Fibroblast mitochondria in idiopathic Parkinson's disease display morphological changes and enhanced resistance to depolarization.

Mitochondrial dysfunction is a hallmark in idiopathic Parkinson's disease (IPD). Here, we established screenable phenotypes of mitochondrial morphology and function in primary fibroblasts derived from patients with IPD. Upper arm punch skin biopsy was performed in 41 patients with mid-stage IPD and 21 age-matched healthy controls. At the single-cell level, the basal mitochondrial membrane potential (Ψm) was higher in patients with IPD than in controls. Similarly, under carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP) stress, the remaining Ψm was increased in patients with IPD. Analysis of mitochondrial morphometric parameters revealed significantly decreased mitochondrial connectivity in patients with IPD, with 9 of 14 morphometric mitochondrial parameters differing from those in controls. Significant morphometric mitochondrial changes included the node degree, mean volume, skeleton size, perimeter, form factor, node count, erosion body count, endpoints, and mitochondria count (all P-values < 0.05). These functional data reveal that resistance to depolarization was increased by treatment with the protonophore FCCP in patients with IPD, whereas morphometric data revealed decreased mitochondrial connectivity and increased mitochondrial fragmentation.

Increased cold and heat pain thresholds influence the thermal grill illusion in schizophrenia.

BACKGROUND: Patients with schizophrenia show decreased sensitivity towards clinical and experimental painful conditions. To date, the exact underlying mechanisms are not completely understood. One method to examine central integrative processes of pain perception is the thermal grill illusion (TGI), in which interlacing cold and warm bars create the illusion of a painful sensation. METHODS: In 18 unmedicated patients with acute paranoid schizophrenia, cold and heat pain thresholds (CPT/HPT) as well as the perception of the TGI were examined and compared to 18 matched controls. In addition, symptom scales were obtained in order to relate pain perception to psychopathology. RESULTS: CPT and HPT were significantly increased in patients compared to controls. In the range of TGI stimuli that were perceived painful by controls, patients did not indicate painful sensations, instead the stimulus response curve of TGI pain perception was shifted towards higher stimulus intensities, i.e., greater temperature differentials between cold and warm bars. This increase was comparable to that seen in CPT and HPT. There was no association with psychopathology for any pain parameter. CONCLUSIONS: CPT and HPT, as well as temperature differentials for the perception of the TGI were increased in patients with schizophrenia as compared to controls. Similar to visual illusions, in which reduced contrast sensitivity has been shown to alter the perception of illusions, the discriminatory somatosensory deficit, which is reflected in higher CPT and HPT as well as the previously reported increased warmth perception thresholds, might account for the attenuation of TGI in patients.

Intragastric acidity during administration of generic omeprazole or esomeprazole - a randomised, two-way crossover study including CYP2C19 genotyping.

BACKGROUND: Generic omeprazole has been approved in many countries for the treatment of acid-related gastrointestinal disorders. However, clinical studies comparing generic to original proton pump inhibitors are limited. AIMS: To compare the effect of generic omeprazole 20 mg/day with esomeprazole 20 mg/day on intragastric acidity and to investigate the influence of the CYP2C19 metabolizer status. METHODS: In this randomised, single-blinded, two-way crossover study, 24 healthy Helicobacter pylori-negative subjects, received generic omeprazole (Omep; Hexal AG, Holzkirchen, Germany) 20 mg once daily or esomeprazole 20 mg once daily for five consecutive days. Twenty-four-hour intragastric pH was recorded on day 5 of each treatment. CYP2C19 status was determined by polymerase chain reaction-restriction fragment length polymorphism. RESULTS: Over all, there were no statistically significant differences between generic omeprazole and esomeprazole with respect to median intragastric pH (3.5 and 3.9, P = 0.07), the total hours with intragastric pH >4 (10.4 and 11.3, P = 0.29), and during upright (9.6 and 9.1, P = 0.77) or supine (2.2 and 2.2, P = 0.94) position. However, in CYP2C19 rapid metabolizers, esomeprazole was superior to omeprazole, with the percentage of time with intragastric pH >3.0 and pH >3.5 being higher with esomeprazole than with generic omeprazole [Δ = 9% (P = 0.026) and Δ = 8% (P = 0.046), respectively]. CONCLUSIONS: Overall, generic omeprazole 20 mg appears to provide a similar intragastric acid control when compared with esomeprazole 20 mg. However, esomeprazole might be advantageous in subjects with a rapid CYP2C19 metabolizer status.

