Helmut Hinghofer-Szalkay

Detection of 4-hydroxynonenal (HNE) as a physiological component in human plasma.

4-Hydroxynonenal (HNE) is a major aldehydic product formed by peroxidation of omega 6-unsaturated fatty acids and is regarded as a specific marker of lipid peroxidation. In this paper we demonstrate that there is a physiological steady-state concentration of HNE in human venous blood plasma. For the quantitative determination of HNE a modified version of an existing, but tedious and time-consuming HPLC method was developed. The extraction of aldehydic hydrazones from plasma was performed using an Extrelut column and the separation step by thin-layer chromatography was replaced by column chromatography on silica gel. The concentration of HNE in human blood plasma was in the same range as the concentration that was found to inhibit the proliferation of cells of the peripheral tissues, i.e., endothelial cells and fibroblasts in vitro. In an experiment with reduced peripheral blood flow a temporary significant increase of HNE was observed during reperfusion. It was concluded that lipid peroxidation occurs in peripheral tissues of humans following temporary congestion of venous blood flow.

A model of artefacts produced by stray capacitance during whole body or segmental bioimpedance spectroscopy

We have developed a novel model for the simulation of artefacts which are produced by stray capacitance during bioimpedance spectroscopy. We focused on whole body and segmental measurements in the frequency range 5-1000 kHz. The current source was assumed to by asymmetric with respect to ground as is the case for many commercial devices. We considered the following stray pathways: 1, cable capacitance; 2, capacitance between neighbouring electrode leads; 3. capacitance between different body segments and earth; 4, capacitance between signal ground of the device and earth. According to our results the pathways 3 and 4 cause a significant spurious dispersion in the measured impedance spectra at frequencies > 500 kHz. During segmental measurements the spectra have been found to be sensitive to an interchange of the electrode cable pairs. The sensitivity was also observed in vivo and is due to asymmetry of the potential distribution along the segment with respect to earth. In contrast to previously published approaches, our model renders possible the simulation of this effect. However, it is unable to fully explain the deviations of in vivo measured impedance spectra from a single Cole circle. We postulate that the remaining deviations are due to a physiologically caused superposition of two dispersions from two different tissues.

Assessing abdominal fatness with local bioimpedance analysis: basics and experimental findings.

OBJECTIVE: Abdominal fat is of major importance in terms of body fat distribution but is poorly reflected in conventional body impedance measurements. We developed a new technique for assessing the abdominal subcutaneous fat layer thickness (SFL) with single-frequency determination of the electrical impedance across the waist (SAI).SUBJECTS AND MEASUREMENTS: The method uses a tetrapolar arrangement of surface electrodes which are placed symmetrically to the umbilicus in a plane perpendicular to the body axis. Twenty-four test subjects (12 male, 12 female) underwent SAI and abdominal magnetic resonance imaging (MRI). The SFL below the sensing electrodes was determined from MRI and correlated with the SAI data at four different frequencies (5, 20, 50 and 204 kHz).RESULTS: A highly significant linear correlation (r2=0.99) between SFL and SAI over a wide range of the abdominal SFL was found. Separate regression models for female and male subjects did not differ significantly, except at 50 kHz.CONCLUSION: SAI represents a good predictor of the SFL and provides an excellent tool for the assessment of central obesity.International Journal of Obesity (2001) 25, 502–511

Ventilation during simulated altitude, normobaric hypoxia and normoxic hypobaria

To investigate the possible effect of hypobaria on ventilation (VE) at high altitude, we exposed nine men to three conditions for 10 h in a chamber on separate occasions at least 1 week apart. These three conditions were: altitude (PB = 432, FIO2 = 0.207), normobaric hypoxia (PB = 614, FIO2 = 0.142) and normoxic hypobaria (PB = 434, FIO2 = 0.296). In addition, post-test measurements were made 2 h after returning to ambient conditions at normobaric normoxia (PB = 636, FIO2 = 0.204). In the first hour of exposure VE was increased similarly by altitude and normobaric hypoxia. The was 38% above post-test values and end-tidal CO2 (PET(CO2) was lower by 4 mmHg. After 3, 6 and 9 h, the average VE in normobaric hypoxia was 26% higher than at altitude (p < 0.01), resulting primarily from a decline in VE at altitude. The difference between altitude and normobaric hypoxia was greatest at 3 h (+ 39%). In spite of the higher VE during normobaric hypoxia, the PET(CO2) was higher than at altitude. Changes in VE and PET(CO2) in normoxic hypobaria were minimal relative to normobaric normoxia post-test measurements. One possible explanation for the lower VE at altitude is that CO2 elimination is relatively less at altitude because of a reduction in inspired gas density compared to normobaric hypoxia; this may reduce the work of breathing or alveolar deadspace. The greater VE during the first hour at altitude, relative to subsequent measurements, may be related to the appearance of microbubbles in the pulmonary circulation acting to transiently worsen matching. Results indicate that hypobaria per se effects ventilation under altitude conditions.

