Biomedical subjects

A Kassner

Publications and source records attributed to A Kassner.

At least 19 recordsLinked to original sources

PubMed2002

Effects of dietary fatty acids on the composition and oxidizability of low-density lipoprotein.

OBJECTIVE: The objective of this study was to compare the effects of dietary monounsaturated fatty acids (MUFA), n-6 and n-3 polyunsaturated fatty acids (PUFA) on LDL composition and oxidizability. DESIGN, SETTING AND SUBJECTS: Sixty-nine healthy young volunteers, students at a nearby college, were included. Six subjects withdrew because of intercurrent illness and five withdrew because they were unable to comply with the dietary regimen. INTERVENTIONS: The participants received a 2-week wash-in diet rich in saturated fatty acids (SFA) followed by diets rich in refined olive oil, rapeseed oil or sunflower oil for 4 weeks. Intakes of vitamin E and other antioxidants did not differ significantly between the diets. RESULTS: At the end of the study, LDL oxidizability was lowest in the olive oil group (lag time: 72.6 min), intermediate in the rapeseed oil group (68.2 min) and highest in the sunflower oil group (60.4 min, P<0.05 for comparison of all three groups). Despite wide variations in SFA intake, the SFA content of LDL was not statistically different between the four diets (25.8-28.5% of LDL fatty acids). By contrast, the PUFA (43.5%-60.5% of LDL fatty acids) and MUFA content of LDL (13.7-29.1% of LDL fatty acids) showed a wider variability dependent on diet. CONCLUSIONS: Enrichment of LDL with MUFA reduces LDL susceptibility to oxidation. As seen on the rapeseed oil diet this effect is independent of a displacement of higher unsaturated fatty acids from LDL. Evidence from this diet also suggests that highly unsaturated n-3 fatty acids in moderate amounts do not increase LDL oxidizability when provided in the context of a diet rich in MUFA.

Adult↗

PubMed2001

Qualitative and quantitative analysis of regional left ventricular wall dynamics using real-time magnetic resonance imaging: comparison with conventional breath-hold gradient echo acquisition in volunteers and patients.

A real-time magnetic resonance imaging (MRI) acquisition sequence was evaluated for the assessment of left ventricular wall motion (WM) and wall thickening (WT). Ten normal volunteers and 21 patients were studied. Short-axis cine images of the left ventricle (LV) were acquired with a fast gradient echo and an ultrafast segmented echo-planar imaging (EPI) sequence. Qualitative and quantitative analysis of WM and WT was performed on a segmental basis. Qualitative scores agreed between the two methods in 691 of 724 segments (95.4%) with good reproducibility. Quantitative measurements of WM and WT were significantly lower (P < 0.001) with the real-time method (WM: mean bias, 0.49 mm; WT: mean bias, 0.61 mm). The largest differences were observed in the anterior and lateral segments and in patients with dilated ventricles. The lower resolution of the real-time sequence and artifacts was probably responsible for these differences. In conclusion, real-time cardiac MRI can be used for qualitative assessment of wall dynamics but is presently insufficient for quantitative analysis.

Adult↗

PubMed2001

Reproducibility of T2* blood volume and vascular tortuosity maps in cerebral gliomas.

The development of anti-angiogenic therapies for tumors has led to a demand for imaging-based surrogate markers of the angiogenic process. The utility of such markers is highly dependent on their test-retest reproducibility. This paper presents a formal assessment of the reproducibility of measurements of relative blood volume (rBV), normalized rBV (rBVnorm), and vascular tortuosity as estimated by measurement of relative recirculation (rR). The study was conducted in 11 patients with glioma who were scanned on two occasions 36-56 hours apart. The observed reliability estimates were used to calculate 95% confidence limits for detection of differences between groups and for changes in individual cases. The results show that measurement of rBV or rBVnorm in consecutive studies is statistically capable of reliably detecting changes in excess of 15% in between group studies and 25% in individual patients. Measurement of vascular tortuosity using is less reproducible but is able to confidently identify changes in excess of 30% in group studies and 35% in individuals.