[Cavernoma of the medulla oblongata mimicking "Anorexia nervosa" - a case report]. / Kavernom der Medulla oblongata unter dem Bild einer "Anorexia nervosa".

An 11-year-old girl presented with recurrent vomiting, reduced food and fluid intake, weight loss and dizziness. In an external hospital she was diagnosed as having habitual vomiting and a beginning eating disorder. The physical examination revealed a very thin, malnourished child with abdominal pain on palpation but without neurologic deficits. Laboratory findings, X-rays, endoscopy and ultrasound revealed no pathological results either. Since the EEG and the cranial computed tomography (CT) were normal, we also suspected the beginning of an eating disorder especially due to some psychical peculiarities. The MRI which was done for ultimate exclusion of an infratentorial tumor showed a well defined displacing structure in the dorsal medulla oblongata and in the upper cervical spinal cord. A corresponding prolongation of the central conduction time was found in the median nerve SSEP whereas the BAEPs were normal. The tumor was excised in toto and the histological examination confirmed the suspected diagnosis of cavernous hemangioma (cavernoma). The post operative phase was without any complications and the intiated nutrition was well tolerated. Neurological deficits were not observed.

Treatment of diglyme containing wastewater by advanced oxidation--process design and optimisation.

Diglyme (CAS No. 111-96-6), a biorefractive ether with teratogenic properties is of considerable importance as a solvent in the synthetic chemical industry. Results of lab-scale investigations into the optimal conditions for the oxidative mineralisation of 0.05 to 1 g/L of diglyme in synthetic process waters by hydrogen peroxide and ozone based advanced oxidation processes are presented. Fenton, photo-assisted Fenton and UV/H2O2 oxidation processes show acceptable TOC removals. At 50 degrees C the initial TOC removal rates varies between 0.07 and 6g TOC/L*h. The rates increase with the initial diglyme and hydrogen peroxide concentration as well as with the UV irradiation energy intensity. For example at a 1 g/l diglyme concentration a stoichiometric H2O2 addition resulted in TOC degradation of 60% to 70% after 30 minutes under the investigated conditions. Treatment with ozone/H2O2 at a pH of 8 and 25 degrees C required at least 100 minutes to achieve comparable mineralisation results. Biodegradability can be reached in far shorter times. Ozone on its own cannot be recommended for diglyme treatment. If complete mineralisation of diglyme is the objective, the UV/H2O2 oxidation process should be favoured. The ozone/H2O2 process might offer a viable alternative in cases where the oxidation step is followed by biological wastewater treatment, so that biodegradability is aimed at.

New ELISA for quantitation of human urokinase receptor (CD87) in cancer.

The serine protease urokinase-type plasminogen activator (uPA), its inhibitor (PAI-1), and its receptor (uPAR; CD87) facilitate cancer cell invasion and metastasis. Whereas uPA and PAI-1 antigen levels determined in tumor tissue extracts of breast cancer patients correlate with disease recurrence and overall survival, the prognostic relevance of uPAR is still a matter of debate. We established two new sandwich-type enzyme-linked immunosorbent assay (ELISA) formats (HU/IIIF10-ELISA and HU/HD13-ELISA) using the epitope-defined monoclonal antibody (mAb) IIIF10 or the conformation-dependent mAb HD13.1, a polyclonal chicken antibody (HU277), and recombinant soluble uPAR (CHO-suPAR) as the standard. The lower detection limit of the assays was at 0.16 ng/ml, with a linear dose-response up to 5 ng/ml of uPAR antigen. Both ELISA formats showed good reproducibility and recovery. The intra-assay and the inter-assay variation coefficients were respectively 4.3% and 11.7% (HU/IIIF10-ELISA) and 4.0% and 10.7% (HU/HD13-ELISA). The recovery rate of uPAR in cell lysates spiked with CHO-suPAR was above 82% and 88%, respectively. With these new ELISA formats, uPAR antigen content in breast cancer tissue extracts and tumor cell lysates was determined and compared to a commercially available ELISA (ADI-ELISA). By all of the three uPAR ELISA formats CHO-suPAR and uPAR present in lysates of non-malignant epithelial cells and stimulated monocytes were quantified with similar sensitivity. Interestingly, in breast cancer cell lines of epitheloid origin a higher uPAR antigen content was determined by the HU/IIIF10-ELISA than the HU/HD13- or ADI-ELISA formats. In lysates of fibroblastic breast cancer cell lines similar uPAR values were obtained with the HU/IIIF10- and ADI-ELISA formats, whereas with the HU/HD13-ELISA significantly lower uPAR concentrations were determined. The prognostic relevance of tumor uPAR antigen was evaluated in 199 primary breast cancer patients with a median follow-up of 24 months. uPAR antigen values above the cut-off of 3.33 ng/mg protein as determined by the HU/IIIF10-ELISA were significantly correlated with short disease-free survival (p=0.025). Results obtained by the other two ELISA formats (HU/HD13-ELISA and ADI-ELISA) were not associated with prognosis. Our findings stress the need of well-characterized antibodies, which detect both uPAR of non-malignant and tumor cells, in setting up a uPAR-ELISA useful for assessing breast cancer patient prognosis.

