors; Cytokines; Heme Oxygenase-1; Intubation, Intratracheal; Lung; Molecular Weight; Rats; Rats, Inbred F344
PubMed: 36142256
DOI: 10.3390/ijms231810345 -
Molecules (Basel, Switzerland) Jul 2021(1) Background: Household humidifier disinfectant (HD) brands containing polyhexamethylene guanidine (PHMG) have been found to cause the most HD-associated lung injuries...
(1) Background: Household humidifier disinfectant (HD) brands containing polyhexamethylene guanidine (PHMG) have been found to cause the most HD-associated lung injuries (HDLIs) in the Republic of Korea. Nevertheless, no study has attempted to characterize the potential association of the health effects, including HDLI, with the physicochemical properties of PHMG dissolved in different HD brands. This study aimed to characterize the molecular weight (MW) distribution, the number-average molecular weight (M), the weight-average molecular weight (M), and the structural types of PHMG used in HD products. (2) Methods: Quantitative measurements were made using matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS). The M, M and MW distributions were compared among various HD products. (3) Results: The mean M and M were 542.4 g/mol (range: 403.0-692.2 g/mol) and 560.7 g/mol (range: 424.0-714.70 g/mol), respectively. The degree of PHMG oligomerization ranged from 3 to 7. The MW distribution of PHMG indicated oligomeric compounds regardless of the HD brands. (4) Conclusions: Based on the molecular weight distribution, the average molecular weight of PHMG, and the degree of polymerization, the PHMG collected from HDLI victims could be regarded as an oligomer. PHMG, as used in household humidifiers, should not be exempted from toxic chemical registration as a polymer. Further study is necessary to examine the association of PHMG oligomeric compounds and respiratory health effects, including HDLI.
Topics: Disinfectants; Guanidines; Humans; Humidifiers; Lung Injury; Molecular Weight; Polymerization; Republic of Korea
PubMed: 34361643
DOI: 10.3390/molecules26154490 -
Biophysical Journal Dec 1973After reaction of DNA with high concentrations of bleomycin, approximately 80% of the DNA becomes trichloroacetic acid (TCA) soluble. The remaining 20% of the DNA... (Comparative Study)
Comparative Study
After reaction of DNA with high concentrations of bleomycin, approximately 80% of the DNA becomes trichloroacetic acid (TCA) soluble. The remaining 20% of the DNA remains TCA insoluble. Upon further treatment of this TCA-insoluble material with high concentrations of the drug, no further drug action can be detected. Drug action is defined as fragmentation of DNA to smaller molecular size, release of free bases, and TCA solubilization. This material which is not attacked by bleomycin has been termed bleomycin-resistant DNA. This bleomycin-resistant DNA does not compete with native DNA in the bleomycin reaction indicating that there is no binding or inactivation of the drug by the resistant DNA. The resistant DNA shows very little hyperchromicity when heated through the melting temperature for the corresponding native DNA, indicating a single-stranded structure. Results of sedimentation and equilibrium analyses yield a molecular weight of about 4,000 daltons. This value is the same regardless of the source of the native DNA. Finally, the bleomycin-resistant DNA exhibits a base composition similar to that of the native DNA from which it was derived.
Topics: Animals; Bacillus subtilis; Binding Sites; Bleomycin; Cell Line; Centrifugation, Density Gradient; Coliphages; Cricetinae; DNA; DNA, Bacterial; DNA, Single-Stranded; DNA, Viral; Deoxyribonucleases; Drug Resistance; Hot Temperature; Lung; Molecular Weight; Nucleic Acid Denaturation; Pancreas; Phosphoric Diester Hydrolases; Phosphorus Radioisotopes; Solubility; Thymidine; Trichloroacetic Acid; Tritium; Ultracentrifugation; Venoms
PubMed: 4128368
DOI: 10.1016/s0006-3495(73)86063-1 -
The Biochemical Journal Oct 1990Human iduronate-2-sulphatase (EC 3.1.6.13), which is involved in the lysosomal degradation of the glycosaminoglycans heparan sulphate and dermatan sulphate, was purified...
Human iduronate-2-sulphatase (EC 3.1.6.13), which is involved in the lysosomal degradation of the glycosaminoglycans heparan sulphate and dermatan sulphate, was purified more than 500,000-fold in 5% yield from liver with a six-step column procedure, which consisted of a concanavalin A-Sepharose-Blue A-agarose coupled step, chromatofocusing, gel filtration on TSK HW 50S-Fractogel, hydrophobic separation on phenyl-Sepharose CL-4B and size separation on TSK G3000SW Ultrapac. Two major forms were identified. Form A and form B, with pI values of 4.5 and less than 4.0 respectively, separated at the chromatofocusing step in approximately equal amounts of recovered enzyme activity. By gel-filtration methods form A had a native molecular mass in the range 42-65 kDa. When analysed by SDS/PAGE, dithioerythritol-reduced and non-reduced form A and form B consistently contained polypeptides of molecular masses 42 kDa and 14 kDa. Iduronate-2-sulphatase was purified from human kidney, placenta and lung, and form A was shown to have similar native molecular mass and subunit components to those observed for liver enzyme. Both forms of liver iduronate-2-sulphatase were active towards a variety of substrates derived from heparin and dermatan sulphate. Kinetic parameters (Km and Kcat) of form A were determined with a variety of substrates matching structural aspects of the physiological substrates in vivo, namely heparan sulphate, heparin and dermatan sulphate. Substrate with 6-sulphate esters on the aglycone residue adjacent to the iduronic acid 2-sulphate residue being attack were hydrolysed with catalytic efficiencies up to 200 times above that observed for the simplest disaccharide substrate without a 6-sulphated aglycone residue. The effect of incubation pH on enzyme activity towards the variety of substrates evaluated was complex and dependent on substrate aglycone structure, substrate concentration, buffer type and the presence of other proteins. Sulphate and phosphate ions and a number of substrate and product analogues were potent inhibitor of form A and form B enzyme activities.