Blood Volume↗

PubMed2001

Measurements of left ventricular dimensions using real-time acquisition in cardiac magnetic resonance imaging: comparison with conventional gradient echo imaging.

This study investigates the use of real-time acquisition in cardiac magnetic resonance imaging (MRI) for measurements of left ventricular dimensions in comparison with conventional gradient echo acquisition. Thirty-one subjects with a variety of left ventricular morphologies to represent a typical clinical population were studied. Short-axis data sets of the left ventricle (LV) were acquired using a conventional turbo-gradient echo and an ultrafast hybrid gradient echo/echo planar sequence with acquisition in real-time. End-diastolic volume (EDV), end-systolic volume (ESV), ejection fraction (EF) and left ventricular mass (LV mass) were measured. The agreement between the two acquisitions and interobserver, intraobserver and interstudy variabilities were determined. The bias between the two methods was 5.86 ml for EDV, 0.23 ml for ESV and 0.94% for EF. LV mass measurements were significantly lower with the real-time method (mean bias 14.38 g). This is likely to be the result of lower spatial resolution and chemical shift artefacts with the real-time method. Interobserver, intraobserver and interstudy variabilities were low for all parameters. In conclusion, real time acquisition in MRI can provide accurate and reproducible measurements of LV dimensions in subjects with normal as well as abnormal LV morphologies, but LV mass measurements were lower than with conventional gradient echo imaging.

Adult↗

PubMed2001

The time course of natriuretic hormones as plasma markers of myocardial recovery in heart transplant candidates during ventricular assist device support reveals differences among device types.

BACKGROUND: The natriuretic hormones ANP and BNP are expressed differently in the myocardium. Both hormones have compensatory diuretic activity during heart failure. Mechanical stretch of the myocardial walls induces the expression of these hormones. In failing human myocardium, both ANP and BNP are transcribed in the ventricular myocardium in high amounts. We measured the plasma concentrations of ANP and BNP in patients supported by various ventricular assist devices (VADs) at various times. We analyzed the time courses of ANP and BNP to determine (1) the time scale of their down-regulation as a marker of putative myocardial recovery, (2) their steady-state levels under VAD support and (3) differences caused by various VAD devices. METHODS: We analyzed ANP and BNP using commercially available radioimmune assays. We analyzed the time courses of patients supported by Thoratec (THO) LVAD (n = 8), TCI Heartmate (TCI) (n = 6), Novacor (NOV) (n = 7), and Lionheart (LIO) (n = 3). RESULTS: Patients supported with NOV and some patients with TCI showed down-regulation of BNP to a steady-state level at 30 to 50 days, following a single exponential decay. In contrast, patients supported by THO or LIO did not reveal a determined time course of the natriuretic hormones. Only a few patients reached normal plasma values during VAD support. CONCLUSION: The time courses of ANP and BNP differ among VAD types because of construction and/or driving mode, which might be important when considering patients for weaning from VAD without heart transplant.

Adult↗

PubMed2000

Abnormalities of the contrast re-circulation phase in cerebral tumors demonstrated using dynamic susceptibility contrast-enhanced imaging: a possible marker of vascular tortuosity.

Dynamic susceptibility contrast-enhanced magnetic resonance (MR) imaging in tumors is restricted by relaxivity effects, which may obscure any abnormality of first-pass kinetics in the re-circulation phase. The purposes of this study were a) to document the magnitude of relaxivity effects with a variety of commonly used MR susceptibility imaging techniques; and b) to determine whether the re-circulation phase of the first-pass curve in tumors differs from that in normal tissue. We have confirmed that residual relaxivity effects can be eliminated from dynamic susceptibility contrast-enhanced data by several techniques. Application of these methods to enhancing vascular tumors allows detection of abnormalities in the re-circulation phase, which would otherwise be obscured. These abnormalities are independent of relative cerebral blood volume (rCBV) and presumably represent deviations from the predicted gamma variat flow pattern seen in normal tissues. We believe that the parameter rR described here provides an indicator of the chaotic nature of neovascular angiogenesis, which may be of benefit in diagnosis and management.

