-
Clinical and Applied... Sep 2016Heparin is a widely used clinical anticoagulant. It is also a linear glycosaminoglycan with an average mass between 10 and 20 kDa and is primarily made up of trisulfated... (Comparative Study)
Comparative Study
Heparin is a widely used clinical anticoagulant. It is also a linear glycosaminoglycan with an average mass between 10 and 20 kDa and is primarily made up of trisulfated disaccharides comprised of 1,4-linked iduronic acid and glucosamine residues containing some glucuronic acid residues. Heparin is biosynthesized in the Golgi of mast cells commonly found in the liver, intestines, and lungs. Pharmaceutical heparin currently used in the United States is primarily extracted from porcine intestines. Other sources of heparin including bovine intestine and bovine lung are being examined as potential substitutes for porcine intestinal heparin. These additional sources are intended to serve to diversify the heparin supply, making this lifesaving drug more secure. The current study examines bovine heparins prepared from both intestines and lung and compares these to porcine intestinal heparin. The structural properties of these heparins are examined using nuclear magnetic resonance, gel permeation chromatography, and disaccharide analysis of heparinase-catalyzed depolymerized heparin. The in vitro functional activities of these heparins have also been determined. The goal of this study is to establish the structural and functional similarities and potential differences between bovine and porcine heparins. Porcine and bovine heparins have structural and compositional similarities and differences.
Topics: Animals; Anticoagulants; Blood Coagulation Tests; Cattle; Heparin; Intestines; Lung; Molecular Structure; Molecular Weight; Swine
PubMed: 27084870
DOI: 10.1177/1076029616643822 -
Matrix Biology : Journal of the... Feb 2023Herein, we tested the hypothesis that low molecular weight hyaluronan (LMW-HA) inhibits lung epithelial ions transport in-vivo, ex-vivo, and in-vitro by activating the...
Herein, we tested the hypothesis that low molecular weight hyaluronan (LMW-HA) inhibits lung epithelial ions transport in-vivo, ex-vivo, and in-vitro by activating the calcium-sensing receptor (CaSR). Twenty-four hours post intranasal instillation of 50-150 µg/ml LMW-HA to C57BL/6 mice, there was a 75% inhibition of alveolar fluid clearance (AFC), a threefold increase in the epithelial lining fluid (ELF) depth, and a 20% increase in lung wet/dry (W/D) ratio. Incubation of human and mouse precision cut lung slices with 150 µg/ml LMW-HA reduced the activity and the open probability (P) of epithelial sodium channel (ENaC) in alveolar epithelial type 2 (ATII) cells, and in mouse tracheal epithelial cells (MTEC) monolayers as early as 4 h. The Cl current through cystic fibrosis transmembrane conductance regulator (CFTR) and the activity of Na,K-ATPase were both inhibited by more than 66% at 24 h. The inhibitory effects of LMW-HA on ion channels were reversed by 1 µM NPS-2143, or 150 µg/ml high molecular weight hyaluronan (HMW-HA). In HEK-293 cells expressing the calcium-sensitive Cl channel TMEM16-A, CaSR was required for the activation of the Cl current by LMW-HA. This is the first demonstration of lung ions and water transport inhibition by LMW-HA, and its mediation through the activation of CaSR.
Topics: Mice; Humans; Animals; Receptors, Calcium-Sensing; Hyaluronic Acid; Sodium-Potassium-Exchanging ATPase; HEK293 Cells; Molecular Weight; Mice, Inbred C57BL; Lung
PubMed: 36758905
DOI: 10.1016/j.matbio.2023.02.002 -
PloS One 2021Optimised pre-clinical models are required for TB drug development to better predict the pharmacokinetics of anti-tuberculosis (anti-TB) drugs to shorten the time taken...
Optimised pre-clinical models are required for TB drug development to better predict the pharmacokinetics of anti-tuberculosis (anti-TB) drugs to shorten the time taken for novel drugs and combinations to be approved for clinical trial. Microdialysis can be used to measure unbound drug concentrations in awake freely moving animals in order to describe the pharmacokinetics of drugs in the organs as a continuous sampling technique. The aim of this work was to develop and optimise the microdialysis methodology in guinea pigs to better understand the pharmacokinetics of rifampicin in the lung. In vitro experiments were performed before progressing into in vivo studies because the recovery (concentration of the drug in the tissue fluid related to that in the collected dialysate) of rifampicin was dependent on a variety of experimental conditions. Mass spectrometry of the dialysate was used to determine the impact of flow rate, perfusion fluid and the molecular weight cut-off and membrane length of probes on the recovery of rifampicin at physiologically relevant concentrations. Following determination of probe efficiency and identification of a correlation between rifampicin concentrations in the lung and skeletal muscle, experiments were conducted to measure rifampicin in the sacrospinalis of guinea pigs using microdialysis. Lung concentrations of rifampicin were estimated from the rifampicin concentrations measured in the sacrospinalis. These studies suggest the potential usefulness of the microdialysis methodology to determine drug concentrations of selected anti-TB drugs to support new TB drug development.
