Allelic variation in HLA-B and HLA-C sequences and the evolution of the HLA-B alleles

Several new HLA-B (B8, B51, Bw62)- and HLA-C (Cw6, Cw7)-specific genes were isolated either as genomic cosmid or cDNA clones to study the diversity of HLA antigens. The allele specificities were identified by sequence analysis in comparison with published HLA-B and -C sequences, by transfection experiments, and Southern and northern blot analysis using oligonucleotide probes. Comparison of the classical HLA-A, -B, and -C sequences reveals that allele-specific substitutions seem to be rare events. HLA-B51 codes only for one allelespecific residue: arginine at position 81 located on the 1 helix, pointing toward the antigen binding site. HLA-B8 contains an acidic substitution in amino acid position 9 on the first central sheet which might affect antigen binding capacity, perhaps in combination with the rare replacement at position 67 (F) on the ul helix. HLA-B8 shows greatest homology to HLA-Bw42, -Bw41, -B7, and-Bw60 antigens, all of which lack the conserved restriction sites Pst I at position 180 and Sac I at position 131. Both sites associated with amino acid replacements seem to be genetic markers of an evolutionary split of the HLA-B alleles, which is also observed in the leader sequences. HLA-Cw7 shows 98% sequence identity to the JY328 gene. In general, the HLA-C alleles display lower levels of variability in the highly polymorphic regions of the 1 and 2 domains, and have more distinct patterns of locus-specific residues in the transmembrane and cytoplasmic domains. Thus we propose a more recent origin for the HLA-C locus.     

Effects of nucleotides and divalent cations on phospholipase activity in Saccharomyces cerevisiae

Divalent cations activate the lysophospholipase and transacylase reactions catalyzed by the same enzymes in the yeast Saccharomyces cerevisiae. The activation was observed at neutral pH, but not at the pH optimum of lysophospholipase/transacylase, near 3.5. Adenine nucleotides, especially AMP and ADP, are strong inhibitors of the same group of enzymes. Half maximal inhibition by AMP was found at a concentration of about 20 M. The inhibition by nucleotides in low concentrations is enhanced by divalent cations.     

Expression of acetylcholinesterase during visual system development in Drosophila

As in other insects acetylcholine (ACh) and acetylcholinesterase (AChE) function in synaptic transmission in the central nervous system of Drosophila. Studies on flies mutant for AChE indicate that in addition to its synaptic function of inactivating acetylcholine, this neural enzyme is required for normal development of the nervous system (J.C. Hall, S.N. Alahiotis, D.A. Strumpf, and K. White, 1980, Genetics 96, 939-965; R.J. Greenspan, J.A. Finn, and J.C. Hall, 1980, J. Comp. Neurol. 189, 741-774). In order to understand what role AChE may play in neural development, it is necessary to know, in detail, where and when the enzyme appears. The use of monoclonal antibodies to localize AChE in the developing visual system of wild type Drosophila has yielded the novel observation that AChE appears in photoreceptor (retinula) cells 4-6 hr after they differentiate and 3 to 4 days before they are functional. Three days later the staining in the cell body of these cells is reduced. Because retinula cells have no functional connections at the time when AChE is first detected, AChE can not be performing its standard synaptic function. Subsequent to the reduction of AChE in the retinula cells, midway through the pupal stage, the enzyme accumulates rapidly in the neuropils of the optic lobes of the brain. Thus, there is a biphasic accumulation of AChE in the developing visual system with the enzyme initially being expressed in the retinula cells and accumulating later in the optic lobes.     

Resource partitioning in a guild of aphid species associated with creeping thistle Cirsium arvense

Four aphid species (Aphis fabae cirsiiacanthoidis Scop., Brachycaudus cardui (L.), Capitophorus carduinus Walker and Uroleucon cirsii (L.)) feed on the creeping thistle Cirsium arvense. They utilize different parts of their host plant and at different times. A wide niche is typical of C. carduinus and U. cirsii, whereas A. f. crisiiacanthoidis and B. cardui, show narrower but overlapping niches. Morphological features such as stylet length and body size as well as colony size and density are associated with the choice of feeding site. C. carduinus, the smallest species with the shortest stylets was able to use leaf veins and lamina, while the other species mainly used the stem and peduncles. Within this group, A. f. cirsiiacanthoidis and B. cardui are restricted to the upper part of the stem because of their short stylets, but adult U. cirsii, the species with the longest stylets, can also feed at the base of the stem.