Brain↗

PubMed2000

Contrast-enhanced 3D MRA using SENSE.

Sensitivity encoding (SENSE) was used to improve the performance of three-dimensional contrast-enhanced magnetic resonance angiography (3D CE-MRA). Utilizing an array of receiver coils for sensitivity encoding, the encoding efficiency of gradient-echo imaging was increased by factors of up to three. The feasibility of the approach was demonstrated for imaging of the abdominal vasculature. On the one hand, using a SENSE reduction factor of two, the spatial resolution of a breath-hold scan of 17 seconds was improved to 1.0 x 2.0 x 2.0 mm(3). On the other hand, using threefold reduction, time-resolved 3D CE-MRA was performed with a true temporal resolution of 4 seconds, at a spatial resolution of 1.6 x 2.1 x 4.0 mm(3). CE-MRA with SENSE was performed in healthy volunteers and patients and compared with a standard protocol. Throughout, diagnostic quality images were obtained, showing the ability of sensitivity encoding to enhance spatial and/or temporal resolution considerably in clinical angiographic examinations.

Aged↗

PubMed2000

The effect of hyperoxia on cerebral blood flow: a study in healthy volunteers using magnetic resonance phase-contrast angiography.

A small decrease in cerebral blood flow (approximately 10%) in response to 100% oxygen (O2) administration is well recognized. This observation was based on human volunteer studies, which employed a nitrous oxide washout method for the measurement of cerebral blood flow. Because this method is now appreciated to be subject to potential errors we have examined the cerebral blood flow response to 100% oxygen using a magnetic resonance imaging technique to quantify changes in carotid and basilar artery flow. The study, was performed in 12 normal male subjects aged 23-42 years. We report decreases in cerebral blood flow ranging from 9 to 31% with a mean value of over 20%. The decrease in cerebral blood flow was greater in seven young subjects (aged 23-26 years) with decreases in cerebral blood flow of 19.3-31.4% (mean 26.8%). In five older subjects (aged 32-42 years), decreases in CBF were smaller (mean 16. 2%). The administration of 100% O2 was accompanied by a small decrease in end-tidal CO2 (3.7-7.1%), insufficient to explain the changes in cerebral blood flow. We conclude that the decrease in cerebral blood flow in response to O2 administration is greater than previously described and appears to be greater in young adults.

Adult↗

PubMed1999

Use of single-slice thick slab phase-contrast angiography for the diagnosis of dural venous sinus thrombosis.

The aim of this study was to examine the reliability of single-slice phase-contrast angiography (SSPCA) as a rapid technique for the investigation of suspected dural venous sinus occlusion. Images were obtained on 25 normal volunteers to document the accuracy of SSPCA in the demonstration of slow flow states. Normal volunteers were imaged using sagittal and coronal SSPCA (slice thickness 13 cm, matrix 256 x 256, TR 14 ms, TE 7 ms, flip angle 20 degrees, peak velocity encoding rate 30 cm/s). Sinus patency and flow rate were confirmed by measurement of flow in the superior sagittal and transverse sinuses using quantified single-slice phase difference images. Imaging was performed in 50 patients undergoing routine brain scans in order to determine the optimal slice orientation for clinical use. Twenty-one patients with suspected dural venous sinus thrombosis were also investigated with SSPCA and the diagnosis confirmed by one or more alternative imaging techniques. Imaging time was 29 s per acquisition and image quality was good in all cases. Variations in dural sinus patency and flow in normal volunteers were accurately predicted by SSPCA (kappa = 0.92). Use of a single angulated slice (130 mm thick, para-sagittal image angled 30 degrees towards coronal and 30 degrees towards transverse) provided sufficient separation of right- and left-sided venous structures to allow use of a single projection. The presence and extent of sinus occlusions in 14 patients and the absence of thrombosis in 7 were accurately identified by SSPCA. Sensitivity and specificity in this limited study were both 100%. The SSPCA technique takes less than 30 s and provides a reliable and rapid technique for the diagnosis of dural venous sinus thrombosis.

