Assessment of absolute metabolite concentrations in human tissue by 31P MRS in vivo. Part II: Muscle,liver, kidney

Absolute metabolite concentrations were assessed in the muscle, the liver, and the kidney of healthy human volunteers by ³¹P MRS. Fully relaxed in vivo spectra were acquired with a surface coil and were localized with an adiabatic lSlS pulse sequence. The spectra were quantified with a subsequent measurement of a calibration phantom and were processed iteratively in the time domain. The following mean metabolite concentrations (mmollliter) were measured in the resting male calf muscle (n = 9), in the fasting liver (n = 12), and in the orthiotopic kidney (n= 5): [PME] = 2.0 ± 0.6, 3.8 ± 0.7, and 2.6 ± 0.9, [Pi] = 2.9 ± 0.3, 1.8 ± 0.3, and 1.6 ± 0.4, [PDE] = 3.8 ± 0.8, 9.7 ± 1.5, and 4.9 ± 1.1, [PCr] = 22.0 ± 1.2, 0, and 0, [NTP] = 5.7 ± 0.4, 2.9 ± 0.4, and 2.0 ± 0.3, respectively. Several interesting findings are to be emphasized: The concentrations of Pi, PCr, and NTP were 20% lower in the muscle of women than of men. In addition, the pH, was significantly lower in female muscle (6.99 ± 0.03) than in male muscle (7.05 ± 0.03). The pH, in the liver (7.12 ± 0.09) and in the kidney (7.09 ± 0.08) were higher than in the muscle of both genders. The free magnesium concentration (mmollliter) was higher in the lliver (1.40 ± 0.64) than in the kidney (0.79 ± 0.39) and in the muscle (0.52 ± 0.10).     

CT-stereotactic fibrinolysis of spontaneous intracerebral hematomas

CT-stereotactic fibrinolysis is an effective alternative to surgical and conservative therapies for intracerebral hematoma. The method consists of stereotactically puncturing and partially evacuating the hematoma. After fibrinolysis using urokinase, the residual hematoma is completely evacuated through a catheter inserted in the cavity of the hematoma. The operation is usually performed under local anesthesia. Stereotactic methods are safer and less invasive than other methods. Since October 1985, a total of 85 patients have been treated with this method in the Department of Stereotaxy and Neuronuclear Medicine at the University of Freiburg Medical School. Although 25 patients died (29.4%) during the mean follow-up period of 20 months, only 16 (18.8%) died in the acute postoperative phase or within the first 60 days after evacuation. Eighteen patients (21.2%) had died six months after the operation. The quality of life of the 60 surviving patients, as measured on the Karnofsky Scale at follow-up, was very good to good in 70% and moderate in 23.3%. Only 6.7% of the patients were so disabled that they required special care and assistance or had to be placed in a nursing home. The long-term results are thus very encouraging.     

Subregions of the ventral striatum show preferential coding of reward magnitude and probability

As shown in non-human primate and human fMRI studies the probability and magnitude of anticipated rewards modulate activity in the mesolimbic dopaminergic system. Importantly, non-human primate data have revealed that single dopaminergic neurons code for both probability and magnitude of expected reward, suggesting an identical system. Using a guessing task that allowed the independent assessment of the factors probability and magnitude we were able to assess the impact of reward probability and magnitude in ventral striatal subregions in a large sample (n=98). We observed more anterior and lateral peak activation foci in the ventral striatum for reward probability and a more posterior and medial activation peak for reward magnitude, suggesting a functional segregation at the mesoscopic level. Importantly, this functional bias observed for the group average was also tested in each individual subject, allowing for proper random effects inference for the spatial dissociation. Taken together, our data point toward a functional dissociation of neuronal assemblies suggesting that certain populations of neurons are more sensitive to expected reward probability and other populations are more sensitive to reward magnitude.     

