sodium-dodecyl-sulfate and Lung-Neoplasms

It is important to detect mediastinal lymph node metastases in patients with lung cancer to improve outcomes, and it is possible that activatable fluorescence imaging with indocyanine green (ICG) can help visualize metastatic lymph nodes. Therefore, we investigated the feasibility of applying this method to mediastinal lymph node metastases in an epidermal growth factor receptor (EGFR)-positive squamous cell carcinoma of the lung. Tumors were formed by injecting H226 (EGFR-positive) and H520 (EGFR-negative) cell lines directly in the lung parenchyma of five mice each. When computed tomography revealed tumors exceeding 8 mm at their longest or atelectasis that occupied more than half of lateral lung fields, a panitumumab (Pan)-ICG conjugate was injected in the tail vein (50 μg/100 μL). The mice were then sacrificed 48 hours after injection and their chests were opened for fluorescent imaging acquisition. Lymph node metastases with the five highest fluorescent signal intensities per mouse were chosen for statistical analysis of the average signal ratios against the liver. Regarding the quenching capacity, the Pan-ICG conjugate had almost no fluorescence in phosphate-buffered saline, but there was an approximate 61.8-fold increase in vitro after treatment with 1% sodium dodecyl sulfate. Both the fluorescent microscopy and the flow cytometry showed specific binding between the conjugate and H226, but almost no specific binding with H520. The EGFR-positive mediastinal lymph node metastases showed significantly higher average fluorescence signal ratios than the EGFR-negative ones (n = 25 per group) 48 hours after conjugate administration (70.1% ± 4.5% vs. 13.3% ± 1.8%; p < 0.05). Thus, activatable fluorescence imaging using the Pan-ICG conjugate detected EGFR-positive mediastinal lymph node metastases with high specificity.

Topics: Animals; Biomarkers, Tumor; Carcinoma, Squamous Cell; Cell Line, Tumor; ErbB Receptors; Female; Fluorescent Dyes; Heterografts; Humans; Indocyanine Green; Lung Neoplasms; Lymph Nodes; Lymphatic Metastasis; Mediastinum; Mice; Mice, Nude; Microscopy, Fluorescence; Optical Imaging; Panitumumab; Photochemistry; Sodium Dodecyl Sulfate; Tomography, X-Ray Computed

Natural rubber Latex (Lax) is a colloidal dispersion of polymer particles in liquid and shows good biodegradable, biocompatibility, and non-toxicity. Natural polymers are the most important materials used in food packaging, micro/nano-drug delivery, tissue engineering, agriculture, and coating. In the present study, natural compounds extracted from plant Lax were designed to function as drug carriers using various surfactants via emulation and solvent evaporation method. Calotropis gigantea belongs to the family Apocynaceae and has received considerable attention in modern medicine, ayurvedeic, siddha, and traditional medicine. Since, we were isolated biodegradable, non-toxic, and biocompatible materials as latex from Calotropis gigantea plant. The Lax was separated as per their solubility nature and it was designed as a carrier using surfactant namely; Sorbitanmonolaurate (Span-20), sodium lauryl sulfate (SLS), and cetyltrimethylammonium bromide (CTAB). The isolated compounds from Lax of Calotropis gigantea were analyzed using high-performance liquid chromatography. To confirm the encapsulation efficiency and in vitro drug release of the carriers, doxorubicin (DOX) was used as a model natural drug. The hybrid nanocarriers were successfully synthesized through simple solvent evaporation using three surfactants, and the morphology was characterized by SEM and TEM technique. The functionality and crystalline nature of the nanocarriers were confirmed using FTIR and XRD, respectively. Within 90min, the maximum amount of DOX was encapsulated in the carriers, and prolonged cumulative drug release by the nanocarriers was observed. The formulated natural carriers were found to have potentially effective cytotoxic effects on lung cancer cells.