Adult↗

PubMed1999

Stepping-table gadolinium-enhanced digital subtraction MR angiography of the aorta and lower extremity arteries: preliminary experience.

PURPOSE: To compare stepping-table digital subtraction gadolinium-enhanced magnetic resonance (MR) angiography of the distal aorta and lower extremity arteries with conventional catheter digital subtraction x-ray angiography in patients with arterio-occlusive disease. MATERIALS AND METHODS: Twenty patients underwent both conventional catheter angiography and fast three-dimensional gadolinium-enhanced MR angiography of the aorta and outflow vessels at 1.5 T; the images were acquired in three consecutive imaging locations during a single infusion of a gadolinium chelate. RESULTS: Compared with catheter angiography, according to the findings of two blinded independent reviewers, MR angiography had sensitivities of 81% and 89% and specificities of 91% and 95%, respectively, for demonstration of insignificant (< or = 50%) stenosis versus significant (51%-100%) stenosis. For demonstration of occlusion, the sensitivity and specificity were 94% and 97%, respectively, by consensus. There was good interobserver correlation between the two readers overall (kappa = 0.65 for reporting the degree of narrowing in all lesions; 0.86, for reporting of insignificant versus significant stenoses; and 0.928, for reporting of occluded versus patient segments). CONCLUSION: Stepping-table digital subtraction contrast material-enhanced MR angiography has high accuracy compared with catheter angiography in patients with arterio-occlusive disease of the aorta and outflow vessels. These preliminary study results suggest that this technique may ultimately provide a safe, noninvasive, and cost-effective alternative to catheter angiography.

Aged↗

PubMed1999

Degradation of the cyclic AMP antagonist prostaglandylinositol cyclic phosphate (cyclic PIP) by dephosphorylation.

The cAMP antagonist, prostaglandylinositol cyclic phosphate (cyclic PIP), is synthesized from prostaglandin E and activated inositol phosphate. From various tissues only that amount of cyclic PIP can be isolated that constitutes the difference between synthesis and degradation. In order to overcome this drawback, the cyclic PIP degrading enzyme or enzymes had to be characterized prior to searching for inhibitors. Cyclic PIP degrading activities have been found in all rat tissues tested, and are lowest in brain (380 pmol x min(-1) x g(-1) wet weight) and highest in liver (1460 pmol x min(-1) x g(-1) wet weight). They are associated primarily with particulate structures of the cells, but not with the plasma membrane. There appear to be at least two different enzymatic activities involved in the degradation of cyclic PIP, because there are two pH-optima, one between pH 7 and 8 and another between pH 4 and 5. It is assumed that these activities are located in microsomes and lysosomes. Because prostaglandylinositol is the final product obtained in the degradation of cyclic PIP, a phosphodiesterase and a phosphatase should be involved, which could not yet be identified individually. Like alkaline phosphatase, cyclic PIP-degrading enzymes require Mg2+ and they are inhibited by heavy metal ions such as mercuric and copper chloride, by sodium fluoride and interestingly, by prostaglandins.

Animals↗

PubMed1999

Comparison of three-dimensional visualization techniques for depicting the scala vestibuli and scala tympani of the cochlea by using high-resolution MR imaging.