Identification of a lectin causing the degeneration of neuronal processes using engineered embryonic stem cells

Unlike the mechanisms involved in the death of neuronal cell bodies, those causing the elimination of processes are not well understood owing to the lack of suitable experimental systems. As the neurotrophin receptor p75(NTR) is known to restrict the growth of neuronal processes, we engineered mouse embryonic stem (ES) cells to express an Ngfr (p75(NTR)) cDNA under the control of the Mapt locus (the gene encoding tau), which begins to be active when ES cell-derived progenitors start elongating processes. This caused a progressive, synchronous degeneration of all processes, and a prospective proteomic analysis showed increased levels of the sugar-binding protein galectin-1 in the p75(NTR)-engineered cells. Function-blocking galectin-1 antibodies prevented the degeneration of processes, and recombinant galectin-1 caused the processes of wild-type neurons to degenerate first, followed by the cell bodies. In vivo, the application of a glutamate receptor agonist, a maneuver known to upregulate p75(NTR), led to an increase in the amount of galectin-1 and to the degeneration of neurons and their processes in a galectin-1-dependent fashion. Section of the sciatic nerve also rapidly upregulated levels of p75(NTR) and galectin-1 in terminal Schwann cells, and the elimination of nerve endings was delayed at the neuromuscular junction of mice lacking Lgals1 (the gene encoding galectin-1). These results indicate that galectin-1 actively participates in the elimination of neuronal processes after lesion, and that engineered ES cells are a useful tool for studying relevant aspects of neuronal degeneration that have been hitherto difficult to analyze.     

Causes of differences in exercise-induced changes of base excess and blood lactate

It has been concluded from comparisons of base excess (BE) and lactic acid (La) concentration changes in blood during exercise-induced acidosis that more H⁺ than La⁻ leave the muscle and enter interstitial fluid and blood. To examine this, we performed incremental cycle tests in 13 untrained males and measured acid–base status and [La] in arterialized blood, plasma, and red cells until 21 min after exhaustion. The decrease of actual BE (−ΔABE) was 2.2 ± 0.5 (SEM) mmol l⁻¹ larger than the increase of [La]_blood at exhaustion, and the difference rose to 4.8 ± 0.5 mmol l⁻¹ during the first minutes of recovery. The decrease of standard BE (SBE), a measure of mean BE of interstitial fluid (if) and blood, however, was smaller than the increase of [La] in the corresponding volume (Δ[La]_if+blood) during exercise and only slightly larger during recovery. The discrepancy between −ΔABE and Δ[La]_blood mainly results from the Donnan effect hindering the rise of [La]_erythrocyte to equal values like [La]_plasma. The changing Donnan effect during acidosis causes that Cl⁻ from the interstitial fluid enter plasma and erythrocytes in exchange for HCO₃⁻. A corresponding amount of La⁻ remains outside the blood. SBE is not influenced by ion shifts among these compartments and therefore is a rather exact measure of acid movements across tissue cell membranes, but changes have been compared previously to Δ[La]_blood instead to Δ[La]_if+blood. When performing correct comparisons and considering Cl⁻/HCO₃⁻ exchange between erythrocytes and extracellular fluid, neither the use of ΔABE nor of ΔSBE provides evidence for differences in H⁺ and La⁻ transport across the tissue cell membranes.     

The effect of electrochemically simulated titanium cathodic corrosion products on ROS production and metabolic activity of osteoblasts and monocytes/macrophages

Nowadays aseptic loosening is the most common cause of orthopaedic implant failure. Some of its reasons have already been described up to now; however, others remain still hypothetical. Besides the inflammatory response to wear particles originating at different sources, the role of reactive oxygen species as products of cellular reactions and/or as a result of the process of corrosion of an implant leading to implant failure has recently been discussed too. In the present study, we used a galvanostatic polarization to simulate the cathodic partial reaction of the corrosion process at a titanium alloy surface. With respect to cells occurring at the interface of a metal implant, the behaviour of osteoblasts and monocytes/macrophages was investigated. It has been found that cathodic polarization of Ti6Al4V induces an increase in the level of intracellular reactive oxygen species as well as suppressing the metabolic activity of cells in a dose-dependent manner. This is in agreement with the results obtained with cells after external addition of hydrogen peroxide as another kind of oxidative stress. In both approaches, monocytes/macrophages show a higher tolerance to oxidative stress than osteoblasts. It could be concluded that the electrochemical setup developed induced intracellular changes occurring during oxidative stress and it could be used for future detailed analysis of the consequences of corrosion processes for cellular reactions.     

