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  • 1
    Keywords: SPECTRA ; Germany ; human ; TOOL ; PROTEIN ; DIFFERENTIATION ; PATIENT ; BIOMARKERS ; IDENTIFICATION ; ARRAYS ; mass spectrometry ; MASS-SPECTROMETRY ; PREDICTION ; sensitivity ; specificity ; CELL CARCINOMA ; ALLOGRAFT-REJECTION ; TECHNOLOGY ; RELEVANCE
    Abstract: Objectives. To develop a noninvasive method for the detection of renal transplant rejection using ProteinChip Arrays (surface-enhanced laser desorption/ionization time-of-flight mass spectrometry). Methods. A total of 23 urine samples were collected from 13 patients showing biopsy-proven renal allograft rejection and from 10 patients without histologic signs of rejection. All 23 patients had clinical symptoms and signs of acute allograft rejection and underwent renal biopsy. Samples were centrifuged, and supernatants were directly spotted onto the ProteinChip arrays with different chromatographic surfaces. The obtained spectra in a range from 2 to 200 kDa were subjected to bioinformatic analysis using the method of Fuzzy c-means, followed by the establishment of rule bases and evaluation using the relevance index according to Kiendl. Results. Several protein peaks were identified allowing differentiation between rejection and no rejection. Using two different ProteinChip surfaces, we found two biomarkers at 25.71 kDa and 28.13 kDa that gave a diagnostic sensitivity of 90% and 93% and a specificity of 80% [SAX2) and 85% (CM 10), respectively. Conclusions. Surface-enhanced laser desorption/ionization time-of-flight mass spectrometry appears to be a promising new diagnostic tool for distinguishing renal transplant patients with no rejection from those with acute rejection
    Type of Publication: Journal article published
    PubMed ID: 16527560
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  • 2
    ISSN: 1434-0879
    Keywords: Renal cell carcinoma ; Renal tubular transport ; Renal cortical slices ; p-Aminohippurate ; Human ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract In vitro accumulation ofp-aminohippurate (PAH) was investigated in “intact” human renal cortical slices of normal kidney tissue and in tissue slices of renal cell carcinoma (RCC). The technique used was established in preliminary experiments on rat kidney tissue slices. In principle, the accumulation capacity is comparable in renal tissue slices of both species (slice to medium accumulation ratios between 4 and 8). In man sex differences in accumulation capacity do not exist. But, as shown in detail for rats, accumulation capacity drops with age. Tissue slices of RCC are unable to accumulate PAH actively; slice to medium ratio reaches about 1 and indicates passive PAH uptake only. Surprisingly, in tumors of stage pTl PAH uptake is lowest, perhaps as a sign of PAH transport out of the cells. There is no difference between peripheral and central parts of RCC. Age and sex are without influence on PAH uptake in RCC tissue slices. Interestingly, the accumulation capacity of “intact” tissue of kidneys infested with RCC also depends on the severity of the tumor (stage, diameter), but not on grading and formation of metastases.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1434-0879
    Keywords: Key words Renal tubular transport ; p-Aminohippurate ; Stimulation ; Renal cell carcinoma ; Dexamethasone ; Triiodothyronine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract This paper is the third of a long-term planned series of papers dealing with ex vivo investigations of drug transport in human kidney. The aims of this study are (a) to investigate whether or not human renal cell carcinoma (RCC) can actively accumulate p-aminohippurate (PAH) and (b) to test the response of RCC on dexamethasone or triiodothyronine (T3) using tissue slices ex vivo. By this approach, the accumulation capacity of RCC should be stimulated as a prerequisite for an increased uptake of anti-tumour drugs. Tissue slices of RCC samples of 30 patients were incubated for 24 h in Williams medium E containing 0.01–50 μM dexamethasone or T3. Thereafter, slices were placed in PAH-containing Cross–Taggart medium, and PAH uptake into kidney tissue was measured for 2h under standardised conditions as described previously. In intact human renal cortical slices, PAH uptake capacity, expressed as slice to medium ratio (Q S/M), was about 2.8 ± 0.16 after 24 h of incubation and increased significantly in dexamethasone-containing medium in a concentration-dependent manner, up to ∼150%, whereas T3 did not influence PAH accumulation. On the other hand, in RCC the PAH accumulation capacity was completely abolished (Q S/M∼1). However, after administration of dexamethasone, the accumulated amount of PAH increased significantly in RCC tissue in a concentration-dependent manner, up to ∼190%. T3 was without effect in RCC, too. Surprisingly, the dexamethasone-mediated stimulation could be differentiated into responders and non-responders, with maximal effects at different concentrations for each patient. Nevertheless, the maximal transport rates remained low in RCC, even under hormone influence. In conclusion, a moderate stimulation of tubular transport capacity can be shown ex vivo in human RCC. This phenomenon is only of a relatively low degree compared with intact renal tissue. However, in principle, the response of RCC on dexamethasone could form a basis for further therapeutic strategies to overcome multi-drug resistance in RCC patients. For this purpose, additional experiments analysing the expression of transporters of the ABC cassette-type are in progress.
    Type of Medium: Electronic Resource
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