The suitability of cytochrome-P4501A1 as a biomarker for PCB contamination in European eel (Anguilla anguilla).

European eel (Anguilla anguilla) elvers were intraperitoneally injected with different doses of 3,3',4,4'-tetrachlorobiyphenyl (PCB77) to examine and characterize the inductive effect of coplanar PCBs on CYP1A1 gene expression in liver and gills by using a semi-quantitative RT-PCR analysis. The influence of PCB77 injection on transcription activity of the housekeeping gene glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) was tested to determine its suitability as a reference gene for further quantitative gene expression analyses. Our results clearly indicate a significant dose-dependent increase in CYP1A1 gene expression in the gills of European eel, while in liver tissues a significant elevation in CYP1A1 gene expression was only detectable at highest contamination rates, indicating the potential of CYP1A1 differential gene expression analysis in gills as a biomarker for PCB contamination in eels. PCB77 contamination did not affect GAPDH transcription in gills but, at highest doses, resulted in a significant elevation in liver, speaking against GAPDH as a reference housekeeping gene after PCB exposure.

Identification of fish species after cooking by SDS-PAGE and urea IEF: a collaborative study.

A collaborative study, to validate the use of SDS-PAGE and urea IEF, for the identification of fish species after cooking has been performed by nine laboratories. By following optimized standard operation procedures, 10 commercially important species (Atlantic salmon, sea trout, rainbow trout, turbot, Alaska pollock, pollack, pink salmon, Arctic char, chum salmon, and New Zealand hake) had to be identified by comparison with 22 reference samples. Some differences in the recoveries of proteins from cooked fish flesh were noted between the urea and the SDS extraction procedures used. Generally, the urea extraction procedure appears to be less efficient than the SDS extraction for protein solubilization. Except for some species belonging to the Salmonidae family (Salmo, Oncorhynchus), both of the analytical techniques tested (urea IEF, SDS-PAGE) enabled identification of the species of the samples to be established. With urea IEF, two laboratories could not differentiate Salmo salar from Salmo trutta. The same difficulties were noted for differentiation between Oncorhynchus gorbuscha and Oncorhynchus keta samples. With SDS-PAGE, three laboratories had some difficulties in identifying the S. trutta samples. However, in the contrast with the previous technique, SDS-PAGE allows the characterization of most of the Oncorhynchus species tested. Only Oncorhynchus mykiss was not clearly recognized by one laboratory. Therefore, SDS-PAGE (Excel gel homogeneous 15%) appears to be better for the identification, after cooking, of fish such as the tuna and salmon species which are characterized by neutral and basic protein bands, and urea IEF (CleanGel) is better for the gadoid species, which are characterized by acid protein bands (parvalbumins). Nevertheless, in contentious cases it is preferable to use both analytical methods.

Fish muscle parvalbumins as marker proteins for native and urea isoelectric focusing.

An isoelectric point (pI) calibration kit containing fish muscle parvalbumins was prepared and tested for its suitability for isoelectric focusing (IEF) in the presence of 8 M urea. The pattern obtained by urea CleanGel IEF consisted of nine bands covering the pI range 4.96-5.64. This range is relevant for species identification of heated fish by urea IEF. The kit may also be used for native IEF in the low pH range, as demonstrated by running an extract made from the kit together with water-soluble fish muscle proteins on Servalyt Precotes 3-6.

A standardized method of identification of raw and heat-processed fish by urea isoelectric focusing: a collaborative study.

A urea-isoelectric focusing (urea-IEF) method of identifying fish species in processed fishery products was investigated as an interlaboratory collaborative study. The technique was optimized with respect to (i) protein extraction conditions, composition of the extraction solution (urea and SDS solutions), determination of protein concentrations of the fish extracts (five tested methods); (ii) nature of gel (with carrier ampholytes and Immobilines), conditions of rehydration of commercial dry gels, urea concentration; (iii) staining conditions, Coomassie blue and silver staining. The results of various experiments were compared to select the most appropriate methodology, with respect to the discrimination power of differentiating species with the minimal influence of heat processing, reproducibility, speed, and ease of application. The method recommended meets the requirements of food control and customs laboratories.

Development of a sodium dodecyl sulfate-polyacrylamide gel electrophoresis reference method for the analysis and identification of fish species in raw and heat-processed samples: a collaborative study.

A collaborative study was carried out in seven European labs with the aim of achieving a sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) standard operation procedure to identify fish species in raw and cooked samples. Urea and SDS-containing solutions were evaluated as extractants. Several preelectrophoretic operations--such as treatment with RNase/DNase, ultrafiltration and desalting--and up to ten types of gels and three SDS-PAGE systems were considered. The SDS-containing solution allowed a higher protein extractability than urea. Unlike urea extraction, SDS extraction seemed not to be influenced so much by the state of the sample (raw, cooked at 60 degrees C, cooked at 85 degrees C). Desalting, ultrafiltration or treatment with RNase/DNase did not improve the discriminatory power of the protein patterns. Commercial homogeneous 15% ExcelGels, especially when they were silver stained, yielded good results and afforded higher reproducibility, thus allowing a better matching of results among the laboratories participating in this collaborative study. Under the optimized technical conditions described above, all the fish species tested, either raw and cooked, yielded reproducible and discriminant species-specific protein patterns.

