Effect of actinomycin D onTrypanosoma cruzi

Summary Viable metacyclic forms ofT. cruzi, Y strain, treated with an adequate dose of actinomycin D (50 g Act-D/ml/10⁷ parasites/ml for 72 h at 28° C) showed the following properties: 1) they lost their ability to replicate in culture medium, in blood and in tissues of normal mice and were no longer able to incorporate tritiated thymidine; 2) they could no penetrate into Vero cells and could not replicate inside normal macrophages; 3) they retained their immunogenicity and the ability to protect mice against a virulent infection; 4) they did not induce histological lesions as described in chronic experimental Chagas' disease.     

Hydrogen peroxide and hydroxyl radical involvement in the activation of caspase-3 in chemically induced apoptosis of HL-60 cells

Apoptosis of HL-60 cells induced by actinomycin D, H7, or daunorubicin was shown to involve the activation of caspase-3-like protease, 2 h after the addition of these drugs, based on microassay of enzyme activity by high-performance liquid chromatography. Catalase and a spin trap, N-t-butyl--phenylnitrone, which effectively inhibited the apoptosis induced by these drugs, also inhibited the activation of caspase-3-like protease. These results suggest that hydrogen peroxide and the hydroxyl radical are common mediators of caspase-3 activation caused by these chemicals, with apparently different functional mechanisms. Based on mitochondrial activity determined by oxygen consumption, complexes I, II, and IV were inhibited by actinomycin D. H7 inhibited complexes I and IV, 1 and 1.5 h respectively, after the addition of the drug to HL-60 cells. Daunorubicin inhibited complex IV, 1.5 h after the addition of the drug to HL-60 cells. Inhibition of complex IV by actinomycin D, H7, and daunorubicin were almost fully restored by the addition of cytochrome c. The release to the cytosol of cytochrome c by these drugs was also demonstrated by Western blot analysis. Addition of catalase inhibited the depression of complex IV activity induced by actinomycin D and H7. These observations indicate a direct relationship between hydrogen peroxide and the release of cytochrome c during apoptosis caused by actinomycin D, H7, and daunorubicin. Received 24 November 2000; received after revision 2 January 2001; accepted 30 January 2001     

The effect of hydrogen peroxide on the cyclin D expression in fibroblasts

Activation of mitogen-activated protein (MAP) kinase is essential for cyclin D1 expression and provides a link between mitogenic signalling and cell cycle progression. Hydrogen peroxide (H₂ O₂ ) activates MAP kinase; however, it is not known whether this leads to cyclin D expression. Sustained expression of cyclin D1 and D2 was observed when Her14 fibroblasts were incu-bated with 3 mM or higher H₂ O₂ concentrations. Similar results were obtained when cells were incubated in the presence of serum (FCS). However, the sustained expres-complex sion of cyclin D1 and D2 upon H₂ O₂ treatment was not due to the MAP kinase pathway, because MAP kinase kinase inhibitors did not inhibit cyclin D expression. Furthermore, cyclin D1 and D2 levels remained constant even after addition of a protein synthesis inhibitor, indicating that the effect of H₂ O₂ was not due to induction of protein synthesis. These results indicate that H₂ O₂ reversibly inhibits the ubiquitin-proteasome dependent degra-dation of cyclin D1 and D2, probably by transiently in-hibiting ubiquitination and/or the proteasome. Received 12 March 2001; received after revision 5 April 2001; accepted 9 April 2001     

Some new data concerning effect of actinomycin D on early chick embryos

Zusammenfassung Actinomycin D führte beim Hühnerembryo zu einer deutlichen Hemmung der Herzentwicklung nach Inkubation im Nährmedium während der Entwicklungsstadien 3 und 3⁺. Nach Vorbehandlung der Stadien 3–7 und anschliessender Kultur in actinomycin D-freiem Medium fanden sich Fehlbildungen von Herz, ZNS und Somiten, die in ihrer Häufigkeit von den betroffenen Entwicklungsphasen abhängig sind.

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This study was supported by a grant from the Rutgers University Research Council No. 07-2189.     

Binding of actinomycin D and divalent cations to lipopolysaccharides ofAgrobacterium tumefaciens as studied by fluorescence spectroscopy

Summary The binding of actinomycin D and divalent cations to lipopolysaccharides ofA. tumefaciens was studied. Fluorimetric titrations revealed 2 binding sites (low and high affinity sites) for divalent cations, and 1 high-affinity site for actinomycin D.Acknowledgment. This project was financed by the Department of Science and Technology, Govt. of India (grant No. D.O. No.11 (19)/77-SERC). To whom reprint requests should be addressed.     