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Räumliche und zeitliche ressourcenaufteilung in der blattlausgilde an der ackerkratzdistel cirsium arvense

Zusammenfassung An der Ackerkratzdistel leben vier Blattlausarten (Aphis fabae cirsiiacanthoidis Scop., Brachycaudus cardui (L.), Capitophorus carduinus Walker und Uroleucon cirsii (L.)), die im Verlauf der Vegetationsperiode verschiedene Strukturen ihrer Wirtspflanze nutzen. Eine breite Nische ist für U. cirsii und C. carduinus typisch, während A. f. cirsiiacanthoidis und B. cardui engere Nischen besitzen, die sich nahezu überlappen. Die Nahrungsplatzwahl wird sowohl durch morphologische Parameter wie Stilettlänge und Körpergewicht als auch durch Koloniegröe und Dispersion innerhalb der Kolonie beeinflut. Die kleinste Art, C. carduinus, die auch das kürzeste Stilett besitzt, ist in der Lage, an Blattadern und auf der Blattspreite zu saugen. Die anderen Arten bevorzugen Stengel, Seitenstengel und Blütenstiele. Innerhalb dieser Gruppe können A. f. cirsiiacanthoidis und C. carduinus wegen ihrer kürzeren Stilette nur am oberen Teil des Stengels saugen, während adulte U. cirsii aufgrund ihrer längeren Stilette auch an der Stengelbasis leben können.

     

Complete secretion of activable bovine prochymosin by genetically engineered L forms of Proteus mirabilis.

To circumvent problems encountered in the synthesis of active chymosin in a number of bacteria and fungi, a recombinant DNA L-form expression system that directed the complete secretion of fully activable prochymosin into the extracellular culture medium was developed. The expression plasmid constructions involved the in-frame fusion of prochymosin cDNA minus codons 1 to 4 to streptococcal pyrogenic exotoxin type A gene (speA') sequences, including the speA promoter, ribosomal binding site, and signal sequence and five codons of mature SpeA. Secretion of fusion prochymosin enzymatically and immunologically indistinguishable from bovine prochymosin was achieved after transformation of two stable protoplast type L-form strains derived from Proteus mirabilis. The secreted proenzyme was converted by autocatalytic processing to chymosin showing milk-clotting activity. In controlled laboratory fermentation processes, a maximum specific rate of activable prochymosin synthesis of 0.57 x 10(-3)/h was determined from the time courses of biomass dry weight and product formation. Yields as high as 40 +/- 10 micrograms/ml were obtained in the cell-free culture fluid of strain L99 carrying a naturally altered expression plasmid of increased segregational stability. The expression-secretion system described may be generally useful for production of recombinant mammalian proteins synthesized intracellularly as aberrantly folded insoluble aggregates.     

Hydrodynamic and circular dichroic analysis of mammary-derived growth inhibitor (MDGI)

The mammary-derived growth inhibitor exists in solution as a monomeric molecule with a molar mass of 14,500 +/- 400 g/mol. The largest diameter and the height of the polypeptide chain were estimated to be 3.75 +/- 0.25 nm and 2.01 +/- 0.13 nm respectively. This is in good agreement with the structurally related bovine peripheral myelin P2 protein (about 70% amino acid sequence homology). CD measurements have revealed MDGI to be a protein with about 50% beta structure and less than 20% alpha helix similarly as in fatty acid-binding proteins. Removal of endogenous long-chain fatty acid by lipidex or storage in the frozen state lead to a destabilization of the active MDGI conformation which is accompanied by a loss of its activity with regard to growth inhibition of Ehrlich Ascites cells.     

Schwermetalle in Organen und Federn von Graureihern (Ardea cinerea) und Kormoranen (Phalacrocorax carbo)

Zusammenfassung Mittels der Atomabsorptionsspektroskopie wurden Lebern, Nieren und Federn von Graureihern und von diesjährigen Kormoranen auf Cadmium und Blei untersucht.Cadmium war in Geweben der Kormorane höher konzentriert als in gleichaltrigen Graureihern (Leber 0,063 bzw. 0,039, Niere 0,113 bzw. 0.104 mg/kg Frischmasse; Federn 0,077 bzw. 0,025 mg/kg Trockenmasse). Dies könnte auf unterschiedliche Nahrung oder unterschiedliche physiologische Situation der Vögel zurückzuführen sein. Als Maximum wurden in der Niere eines Kormorans 0,375 mg/kg gemessen. Bei den Graureihern konnte in Lebern und Nieren eine Altersakkumulation erkannt werden. Blei war in den Geweben beider Arten unterschiedlich hoch konzentriert. Kormorane enthielten in Lebern 0,124 und in Nieren 0,147 mg/kg Frischmasse sowie in Federn 0,442 mg/kg Trockenmasse, Graureiher 0,110, 0,157 bzw. 0,739 mg/kg. Die höchsten Konzentrationen wurden in Federn gemessen und betrugen bei einem Graureiher 4,525 mg/kg. Eine Altersakkumulation konnte bei Graureihern in Lebern nachgewiesen werden. Zusammenhänge zwischen Feder- und Organbelastungen wurden nur bei diesjährigen Graureiher- erkannt: Federcadmium und Lebercadmium korrelierten positiv miteinander.

