A gene cluster for amylovoran synthesis in Erwinia amylovora: characterization and relationship to cps genes in Erwinia stewartii

A large ams gene cluster required for production of the acidic extracellular polysaccharide (EPS) amylovoran by the fire blight pathogen Erwinia amylovora was cloned. Tn5 mutagenesis and gene replacement were used to construct chromosomal ams mutants. Five complementation groups, essential for amylovoran synthesis and virulence in E. amylovora, were identified and designated amsA-E. The ams gene cluster is about 7 kb in size and functionally equivalent to the cps gene cluster involved in EPS synthesis by the related pathogen Erwinia stewartii. Mucoidy and virulence were restored to E. stewartii mutants in four cps complementation groups by the cloned E. amylovora ams genes. Conversely, the E. stewartii cps gene cluster was able to complement mutations in E. amylovora ams genes. Correspondence was found between the amsA-E complementation groups and the cpsB-D region, but the arrangement of the genes appears to be different. EPS production and virulence were also restored to E. amylovora amsE and E. stewartii cpsD mutants by clones containing the Rhizobium meliloti exoA gene.     

Polymorphism of the tumor necrosis factor beta gene in systemic lupus erythematosus: TNFB-MHC haplotypes

We investigated the Nco I restriction fragment length polymorphism (RFLP) of the tumor necrosis factor beta (TNFB) gene in 173 patients with systemic lupus erythematosus (SLE), 192 unrelated healthy controls, and eleven panel families, all of German origin. The phenotype frequency of the TNFB*1 allele was significantly increased in patients compared to controls (63.6% vs 47.1%, RR = 1.96, p <0.002). The results of a two-point haplotype statistical analysis between TNFB and HLA alleles show that there is linkage disequilibrium between TNFB*1 and HLA-A1, Cw7, B8, DR3, DQ2, and C4A DE. The frequency of TNFB*1 was compared in SLE patients and controls in the presence or absence of each of these alleles. TNFB*1 is increased in patients over controls only in the presence of the mentioned alleles. Therefore, the whole haplotype A1, Cw7, B8, TNFB*1, C4A DE, DR3, DQ2 is increased in patients and it cannot be determined which of the genes carried by this haplotype is responsible for the susceptibility to SLE. In addition, two-locus associations were analyzed in 192 unrelated healthy controls for TNFB and class I alleles typed by serology, and for TNFB and class II alleles typed by polymerase chain reaction/oligonucleotide probes. We found positive linkage disequilibrium between TNFB*1 and the following alleles: HLA-A24, HLA-B8, DRB1*0301, DRB1*1104, DRB1*1302, DQA1*0501, DQB1*0201, DQB1*0604, and DPB1*0101. TNFB*2 is associated with HLA-B7, DRB1*1501, and DQB1*0602.This study was supported by grants from the Federal Ministry of Research and Technology (BMFT/DFVLR, 01 VM 8608/9), the German Academic Exchange Service (DAAD, 322/501/014/0), and SFB (217).This work is part of the doctoral thesis of M. P. Bettinotti.     

Polymorphism of the DQA1 promoter region (QAP) and DRB1, QAP,DQA1, DQB1 haplotypes in systemic lupus erythematosus

We have investigated the DNA polymorphism for the DQA1 promoter region (QAP) and HLA-class II DRB1, DQA1, and DQB1 genes in 178 central European patients with Systemic lupus erythematosus (SLE) using polymerase chain reaction and Dig-ddUTP labeled oligonucleotides. Increased frequencies of DRB1*02 and *03 are confirmed by DNA typing. In addition, the frequencies of DQA1*0501, *0102 and DQB1*0201, *0602 alleles are increased in the patients as compared to controls. The strongest association to SLE is found with DRB1*03 and DQB1*0201 alleles (p<10^–7, p corr. <10^–5 and p<10^–6, p corr. <10^–4, respectively). By investigating the DQA1 promoter region in the SLE patients we have detected nine different QAP variants. Increased frequencies of QAP1.2 and QAP4.1 are observed in patients as compared to controls (p <0.05, p corr. = n. s.). Analysis of linkage disquilibria demonstrates a very strong association between QAP variants and DQA1, DRB1 alleles. Certain QAP variants are completely associated with DQA1 and DRB1 alleles, whereas others can combine with different DQA1 and DRB1 alleles. All DRB1*02-positive patients and controls carry QAP1.2, and all DRB1*03-positive patients and controls carry QAP4.1. Conversely, the QAP1.2 variant appears only in DRB1*02 haplotypes, while the QAP4.1 variant can be observed in DRB1*03, *11, and *1303 haplotypes. Based on the strong linkage disequilibria between DRB1-DQA1-DQB1 genes and between DRB1-QAP-DQA1, we have deduced the four-point haplotypes for DRB1-QAP-DQA1-DQB1 in patients and controls. Two haplotypes DRB1*02-QAP1.2-DQA1*0102-DQB1*0602-and DRB1*03-QAP4.1-DQA1*0501-DQB1*0201 are significantly increased in patient as compared to controls (p<0.01, p corr. = n.s., RR = 1.8 and p <10^–7, p corr. <10^–5, RR = 3.1, respectively). The analysis of relative risks attributed to the various alleles of QAP, DQA1, and DQB1 as well as the investigation of the deduced DRB1-QAP-DQA1-DQB1 haplotypes leads to the conclusion that QAP4.1 and DQA1*0501 on the DR3 haplotypes are probably not involved in SLE susceptibility. There is no evidence for the involvement of DQ2 / dimers coded in transposition. Thus, susceptibility to SLE is on the DR3 haplotype most probably localized at DRB1 or telomeric of DRB1, while for the DR2 haplotype such orientation cannot be given. SLE study group members: M. Baur, A. Corvetta, H. Ehrfeld, J. Frey, J. R. Kalden, F. Krapf, B. Lang, G. G. Lange, K. Pirner, C. Rittner, E. Röther, P. Schneider, H. P. Seelig, S. Seuchter, W. Stangel, C. Specker, P. Späth, H. Deicher. Correspondence to: Z. Yao.     

