Expression of renal cell protein markers is dependent on initial mechanical culture conditions.

The rotating wall vessel is optimized for suspension culture, with laminar flow and adequate nutrient delivery, but minimal shear. However, higher shears may occur in vivo. During rotating wall vessel cultivation of human renal cells, size and density of glass-coated microcarrier beads were changed to modulate initial shear. Renal-specific proteins were assayed after 2 days. Flow cytometry antibody binding analysis of vitamin D receptor demonstrated peak expression at intermediate shears, with 30% reduction outside this range. Activity of cathepsin C showed the inverse pattern, lowest at midshear, with twofold increases at either extreme. Dipeptidyl-peptidase IV had no shear dependence, suggesting that the other results are specific, not universal, changes in membrane trafficking or protein synthesis. On addition of dextran, which changes medium density and viscosity but not shear, vitamin D receptor assay showed no differences from controls. Neither cell cycle, apoptosis/necrosis indexes, nor lactate dehydrogenase release varied between experiments, confirming that the changes are primary, not secondary to cell cycling or membrane damage. This study provides direct evidence that mechanical culture conditions modulate protein expression in suspension culture.     

The carbon-13 nuclear magnetic resonance spectra of four eudesmane sesquiterpenols

The ¹³C NMR spectra of geosmin, selina-4(14),7(11)-diene-99-ol and two dihydroeudesmol isomers have been obtained and the individual resonances assigned. Several different empirical correlations developed by others have been combined in simple calculations to predict chemical shift values for sesquiterpenols of the eudesmane group.     

CHROMOSOME NUMBERS OF CAMPANULACEAE. II. THE LOBELIA TUPA COMPLEX OF CHILE

Gametic chromosome numbers are reported for 27 collections representing the four species of the Lobelia tupa complex (Campanulaceae, Lobelioideae) in Chile; all are n = 21. This represents the first report of chromosome numbers for L. bridgesii Hook. & Arn., L. excelsa Bonpl., and L. polyphylla Hook. & Arn., and confirms previous reports of this number in L. tupa L. As the basic chromosome number of Lobelioideae is x = 7, these species are interpreted as hexaploids. Higher polyploids are extremely rare among Lobelioideae; most of those previously reported have been either sporadic individuals or populations within an otherwise diploid or tetraploid species, or occasional species within an otherwise diploid and tetraploid lineage. This is the first report of an entire complex of lobelioid species that is uniformly hexaploid. This suggests that the Chilean endemics are relatively derived within Lobelia, and offers some support for the monophyly of the complex.