[Complications in 4,000 arthroscopies]. / Komplikationen bei 4,000 Arthroskopien.

From 1983 to 1989 we performed about 4,000 arthroscopies of the knee in our hospital. Iatrogen lesions of the cartilage were the most frequent complication. Other complications were edemas of the subcutaneous tissue, the necessary of arthrotomy, long operating time, subcutaneous hematomas, postoperative effusion or hemarthrosis and thrombosis. The rate of severe complications was 0.5% as described by other authors. Arthroscopy has a low rate of complications, but it isn't a method without risks.

[Arthroscopic treatment of chondropathia patellae with lateral release and motor driven instruments]. / Die arthroskopische Behandlung der Chondropathia patellae mit dem lateral release und motorgetriebenen Instrumenten.

The treatment of chondromalacia of the patella is a medical problem. There are no sure concepts of therapy. We have analysed the value of the only lateral release or shaving by motorizised instruments or the combination of both methods. After lateral release have many patients no pain but the other methods also helped many patients. The comparison of these groups is impossible because of the different structure. A prospective randomised study has to bring the answer to this question.

[Indication for and technic in arthroscopic retinaculum division]. / Indikation und Technik der arthroskopischen Retinakulumspaltung.

We reviewed 57 arthroscopic lateral release of the knee-joint during the period from January 1985 to December 1988. A high percentage of chondropathies affecting the patella and the patellofemoral joint was the mainly indication to this surgical operation. Included in the above mentioned cases were 8 traumatic patella luxations. Therefore, the necessity of strict indication and adequate postoperative therapy is emphasized.

[Arthroscopic surgery of the knee joint cartilage]. / Arthroskopische Operationen am Knorpel des Kniegelenkes.

The arthroscopic surgery of articular cartilage of the knee is beside conservative therapy and invasive surgical procedures an important enrichment in treatment of chondromalacia stage II and III. In the Department of Surgery of the Municipal Hospital of Berlin-Weissensee 3250 arthroscopic operations were performed between January 1983 and December 1988. In 227 patients chondromalacia stage II and III was found which had to be treated by arthroscopic surgery. Four different operation methods were used.

[82 arthroscopic meniscus refixations and results of follow-up]. / 82 arthroskopische Meniskusrefixationen und Nachuntersuchungsergebnisse.

In 3,126 arthroscopies of the knee 82 menisci were sutured by the inside-outside-technique. 52 patients were followed up. 42 patients had no or small disabilities. In 10 patients we performed a rearthroscopy. The suture of the meniscus includes more social and economic problems then the partial meniscectomy. It's well known, that the menisectomy induces arthrosis, and so should be tried to suture possible tears of the meniscus.

[Arthroscopy of the knee joint. Analysis of 1,200 studies]. / Die Arthroskopie des Kniegelenks. Analyse von 1,200 Untersuchungen.

Knee-joints were arthroscopically examined in 1,200 cases at Berlin-Weissensee Hospital, between January 1983 and February 1986. A very high number of patients was above 40 years of age. A high percentage of chondropathies affecting the median condyle of the femur was one of the conspicuous findings. These lesions were causatively attributed to median meniscus damage or instability of ligaments. Included in the above cases were 221 total ruptures, 72 partial ruptures of the LCA, 13 full and 32 partial ruptures of the LCP. Partial meniscectomy had to be performed on 190 patients. Cartilage surgery proved to be problematic. Long-time results are still missing. Two infections of joints were recorded among the above 1,200 knee-joint examinations by arthroscopy.

[Indications and technic of arthroscopic meniscus refixation]. / Indikation und Technik der arthroskopischen Meniskusrefixation.

Arthroscopy was applied in 792 instances to 47 knee joints, between 5 and 20 years after meniscectomy. High rates of cartilage damage, some of them accompanied by instability of the knee joint were among the most conspicuous findings. Total, subtotal, and partial meniscectomy and their effects upon the mechanism and functionality of the knee joint are discussed in some detail. An account is given of indication and arthroscopic technique for peripheral meniscus fixation.

[Arthroscopic meniscus surgery]. / Die arthroskopische Meniskuschirurgie.

Reported in this paper are 142 cases of arthroscopic meniscectomy treated in the Surgical Department of Berlin-Weissensee Hospital, between 1983 and 1985. An account is given of techniques applied to different types of rupture, and reference is made to both advantages and potential complications of arthroscopic surgery. The video technique is considered to be of great importance to endoscopic operations on the knee joint.