The European intercomparison of in-vivo monitoring laboratories: the EIVIC-2020 project.

The EIVIC project was launched in 2020, and the main goal was the organisation of a European intercomparison of in-vivo monitoring laboratories dealing with direct measurements of gamma-emitting radionuclides incorporated into the body of exposed workers. This project was organised jointly by members of EURADOS Working Group 7 on internal dosimetry (WG7), the Federal Office for Radiation Protection (BfS, Germany) and the Radioprotection and Nuclear Safety Institute (IRSN, France). The objective was to assess the implementation of individual-monitoring requirements in EU Member States on the basis of in-vivo measurements and to gain insight into the performance of in-vivo measurements using whole-body counters. In this context, a total of 41 in-vivo monitoring laboratories from 21 countries, together with JRC (EC) and IAEA participated. The results were submitted in terms of activity (Bq) of the radionuclides identified inside phantoms that were circulated to all participants. The measured data were compared with reference activity values to evaluate the corresponding bias according to the standards ISO 28218 and ISO 13528. In general, the results of the different exercises are good, and most facilities are in conformity with the criteria for the bias and z-scores in the ISO standards. Furthermore, information about technical and organisational characteristics of the participating laboratories was collected to test if they had a significant influence on the reported results.

Molecular basis for the auxin-independent phenotype of crown gall tumor tissues.

The transfer of specific Ti (tumor-inducing) plasmid sequences, the T-DNA, from Agrobacterium tumefaciens to a wide range of plants results in the formation of crown gall tumors. These tissues differ from most plant cells in that they can be grown in vitro in the absence of added phytohormones. Here, data are presented that offer an explanation for the auxin-independent phenotype of crown gall tissues. It is shown that crude cell-free extracts prepared from three bacterial species harboring pTiA6 gene 1 could convert L-tryptophan to indole-3-acetamide; control extracts lacking gene 1 could not carry out the reaction. Other reports indicate that the pTiA6 gene 2 product can convert indole-3-acetamide to indole-3-acetic acid, a naturally occurring auxin of plants. It is concluded that the auxin-independent phenotype of crown gall tissue involves the introduction of Ti plasmid sequences encoding a two-step pathway for auxin synthesis.

Mechanisms contributing to myocardial accumulation of technetium-99m stannous pyrophosphate after coronary arterial occlusion.

The relation between the accumulation of pyrophosphate and technetium-99m in myocardium with reversible and irreversible ischmic injury was studied in dogs subjected to transitory or persistent coronary arterial occlusion. Among four dogs with coronary occlusion maintained for less than 20 minutes, none had either increased MB creatine kinase (CK) (the "myocardial" CK isoenzyme) activity serum or a positive 99mTc stannous pyrophosphate image. Seven dogs with coronary occlusion maintained for 30 or more minutes had elevated serum MB CK activity, and five of the seven had positive (abnormal) images. Thus, although false negative images may occur occasionally despite myocardial damage, both increased serum MB CK and abnormal images generally accompanied prolonged coronary occlusion. In contrast, ischemia without infarction was not associated with abnormal images. Both 99mTc and 32P labeled pyrophosphate were accumulated extensively and proportionally in myocardium from zones of infarction, and uptake of both tracers was comparable although modest in isolated mitochondria. Similar results were obtained after myocardial infarction in animals with induced profound leukopenia. Thus, phagocytosis of the radiopharmaceutical agent by leukocytes migrating into the infarct is not an essential mechanism accounting for uptake. These results indicate that abnormal images reflect uptake of pyrophosphate, associated with 99mTc, by irreversibly injured myocardium rather than leukocytic infiltration involved in the inflammatory response in the heart.

When the body and appendage of the left atrium disagree: "Focal" atrial fibrillation-implications for atrial thrombus formation and risk of thromboembolism.

A case is presented of a man who had 5 hours of atrial fibrillation followed by spontaneous conversion and maintained sinus rhythm that persisted as shown by surface electrocardiography. Transesophageal echocardiography performed 24 hours after electrocardiographic conversion documented an atrial fibrillation pattern within the left atrial appendage, with a normal sinus Doppler pattern in the body of the left atrium. This apparent regional discrepancy in atrial function may partially explain the increased risk for "late" thromboembolism among patients with atrial fibrillation who appear to be successfully converted with sustained sinus rhythm.

The consequences of the Odra flood (summer 1997) for the Odra lagoon and the beaches of Usedom: what can be expected under extreme conditions?

