Isolating bronchial epithelial cell preparations from gross lung specimens

Tissue microdissection is appropriate for separating pure cells from heterogeneous tissues. Recently, we have focused on whole genome DNA methylation patterns of lung squamous cell carcinoma (SCC) and needed to obtain the appropriate counterpart cells of lung SCC. However, in some regions of human tissues, such as in bronchial epithelium, it is difficult to apply tissue microdissection as a means to isolate pure cells from a heterogenous mixture of cells. Accordingly, we developed the pop brush method to retrieve sufficient amounts of pure bronchial epithelium from gross lung specimens, and this method enables us to study epigenetic variations of lung SCC.     

Cytology of plasma cell myeloma in bronchial washing

A 60-year-old man presenting with cough, dyspnea and chest pain was found to have plasma cell myeloma with pulmonary involvement. Cytologic smears of a bronchial washing showed clusters and sheets of cells with vague plasmacytoid features. Prominent nucleoli, which were present in most of the cells, and occasional glandlike patterns suggested an adenocarcinoma, thus causing a differential diagnostic problem.     

Role of special stains in the diagnosis of Pneumocystis carinii infection from bronchial washing specimens in patients with the acquired immune deficiency syndrome

Fifty bronchial washing specimens from 36 patients with acquired immune deficiency syndrome (AIDS) were retrospectively reviewed to assess the sensitivity of the various special stains used to diagnose Pneumocystis carinii. In 76% of the cases, the Diff-Quik stain was positive; it was the easiest and most rapid of the special stains used. The sensitivity was increased to 92%, 96% and 100%, respectively, by also doing cresyl echt violet, Grocott's Gomori methenamine silver and both the cresyl violet and Grocott stains in addition to the Diff-Quik stain. We conclude that the Diff-Quik stain is a fairly reliable and rapid screening procedure for making the diagnosis of Pneumocystis infection in bronchial washings from AIDS patients. The routine Papanicolaou stain gave less sensitive results in the smears of the washing specimens, but does give a markedly improved yield in bronchoalveolar lavage specimens.     

Thin-layer chromatography of ceramides

Ceramides with mono-, di-, and trihydroxy long-chain bases, and normal (saturated and unsaturated), branched-chain, and 2-hydroxy fatty acids have been analyzed by thin-layer chromatography. In most cases the compounds were also run as acetates. Borate, arsenite, and silver ions were used as complexing agents, and the effects of number, position, and stereochemistry of hydroxy groups, and of unsaturation, were studied. The results are discussed in view of analysis of natural ceramide species.     

Thin-layer Chromatography of Cyclodextrins

N-methyl-2-pyrrolidone (NMP) is known for its multi-solvent properties. However, its biological, especially immunological significance still remains to be elucidated. In this study, we show for the first time that NMP stimulates the skin immune system by activating epidermal Langerhans cells (LCs). In contrast with the placebo tape, when the NMP-containing adhesive tape was applied on murine skin, LCs were stimulated immediately. Activated LCs not only exhibited enhanced expression of major histocompatibility complex class II and morphological changes, including the loss of dendrites, but also migrated effectively to draining lymph nodes. In addition, application of the tyrosine-related protein-2 peptide, which is the cytotoxic T lymphocyte (CTL) epitope against B16 melanoma, in combination with the NMP tape, resulted in explosive expansion of specific CTLs in mouse spleens. Taken together, these results demonstrate a novel role of NMP as an adjuvant in percutaneous peptide immunization.     

Metabolic activation and deactivation of mutagens by preparations of human lung parenchyma and bronchial tree

69 S12 preparations of human lung (43 of peripheral lung parenchyma and 26 bronchial preparations) were assayed for their ability to activate procarcinogens to mutagenic metabolites in the Ames test or to lower the mutagenic response of direct-acting compounds. No sample activated polycyclic aromatic hydrocarbons, such as 3-methylcholanthrene, benzo[a]pyrene or its metabolites 3-hydroxy-B(a)P and B(a)P-trans-7,8-diol, and only borderline effects were observedwith cigarette-smoke condensates. Conversely, some activating ability was detected in the case of 2-aminofluorene and of cyclophosphamide. The same samples produced a slight decrease of mutagenicity of epichlorohydrin and ICR-191 and a more pronounced loss of activity of sodium dichromate and 4-nitroquinoline N-oxide.     