The continuous high-precision measurement of the density of flowing blood

SummaryThe “mechanical oscillator” technique for the measurement of the density of fluids is based on the influence of mass on the natural frequency of a mechanical oscillator. The practical application of this principle was worked out by Kratky et al. (1969) and Leopold (1970). It is demonstrated in this study that the method permits the continuous high-precision measurement of the density of flowing blood in anesthetized animals. The accuracy is 10−5 g/ml, the maximum sampling rate 20/min.As found in rabbits and cats during the control state, physiological blood density changes related to spontaneous blood pressure variations are up to 2·10−4 g/ml. The method can be combined with i.v. injections of isotonic and iso-oncotic solutions to determine cardiac output and blood volume on the basis of a “density dilution” principle. Since the density of the interstitial fluid is lower than that of blood, fluid shifts through the capillary walls can be detected. The effects of hypertonic glucose and of hyperoncotic dextran have been examined. Changes in the density of the arterial blood appear within 10 s after i.v. injection of these fluids. Similarly, density changes result from hemorrhage and reinfusion. During and after i.v. administration of vasoactive drugs (noradrenaline, angiotensin II, acetylcholine), marked transient changes in blood density are seen which obviously reflect the effects of fluid shifts through the capillary walls. During hemorrhagic hypotension we found periodic variations in the blood density synchronous with spontaneously occurring Mayer waves. The new method seems to be a promising tool for investigations on physiological and pathological capillary fluid dynamics.

Skeletal muscle is protected from disuse in hibernating dauria ground squirrels

The purpose of this study is to test the hypothesis that muscle fibers are protected from undue atrophy in hibernating dauria ground squirrels (Spermophilus dauricus, Brandt). Muscle mass, fiber cross sectional area (CSA, video analysis) and fiber type distribution (m-ATPase staining) were determined in extensor digitorum longus (EDL) muscle from non-hibernating control animals (Pre-H), from animals who hibernated for one (H1) or two (H2) months, and from animals 2-4days after arousal (Post-H; N=8 each). Muscle wet weight decreased less than body weight in hibernating animals, resulting in a steady increase in muscle-to-body mass ratio (+37% in Post-H compared to Pre-H, p<0.001). In the Pre-H group, Type I (6.3±2.0%) and II (93.7±2.0%) fiber CSAs were 1719±201 and 2261±287μm(2), respectively. There was a tendency (n.s.) of larger CSA of type I in hibernators compared to pre-H. In the Post-H group, fiber CSA and type distribution were not different from Pre-H. We are the first to report data on EDL fiber type distribution and confirm a protective effect that prevents muscle atrophy in spite of prolonged disuse during hibernation in dauria ground squirrels.

Orthostatic Stimuli Rapidly Change Plasma Adrenomedullin in Humans

The aim of this study was to evaluate the effect of orthostasis on the time course of plasma adrenomedullin concentration. On 5 different days, normotensive subjects were randomized to undergo for 30 minutes either 12 degrees, 30 degrees, 53 degrees, or 70 degrees passive head-up tilt or to remain supine. Venous blood was collected from each subject in the supine position before tilting, at 3 and 27 minutes during tilting, and at 2 and 50 minutes after orthostasis. Plasma adrenomedullin increased significantly with tilt of >/=30 degrees in a stimulus-dependent manner. Approximately half of the increase seen at 27 minutes occurred during the first 2 minutes of upright positioning; the maximum effect with 70 degrees tilt was +70%. Elevations in norepinephrine, epinephrine, aldosterone, plasma renin activity, vasopressin, heart rate, and mean arterial pressure were also significant. Hematocrit, blood density, plasma density, and plasma volume loss rose (P<0.05) at 53 degrees and 70 degrees tilt. Our results indicate that adrenomedullin may play an important role in stabilization of hemodynamics during passive orthostasis. In conclusion, plasma adrenomedullin rapidly increases with orthostatic challenge in a stimulus-dependent manner and also swiftly returns to baseline levels after the subject resumes the supine position.