Topics: A549 Cells; Calotropis; Cell Line, Tumor; Cetrimonium; Cetrimonium Compounds; Chemistry, Pharmaceutical; Delayed-Action Preparations; Doxorubicin; Drug Carriers; Hexoses; Humans; Latex; Lung Neoplasms; Nanoparticles; Plant Extracts; Sodium Dodecyl Sulfate; Solubility

Dansyl fluoride (Dan-F), an active site directed fluorescent inhibitor of guanidinobenzoatase (GB), has been used for the location of tumour cells in frozen sections of human squamous cell carcinoma and colonic carcinoma tissues. The tumour cell surfaces having active GB bind Dan-F and fluoresce blue. The surrounding normal epithelial lung cell surfaces fail to bind Dan-F and hence lack fluorescence, whilst the normal colon cell surfaces have another isoenzymic form of GB, bind Dan-F and fluoresce blue. Kinetic studies have shown that Dan-F is an irreversible inhibitor of GB, and Dan-GB complexes are not dissociated with SDS and high salt concentration. However hydroxylamine (1 M) can dissociate Dan-GB complexes in the presence of 0.1% SDS, both on membrane-bound and in free solution. These studies suggest that Dan-F is a potent inhibitor of GB, and in very low concentration (3 x 10(-8) M) can be used as a novel fluorescent probe for the location of tumour cells in histological sections of human tissues.

Topics: Aminacrine; Binding Sites; Carboxylic Ester Hydrolases; Colonic Neoplasms; Coloring Agents; Dansyl Compounds; Endopeptidases; Enzyme Activation; Histocytochemistry; Humans; Hydroxylamine; Hydroxylamines; Hymecromone; Isoenzymes; Lung Neoplasms; Microscopy, Fluorescence; Serine Proteinase Inhibitors; Sodium Dodecyl Sulfate; Spectrometry, Fluorescence

Guanidinobenzoatase (GB) is a cell surface proteolytic enzyme capable of degrading fibronectin, and is associated with tumour cells and cells capable of migration. The location of active GB in sections has been demonstrated with 9-aminoacridine (9-AA), a competitive inhibitor of GB. 3,4-Dichloroisocoumarin (3,4-DCI) and pentamidine isethionate (PI) are inhibitors of trypsin-like enzymes. It has now been demonstrated that 3,4-DCI, PI, and guanidino derivative compounds are significant inhibitors of GB, on the surfaces of lung squamous cell carcinoma cells in frozen sections and free GB in solution. Dexamethasone acetate (DMA) and medroxy-progesterone (MP) did not show any significant inhibition of GB activity. These molecules lack a reactive chloride or guanidino groups and are thought to react at the nuclear level, rather than directly on this cell surface protease. Kinetic studies have shown that 3,4-DCI, PI and guanidino derivatives are reversible competitive inhibitors of GB, as determined in vitro on the purified enzyme. The inhibition resulting with 3,4-DCI was a time-dependent process. It is suggested that these inhibitors interact with GB by binding to its active site, resulting in the formation of enzyme-inhibiter complexes (GB-I). The GB-I complexes can be dissociated with SDS treatment, resulting in the regain of GB activity.

Topics: Benzoates; Binding, Competitive; Carboxylic Ester Hydrolases; Carcinoma, Squamous Cell; Cell Membrane; Coumarins; Dexamethasone; Endopeptidases; Guanidines; Humans; Isocoumarins; Kinetics; Lung Neoplasms; Medroxyprogesterone; Microscopy, Fluorescence; Pentamidine; Protease Inhibitors; Sodium Dodecyl Sulfate; Sulfaguanidine

