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Biomedical subjects

A Schroeter

Publications and source records attributed to A Schroeter.

At least 19 recordsLinked to original sources

[Prevalence of motile salmonellae in egg-laying hens at the end of the laying period].

A total of 3504 hens of the layer-type from 122 flocks (belonging to 89 farms), each with more than 10,000 animals, were culturally examined at the time of slaughter. Of these hens, 2112 (60.3%) from 74 flocks (60.7%) were obtained from 21.3% of the laying-hen farms in a selected region of Lower Saxony in Germany. The other hens came from the remaining part of Lower Saxony and seven other German states (Brandenburg, Mecklenburg Vorpommern, North Rhine Westphalia, Schleswig Holstein, Saxony, Saxony Anhalt, and Thuringia). After arrival at the slaughter house, a random sample of 29 layers was collected from each of the flocks, and liver and spleen, as well as cecal samples, were separately cultured for each bird. Motile salmonellae could be proved in 365 (10.4%) layers from 67 flocks (54.9%). In the selected region, 48 out of 74 flocks (64.9%) and 289 out of 2112 layers (13.7%) were Salmonella-positive. However, the isolation frequency of salmonellae did not differ significantly between flocks of brown and white layers. These Salmonella (S.) isolates could be serologically assigned to 6 different serovars, namely S. enteritidis (SE), S. infantis (SI), S. livingstone (SL), S. typhimurium (ST), S. indiana (SID) and S. cerro; only one isolate of serogroup D1 was incompletely serotyped. SE was detected in 5.8% of the hens from 47.5% of the tested flocks, of which 4.6% of the animals and 32.8% of the flocks came from the selected region in Lower Saxony. The SE isolates were classified into 12 different lysotypes. In 41 out of 58 SE-positive flocks (70.7%), the isolates belonged to lysotype (lt) 4, in 12 flocks (20.7%) to lt 8, in 5 flocks (8.6%) to lt 7, and in 3 flocks (5.2%) to lt 11. A total of 190 (93.1%) out of 204 isolates of the serovar SE carried plasmids. All the plasmid-positive SE-strains harboured the serovar-specific 37 MD virulence-plasmid, nine of them (4.4%) in conjunction with a second and eight strains (3.9%) with a second and a third smaller plasmid.

Animals

[Immunization with potential Salmonella enteritidis mutants--1. Production and in vitro characterization].

Production and in vitro characterization of potential vaccine strains are the first steps leading to an efficient Salmonella Enteritidis oral live vaccine for homologous immunization of poultry. The paper presents the results of the production of adenine-amino acid auxotrophic mutants using N-methyl-N'-nitro-N-nitrosoguanidine mutagenesis. The mutant strains were characterized using the following properties: auxotrophy, stability of mutation, reversion rate, generation time, metabolic properties, serotype, motility, plasmid content, phage type, SDS-PAGE patterns, as well as cell culture adhesion and invasion. Ten S. Enteritidis double auxotrophic mutants were obtained which are stable auxotrophically and where the risk of reversion was minimal. All strains were found to be plasmid-free. 5 mutants were selected for further investigations concerning their attenuation and immunological value.

Animals

Salmonella enteritidis phage types in Germany.

In order to monitor the epidemiological situation of S. enteritidis in Germany, in 1990-91 1138 isolates from more than 180 locations in West Germany were phage typed. 1124 strains (98.8%) from all sources were typeable, belonging to 21 different phage types (PT). PT4 strains were isolated most frequently (70.8%). In addition, PT7, 25, 34 and 8 were of epidemiological relevance with incidences of 7.2 to 4.5%. The comparison of data shows that in Germany, like in other parts of Europe, PT4 predominates. This phage type is, however, infrequent in North America, where PT8 has the highest incidence.

Animals

Molecular typing methods for S. enteritidis.