BACKGROUND AND PURPOSE: Cochlear implantation requires introduction of a stimulating electrode array into the scala vestibuli or scala tympani. Although these structures can be separately identified on many high-resolution scans, it is often difficult to ascertain whether these channels are patent throughout their length. The aim of this study was to determine whether an optimized combination of an imaging protocol and a visualization technique allows routine 3D rendering of the scala vestibuli and scala tympani. METHODS: A submillimeter T2 fast spin-echo imaging sequence was designed to optimize the performance of 3D visualization methods. The spatial resolution was determined experimentally using primary images and 3D surface and volume renderings from eight healthy subjects. These data were used to develop the imaging sequence and to compare the quality and signal-to-noise dependency of four data visualization algorithms: maximum intensity projection, ray casting with transparent voxels, ray casting with opaque voxels, and isosurface rendering. The ability of these methods to produce 3D renderings of the scala tympani and scala vestibuli was also examined. The imaging technique was used in five patients with sensorineural deafness. RESULTS: Visualization techniques produced optimal results in combination with an isotropic volume imaging sequence. Clinicians preferred the isosurface-rendered images to other 3D visualizations. Both isosurface and ray casting displayed the scala vestibuli and scala tympani throughout their length. Abnormalities were shown in three patients, and in one of these, a focal occlusion of the scala tympani was confirmed at surgery. CONCLUSION: Three-dimensional images of the scala vestibuli and scala tympani can be routinely produced. The combination of an MR sequence optimized for use with isosurface rendering or ray-casting algorithms can produce 3D images with greater spatial resolution and anatomic detail than has been possible previously.

Adult↗

PubMed1998

Electrogenicity of hepatocellular fatty acid uptake.

Sensitivity of cellular fatty acids uptake to the membrane potential difference is still a matter of controversy. For direct evaluation of potential sensitivity the effect of changing membrane potential on uptake of a fluorescent long chain fatty acid derivative, 12-NBD-stearate, in isolated rat hepatocytes, was examined. Changes in membrane potential were achieved by patch clamp procedures. Fatty acid influx was simultaneously determined by recording of cell fluorescence. Hyperpolarization from -30 to -70 mV accelerated fatty acid influx whereas depolarization to +50 mV reduced uptake. After obtaining equilibrium hyperpolarization increased cell fluorescence, whereas depolarization pushed NBD-stearate out of cells. Potential sensitivity of uptake was dependent on the fatty acid concentrations in the medium with most prominent effects at low unbound concentrations. These data show that, at low fatty acid concentrations, uptake is, in part, driven by an intracellular negative electric membrane potential.

4-Chloro-7-nitrobenzofurazan↗

PubMed1998

Anatomical variation of cerebral venous drainage: the theoretical effect on jugular bulb blood samples.

Recent studies have demonstrated significant variation in bilateral jugular venous oxygen saturation measurements which may be of clinical significance. We have therefore measured variations in normal dural sinus venous drainage to assess the possible effects of normal anatomical variations on measured jugular venous oxygen saturation. Normal volunteers (n = 25) were imaged using magnetic resonance venography to demonstrate variations in venous anatomy. Flow was measured in the superior sagittal sinus and bilaterally in the transverse sinus, sigmoid sinus proximal to the jugular bulb and proximal jugular vein using phase difference magnetic resonance imaging. Examination of magnetic resonance venogram images showed considerable variability in the symmetry of transverse sinus flow. Complete absence of one transverse sinus was seen in four cases and significant asymmetry in the size of the transverse sinuses was present in 13. Quantitative flow studies demonstrated that the ratio of superior sagittal sinus to combined jugular bulb flow showed remarkably little variation (0.46 +/- 0.06). Measurements of transverse sinus flow showed significant asymmetry (< 40% of superior sagittal sinus flow in one transverse sinus) in 21 of 25 volunteers. The effect of the observed asymmetry on jugular venous oxygen saturation was modelled based on the assumption of either a supratentorial or infratentorial lesion. This model predicted significant asymmetry in jugular venous oxygen saturation measurements (> 10%) in 65% of cases with a supratentorial lesion which is in close agreement with clinical observations. This study suggests that normal variations in venous drainage may account for observed asymmetry in jugular venous oxygen saturation measurements.

