The Na+/H+ exchanger isoforms NHE1, NHE2, and NHE3 were all found to be expressed in Ehrlich ascites tumor cells, as evaluated by Western blotting and confocal microscopy. Under unstimulated conditions, NHE1 was found predominantly in the plasma membrane, NHE3 intracellularly, and NHE2 in both compartments. Osmotic cell shrinkage elicited a rapid intracellular alkalinization, the sensitivity of which to EIPA (IC50 0.19 microM) and HOE 642 (IC50 0.85 microM) indicated that it predominantly reflected activation of NHE1. NHE activation by osmotic shrinkage was inhibited by the protein kinase C inhibitors chelerythrine (IC50 12.5 microM), Gö 6850 (5 microM), and Gö 6976 (1 microM), and by the p38 MAPK inhibitor SB 203580 (10 microM). Furthermore, hypertonic cell shrinkage elicited a biphasic increase in p38 MAPK phosphorylation, with the first significant increase detectable 2 minutes after the hypertonic challenge. Neither myosin light chain kinase-specific concentrations of ML-7 (IC50 40 microM) nor ERK1/2 inhibition by PD 98059 (50 microM) had any effect on NHE activation. Under isotonic conditions, the serine/threonine protein phosphatase inhibitor calyculin A elicited an EIPA- and HOE 642-inhibitable intracellular alkalinization, indicating NHE1 activation. Similarly, shrinkage-induced NHE activation was potentiated by calyculin A. The calyculin A-induced alkalinization was not associated with an increase in the free, intracellular calcium concentration, but was abolished by chelerythrine. It is concluded that shrinkage-induced NHE activation is dependent on PKC and p38 MAPK, but not on MLCK or ERK1/2. NHE activity under both iso- and hypertonic conditions is increased by inhibition of serine/threonine phosphatases, and this effect appears to be PKC-dependent.

The influence of isolation method on the determination of important aroma compounds in black currant juice was investigated by surface of nasal impact frequency (SNIF) gas chromatography-olfactometry (GC-O). The applied methods were solvent extraction, static headspace, and purge and trap using 15 and 60 min of purge time. By the four methods, a total of 59 odors were observed, and, of these, 44 corresponded to compounds that could be identified. For the headspace methods increasing purge volumes resulted in recoveries of additional, less volatile compounds. The main compound groups recovered by the headspace methods were esters and terpenes, whereas compounds recovered by solvent extraction were not as dominated by fruity odors. For most compounds there was agreement between the size of the SNIF value obtained by GC-O and the amount of the measurable compound found by gas chromatography-mass spectrometry.

The influence of thermal treatment on black currant juice aroma was investigated in temperature and time ranges relevant for black currant juice concentration processes, namely, 45, 60, 75, and 90 degrees C. Forty-nine aroma compounds were quantified, and the thermal treatment resulted in concentration increases of most terpenes, aldehydes, furans, and phenols, whereas the concentration of esters slightly decreased. Higher temperatures and longer exposure times had larger effects on the aroma compounds. Odor triangle tests showed no sensory difference between pasteurized juice and juice heated at 60 degrees C, whereas juice heated at 90 degrees C differed significantly from pasteurized juice. It is concluded that a 90 degrees C thermal treatment of black currant juice, which is in the temperature range used for conventional evaporation of black currant juice, has an effect on the aroma and sensory properties.

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Volatile compounds in skim milk and nonstandardised milk subjected to instant infusion pasteurisation at 80°C, 100°C and 120°C were compared with raw milk, high temperature short time pasteurised milk and milk pasteurised at 85°C ⁄ 30 s. The composition of volatile compounds differed between infusion pasteurisation treated samples and the reference pasteurisations. The sensory properties of skim milk subjected to instant infusion pasteurisation were described by negative attributes, such as cardboard sour and plastic flavours, which are not associated normally with fresh milk. Partial least squares modelling showed good correlation between the volatile compounds and the sensory properties, indicating the predictive and possible causal importance of the volatile compounds for the sensory characteristics.