The predominance of certain phage types of Salmonella enteritidis in various countries makes further epidemiological subgrouping necessary. Today this is achieved by using molecular typing methods. For various bacterial species, plasmid profiling, the pattern of outer membrane proteins and lipopolysaccharides, the fingerprinting of total genomic DNA including ribotyping, and multilocus enzyme electrophoretic typing, have proven very useful. When such methods have been applied to S. enteritidis, they revealed a homogeneous, clonal structure in contemporary PT4 isolates. Furthermore they indicate that the clone observed today emerged from a heterogeneous population before the onset of the epidemic.

Animals

[Plasmid profiles of different Salmonella serovars from poultry flocks in Germany].

151 salmonella isolates belonging to 6 serotypes and originating from diverse material collected in broiler chicken farms, hatcheries and poultry slaughteries in Northern Germany during 1984 and 1990 were investigated for their plasmid DNA content. This is the first report describing plasmids of S. saint-paul, isolated from poultry lines. The different plasmids of 22 Md, 3.1 Md, 2.4 Md and 1.1 Md molecular weight can be used for further epidemiological studies of salmonellosis in poultry. The 62 Md plasmid of S. typhimurium and the 36 Md plasmid of S. enteritidis have been found to be serovar specific. The smaller plasmids (m. w. less than 5.0 Md), found in S. virchow and S. blockley can be used for further epidemiological studies.

Animals

RelA mutation and pBR322 plasmid amplification in amino acid-starved cells of Escherichia coli.

Plasmid pBR322 is amplified following amino-acid limitation in Escherichia coli relA hosts. In relA+ hosts there was no significant amplification or a much smaller one. Plasmid amplification is due to the relA mutation; when the relA+ allele is transferred into the relA mutant CP79 this strain no longer amplifies plasmid DNA during amino acid starvation. It is concluded that ppGpp is a negative effector of plasmid replication. Amplification is temperature dependent, being maximal at 32 degrees C and negligible at 37 degrees C.

Amino Acids

Cutaneous T cell lymphoma with suppressor phenotype and function.

Cutaneous T cell lymphomas (CTCL) are typically malignancies of postthymic helper T cells which retain helper function when assayed in vitro. We have described a case clinically and morphologically consistent with CTCL, but in which the tumor cells unequivocally had a suppressor T cell phenotype and suppressor function. This case could be differentiated from other varieties of peripheral T cell lymphomas by multiple clinical and laboratory parameters. The increasingly routine use of immune phenotyping in the evaluation of malignant lymphomas will likely result in the identification of additional cases of this entity, which should be studied to assess its clinical and prognostic relationship to the more common helper cell CTCL.

Humans

Amplification of different ColE1 plasmids in an Escherichia coli relA strain.

Amino acid starved cells of an E. coli relA strain accumulate a large amount of pBR322 plasmid DNA. In this study ColE1 related plasmids of different copy number and size including a high copy number plasmid mutant of pBR322 were amplified in a relA strain of E. coli K-12 under amino acid limitation in order to determine the upper plasmid level in amino acid starved cells. In all cases we measured a 4 to 6 fold increase of the plasmid copy number in comparison to log-phase cells independent of the size, the number of origins per plasmid molecule or the copy number in log-phase cells. The plasmid copy number in amino acid starved cells varies from about 200 (pBR322-dimer) to about 2000 (high copy number plasmid pERIII-BPL4, see Boros et al. 1986). Rop+ and rop- plasmids show the same amplification rate under the used conditions.

Amino Acids

Expression of a cloned beta-glucanase gene from Bacillus amyloliquefaciens in an Escherichia coli relA strain after plasmid amplification.