Electric birefringence imaging of DNA in agarose electrophoresis gels: noise limits and prolonged electric field effects.

Electric birefringence imaging (EBI) maps stresses in agarose electrophoresis gels. Both DNA and gel alignment can be monitored using this technique. Previously, we have reported birefringence images associated with resolved small (< 48 kbp) and partially resolved large 48 kb-(2 Mbp) DNA fragments oriented with relatively short electric field pulses. In this paper, differences between images acquired using short and long (DC) voltage pulses are presented. It is shown that stress signals distribute over long distances when exposed to prolonged electric fields. Resolution also affects band shapes. Refinements in instrumental design and evaluation of limiting noise sources of the technique are also described.

Noninvasive imaging of nanogram quantities of DNA in agarose electrophoresis gels.

Image processing techniques which separate the DNA Kerr effect and induced electrokinetic distortion contributions to electric birefringence images of agarose nucleic acid electrophoresis gels are described. Under standard electrophoresis conditions, detection limits of 10 ng of DNA per well are obtained in hydroxyethylated agarose without signal averaging or 7.5 ng with averaging of four measurements. Maintaining constant gel temperature is shown to improve the quality of the images. Monochromatic light (589 nm) is shown to give a small increase in sensitivity compared to polychromatic (650 +/- 20 nm) light.

Uptake of corticosterone into isolated rat liver cells: possible involvement of Na+/K(+)-ATPase.

Isolated rat hepatocytes possess a saturable glucocorticoid uptake system with high affinity (Kd value = 2.8 +/- 0.7 x 10(-8) M; 318,000 +/- 80,000 binding sites per cell; 317 fmol/mg protein). The initial rates of uptake decrease by about 30-40% if the cells are incubated simultaneously with [3H]corticosterone and either SH-reagents (N-ethylmaleimide and p-chloromercuriphenylsulphonate, 1 mM), metabolic inhibitors (2,4-dinitrophenol, 1 mM; and antimycin, 0.1 mM) or the Na+/K(+)-ATPase-inhibitors, ouabain and quercetine. These Na+/K(+)-ATPase-blockers exert half-maximal inhibition at 3 x 10(-7) and 3 x 10(-6) M, respectively. A slight increase in K+ concentration and a corresponding decrease in Na+ in the medium leads to a significant reduction in the initial uptake rate. The uptake system from the rat hepatocytes shows a clear steroid specificity, being different from the intracellular receptor. Corticosterone and progesterone are the strongest competitors, cortisol, 5 alpha- and 5 beta-dihydrocorticosterone, 11-deoxycorticosterone, cortisone and testosterone have an intermediate effect and only weak competition is exerted by dexamethasone and by the mineralocorticoid, aldosterone. Estradiol and estrone sulphate as well as the synthetic glucocorticoid triamcinolone acetonide are unable to inhibit initial corticosterone uptake.

Uptake and retention of moulting hormones by the integument of crayfish in vitro. III. The possible involvement of Na+/K+-ATPase.

The specific uptake of the ecdysteroid [3H]ponasterone A into crayfish hypodermis in vitro is inhibited by increasing concentrations of potassium in the medium. The Na+/K+-ATPase blocker ouabain also decreases uptake of the ecdysteroid. Maximal inhibition of uptake is reached at 5 X 10(-5) M ouabain.

[Immunostimulation in the lymphadenopathy syndrome and in AIDS--Inosiplex (inosin pranobex, methisoprinol]. / Immunstimulation beim Lymphadenopathiesyndrom und bei AIDS--Inosiplex (Inosin pranobex, Methisoprinol).

In patients with AIDS, LAS and ARC a defect in producing lymphokines such as Interleukin 2 and Gamma-interferon is found. Possibilities of therapy are reported. Especially the immunostimulant properties of inosiplex are commented, and the first successful results of therapy are discussed.