Characterization of rat alveolar type II cells in vitro by immunological, biochemical, and morphological criteria

Using an anti-rat surfactant apoprotein antiserum which specifically reacts with cytoplasmic structures in alveolar type II cells on histopathology sections of rat lung, we have examined the immunoreactivity of pulmonary type II cells in vitro. Single cell suspensions of lung tissue were prepared from male Fischer 344 rats by intratracheal elastase digestion according to standard published methods. Cytocentrifuged preparations of the resulting cell suspensions revealed that approximately 40% of the cells stained positive for surfactant apoprotein using an immunoperoxidase staining technique. Without further cell fractionation steps, the cell suspensions were plated at colonial densities in growth medium. The cells that attached after 24 hours of incubation and at daily intervals were analyzed for surfactant apoprotein immunoreactivity as well as for proliferation, morphology, and phospholipid biosynthesis. The percentage of immunopositive cells increased with time from 75% at day 1 to 94% at 4 days after plating. This increase was paralleled by a linear increase in the number of immunopositive cells, which expanded into cell colonies. During the initial 5 days in vitro, the immunopositive cells retained their epithelial morphology and contained cytoplasmic osmiophilic bodies. Phospholipid biosynthesis by the isolated lung cells was analyzed and the data revealed that the rate of incorporation of 14C-choline into phosphatidylcholine increased with time in culture. These studies indicated that the anti-rat surfactant apoprotein antisera can be used to identify and quantitate functional alveolar type II cells in vitro. Thus the specific antisera may facilitate studies of type II cells undergoing various environmental alterations both in vivo and in vitro.     

Identification, isolation, and partial characterization of a 7.5-kDa surfactant-associated protein

Human surfactant was analyzed for proteins associated with the lipids. Surfactant was isolated from lung lavage by the salt gradient centrifugation method, and the soluble proteins binding to the lipids were recovered by extraction with a low pH buffer. Antiserum to this preparation reacted with surfactant apoprotein A and a 7.5-kDa protein. The 7.5-kDa protein was isolated from reduced and alkylated lung lavage pellet by chromatography. The N-terminal amino acid sequence of the protein indicates that it is a novel protein.     

The pathobiologic features of carcinomas of type II pneumocytes. An immunocytologic study

Adenocarcinomas of the lung without squamous differentiation or obvious mucus production from 51 patients were studied. Twenty-eight tumors (55%), when stained with rabbit anti-human surfactant apoprotein antiserum by the peroxidase-antiperoxidase method, demonstrated characteristic nuclear inclusions. Most of these tumors could be identified histochemically by the presence of eosinophilic, periodic acid-Schiff (PAS)-positive nuclear inclusions. In patients with apoprotein-immunoreactive tumors there were nine deaths due to tumor (32%) within 5 years of diagnosis. Eight of the nine deaths occurred in patients whose tumors exceeded 3 cm in diameter; an equal number of patients were smokers. The average age of patients with apoprotein-positive tumors was 67.3 years, a figure greater than that for patients with apoprotein-negative tumors (61.4 years). Five of the 23 patients with apoprotein-negative tumors died of their neoplasms; in two of the subjects the tumors exceeded 3 cm in diameter. In that no significant difference in survival was observed in patients with apoprotein-positive and apoprotein-negative tumors, it was concluded that subclassification of adenocarcinomas of the lung as apoprotein-positive, i.e., type II pneumocytic, or as apoprotein-negative, is of no clinical or prognostic significance. Nonetheless, peripheral tumors measuring less than 3 cm in size and showing neither squamous differentiation nor obvious mucus production should be recognized as a prognostically favorable group in comparison with other types of lung carcinomas.     

Cyclosporine stimulates hepatocyte proliferation and accelerates development of hepatocellular carcinomas in rats

A recent study from our laboratory demonstrated that cyclo-sporine(CsA), a prototype immunosuppressant, enhanced the growth ofcarcinogen-induced enzyme altered foci in rat liver, suggestingthat CsA may stimulate development of hepato-cellular carcinomas.In the present study, we examined (i) whether CsA acceleratesdevelopment of hepatocellular carcinomas in experimental animals,(ii) whether CsA stimulates the proliferation of resting hepatocytein vivo and (iii) whether CsA modulates the production of growthfactors implicated in liver cell growth, hepatocyte growth factor(HGF), transforming growth factor a (TGF     

Long-circulating poly(ethylene glycol)-grafted gelatin nanoparticles customized for intracellular delivery of noscapine: preparation, in-vitro characterization, structure elucidation, pharmacokinetics, and cytotoxicity analyses