Amino acid starvation of cells of the Escherichia coli relA strain, CP79, which cannot accumulate guanosine tetraphosphate (ppGpp) in response to amino acid limitation, increased the pEG1 plasmid content about 5- to 7-fold in comparison with exponentially growing cells (pEG1:pBR322 with an insertion of Bacillus amyloliquefaciens DNA coding for beta-glucanase). In contrast, no pEG1 amplification occurred in E. coli CP78, the stringently controlled counterpart, after amino acid starvation. In order to verify these results, the plasmid DNA content was monitored by measuring the expression of pEG1-encoded beta-glucanase from B. amyloliquefaciens both before and after plasmid amplification. When amino acid starved CP79 cells were given an additional dose of amino acids, a more than 10-fold increase in pEG1-encoded beta-glucanase activity (per cell mass) was measured. This increase in enzyme activity correlates with pEG1 amplification during amino acid limitation. Under comparable conditions the activity of beta-glucanase was not increased in strain CP78, which did not amplify the plasmid. We suggest that the replication of pEG1 in amino acid starved E. coli cells is somehow under negative control by ppGpp. Moreover, we found the Bacillus beta-glucanase in E. coli relA cells to be excreted into the growth medium after starvation and overexpression.

Amino Acids

Formation of extracellular neutral proteinase and the stringent response in Bacillus subtilis.

The kinetics of extracellular neutral proteinase synthesis by an isogenic stringent (IS58) and a relaxed (IS56) strain of B. subtilis were compared. The specific enzyme formation rate by the stringent strain was higher than that of the relaxed one. Norvaline addition (1 mg/mL) induced the formation of pppGpp and ppGpp, respectively, as well as the appearance of extracellular neutral proteinase activities in cultures of the stringent strain IS58 and a strain with high proteinase production (ZF-178) only. These correlations support the suggestion that (p)ppGpp are involved in the regulation processes responsible for production of extracellular neutral proteinases by B. subtilis.

Bacillus subtilis

Physiological studies on pBR 322 DNA amplification in an Escherichia coli relA strain.

Amino acid limitation leads in E. coli relA cells which cannot synthesize guanosine tetraphosphate (ppGpp) under these conditions to an amplification of pBR 322 DNA. We previously proposed that ppGpp produced in E. coli relA+ cells subjected to amino acid limitation inhibits pBR 322 DNA replication (Hecker et al. 1983). In further experiments it was established that an E. coli relA strain shows plasmid amplification during amino acid limitation (arginine, threonine, leucine or histidine) only in the presence of sufficient concentrations of phosphate, ammonia and glucose. Plasmid amplification does not occur if ammonia or phosphate is depleted. We suggest that glucose, ammonia and phosphate are needed for nucleotide and deoxynucleotide synthesis as the essential prerequisite for plasmid amplification. The activity of beta-lactamase was determined as an indicator for the expression of plasmid-encoded genes. The enzyme activity remains on a low level during plasmid amplification because of arginine exhaustion. A remarkable increase in the activity of beta-lactamase was observed after resumption of growth of relA cells containing amplified plasmid DNA. The plasmid content decreased as the cells continued to grow. We found that plasmid amplification and expression of plasmid-localized genes are opposite reactions which do not occur at the same time.

Amino Acids

Role of relA mutation in the survival of amino acid-starved Escherichia coli.

Amino acid-starved cells of Escherichia coli relA+, which contain a large number of glycogen particles, are able to survive in phosphate buffer for a longer time period than their relaxed counterparts. With regard to NH4+ starvation differences in the survival of both strains were not found. NH4+ starved cells of E. coli relA are able to synthesize glycogen but amino acid-starved cells of the relA strain are not. We suggest that the synthesis of glycogen triggered by guanosine tetraphosphate during amino acid starvation is responsible for the prolonged viability of the E. coli relA+ strain.

Amino Acids

[Synthesis of alkaline phosphatase in a stringent and a relaxed strain of Escherichia coli under amino acid and phosphate limitation].

Synthesis of alkaline phosphatase in Escherichia coli is derepressed under phosphate starvation in the stringent strain CP78 as well as in its relaxed counterpart CP79. During limitation of phosphate as well as of amino acids a decrease of enzyme activity is observed, especially in the relaxed strain, which can not produce ppGpp under this conditions. After phosphate limitation synthesis of ppGpp is not stimulated and the kinetics of RNA synthesis is similar in both strains. We suggest that ppGpp is not directly involved in the regulation of gene expression during phosphate starvation.