Topics: Animals; Antitubercular Agents; Drug Development; Female; Guinea Pigs; Lung; Microdialysis; Rifampin
PubMed: 33481939
DOI: 10.1371/journal.pone.0245922 -
PloS One 2011Infection by Burkholderia cenocepacia in cystic fibrosis (CF) patients is associated with poor clinical prognosis. Previously, we demonstrated that one of the highly...
BACKGROUND
Infection by Burkholderia cenocepacia in cystic fibrosis (CF) patients is associated with poor clinical prognosis. Previously, we demonstrated that one of the highly transmissible strains, BC7, expresses cable pili and the associated 22 kDa adhesin, both of which contribute to BC7 binding to airway epithelial cells. However, the contribution of these factors to induce inflammation and bacterial persistence in vivo is not known.
METHODOLOGY/PRINCIPAL FINDINGS
Wild-type BC7 stimulated higher IL-8 responses than the BC7 cbl and BC7 adhA mutants in both CF and normal bronchial epithelial cells. To determine the role of cable pili and the associated adhesin, we characterized a mouse model of B. cenocepacia, where BC7 are suspended in Pseudomonas aeruginosa alginate. C57BL/6 mice were infected intratracheally with wild-type BC7 suspended in either alginate or PBS and were monitored for lung bacterial load and inflammation. Mice infected with BC7 suspended in PBS completely cleared the bacteria by 3 days and resolved the inflammation. In contrast, mice infected with BC7 suspended in alginate showed persistence of bacteria and moderate lung inflammation up to 5 days post-infection. Using this model, mice infected with the BC7 cbl and BC7 adhA mutants showed lower bacterial loads and mild inflammation compared to mice infected with wild-type BC7. Complementation of the BC7 cblS mutation in trans restored the capacity of this strain to persist in vivo. Immunolocalization of bacteria revealed wild-type BC7 in both airway lumen and alveoli, while the BC7 cbl and BC7 adhA mutants were found mainly in airway lumen and peribronchiolar region.
CONCLUSIONS AND SIGNIFICANCE
B. cenocepacia suspended in alginate can be used to determine the capacity of bacteria to persist and cause lung inflammation in normal mice. Both cable pili and adhesin contribute to BC7-stimulated IL-8 response in vitro, and BC7 persistence and resultant inflammation in vivo.
Topics: Adhesins, Bacterial; Alginates; Animals; Burkholderia Infections; Burkholderia cenocepacia; Disease Models, Animal; Epithelial Cells; Fimbriae, Bacterial; Genes, Bacterial; Glucuronic Acid; Hexuronic Acids; Humans; Interleukin-8; Lung; Mice; Mice, Inbred C57BL; Molecular Weight; Mutation; Neutrophil Infiltration; Pneumonia
PubMed: 21811611
DOI: 10.1371/journal.pone.0022435 -
The Journal of Biological Chemistry Jul 1982To elucidate mechanisms involved in the regulation of lung collagen content we studied hamsters with bleomycin-induced pulmonary fibrosis. Lung collagen in this model is...
Modulation of collagen production following bleomycin-induced pulmonary fibrosis in hamsters. Presence of a factor in lung that increases fibroblast prostaglandin E2 and cAMP and suppresses fibroblast proliferation and collagen production.
