Biochemical characterization of chromatin fractions isolated from induced and uninduced friend erythroleukemia cells

Summary Chromatin fractions from Friend erythroleukemia cells after induction of differentiation by dimethylsulfoxide (DMSO) were compared in their biochemical characteristics to fractions from uninduced cells. Fractions were prepared by extracting chromatin from nuclei after mild micrococcal nuclease treatment with increasing concentrations of NaCl according to Sanders [1]. This procedure has been found to release chromatin containing hyperacetylated histones preferentially [2]. The fractions obtained by this procedure were analysed in respect to the amount of chromatin released, the amount of histone H1, the degree of acetylation of histone H4, the presence of non-histone proteins and the concentration of transcribed and non-transcribed sequences. It was found that the fractions differ in the amount of histone H1 present, in several non-histone proteins and in the acetylation of histonie H4, regardless whether induced or uninduced cells were analysed. The distribution of transcribed sequences versus non-transcribed sequences among the fractions was the same, demonstrating that this fractionation procedure, although leading to fractions with biochemical differences, is not able to discriminate functional states of chromatin and that the biochemical characteristics of the fractions may be common to both, active as well as inactive states of chromatin.     

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.     

Sporomusa malonica sp. nov., a homoacetogenic bacterium growing by decarboxylation of malonate or succinate

A new strictly anaerobic bacterium was isolated from an enrichment culture with glutarate as sole substrate and freshwater sediment as inoculum, however, glutarate was not metabolized by the pure culture. The isolate was a mesophilic, spore-forming, Gram-negative, motile curved rod. It fermented various organic acids, alcohols, fructose, acetoin, and H₂/CO₂ to acetate, usually as the only product. Other acids were fermented to acetate and propionate or acetate and butyrate. Succinate and malonate were decarboxylated to propionate or acetate, respectively, and served as sole sources of carbon and energy for growth. No inorganic electron acceptors except CO₂ were reduced. Yeast extract (0.05% w/v) was required for growth. Small amounts of cytochrome b were detected in membrane fractions. The guanine-plus-cytosine content of the DNA was 44.1±2 mol%. The isolate is described as a new species of the genus Sporomusa, S. malonica.     

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.

     

Effects of serum type on growth and permeability properties of cultured endothelial cells

Serum is frequently added to defined basal media as a source of certain nutrients and macromolecular growth factors essential for cell growth. The many different sera commercially available may not be equally suitable for all cell types. The effects of four sera, fetal bovine serum (FBS), calf bovine serum (CS), equine serum (ES-1), and plasma-derived equine serum (ES-2), on growth and permeability properties of cultured porcine endothelial cells were determined. The rate of DNA synthesis, measured as [3H]thymidine incorporation, reached a peak at around 24 h, regardless of serum type, and was most marked with ES-1- or ES-2-treated cells. However, when estimated by total DNA, FBS, CS, or ES-1 treatment resulted in greater cell proliferation than ES-2. Based on protein synthetic rate and total cell protein, both FBS and CS appeared to be most growth supporting. At 72 h after cell plating, albumin passage across cultured endothelial monolayers was elevated in ES-1- and ES-2-treated cells compared with FBS- or CS-treated cells. "Leaky" cell monolayers were most marked with ES-1-treated cells. Cells grown in ES-2- and particularly in ES-1-enriched media were larger and more spindle-shaped compared with the typical cobblestone appearance of cells cultured in media enriched with either FBS or CS. These data suggest that CS, but not ES-1 or ES-2, is an excellent substitute for FBS to support desirable growth properties of macrovascular endothelial cells in culture.     

Noncovalent modulation by ATP of the acyl transfer from acyl-glyceraldehyde-3-phosphate dehydrogenase to phosphate

The effect of ATP on the formation, spectral properties, and reactions of [beta-(2-furyl)acryloyl]glyceraldehyde-3-phosphate dehydrogenase (FA-GPDH) has been investigated. The chromophoric FA-GPDH has the advantage of providing spectrophotometric signals of the interaction of acyl enzyme with nucleotides and dinucleotides. The results are consistent with the exclusive existence of two acyl-enzyme conformations previously inferred from the interaction of the acyl enzyme with NAD+ and NADH. ATP interaction stabilizes a conformation different from that stabilized by NAD+. The inhibitory effects of ATP on these reactions are consistent with the reported inhibitory effect of ATP on the steady-state reaction with the true substrate. The physiological significance of these results to the regulation of glycolysis, via the ligand-dependent fate of 3-phosphoglycerol-GPDH, is discussed.     

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.

     

New motile anaerobic bacteria growing by succinate decarboxylation to propionate

Three strains of new anaerobic, gram-negative bacteria which grew with succinate as sole source of carbon and energy were isolated from anoxic marine and freshwater mud samples. Cells of the three strains were small, non-spore-forming, motile rods or spirilla. The guanine-plus-cytosine content of the DNA of strain US2 was 52.6±1.0 mol%, of strain Ft2 63.5±1.4 mol%, and of strain Ft1 62.6±1.0 mol%. Succinate was fermented stoichiometrically to propionate and carbon dioxide. The growth yields were 1.2–2.6 g dry cell mass per mol succinate degraded. Strains US2 and Ft2 required 0.05% w/v yeast extract in addition to succinate for reproducible growth. Optimal growth occurred at 30°–37°C and pH 6.8–8.0. Addition of acetate as cosubstrate did not stimulate growth with any strain. Strain Ft2 grew only under strictly anaerobic conditions, whereas strains US2 and Ft1 tolerated oxygen up to 20% in the headspace. Strains US2 and Ft2 grew only with succinate. Strain Ft1 also converted fumarate, aspartate, and sugars to propionate and acetate. This strain also oxidized propionate with nitrate to acetate. Very low amounts of a c-type cytochrome were detected in propionate plus nitrate- or glucose-grown cells of this strain (0.4 g x g protein^-1). Moderate activities of avidin-sensitive methylmalonyl-CoA decarboxylase were found in cell-free extracts of all strains.     

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.