Adult↗

PubMed1998

Optic neuritis: MR imaging with combined fat- and water-suppression techniques.

PURPOSE: To examine the benefits of combined fat- and water-suppressed T2-weighted magnetic resonance (MR) images in the diagnosis of optic neuritis. MATERIALS AND METHODS: MR imaging was performed with a 1.5-T unit in five healthy volunteers and 18 patients (21 nerves). All patients had abnormalities of visual evoked potentials and fulfilled the clinical criteria for the diagnosis of optic neuritis. Imaging was performed within 4 weeks of diagnosis (n = 12) or between 3 and 6 months after diagnosis (n = 6). Coronal images were obtained throughout the course of the optic nerve with use of three sequences: (a) short inversion time inversion recovery with fast spin-echo (SE) acquisition, (b) selective partial inversion-recovery (SPIR) prepared T2-weighted fast SE acquisition, and (c) SPIR-fluid-attenuated inversion recovery (FLAIR) with fast SE acquisition. RESULTS: Neuritic segments were demonstrated in all 21 symptomatic nerves. The extent of neuritic involvement (number of images showing abnormality) was significantly greater with the SPIR-FLAIR sequence (P < .01). The contrast ratio between neuritic optic nerve and orbital fat, normal nerve, and cerebral spinal fluid was significantly greater with SPIR-FLAIR than with the other sequences (P < .001). SPIR-FLAIR images also improved demonstration of optic nerve atrophy in chronic neuritis when compared with the other sequences. CONCLUSION: The SPIR-FLAIR sequence offers important advantages over current methods in the demonstration of optic neuritis.

Adult↗

PubMed1997

Muscarinic acetylcholine receptor stimulation of biliary epithelial cells and its effect on bile secretion in the isolated perfused liver [corrected].

The aim was to explore whether biliary epithelial cells show muscarinic acetylcholine receptors and to investigate their role in ductular bile formation. In both, isolated rat biliary epithelial cells and Mz-Cha-1 cells, a biliary epithelial cell line, binding of [3H]N-methyl-scopolamine occurred with 0.718 +/- 0.08 and 0.482 +/- 0.05 fmol per 10(6) cells, respectively. To characterize the involved second messenger, intracellular Ca2+ levels were monitored by confocal microscopy. Stimulation of biliary epithelial cells with carbachol produced an increase in free cytosolic Ca2+ levels that declined to baseline values describing a sinusoidal oscillation curve. Increasing concentrations of the agonist decreased latency of the response and increased oscillation frequency. Similar results were obtained in Mz-Cha-1 cells. The intracellular Ca2+ originated from IP3 sensitive intracellular stores and from the extracellular medium. The Ca2+ response could partially be blocked by atropine and completely by pirenzepine, a specific muscarinic receptor-type M1 antagonist. The presence of M1 receptor messenger RNA (mRNA) in biliary epithelial cells was confirmed by reverse transcriptase polymerase chain reaction. In the isolated perfused guinea pig liver, a model with high ductular bile flow, carbachol induced a dose dependent decrease of bile flow by 79.6% +/- 9.8% at 50 mumol/L carbachol (P < .001), without affecting perfusion pressure or biliary electrolyte concentrations. It is concluded that biliary epithelial cells express muscarinic acetylcholine receptors. Stimulation of this receptor leads to cholestasis. This could be because of changes in peribiliary permeability and/or inhibition of biliary epithelial cell secretory function.

Animals↗

PubMed1996

Sodium, hydrogen antiporter activation by extracellular adenosine triphosphate in biliary epithelial cells.