To elucidate mechanisms involved in the regulation of lung collagen content we studied hamsters with bleomycin-induced pulmonary fibrosis. Lung collagen in this model is increased as the result of greatly increased lung collagen synthesis rates. However, collagen synthesis rates are subsequently restored to normal. Hamster lung explants from both normal and bleomycin-exposed hamsters were cultured, and the effects of explant conditioned medium (CM) on lung fibroblast (IMR-90) proliferation and collagen production in vitro were determined. Lung explant CM increased fibroblast prostaglandin (PG)E2 production and intracellular cAMP, and decreased both fibroblast proliferation and collagen production in a dose-dependent manner. Greater activity was observed with lung explant CM from bleomycin-exposed lungs. Incubation of fibroblasts with indomethacin prior to addition of CM blocked CM-mediated changes in PGE2 and cAMP and inhibited changes in fibroblast proliferation and collagen production. Exogenous PGE2 or dibutyryl cAMP also suppressed fibroblast proliferation and collagen production. The suppressive activity in lung-conditioned medium is nondialyzable, has an apparent molecular weight of 15,000-20,000 by gel filtration, and is heat-stable. It is not species-restricted since CM from hamster lung affected human and hamster lung fibroblasts similarly. Activity is present preformed in lung and bronchoalveolar lavage fluid, although bronchoalveolar macrophages produce a nondialyzable factor in culture which suppresses fibroblast proliferation. The suppressive activity identified in fibrotic lung may represent a means for limiting collagen accumulation following tumor injury.
Topics: Animals; Bleomycin; Cell Division; Cells, Cultured; Collagen; Cricetinae; Cyclic AMP; DNA Replication; Dinoprostone; Female; Fetus; Fibroblasts; Humans; Kinetics; Lung; Macrophages; Male; Mesocricetus; Pregnancy; Prostaglandins E; Pulmonary Fibrosis
PubMed: 6177695
DOI: No ID Found -
American Journal of Physiology. Lung... Feb 2016We tested the hypothesis that Pseudomonas aeruginosa type 3 secretion system effectors exoenzymes Y and U (ExoY and ExoU) induce release of a high-molecular-weight...
We tested the hypothesis that Pseudomonas aeruginosa type 3 secretion system effectors exoenzymes Y and U (ExoY and ExoU) induce release of a high-molecular-weight endothelial tau, causing transmissible cell injury characteristic of an infectious proteinopathy. Both the bacterial delivery of ExoY and ExoU and the conditional expression of an activity-attenuated ExoU induced time-dependent pulmonary microvascular endothelial cell gap formation that was paralleled by the loss of intracellular tau and the concomitant appearance of high-molecular-weight extracellular tau. Transfer of the high-molecular-weight tau in filtered supernatant to naïve endothelial cells resulted in intracellular accumulation of tau clusters, which was accompanied by cell injury, interendothelial gap formation, decreased endothelial network stability in Matrigel, and increased lung permeability. Tau oligomer monoclonal antibodies captured monomeric tau from filtered supernatant but did not retrieve higher-molecular-weight endothelial tau and did not rescue the injurious effects of tau. Enrichment and transfer of high-molecular-weight tau to naïve cells was sufficient to cause injury. Thus we provide the first evidence for a pathophysiological stimulus that induces release and transmissibility of high-molecular-weight endothelial tau characteristic of an endothelial proteinopathy.
Topics: Animals; Cyclic AMP; Endothelial Cells; Lung; Microvessels; Pseudomonas Infections; Pseudomonas aeruginosa; Rats
PubMed: 26637633
DOI: 10.1152/ajplung.00103.2015 -
Archives of Razi Institute Jul 2021In the last couple of years, a number of new and rapid tests for the diagnosis of Tuberculosis (TB) have been developed based on the low molecular weight antigens from...
In the last couple of years, a number of new and rapid tests for the diagnosis of Tuberculosis (TB) have been developed based on the low molecular weight antigens from Mycobacterium tuberculosis (Mtb) culture supernatant. This study aimed to isolate and purify low molecular weight antigens secreted by Mtb strain C for diagnostic purpose. The secretory proteins from culture filtrate of Mtb were extracted using ammonium sulphate precipitations and sephadex-G50 gel chromatography. The obtained antigen fractions were analyzed for their protein concentrations and approximate molecular weight using Lowry method and SDS-PAGE (12.5%), respectively. DOT-ELISA and Western blot assay was performed to confirm the presence of purified low molecular weight proteins isolated from Mtb using sera from pulmonary tuberculosis patients (polyclonal antibodies). During chromatography, low molecular weight proteins were separated, that was approximately 0.7 mg/ml of the total proteins (1.662 mg/ml). The purified protein fractions in molecular weight range of 14 kDa-41kDa appeared during SDS-PAGE analysis. The chromatographic band fraction in the weight range of 30-41 kDa was identified in the TB patients’ sera using Western blotting. The low molecular weight proteins in the culture filtrate of Mtb strain C were purified using ammonium sulphate and chromatography. These fractions were confirmed using Western blotting. The obtained results might support the hypothesis that the Mtb culture filtrate antigens could be used as a rapid and sensitive assay for the detection of patients with pulmonary TB.