Expression of heat shock proteins during development of barley

Barley heat shock proteins have been cloned, characterized by hybrid release translation and sequenced. Clones coding for proteins of 17, 18, 30, 32 and 70 kDa have been obtained. Out of these the 32 and 30 kDa proteins have been characterized as precursors to plastidic proteins of 26 kDa by posttranslational transport and by cDNA sequencing. The coding regions of these two transcribed genes are highly homologous. Accumulation of the plastid HSP as well as of HSP 70 as well as their corresponding mRNAs has been studied in 2- to 6-day old seedlings and in the 7-day old barley leaf. The mRNA for all investigated proteins were only found after a heat shock; the mRNA levels increase towards the tip of the leaf and with development. Furthermore, under the conditions used the mRNAs for all investigated heat shock proteins accumulate in parallel. Unexpectedly, both proteins, HSP 70 and HSP 26, are found by western blotting in the 2-day old control plants in the absence of any inducing heat shock. At later stages of development and in the leaf gradient only immunoreactivity with HSP 70 was observed. In contrast to the levels of their mRNAs the highest levels of HSP 30–26 and 70 have been observed in the basal segments indicating that translational control plays a role during HSP expression. Under severe heat shock a protein of 30 kDa is induced whose identity is not known but which reacts with the antibody to HSP 30–26 and might represent the accumulating precursors of the plastidic proteins.     

F?nge von Habichten (Accipiter gentilis) im Wurzacher Ried: Kritische Fragen zu einem beh?rdlich genehmigten Wiedereinbürgerungsprojekt

Zusammenfassung Während des unter der Trägerschaft des Landesjagdverbandes Baden-Württemberg e. V. seit 1978 laufenden Projektes zur Wiedereinbürgerung des Birkhuhnes im Wurzacher Ried wurde behördlicherseits der Wegfang von Habichten unter bestimmten Auflagen genehmigt. Nach Beringung mußten die Vögel wieder freigelassen werden. Eine vergleichende Untersuchung der von den Projektbetreibern veröffentlichten Berichte und der Beringungsdokumente ergab erhebliche Ungereimtheiten. Die aus den Berichten der Projektbetreiber zu entnehmende Zahl von 168 gefangenen, angeblich beringten und angeblich wieder freigelassenen Habichten steht der Zahl von 98 in den Beringungslisten dokumentierten Vögel gegenüber. Bereits ein Vergleich zwischen der Wiederfundrate der dokumentierten Vögel (1 %) mit der Fänglingfundrate der Vogelwarte Radolfzell (13 %) ergab einen hochsignifikanten Unterschied. Auch der Vergleich mit der Fundrate (15 %) einer anderen, parallel laufenden Verfrachtungsstudie zeigte, daß die Wiederfundrate der Wurzacher Habichte hochsignifikant geringer ist. Die Darstellung der Projektbetreiber, wonach 1978–1981 insgesamt 90 Habichte durch die Landesanstalt für Umweltschutz verfrachtet worden sein sollen, wurde durch die Landesanstalt nicht bestätigt. Die Landesanstalt bestätigte nur fünf verfrachtete Habichte aus dem Wurzacher Projekt. Obwohl der Habichtfang im März nicht erlaubt war, haben die Projektbetreiber insgesamt vier Habichte im März gefangen und zumindest einen (nach eigenen Angaben jedoch alle!) auch noch verfrachtet. Vermutlich im Auftrag von Geflügelzüchtern wurden sieben Habichte, darunter vier Altvögel, innerhalb eines kleinen Gebietes in einer Entfernung von 13–18,5 km vom Wurzacher Ried gefangen. So besteht begründeter Verdacht, daß ein beträchtlicher Teil der im oder um das Wurzacher Ried gefangenen Habichte nicht wieder freigelassen, sondern getötet wurde. Eine Reihe der in den Projektberichten enthaltenen Angaben lassen zudem ganz erhebliche Zweifel an der Fachkompetenz der Projektbetreiber aufkommen.