The exceptional flood of the river Odra in July/August 1997 caused severe damage, especially on the Polish side of the Odra valley. An additional 5 km3 of water were discharged during the flood. This represents about a third of the normal annual Odra discharge of 17 km3. Large agricultural and industrial areas were submerged, as well as towns and villages. However, as regards the Odra lagoon and the beaches of the Isle of Usedom, the substances transported, such as nutrients and pollutants, did not cause much damage, due to strong dilution. Hygienic investigations (human pathogenic bacteria and viruses) showed that the water had bathing quality during the whole flood.

[Colposcopy and early changes of cervical carcinoma (author's transl)]. / Kolposkopie und Frühveränderungen des Zervixkarzinoms.

Cytology and colposcopy are well recognized methods in the early diagnosis of pre-malignant and early invasive changes of the uterine cervix. Since 1971 the University Department for Women in Leipzig has a cytology clinic. The results of repeat smears before biopsies and the follow-up examinations on 202 cone biopsies are reviewed. The incidence of negative findings decreases with increasing atypia of the squamous epithelium. The highest incidence of false negative smears was 73.3% in mild and moderate dysplasias. The more pronounced the atypia the smaller is the number of inadequate pap smears. In cases with negative cytology and colposcopic atypia the colposcopic findings must be clarified. Only the combination of colposcopy and cytology leads to optimal early diagnosis of cervical intra-epitheal neoplasia and invasive carcinoma of the cervix. The false positive cytosmears in the screening are discussed and a treatment with estrogens prior to repeat examination is recommended. Diagnostic problems arise from a discrepancy in the colposcopic and cytologic findings. Discrepancies depend on the colposcopic experience of the examiner. Improvement of the colposcopic diagnosis is obtained by directed colposcopic biopsy.

Comparison of T-DNA oncogene complements of Agrobacterium tumefaciens tumor-inducing plasmids with limited and wide host ranges.

The T-DNA oncogene complements of the limited-host-range tumor-inducing plasmid pTiAg63 and the wide-host-range plasmid pTiA6 were compared. The resulting data indicate that pTiAg63 has DNA sequences related to most of the genes encoded by the oncogene region, the TL-DNA, of pTiA6 and that these sequences are divided between two T-DNA regions, the TA-DNA, which encoded sequences related to pTiA6 genes 4 (the cytokinin independence gene) and 6a, as well as to a pTiA6 TL-DNA fragment that encoded gene 6b and a portion of gene 3, and the TB-DNA, which encoded sequences related to genes 1 and 2 (the auxin independence genes). Tumor tissues of Nicotiana rustica incited by Agrobacterium tumefaciens harboring either pTiA6 or pTiAg63 grew axenically in vitro on phytohormone-free medium. The morphologies of the tissues, however, differed; whereas those incited with pTiA6 grew as loose, friable, unorganized callus, the tumors incited by pTiAg63 grew as clumps of rootlike structures. Thus, the T-DNA oncogene complements of these plasmids were not equivalent. The results are discussed in relation to the A. tumefaciens host range.

Host range encoded by the Agrobacterium tumefaciens tumor-inducing plasmid pTiAg63 can be expanded by modification of its T-DNA oncogene complement.

Agrobacterium tumefaciens harboring pTiA6 incite unorganized tumors on Nicotiana rustica, sunflowers, carrots, and tomatoes, whereas isogenic strains of agrobacteria harboring pTiAg63 form "rooty" tumors on N. rustica and are essentially avirulent on sunflowers, carrots, and tomatoes. In this report we show that the different host range characteristics of these two plasmids were due, in part, to differences in the T-DNA oncogene complements of the plasmids. Specifically, we constructed derivatives of pTiAg63 that contained pTiA6 oncogenes 4, 6a, and 6b inserted into the TB-DNA region and found that agrobacteria harboring these plasmids could incite unorganized tumors on N. rustica, tomatoes, carrots, and the inbred sunflower line HA202R. Undefined host factors, however, also appeared to be involved in determining A. tumefaciens host range since three inbred sunflower lines, HA303B, HA89B, and HA290B, were susceptible to tumor formation by agrobacteria harboring pTiA6 but not by strains harboring pTiAg63 or the modified pTiAg63 plasmids.

[Severe iatrogenic lesions caused by Huneke's neural therapy]. / Schwere iatrogene Schädigung durch Neuraltherapie nach Huneke.

During a combined acupuncture-neural therapy-treatment on account of a gastric ulcer several lesions with vast haemorrhages in the abdominal cavity and in the retroperitoneal space on the right (lesions of the kidney) were produced. At the operation a duodenal ulcer without perforation was found. The competence of the neural therapy in its relation to the complications is discussed, the indication to the application of these forms of therapy in such cases is refused.

[Discussion of acupuncture as a therapeutic method]. / Kritik der Akupunktur als Heilbehandlungsmethode.