Dynamic force microscopy imaging of plasmid DNA and viral RNA

Plasmid DNA and viral RNA were imaged in a liquid environment by dynamic force microscopy (DFM) and fine structures of DNA with heights of 1.82+/-0.66 nm were obtained in topographical images. In simultaneously acquired phase images, DNA could be imaged with better contrast at lower imaging forces. By splitting the cantilever oscillation signal into lower and upper parts, the contribution of the adhesion between tip and sample to the topographical images was eliminated, resulting in better signal-to-noise ratio. DFM of the single stranded RNA genome of a human rhinovirus showed loops protruding from a condensed RNA core, 20-50 nm in height. The mechanical rigidity of the RNA was determined by single molecule pulling experiments. From fitting RNA stretching curves to the Worm-Like-Chain (WLC) model a persistence length of 1.0+/-0.17 nm was obtained.     

Inter-laboratory validation of PCR-based HPV detection in pathology specimens

The detection and typing of human papilloma virus (HPV) in pathology specimens is gaining increasingly in importance. In the context of the initiative for quality assurance in pathology (QuIP) of the German Society of Pathology and the Professional Association of German Pathologists, four panel laboratories with experience and expertise in polymerase chain reaction (PCR)-based HPV detection were selected to establish an inter-laboratory trial. In a first step, these laboratories performed an internal testing of their own methodologies, which comprised DNA sequencing, multiplex nested PCR and hybridization techniques. Material from 39 samples including paraffin sections and DNA preparations of tissues and plasmids were evaluated by each panel institute according to their own protocols. Despite the different methodologies, a high degree of inter-laboratory reliability was achieved. In this report, we summarise the results. Pretested specimens are available for the external trail and can be ordered from the steering institute via provitro GmbH Berlin (). Supplementary data are online available at (rubric "Forschung"), which includes a web-based photo gallery of HPV-associated lesions and their potential association with specific virus types. The initiative is intended to foster the quality assurance of molecular HPV analysis in pathology and its correlation with morphological changes.     

Immune response in the tilapia, Oreochromis mossambicus on exposure to tannery effluent

The objective of the study was to investigate the effect of chronic exposure to sublethal concentrations of tannery effluent (TE) on the specific immune response and nonspecific immunity in tilapia, Oreochromis mossambicus. The effluent from the tannery was collected directly from a chrome-tanning factory situated in Dindigul district, Tamil Nadu, India. Apart from chromium (88.2 ppm), the effluent contained appreciable amount of calcium carbonate and sodium sulphate. Groups of fish (45-50 g) were exposed to 0.0053, 0.053 or 0.53% [0.1%, 1% or 10% LC50] of TE for 28 days. The specific immune response of fish was assessed by antibody response to heat-killed Aeromonas hydrophila by ELISA and bacterial agglutination assay. Nonspecific immune mechanisms were assessed in terms of serum lysozyme activity, production of intracellular reactive oxygen species (ROS) and reactive nitrogen intermediates (RNI) by peripheral blood leucocytes (PBL). The results indicate that chronic exposure of fish to 0.53% of TE, significantly suppressed antibody response, nonspecific serum lysozyme activity, and ROS and RNI production. Exposure to 0.053% (1% LC50) of TE also caused a similar suppressive effect though at a lesser degree. In conclusion, the study shows, that exposure to sublethal concentrations of TE, can lead to adverse effects on selected immune reactions in tilapia. Further, these findings may be important in terms of monitoring fish health and risk assessment during periods of fluctuating levels of pollutants in the natural and farm environments.