Diagnostic significance of flow cytometric DNA analysis applied for the detection of cancer cells in bronchial washing fluid

Since both DNA aneuploidy and increased proliferative activity are important characteristics of malignant neoplasms, flow cytometric (FCM) analysis was used to examine the cell content in bronchial washing samples obtained via fiberoptic bronchoscopy from 73 patients. The results were compared with the results of histology and conventional cytology. The patients included 30 with bronchial carcinomas, 12 with bronchopneumonia and 31 with no evidence of lung disease. Of the 30 patients with histologically confirmed lung cancers, 25 showed either aneuploid stem lines (19 cases) or high levels of proliferation (6 cases) as determined by analysis of cell-cycle stages. The same rate of cancer cell detection was obtained by cytology. In the 43 cases with neither histologic nor clinical evidence of malignancy, FCM data yielded 5 false-positive results, as compared to 4 erroneous suspicions of cancer by cytology. From these data, it is concluded that FCM measurements of both DNA ploidy and proliferative activity may complement conventional cytology in the recognition of bronchial carcinomas.     

Human isolated bronchial muscle preparations from asthmatic patients: effects of indomethacin and contractile agonists

Airway reactivity to histamine was determined in a group of non-asthmatic and asthmatic patients prior to thoracotomy. The latter group was more reactive to histamine provocation than the former (PC40: 28.40 +/- 6.27 mg/ml and 1.15 +/- 0.19 mg/ml, respectively). Subsequent to the surgical intervention, isolated human bronchial muscle preparations were obtained from both groups (15 non-asthmatic and 5 asthmatic subjects). Histamine concentration-effect curves were generated both in the absence and in the presence of indomethacin (1.7 microM; 30 min). Neither the basal tone nor the histamine response and sensitivity of the preparations were altered by the antiinflammatory drug. In bronchial preparations from one asthmatic subject, indomethacin significantly reduced the prostaglandin production during histamine contraction. Prostaglandin E2 and F2 alpha contracted isolated human bronchial muscle preparations from these asthmatic individuals. These data suggest that endogenous prostaglandins may not regulate the contractile response to histamine in vitro.     

Thin-layer chromatography of the phosphoinositides

Thin-layer chromatography for the separation of mono-, di-, and triphosphoinositides (0.3-3 micro g total phosphorus) is described.     

Thin-layer chromatographic collector-eluter.

A combination thin-layer chromatographic collector and elution apparatus is described. The device is designed to minimize manipulations and maximize accuracy and can be used for both analytical and small-scale preparative separations.     

Circular Thin-layer Chromatography of Oligosaccharides

The physicochemical properties and chemical constituents of the blue protein from rice bran were investigated. The blue protein was a copper-containing glycoprotein, the molecular weight of which was found to be 18,300 Daltons by the sedimentation equilibrium method assuming the partial specific volume 0.72 cm³ g^?1. The hexose and pentose contents were 5.49 and 4.01 g per 100 g protein respectively. The copper content was 0.38% which corresponded to 1.09 atoms per one molecule of the protein. The electron spin resonance spectrum showed that the copper was in a cupric state. The standard oxidation-reduction potential of the copper was found to be +275 mV at 20°C and at pH 7.39. The visible and near infrared absorption maxima were found at 450, 600 and 890 mμ, and the 450 mμ band was optically active in the optical rotatory dispersion exhibiting a large Cotton effect.     

Thin-layer chromatography of hydroxamic acids

The separation of very short-chain from long-chain fatty acyl hydroxamates by thin-layer chromatography is described. The detection limit was 2 micrograms.