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Heavy metals in tissue and feathers of Grey Herons (Ardea cinerea) and Cormorants (Phalacrocorax carbo sinensis)

Summary The livers, kidneys and feathers of 40 Grey Herons and 20 Cormorants were investigated by AAS to determine the extent of contamination with cadmium and lead. The birds had been shot in autumn 1979–1986 in Schleswig-Holstein. All results are given in mg/kg (ppm) wet mass (liver, kidney) or dry weight (feather). On the average, the contamination by cadmium in the livers (0.063), kidneys (0.113) and feathers (0.077) was higher in Cormorants than in Grey Herons (0.039, 0.104 and 0.025). These differences may be the result of different food or physiology. The livers and kidneys of young Grey Herons contained less cadmium than the tissue of older birds. The feathers of male Grey Herons contained more cadmium than feathers of females do. This was also the case in kidneys of male Cormorants as compared to the females. Cadmium values in livers and kidneys were positive correlated. No general tendency was detectable in regards to lead contamination. The results concerning Cormorants were 0.124 (liver), 0.147 (kidney), 0.442 (feather) and concerning Grey Herons 0.110, 0.157 and 0.739. The livers of old Grey Herons were more highly contaminated by lead than livers of young ones. The feathers of male Herons contained two times more lead than the feathers of females do.In general, a concentration of heavy metals in feathers does not indicate a contamination of livers or kidneys. A correlation between the values in feathers and tissue was evident only in one case, where a positive correlation was found between feather- and liver cadmium in young male Grey Herons.

     

Core substructure in Mastigocladus laminosus phycobilisomes

A native high molecular complex (M_r 850000) containing about 50% of the allphycocyanin of the phycobilisome but lacking allophycocyanin B was separated from isolated phycobilisomes by gel electrophoresis. It was designated AP_CM since the large linker polypeptide L_CM was exclusively localized in this complex. The complex exhibited a ?196°C fluorescence emission maximum at 673 nm (671 nm at 25°C). In addition, a core complex (designated AP_C, M_r≥1000000) consisting of both AP_CM and AP 680 was isolated by combined gel filtration and linear gradient centrifugation. At 25°C this complex showed dual emission peaks at 670 and 680 nm demonstrating functional independence of the terminal emitters. A complex similar to AP_CM can be isolated from phycobilisomes of Anabaena variabilis. This is evidence that AP_CM is the constitutive center of the tricylindrical core of hemidiscoidal cyanobacterial phycobilisomes. Two models summarizing the structural and functional consequences of the results are presented in the discussion.     

Heterozygous expression of X-linked chondrodysplasia punctata

Summary Two females showing partial expression of X-linked chondrodysplasia punctata were identified in a family. Bone dysplasia was caused by an aberrant X chromosome that had an inverse duplication of the segment Xp21.2–Xp22.2 and a deletion of Xp22.3-Xpter. To characterise the aberrant X chromosome, dosage blots were performed on genomic DNA from a carrier using a number of X-linked probes. Anonymous sequences from Xp21.2–Xp22.2 to which probes D2, 99.61, C7, pERT87-15, and 754 bind were duplicated on the aberrant X chromosome. The proposita was heterozygous for all these markers. Dosage blots also showed that the loci for steroid sulfatase and the cell surface antigen 12E7 (MIC2) were deleted as expected from the cytogenetic results. Mouse human cell hybrids were constructed that retained the normal X in the active state. Analysis of these hybrid clones for the markers from Xp21.2–Xp22.2 revealed that all the alleles of the informative markers, present in a single dosage in the genomic DNA, were carried on the normal X chromosome of the proposita. The duplicated X chromosome therefore had two identical alleles, indicating that the aberration resulted from an intrachromosomal rearrangement.