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Goshawk (Accipiter gentilis) trapping in the nature reserve Wurzacher Ried: critical questions on a Black Grouse (Tetrao tetrix) reintroduction project

Summary A Black Grouse reintroduction project was established in 1978 in the nature reserve Wurzacher Ried in southwestern Germany. Since the beginning of this project the staff had a licence to catch, ring and to release but not to kill goshawks. A comparison of the very low Euring-recovery-rate of 1 percent from this project with the Euring-recovery-rate of the Vogelwarte Radolfzell (13 %) showed a significant difference (p=0.00017). In contrast, we found no difference between the recovery-rate (15 %) of a study with almost similar design conducted by the University of Tübingen (Schmidt-Koenig 1982) only 100 km apart from the Wurzacher Ried and the Euring-recovery-rate of the Vogelwarte Radolfzell (p=0.624). The recovery-rate of the reintroduction project differed, however, significantly from the recovery-rate ofSchmidt-Koenig (p=0.006). Furthermore, we found serious inconsistencies between the project reports and the ringing documents concerning this project. Our results indicate, that most of the goshawks caught in the project Wurzacher Ried may not have been released but illegally killed by the project staff.

     

Short-term photosynthesis measurements predict leaf carbon balance in tropical rain-forest canopy plants

Diel (24 h) courses of net CO₂ exchange of leaves were determined in eight species of tropical rainforest plants on Barro Colorado Island, Panama, during 1990 and 1991. The species included three canopy trees, one liana, two epiphytes and one hemiepiphyte. One of the species studied was growing in a rain-forest gap. Daily carbon gain varied considerably across species, leaf age, and season. The analysis of data for all plants from 64 complete day/night cycles revealed a linear relationship between the diurnal carbon gain and the maximum rate of net CO₂ uptake, A_max. Nocturnal net carbon loss was about 10% of diurnal carbon gain and was positively related to A_max. We conclude that short-term measurements of light-saturated photosynthesis, performed at periodic intervals throughout the season, allow the annual leaf carbon balance in these rain-forest plants to be predicted.     

Dynamic properties of cortical evoked (10 Hz) oscillations: theory and experiment

Experiments probed the dynamic properties of stimulus-evoked (10 Hz) oscillations in somatosensory cortex of anesthetized rats. Experimental paradigms and statistical time series analysis were based on theoretical ideas from a dynamic approach to temporal patterns of neuronal activity. From the results of a double-stimulus paradigm we conclude that the neuronal response contains two components with different dynamics and different coupling to the stimulus. Based on this result a quantitative dynamic model is derived, making use of normal form theory for bifurcating vector fields. The variables used are abstract, but measurable, dynamic components. The model parameters capture the dynamic properties of neuronal response and are related to experimental results. A structural interpretation of the model can be given in terms of the collective dynamics of neuronal groups, their mutual interaction, and their coupling to peripheral stimuli. The model predicts the stimulusdependent lifetime of the oscillations as observed in experiment. We show that this prediction relies on the basic concept of dynamic bistability and does not depend on the modeling details.     

Immunophenotyping of mitotic cells from long-term cultures of chorionic villi

Chromosomal mosaicism in chorionic villus samples (CVS) may arise from different sources, such as clonal diversity within the chorionic tissue or contamination with maternal cells. To determine the origin of karyotyped cells, we compared the immunocytochemical features of mitotic cells in CVS long-term cultures with histological sections of their tissue of origin, i.e. chorionic villi. Immunolabelling of intermediate filaments specific for epithelial cells (cytokeratin) and mesenchymal cells (vimentin) established that mitoses yielded from CVS long-term cultures indeed stem from independently growing clones derived from both the epithelial and mesenchymal parts of the chorionic villi. Thus, mosaicism in CVS cultures may reflect true genetic heterogeneity within the biopsy. However, epithelial chorionic cells undergo in vitro metaplasia leading to co-expression of cytokeratins and vimentin. Fetal-specific immune markers (-HCG and SP1-glycoprotein) are not reliably expressed in CVS cell culture.