Elevated sputum BPIFB1 levels in smokers with chronic obstructive pulmonary disease: a longitudinal study.

A previous study involving a proteomic screen of induced sputum from smokers and patients with chronic obstructive pulmonary disease (COPD) demonstrated elevated levels of bactericidal/permeability-increasing fold-containing protein B1 (BPIFB1). The aim of the present study was to further evaluate the association of sputum BPIFB1 levels with smoking and longitudinal changes in lung function in smokers with COPD. Sputum BPIFB1 was characterized by two-dimensional gel electrophoresis and mass spectrometry. The expression of BPIFB1 in COPD was investigated by immunoblotting and immunohistochemistry using sputum and lung tissue samples. BPIFB1 levels were also assessed in induced sputum from nonsmokers (n = 31), smokers (n = 169), and patients with COPD (n = 52) via an ELISA-based method. The longitudinal changes in lung function during the 4-year follow-up period were compared with the baseline sputum BPIFB1 levels. In lung tissue samples, BPIFB1 was localized to regions of goblet cell metaplasia. Secreted and glycosylated BPIFB1 was significantly elevated in the sputum of patients with COPD compared with that of smokers and nonsmokers. Sputum BPIFB1 levels correlated with pack-years and lung function as measured by forced expiratory volume in 1 s (FEV1) % predicted and FEV1/FVC (forced vital capacity) at baseline and after the 4-year follow-up in all participants. The changes in lung function over 4 years were significantly associated with BPIFB1 levels in current smokers with COPD. In conclusion, higher sputum concentrations of BPIFB1 were associated with changes of lung function over time, especially in current smokers with COPD. BPIFB1 may be involved in the pathogenesis of smoking-related lung diseases.

Development of in vitro procedures for regeneration of petiole and leaf expiants and production of protoplast-derived callus in Tanacetum vulgare L. (Tansy).

Tanacetum vulgare (Tansy) was established in vitro on Murashige and Skoog (MS) medium supplemented with αnaphthaleneacetic acid (NAA) and 6-benzylaminopurine (BAP) using shoot tips and embryos. From petiole expiants 93% formed callus, and 27% produced shoots on MS medium containing 4.5 mg l(-1) NAA and BAP. NAA alone induced root formation from leaf expiants. Up to 7 ×10(6) viable protoplasts were obtained by macerating 1 g of leaves in 0.5 % Macerozyme R-10, 1.0% Cellulase R10, and 1.0% Cellulysin. Cell division was observed 3-4 days after protoplast isolation at the optimum plating density of 0.2-0.4×10(6) cells ml(-1). A total of 350 protoplast-derived calluses were produced on which nodules with meristematic zones developed. Roots regenerated on MS medium supplemented with BAP 3.0 mg 1(-1), NAA 2.0 mg l(-1), and 250 mg l(-1) casein hydrolysate, however no shoots have been obtained yet.