Noscapine, the tubulin-binding anticancer agent, when administered orally, requires high ED(50) (300-600 mg/kg), whereas intravenous administration (10 mg/kg) results in rapid elimination of the drug with a half-life of 0.39 h. Hence, the development of long-circulating injectable nanoparticles can be an interesting option for designing a viable formulation of noscapine for anticancer activity. Noscapine-enveloped gelatin nanoparticles and poly(ethylene glycol)-grafted gelatin nanoparticles were constructed and characterized. Data indicate that smooth and spherical shaped nanoparticles of 127 ± 15 nm were engineered with maximum entrapment efficiency of 65.32 ± 3.81%. Circular dichroism confirms that nanocoacervates retained the α-helical content of gelatin in ethanol whereas acetone favored the formation of a random coil. Moreover, the Fourier transform infrared and powder X-ray diffraction pattern prevents any significant change in the noscapine-loaded gelatin nanoparticles in comparison with individual components. In-vitro release kinetic data suggest a first-order release of noscapine (85.1%) from gelatin nanoparticles with a release rate constant of 7.611×10(-3). It is to be noted that there is a 1.43-fold increase in the area under the curve up to the last sampling point for the noscapine-loaded poly(ethylene glycol)-grafted gelatin nanoparticles over the noscapine-loaded gelatin nanoparticles and a 13.09-fold increase over noscapine. Cytotoxicity analysis of the MCF-7 cell line indicated that the IC(50) value of the noscapine-loaded poly(ethylene glycol)-grafted gelatin nanoparticles was equivalent to 20.8 μmol/l, which was significantly (P<0.05) lower than the IC(50) value of the noscapine-loaded gelatin nanoparticles (26.3 μmol/l) and noscapine (40.5 μmol/l).Noscapine-loaded poly(ethylene glycol)-grafted gelatin nanoparticles can be developed as a promising therapeutic agent for the management of breast cancer.     

Antidepressant and anxiolytic like effects of Urtica dioica leaves in streptozotocin induced diabetic mice

Metabolic Brain Disease - The present study was&nbsp;aimed to investigate the effect of Urtica dioica Linn. (UD) extract against chronic diabetes mediated anxiogenic and depressive like...     

A newborn with transposition of the great vessels and long-segment tracheal stenosis

Congenital tracheal stenosis is an uncommonly associated malformation in children with congenital heart defects. Airway symptomatology should prompt immediate work-up to prevent a delay in diagnosis. Although results to date show acceptable mortality and morbidity for simultaneous repair of tracheal stenosis and cardiac anomalies there are cases where the severity of the lesion in association with other anomalies increases the mortality significantly.We report about a newborn diagnosed with transposition of the great vessels and several other anomalies who developed progressively worsening respiratory distress. Imaging revealed long-segment tracheal stenosis. The family was offered the option of slide tracheoplasty but decided not to pursue any further surgical intervention. The patient soon thereafter died.     

Modulatory influence of noscapine on the ethanol-altered hepatic biotransformation system enzymes, glutathione content and lipid peroxidation in vivo in rats.

The modulatory potential of noscapine, an opium alkaloid was assessed on the ethanol-induced changes in hepatic drug metabolizing enzyme systems, glutathione content and microsomal lipid peroxidation. Noscapine was administered orally to male Wistar rats at a dose level of 200 mg/kg bw alone as well as in combination with 50% ethanol (v/v) for 5 days. Noscapine administration was associated with a approximately 91% decrease in hepatic microsomal cytochrome P-450 content. A decline of approximately 36% was observed in the NADPH-cytochrome c reductase activity on noscapine administration. The lowering of cytochrome P-450 levels on noscapine administration was accompanied by a concomitant increase in heme oxygenase activity as well as serum bilirubin levels. Our results indicate that the combination dosage of noscapine and ethanol antagonised the ethanol-induced elevation of cytochrome P-450 levels. Noscapine fed rats had decreased glutathione (GSH) content and enhanced lipid peroxidation compared to control rats as indexed by MDA method. Further, noscapine and ethanol coexposure produced a more pronounced elevation in lipid peroxidation and the glutathione levels also decreased significantly. We speculate on the basis of our results that the significant enhancement of lipid peroxidation on combination dosage of noscapine and ethanol is a consequence of depletion of glutathione to certain critical levels. The inhibition of glutathione-S-transferase (GST) as well as lowering of cytochrome P-450 suggests that the biotransformation of noscapine and ethanol is significantly altered following acute coexposures.