Only relatively few objective critical controls of the acupuncture with the help of double-blind studies face the numerous publications on the successes of this form of treatment. These examinations led to the results that the successes of acupuncture in the same way may be reproduced by means of sham acupuncture. Since a (specific) effect transgressing the placebo or suggestive effect could not be proved on any part for the application of acupuncture the medico-juridicial demands must be put as in experiments on man.

Direct analysis of RNA transcripts in electroporated carrot protoplasts.

We describe a method for direct analysis of RNA transcribed from DNA introduced into carrot cells by electroporation. Octopine synthase RNA transcribed from the plasmid p35SOcs was detected in total and poly A(+) RNA on Northern blots and in RNA protection assays. The highest level of octopine synthase transcript was detected at approximately 8 hrs post-electroporation, although RNA could still be detected after 48 hrs. This method allows detection of foreign gene expression in a plant system and bypasses the need for reporter genes.

Quantitation of chloramphenicol acetyl transferase in transgenic tobacco plants by ELISA and correlation with gene copy number.

A monoclonal antibody to chloramphenicol acetyl transferase (CAT) was used in an indirect competitive enzyme immunoassay (ELISA) for the quantitation of CAT in leaf extracts of eighteen transgenic tobacco plants containing the CAT gene fused to the cauliflower mosaic virus 35S promoter. The ELISA could be used to quantify CAT when present in extracts at 20 ng/ml. Enzymatic activity and electrophoretic mobility of CAT in these extracts was not different from CAT from Escherichia coli. Concentrations of CAT in these transgenic plants ranged from 79 to 732 ng CAT/mg protein. The average coefficient of variation among three replicate samples was 15%. All plants were sampled on two separate occasions. The CAT concentrations often varied between the two sampling dates. We determined the CAT gene copy number and the number of independently segregating loci in each plant by Southern blot analysis and progeny testing. We found no significant differences in CAT expression among all ten plants with a single CAT gene. We also found a significant correlation between CAT gene copy number and the level of CAT expressed in each plant, although plants with one gene copy sometimes had more CAT than plants with more than one gene copy. In this population, therefore, gene copy number contributed more to the variation in CAT expression than did position effects.

Epigenetic transcriptional silencing and 5-azacytidine-mediated reactivation of a complex transgene in rice.

Despite a growing number of reports indicating non-Mendelian inheritance of transgene expression in monocots, no detailed description of the structure and stability of the transgene exists for transformants generated by direct DNA-transfer techniques, making the cause for these observations difficult to determine. In this paper we describe the complex organization of Btt cryIIIA and bar transgenes in rice (Oryza sativa L.) that displayed aberrant segregation in R1 progeny. Silencing rather than rearrangement of the bar gene was implicated because the herbicide-sensitive R1 plants had a DNA hybridization profile identical to that of the resistant R0 parent and R1 siblings. Genomic DNA analysis revealed substantial methylation of the Ubi1/bar sequences in silenced plants and, to a lesser degree, in herbicide-resistant plants, suggesting that the transgene locus was potentiated for silencing. Nuclease protection and nuclear run-on assays confirmed that silencing was due to transcriptional inactivation. Treatment of R2 progeny of silenced plants with 5-azacytidine resulted in demethylation of the Ubi1 promoter and reactivation of bar gene expression, demonstrating a functional relationship for methylation in gene silencing. These findings indicate that methylation-based silencing may be frequent in cereals transformed by direct DNA protocols that insert multiple, often rearranged sequences.

Cyclophilins are encoded by a small gene family in rice.

cDNA clones were isolated and sequenced that encode two related but distinct rice cyclophilins, Cyp1 and Cyp2. The predicted amino acid sequences of each are 72% identical to human T-cell cyclophilin. Genomic DNA gel blot analysis suggests cyclophilins in rice are encoded by a small, 6-10-member gene family. Both Cyp1 and Cyp2 have seven extra amino acid residues in the N-terminal portion of the proteins that are not found in human or other non-plant cyclophilins, suggesting that this is a characteristic of plant cyclophilins. Cyp2 was expressed as 1000 nt transcripts in leaf and root tissues. Cyp1 was expressed as 800 and 900 nt transcripts. Whereas the 900 nt transcript was present in both root and leaf mRNA, the 800 nt transcript was only detectable in root mRNA. A genomic clone of Cyp2 was isolated, sequenced and shown to lack introns. A single transcriptional start site was identified 27 residues downstream of a putative TATA box. The 5' end of the transcript was shown to contain a region rich in adenyl residues (27 of 35). This region would not be conducive to secondary structure formation, which raises the possibility that Cyp2 might be preferentially translated during stress conditions.