Reactive hyperemia in the human liver

We tested whether hepatic blood flow is altered following central hypovolemia caused by simulated orthostatic stress. After 30 min of supine rest, hemodynamic, plasma density, and indocyanine green (ICG) clearance responses were determined during and after release of a 15-min 40 mmHg lower body negative pressure (LBNP) stimulus. Plasma density shifts and the time course of plasma ICG concentration were used to assess intravascular volume and hepatic perfusion changes. Plasma volume decreased during LBNP (-10%) as did cardiac output (-15%), whereas heart rate (+14%) and peripheral resistance (+17%) increased, as expected. On the basis of ICG elimination, hepatic perfusion decreased from 1.67 +/- 0.32 (pre-LBNP control) to 1.29 +/- 0.26 l/min (-22%) during LBNP. Immediately after LBNP release, we found hepatic perfusion 25% above control levels (to 2.08 +/- 0.48 l/min, P = 0.0001). Hepatic vascular conductance after LBNP was also significantly higher than during pre-LBNP control (21.4 +/- 5.4 vs. 17.1 +/- 3.1 ml.min(-1).mmHg(-1), P < 0.0001). This indicates autoregulatory vasodilatation in response to relative ischemia during a stimulus that has cardiovascular effects similar to normal orthostasis. We present evidence for physiological post-LBNP reactive hyperemia in the human liver. Further studies are needed to quantify the intensity of this response in relation to stimulus duration and magnitude, and clarify its mechanism.

Fluid volume distribution within superficial shell tissues along body axis during changes of body posture in man

In up to six different sides along the body axis during tilting manoeuvres, volume shifts into or out off superficial tissues were followed with a newly developed miniature plethysmograph. It was possible to localized a region where no or only minor volume changes during the tilt table experiments occurred. This region is identical with the Hydrostatic Indifferent Point (HIP) being localized below the apex of the heart in the upper third of abdominal vena cava. Above the HIP fluid is drained out off the tissues during assumption of upright posture whereas below the HIP fluid volume is pooled. The volume changes occurred in two phases. Within the first 5 s in the cephalad parts of the body a rapid decrease occurred, thereafter the volume remained unchanged or even increased; below the HIP within the first 5 s a large volume increase was followed by a slow continuous volume increment.The functional peculiarities of the low pressure system as a whole were visible studying only the superficial shell tissues of the body with the non invasive miniature plethysmographic technique.

Oxytocin levels in the posterior pituitary and in the heart are modified by voluntary wheel running

We hypothesized that voluntary wheel running results in increased secretion of oxytocin, a peptide involved in the stress response. An additional hypothesis was that prolonged exercise affects oxytocin levels in the heart, which is in line with the potential role of oxytocin in cardiovascular functions. Voluntary wheel running lasted 3 weeks and daily running distances increased progressively reaching maximum levels about 8 km (Sprague-Dawley rats) and 4 km (Lewis strain). The exercise resulted in significant reduction of epididymal fat, slight increase in glucose transporter GLUT4 mRNA levels and significant enhancement of plasma density. Voluntary exercise failed to influence plasma oxytocin levels either in Lewis or Sprague-Dawley rats, but it resulted in a significant decrease of oxytocin concentrations in the posterior pituitary. Plasma oxytocin concentrations were not modified even if the measurements were made in the dark phase of the day. In voluntary wheel running Sprague-Dawley rats, the content of oxytocin in the right heart atrium was lower than in controls. Thus, the present findings demonstrate that prolonged voluntary wheel running results in a decrease in pituitary oxytocin content without evident changes in hormone concentrations in peripheral blood. However, prolonged exercise used has a significant impact on oxytocin levels in the heart.