A drug-resistant human small cell lung cancer cell line, H209/V6, selected in the presence of increasing concentrations of 9-(4,6-O-ethylidene-beta-D-glucopyranosyl)-4'-demethylepipodophylloto xin (VP-16) from parental H209 cells, is 22-, 9-, and 4-fold resistant to VP-16, 4'-(9-acridinyl-amino)methanesulfon-m-anisidide, and doxorubicin, respectively, but not cross-resistant to 1,4-dihydroxy-5,8-bis((2-[(2-hydroxyethyl)amino] ethyl]-amino)-9,10-anthracenedione. These cells do not overexpress P-glycoprotein or the multidrug resistance-associated protein. Immunoblotting demonstrates that H209 cells contain the M(r) 170,000 isoform of topoisomerase II (topo II), while H209/V6 cells have a M(r) 160,000 enzyme but none of the M(r) 170,000 isoform. The cell lines have equal amounts of topo II beta. The H209/V6 cells have a 5-fold decrease in total immunoreactive topo II alpha. The catalytic and VP-16-induced DNA cleavage activities of the topo II present in 0.35 M NaCl nuclear extracts are decreased 2- to 3-fold in the drug-resistant cell line. This decrease in enzymatic activity is not consistent with either the 22-fold VP-16 resistance of the H209/V6 cell line or the approximately 5-fold decrease in immunoreactive topo II alpha in the cells. The M(r) 160,000 isoform from the H209/V6 cell line and the M(r) 170,000 enzyme from the parental cell line were purified so that the enzymatic activity of the 2 isoforms could be evaluated. The catalytic activities of the purified isoforms were found to be very similar. The drug-induced DNA cleavage activity of the M(r) 160,000 enzyme was reduced compared to the M(r) 170,000 enzyme. However, as with the nuclear extracts, the differences in enzymatic activity of the purified enzymes are considerably less than the level of drug resistance. Investigations of the subcellular localization of topo II by immunocytochemical techniques and cytoplasm/nuclear fractionation studies demonstrated that the M(r) 160,000 topo II alpha-related enzyme is primarily localized in the cytoplasm, while the M(r) 170,000 topo II alpha enzyme and topo II beta are located in the nucleus. These data imply that the deleted sequence in the M(r) 160,000 enzyme is not necessary for catalytic activity but is required to facilitate nuclear localization.

Topics: Antineoplastic Agents; Blotting, Western; Carcinoma, Small Cell; Cell Nucleus; DNA Topoisomerases, Type II; Drug Resistance; Electrophoresis, Polyacrylamide Gel; Flow Cytometry; Humans; Immunohistochemistry; Isoenzymes; Lung Neoplasms; Molecular Weight; Sodium Dodecyl Sulfate; Subcellular Fractions; Tumor Cells, Cultured

Using ELISA, we studied anti-PPD antibody values in 31 tuberculous and 39 carcinomatous pleural effusions. Mean ODI values of anti-PPD IgG, IgA and IgM antibodies in tuberculous pleural effusions were higher than those in carcinomatous pleural effusions (p less than 0.01 in IgG and IgA, p less than 0.05 in IgM antibodies). We also analyzed the detected antigens in PPD and BCG whole cell fraction recognized by IgG antibody in tuberculous pleural effusions. Among heteromolecular components, the antigen band from 25 to 40 Kd region was most frequently stained. This antigen was heat-stable, contained sugar components, and included no disulfide bonds.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Antibodies, Bacterial; Antibodies, Neoplasm; Antigens, Bacterial; Antigens, Neoplasm; Electrophoresis, Polyacrylamide Gel; Enzyme-Linked Immunosorbent Assay; Humans; Immunoblotting; Immunoglobulin G; Lung Neoplasms; Middle Aged; Mycobacterium tuberculosis; Pleural Effusion; Sodium Dodecyl Sulfate; Tuberculin; Tuberculosis, Pleural

Topics: Adenocarcinoma; Antigen-Antibody Complex; Carcinoembryonic Antigen; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Cell Membrane; Cross Reactions; Electrophoresis, Starch Gel; Epitopes; Fluorescent Antibody Technique; Glycoproteins; Humans; Immunodiffusion; Immunoelectrophoresis; Lung Neoplasms; Microsomes; Salicylates; Sodium Dodecyl Sulfate

Topics: Adolescent; Adult; Aged; Benzalkonium Compounds; Breast Neoplasms; Croton Oil; Dermatitis, Contact; Female; Humans; Lung Neoplasms; Male; Middle Aged; Neoplasm Metastasis; Neoplasms; Skin Tests; Sodium Dodecyl Sulfate

Topics: Adenosine; Aminohydrolases; Animals; Autopsy; Chromatography, DEAE-Cellulose; Chromatography, Gel; Electrophoresis, Disc; Electrophoresis, Polyacrylamide Gel; gamma-Globulins; Humans; Immune Sera; Immunodiffusion; Lung; Lung Neoplasms; Macromolecular Substances; Male; Molecular Weight; Precipitin Tests; Rabbits; Sodium Dodecyl Sulfate; Spectrophotometry, Ultraviolet; Tetrazolium Salts; Time Factors