Topics: Humans; Antigens, Bacterial; Enzyme-Linked Immunosorbent Assay; Molecular Weight; Mycobacterium tuberculosis; Tuberculosis, Pulmonary
PubMed: 34223726
DOI: 10.22092/ari.2020.127691.1390 -
Annals of the Royal College of Surgeons... Jan 2021Pulmonary sequestration is a congenital abnormality of a non-functional pulmonary mass with anomalous systemic arterial supply. Surgical resection is the gold standard...
Pulmonary sequestration is a congenital abnormality of a non-functional pulmonary mass with anomalous systemic arterial supply. Surgical resection is the gold standard treatment, but it carries a risk of life-threatening haemorrhage from accidental injury of the anomalous artery. Endovascular embolisation has been introduced as a safe alternative, but does not eliminate the possibility of symptom recurrence. We report a case of a 61-year old woman with intralobar pulmonary sequestration treated with a combination of endovascular coil embolisation and surgical resection.
Topics: Bronchopulmonary Sequestration; Combined Modality Therapy; Embolization, Therapeutic; Endovascular Procedures; Female; Humans; Lung; Middle Aged; Pneumonectomy; Tomography, X-Ray Computed; Treatment Outcome; Vascular Access Devices
PubMed: 32969253
DOI: 10.1308/rcsann.2020.0201 -
Clinical and Applied... Sep 2017Heparin and its low-molecular-weight heparin (LMWH) derivatives are widely used clinical anticoagulants. These drugs are critical for the practice of medicine in... (Comparative Study)
Comparative Study
Heparin and its low-molecular-weight heparin (LMWH) derivatives are widely used clinical anticoagulants. These drugs are critical for the practice of medicine in applications including kidney dialysis, cardiopulmonary bypass, and in the management of venous thromboembolism. Currently, these drugs are derived from livestock, primarily porcine intestine. The worldwide dependence on a single animal species has made the supply chain for this critical drug quite fragile, leading to the search for other sources of these drugs, including bovine tissues such as bovine intestine or lung. A number of laboratories are currently examining the similarities and differences between heparins prepared from porcine and bovine tissues. The current study is designed to compare LMWH prepared from bovine heparins through chemical β-elimination, a process currently used to prepare the LMWH, enoxaparin, from porcine heparin. Using top-down, bottom-up, compositional analysis and bioassays, LMWHs, derived from bovine lung and intestine, are shown to closely resemble enoxaparin.
Topics: Animals; Anticoagulants; Cattle; Clinical Laboratory Techniques; Enoxaparin; Heparin, Low-Molecular-Weight; Intestines; Lung; Swine
PubMed: 28056526
DOI: 10.1177/1076029616686422 -
Cancer Reports (Hoboken, N.J.) Nov 2022Low-grade fibromyxoid sarcoma is a rare painless neoplasm that primarily grows in young adults' proximal extremities and trunks. The lungs are infrequent sites for this... (Review)
Review
BACKGROUND
Low-grade fibromyxoid sarcoma is a rare painless neoplasm that primarily grows in young adults' proximal extremities and trunks. The lungs are infrequent sites for this type of sarcoma.
CASE PRESENTATION
We reported a 26-year-old female that presented with a chief complaint of chest pain from a few months ago to Kasra hospital, Tehran, Iran, in August 2021. Chest computed tomography (CT) showed a hypodense mass with a well-defined margin measuring 9.3 cm in the left upper lobe and multiple hypodense lesions with a lobulated appearance with a total diameter of 15.5 × 13.5 cm in the left lower lobe of the lung.
CONCLUSION
This is the largest case of primary pulmonary low-grade fibromyxoid sarcoma (30 × 28 × 7 cm), which seemed unresectable at first evaluation. Due to the extent of the tumor, left pneumonectomy was performed, leading to attenuation of symptoms and no recurrence at a six-month follow-up.
Topics: Young Adult; Female; Humans; Adult; Iran; Fibrosarcoma; Sarcoma; Soft Tissue Neoplasms; Lung
PubMed: 36148539
DOI: 10.1002/cnr2.1718