Topics: Carbohydrate Sequence; Catalysis; Chromatography; Dermatan Sulfate; Dithioerythritol; Electrophoresis, Polyacrylamide Gel; Heparitin Sulfate; Humans; Hydrogen-Ion Concentration; Iduronate Sulfatase; Isoelectric Point; Kidney; Kinetics; Liver; Lung; Molecular Sequence Data; Molecular Weight
PubMed: 2222422
DOI: 10.1042/bj2710075 -
The Journal of Biological Chemistry Sep 1985Both immature and adult rat lungs contain three prominent soluble beta-galactoside-binding proteins with subunit Mr approximately 14,500, 18,000, and 29,000 rather than...
Both immature and adult rat lungs contain three prominent soluble beta-galactoside-binding proteins with subunit Mr approximately 14,500, 18,000, and 29,000 rather than only the one noted previously. They are readily resolved by ion-exchange chromatography, and antibodies raised against them show little cross-reaction. The three proteins were also found in immature heart, skeletal muscle, and liver, but only the protein with subunit Mr approximately 14,500 was found in these tissues in young adults.
Topics: Aging; Amino Acids; Animals; Asialoglycoproteins; Carbohydrates; Chromatography, Affinity; Electrophoresis, Polyacrylamide Gel; Fetuins; Galactosides; Galectins; Glycosides; Hemagglutinins; Lung; Macromolecular Substances; Molecular Weight; Rats; alpha-Fetoproteins
PubMed: 2411725
DOI: No ID Found -
International Journal of Molecular... Feb 2023Pharmacological therapies in lung diseases are nowadays useful in reducing the symptomatology of lung injury. However, they have not yet been translated to effective...
Pharmacological therapies in lung diseases are nowadays useful in reducing the symptomatology of lung injury. However, they have not yet been translated to effective treatment options able to restore the lung tissue damage. Cell-therapy based on Mesenchymal Stem Cells (MSCs) is an attractive, as well as new therapeutic approach, although some limitations can be ascribed for therapeutic use, such as tumorigenicity and immune rejection. However, MSCs have the capacity to secrete multiple paracrine factors, namely secretome, capable of regulating endothelial and epithelial permeability, decrease inflammation, enhancing tissue repair, and inhibiting bacterial growth. Furthermore, Hyaluronic acid (HA) has been demonstrated to have particularly efficacy in promoting the differentiation of MSCs in Alveolar type II (ATII) cells. In this frame, the combination of HA and secretome to achieve the lung tissue regeneration has been investigated for the first time in this work. Overall results showed how the combination of HA (low and medium molecular weight HA) plus secretome could enhance MSCs differentiation in ATII cells (SPC marker expression of about 5 ng/mL) compared to the only HA or secretome solutions alone (SPC about 3 ng/mL, respectively). Likewise, cell viability and cell rate of migration were reported to be improved for HA and secretome blends, indicating an interesting potentiality of such systems for lung tissue repair. Moreover, an anti-inflammatory profile has been revealed when dealing with HA and secretome mixtures. Therefore, these promising results can allow important advance in the accomplishment of the future therapeutic approach in respiratory diseases, up to date still missing.
Topics: Hyaluronic Acid; Secretome; Mesenchymal Stem Cells; Cell- and Tissue-Based Therapy; Lung
PubMed: 36835068
DOI: 10.3390/ijms24043642 -
Scientific Reports Mar 2020Lungs of the rodent species, the African giant pouched rat (Cricetomys gambianus) and the Nigerian mole rat (Cryptomys foxi) were investigated. Significant morphometric... (Comparative Study)
Comparative Study
Lungs of the rodent species, the African giant pouched rat (Cricetomys gambianus) and the Nigerian mole rat (Cryptomys foxi) were investigated. Significant morphometric differences exist between the two species. The volume of the lung per unit body mass was 2.7 times larger; the respiratory surface area 3.4 times greater; the volume of the pulmonary capillary blood 2 times more; the harmonic mean thickness of the blood-gas (tissue) barrier (τht) ~29% thinner and; the total pulmonary morphometric diffusing capacity (DLo) for O 2.3 times more in C. foxi. C. gambianus occupies open burrows that are ventilated with air while C. foxi lives in closed burrows. The less morphometrically specialized lungs of C. gambianus may be attributed to its much larger body mass (~6 times more) and possibly lower metabolic rate and its semifossorial life whereas the 'superior' lungs of C. foxi may largely be ascribed to the subterranean hypoxic and hypercapnic environment it occupies. Compared to other rodents species that have been investigated hitherto, the τht was mostly smaller in the lungs of the subterranean species and C. foxi has the highest mass-specific DLo. The fossorial- and the subterranean rodents have acquired various pulmonary structural specializations that relate to habitats occupied.