Fluorescence replication banding of frog chromosomes

To identify individual chromosomes of a frog karyotype by their fluorescence banding patterns, chromosomes were stained with actinomycin D and 4,6-diamidino-2-phenylindole (DAPI) after incorporation of BrdU during the late S-phase. The chromosomes of three Rana species which were selected for this study (R. ridibunda, R. lessonae and R. japonica) showed well-defined late replication bands. The fluorescence patterns obtained were the reverse of those produced by a 4Na-EDTA Giemsa-staining technique. Fluorescence patterns of the two water frog species (R. ridibunda and R. lessonae) were similar to each other, except for the different fluorescence of the centromeric heterochromatin, which gave extremely bright signals in R. ridibunda but no signal in R. lessonae. Experiments also showed differences between the fluorescence patterns of R. lessonae chromosome 13 in the Italian and Luxembourgian populations. These results sho w that the fluorescence replication banding using actinomycin D and DAPI is very effective in identifying individual frog chromosomes and detecting their structural changes. Received 7 June 1996; received after revision 23 July 1996; accepted 21 August 1996     

Effect of actinomycin D on Trypanosoma cruzi

Viable metacyclic forms of T. cruzi, Y strain, treated with an adequate dose of actinomycin D (50 micrograms Act-D/ml/10(7) parasites/ml for 72 h at 28 degrees C) showed the following properties: 1) they lost their ability to replicate in culture medium, in blood and in tissues of normal mice and were no longer able to incorporate tritiated thymidine; 2) they could not penetrate into Vero cells and could not replicate inside normal macrophages; 3) they retained their immunogenicity and the ability to protect mice against a virulent infection; 4) they did not induce histological lesions as described in chronic experimental Chagas' disease.     

基于高光谱的柑橘树红边特征及叶绿素和LAI的监测

为从光谱信息中寻找最能反映叶面积指数（LAI）的参数并建立模型,文中以砂糖橘树与年橘树为实验对象,通过室外实验,同步测定柑橘树冠层在不同生育期的高光谱反射率、红边参数和叶片的叶绿素含量及LAI.结果表明,柑橘树冠层光谱红边具有"双峰"和"红边平台"现象,红边位置λ_red位于695～724 nm之间.红边幅值Dλ_red和红边面积S_red有"红移"和"蓝移"现象;叶面积指数与冠层光谱红边参数λ_red,Dλ_red,S_red之间显著相关.利用柑橘树冠层的五种光谱形式,分别与叶面积指数进行逐步回归相关分析并与LAI建立模型;叶片叶绿素含量与其反射光谱的λ_red,Dλ_red,S_red也显著相关.     

Signalling roles of mammalian phospholipase D1 and D2

Phospholipase D (PLD) catalyses the hydrolysis of phosphatidylcholine to generate the lipid second messenger, phosphatidate (PA) and choline. PLD activity in mammalian cells is low and is transiently stimulated upon activation by G-protein-coupled and receptor tyrosine kinase cell surface receptors. Two mammalian PLD enzymes (PLD1 and PLD2) have been cloned and their intracellular regulators identified as ARF and Rho proteins, protein kinase Cα as well as the lipid, phosphatidylinositol [4, 5] bisphosphate (PIP₂). I discuss the regulation of these enzymes by cell surface receptors, their cellular localisation and the potential function of PA as a second messenger. Evidence is presented for a role of PA in regulating the lipid kinase activity of PIP 5-kinase, an enzyme that synthesises PIP₂. A signalling role of phospholipase D via PA and indirectly via PIP₂ in regulating membrane traffic and actin dynamics is indicated by the available data. Received 25 April 2001; received after revision 15 June 2001; accepted 15 June 2001     

Effets de l'actinomycine D sur le développement normal et sur les modifications expérimentales de la morphogénèse de l'oeuf de l'oursinParacentrotus lividus

Summary In the presence of actinomycin D (20–40 µg/ml), the development of the eggs of the sea urchin,Paracentrotus lividus, is slowed from the late morula and stopped at the blastula stage. The development is immediately stopped in the blastula treated with actinomycin D (20–40 µg/ml). The inhibitory effects of actinomycin D are prevented by deoxyribonucleic acid. Actinomycin D does not exert animalizing or vegetalizing effects. However, the enhancing of vegetalizing action of lithium and the weakening of animalizing effects of zinc ions and Evans blue have been observed in the presence of actinomycin D. These observations may reflect some difference in the state of dependence of differentiation of entomesodermic and ectodermic structures towards the nucleus.     