Alkaline Phosphatase

[3H-thymidine incorporation following isoleucine limitation in stringent and relaxed controlled strains of Escherichia coli].

Stringent and relaxed strains of E. coli subjected to isoleucine starvation were examined by follow-wing the incorporation of 3H-thymidine into chromosomal DNA. After valine treatment to trigger an isoleucine deprivation (p)ppGpp is synthesized in the stringent strain only. Remarkable differences in the morphology of the amino acid starved cells of the stringent and relaxed strains can be observed. Upon isoleucine limitation 3H-thymidine incorporation into DNA is reduced in both strains, but this inhibition is remarkably delayed in the relaxed strain. Our result show that the reduction of chromosomal DNA synthesis during amino acid limitation occurs also without ppGpp, but in the presence of ppGpp this process is accelerated.

Chloramphenicol

[Replication and expression of plasmid pBR 322 during discontinuous culture of a stringent and relaxed Escherichia coli strain].

The E. coli strains CP78 and CP79 carrying the plasmid pBR 322 display similar growth kinetics in discontinuous culture. During limitation of amino acids the stringent strain CP78 is able to synthesize guanosine-5'diphosphate-3'-diphosphate (ppGpp) and guanosine-5'-triphosphate-3' diphosphate, but the relaxed strain can not produce highly phosphorylated guanosine nucleotides. During the logarithmic phase of growth both strains contain similar amounts of plasmid DNA. During amino acid starvation plasmid DNA is amplified in the relaxed strain only, whereas in the stringent strain the plasmid content per cell remains constant. In stationary phase cells of CP78 a higher activity of plasmid-coded beta-lactamase than in CP79 cells was detected. Furthermore, remarkable differences between both strains were observed in the composition of proteins derived from the periplasmic fraction and separated by polyacrylamide gel electrophoresis. Our results might indicate a negative control of pBR 322 DNA replication by ppGpp during amino acid starvation.

Amino Acids

Replication of pBR322 DNA in stringent and relaxed strains of Escherichia coli.

Synthesis of both chromosomal and plasmid (pBR322) DNA was measured in E. coli strains differing in their relA allele (relA+:CP78; relA:CP79). It was found that upon limitation of a required amino acid or after valine treatment to trigger a stringent response synthesis of pBR322 DNA was stimulated only in the relaxed strain and was inhibited in its stringent counterpart. The results suggest that replication of plasmid DNA is negatively controlled by the relA+ allele.

Chloramphenicol

Characterization of the glucocorticoid receptor in human skin.

Because of the profound importance glucocorticoids have in dermatologic therapy, we studied the glucocorticoid receptor in human skin. A cytosol fraction was prepared from frozen skin by homogenization and centrifugation. When reacted with [3H]dexamethasone, this cytosol contained saturable, low-capacity binding. The glucocorticoid binding was stabilized by a protease inhibitor, phenylmethylsulfonylfluoride, and by sodium molybdate and was destroyed by trypsin. Sedimentation analysis of the glucocorticoid binding protein showed an 8S to 4S transition in high salt, a property of many known steroid hormone receptors. The binding was steroid specific, supporting the conclusion that this binding protein was a glucocorticoid receptor. The receptor molecule had a frictional ratio of 1.60 and a Mr of about 226,000 under low-salt conditions (0.05 M KCl) and a frictional ratio of 1.86 and a Mr of about 100,000 under high-salt conditions (0.3 M KCl) consistent with a nonglobular, elongated molecule. Isoelectric focusing showed that the receptor had 2 molecular species with isoelectric points of approximately 5.8 and 7.5. Quantitation of receptor in human skin showed 4-7 times more receptors in the epidermis and papillary dermis than in the lower dermis and nearly equal numbers in epidermis and papillary dermis. The concentration of receptors varied in different anatomic areas, with male foreskin showing the highest concentration, followed by female face, breast, and abdominal skin. Interestingly, the concentration of glucocorticoid receptors also varied with age; the highest levels were present at the extremes of life and a significantly lower level at midlife.

Aging