Topics: Animals; Ecosystem; Lung; Mole Rats; Muridae
PubMed: 32251351
DOI: 10.1038/s41598-020-61873-8 -
Respiratory Research Sep 2010Idiopathic pulmonary fibrosis (IPF) and chronic obstructive pulmonary disease (COPD) are disorders of the lung parenchyma. They share the common denominators of a... (Comparative Study)
Comparative Study
BACKGROUND
Idiopathic pulmonary fibrosis (IPF) and chronic obstructive pulmonary disease (COPD) are disorders of the lung parenchyma. They share the common denominators of a progressive nature and poor prognosis. The goal was to use non-biased proteomics to discover new markers for these diseases.
METHODS
Proteomics of fibrotic vs. control lung tissue suggested decreased levels of several spots in the lung specimens of IPF patients, which were identified as Hemoglobin (Hb) α and β monomers and Hbα complexes. The Hbα and β monomers and complexes were investigated in more detail in normal lung and lung specimens of patients with IPF and COPD by immunohistochemistry, morphometry and mass spectrometry (MS).
RESULTS
Both Hb monomers, in normal lung, were expressed especially in the alveolar epithelium. Levels of Hbα and β monomers and complexes were reduced/lost in IPF but not in the COPD lungs when compared to control lung. MS-analyses revealed Hbα modification at cysteine105 (Cysα105), preventing formation of the Hbα complexes in the IPF lungs. Hbα and Hbβ were expressed as complexes and monomers in the lung tissues, but were secreted into the bronchoalveolar lavage fluid and/or induced sputum supernatants as complexes corresponding to the molecular weight of the Hb tetramer.
CONCLUSIONS
The abundant expression of the oxygen carrier molecule Hb in the normal lung epithelium and its decline in IPF lung are new findings. The loss of Hb complex formation in IPF warrants further studies and may be considered as a disease-specific modification.
Topics: Adult; Aged; Biomarkers; Bronchoalveolar Lavage Fluid; Diagnosis, Differential; Female; Hemoglobins; Humans; Idiopathic Pulmonary Fibrosis; Lung; Male; Middle Aged; Peptide Fragments; Pulmonary Disease, Chronic Obstructive; Respiratory Mucosa; beta-Globins
PubMed: 20836851
DOI: 10.1186/1465-9921-11-123 -
BMC Pulmonary Medicine May 2011KL-6 is a high-molecular-weight glycoprotein classified as a human MUC1 mucin. It was hypothesized that KL-6 could be detectable in the circulating blood and especially...
BACKGROUND
KL-6 is a high-molecular-weight glycoprotein classified as a human MUC1 mucin. It was hypothesized that KL-6 could be detectable in the circulating blood and especially in airway secretions in lung diseases associated with mucus production such as chronic obstructive pulmonary disease (COPD). Additional aims of this study were to investigate whether the levels of KL-6 in plasma and sputum are related to ageing and smoking history.
METHODS
The concentrations of KL-6 in plasma and induced sputum supernatants from young and/or middle aged/elderly non-smokers, smokers and patients with COPD were assayed by ELISA (n = 201). The subjects were classified into five groups according to age, smoking status and presence of COPD. In addition, KL-6 expression in control and diseased lung i.e. samples from patients with COPD (n = 28), were analyzed by immunohistochemistry and digital image analysis.
RESULTS
The plasma levels of KL-6 increased with age both in non-smokers and smokers. Among middle aged/elderly subjects, plasma KL-6 levels in all smokers regardless of COPD were significantly higher than in non-smokers, whereas sputum levels of KL-6 were significantly higher in COPD compared not only to non-smokers but also to smokers. KL-6 was more prominently expressed in the bronchiolar/alveolar epithelium in COPD than in the control lungs. Plasma and sputum KL-6 levels correlated inversely with obstruction and positively with smoking history and ageing. The linear multiple regression analysis confirmed that age and cigarette smoking had independent effects on plasma KL-6.
CONCLUSIONS
KL-6 increases with ageing and chronic smoking history, but prospective studies will be needed to elucidate the significance of KL-6 in chronic airway diseases.
Topics: Adult; Aged; Aging; Bronchi; Case-Control Studies; Epithelium; Female; Finland; Humans; Lung; Male; Middle Aged; Mucin-1; Pulmonary Alveoli; Pulmonary Disease, Chronic Obstructive; Smoking; Sputum
PubMed: 21569324
DOI: 10.1186/1471-2466-11-22