BACKGROUND & AIMS: Extracellular nucleotides are secretagogues and influence ion permeability. The aim of this study was to investigate whether secretagogues activate transmembrane acid-base carriers, e.g., sodium, hydrogen antiporter in cholangiocytes. METHODS: Cells were loaded with pH and Ca(2+)-sensitive dyes. Intracellular changes in ion concentrations were monitored by confocal laser scanning microscopy. Adenosine 3', 5'-cyclic monophosphate was determined by standard methods. RESULTS: Baseline intracellular pH (pH1) averaged 7.28 +/- 0.17 pH units. Adenosine triphosphate (ATP; 10 mumol/L) increased baseline pH1 by 0.28 +/- 0.06 pH units/10 min (P < 0.01). Ten and 100 mumol/L ATP increased Na+, H+ antiporter-mediated proton flux from 9.4 +/- 4.9 mmol. L-1 min-1 to 17.2 +/- 11.8 and 16.7 +/- 10.3 mmol. L(-1)-min(-1) (P < 0.001), respectively. Under control and ATP-stimulated conditions, 1 mmol/L amiloride blocked pH1 recovery, indicating true activation of Na+, H+ antiporter by extracellular ATP. Inhibition of basolateral Na+, H+ antiporter isoform inhibited stimulation by ATP. Na+, H+ antiporter-mediated proton flux was stimulated by adenosine, uridine triphosphate, adenosine-5'-0-(3-thiotriphosphate), alpha, beta-methylene-ATP, and 2-methylthio-ATP but not prevented by adenosine receptor blocking. Activation by ATP was not influenced by the Ca2+/protein kinase C/calmodulin system but could be mimicked by addition of N6,2'-O-dibutyryladenosine-3',5'-cyclic monophosphate and was inhibited by pertussis toxin. CONCLUSIONS: Extracellular nucleotides may modulate secretory and absorptive function of cholangiocytes by activating Na+/H+ exchange mechanisms.

Adenosine↗

PubMed1996

Effect of surface and intracellular pH on hepatocellular fatty acid uptake.

Fatty acids enter hepatocytes, at least in part, by a carrier-mediated uptake mechanism. The importance of driving forces for fatty acid uptake is still controversial. To evaluate possible driving mechanisms for fatty acid transport across plasma membranes, we examined the role of transmembrane proton gradients on fatty acid influx in primary cultured rat hepatocytes. After hepatocytes were loaded with SNARF-1 acetoxymethyl ester, changes in intracellular pH (pHi) under different experimental conditions were measured and recorded by confocal laser scanning microscopy. Fatty acid transport was increased by 45% during cellular alkalosis, achieved by adding 20 mM NH4Cl to the medium, and a concomitant paracellular acidification was observed. Fatty acid uptake was decreased by 30% during cellular acidosis after withdrawal of NH4Cl from the medium. Cellular acidosis activates the Na+/H+ antiporter to export excessive protons to the outer cell surface. Inhibition of Na+/H+ antiporter activity by amiloride diminishes pHi recovery and thereby accumulation of protons at the outer surface of the plasma membrane. Under these conditions, fatty acid uptake was further inhibited by 57% of control conditions. This suggests stimulation of fatty acid influx by an inwardly directed proton gradient. The accelerating effect of protons at the outer surface of the plasma membrane was confirmed by studies in which pH of the medium was varied at constant pHi. Significantly higher fatty acid influx rates were observed at low buffer pH. Recorded differences in fatty acid uptake appeared to be independent of changes in membrane potential, because BaCl2 did not influence initial uptake velocity during cellular alkalosis and paracellular acidosis. Moreover, addition of oleate-albumin mixtures to the NH4Cl incubation buffer did not change the observed intracellular alkalinization. In contrast, after cells were acid loaded, addition of oleate-albumin solutions to the recovery buffer increased pHi recovery rates from 0.21 +/- 0.02 to 0.36 +/- 0.05 pH units/min (P < 0.05), indicating that fatty acids further stimulate Na+/H+ antiporter activity during pHi recovery from an acid load. It is concluded that carrier-mediated uptake of fatty acids in hepatocytes follows an inwardly directed transmembrane proton gradient and is stimulated by the presence of H+ at the outer surface of the plasma membrane.

Animals↗