On the synthesis of serine and homoserine samples asymmetrically labelled with tritium and deuterium in the hydroxymethylene group

Summary (1 R) [1-³H,²H₁] 3-Phenylpropanol, the key intermediate in the synthesis of (4 R) [4-³H,²H₁] D, L-homoserine and of the (4 S)-isomer, is obtained from (1 S) [1-²H₁] 3-phenylpropanol and (1 RS) [1-³H] ethanol upon incubation with yeast alcohol dehydrogenase and NAD⁺; under similar conditions 2-phenylethanol undergoes very small exchange with [1-²H₂] ethanol.     

Time reversal operations,representations of the Lorentz group,and the direction of time

A theory is usually said to be time reversible if whenever a sequence of states S₁(t₁), S₂(t₂), S₃(t₃) is possible according to that theory, then the reverse sequence of time reversed states S₃^T(t₁), S₂^T(t₂), S₁^T(t₃) is also possible according to that theory; i.e., one normally not only inverts the sequence of states, but also operates on the states with a time reversal operator T. David Albert and Paul Horwich have suggested that one should not allow such time reversal operations T on states. I will argue that time reversal operations on fundamental states should be allowed. I will furthermore argue that the form that time reversal operations take is determined by the type of fundamental geometric quantities that occur in nature and that we have good reason to believe that the fundamental geometric quantities that occur in nature correspond to irreducible representations of the Lorentz transformations. Finally, I will argue that we have good reason to believe that space-time has a temporal orientation.     

Distribution of the 1,25 dihydroxy-vitamin D3 receptor in the bursa of Fabricius of chicken

The vitamin D₃ metabolite 1,25(OH)₂D₃ is probably involved in B lymphocyte ontogeny. We therefore determined the distribution of the 1,25(OH)₂D₃ receptor in the bursa of Fabricius and spleen cells of 7-day-old chicks, by immunohistochemistry using a monoclonal antibody against the chick intestinal cell 1,25(OH)₂D₃ receptor. The bursa cells of young (7-day-old) chicks contained large amounts of receptor while the spleen cells did not. The bursa cells of older (35-day-old) chicks contained fewer receptors, but the number of receptors in the spleen increased.     

Vitamin D receptors in heart: Effects on atrial natiuretic factor

We report that receptors for vitamin D exist in distinct regions of the heart in female and male mice, predominantly in the right atrium where most of the cardial atrial natriuretic peptide (ANF) is produced. Tritiated 1,25-dihydroxyvitamin D₃ (1,25-D₃, vitamin D, soltriol) and ANF are colocalized in nuclei and cytoplasm respectively in identical cardiomyocytes. Changes of ANF tissue and blood levels under dietary deficiency and treatment with 1,25-D₃ suggest direct genomic actions of vitamin D on myoendocrine cells of the atrium for the regulation of ANF manufacture and secretion. These results were obtained by combining thaw-mount autoradiography with immunocytochemistry using tritiated 1,25-D₃ and an antibody against rat ANF. This antibody was also used in a radioimmunoassay to determine atrial natriuretic factor in plasma, atria and ventricles of normal or vitamin D-deficient mice.     

The molecular composition of a photographic fixing bath

Zusammenfassung In Thiosulfatlösungen von mäßiger Stärke bildet Silber die zwei Ionen Ag(S₂O₃) ₂ ^3– und Ag(S₂O₃) ₃ ^5– , deren Komplexkonstanten bestimmt wurden.     

Glucose-evoked recovery of hepatic thyroxine 5′-deiodinase independent of de novo protein synthesis in fasted rat

Summary The glucose-evoked recovery of Type I thyroxine 5-deiodinase activity in the hepatic microsomes of fasted rat was not inhibited by either cycloheximide, puromycin or actinomycin D during 3 h after glucose feeding; however, [³H]-leucine uptake by the liver or the hepatic microsomal fraction was significantly inhibited by cycloheximide and puromycin but not by actinomycin D. These results indicate that the glucose-evoked recovery of deiodinase activity may be independent of de novo protein synthesis.     

Opposite effect of PGE2 on cAMP levels in human adrenal medulla and pheochromocytoma

Summary PGE₂ (10^–7 M) caused increased cAMP accumulation in 5 pheochromocytomas, while in 3 human adrenal medullae PGE₂ caused a significant decrease of cAMP level on incubating slices in vitro. This finding is discussed in relation to the opposite effect of PGE₂ on catecholamine release from human medulla and pheochromocytoma slices in vitro.Acknowledgment. This paper is part of a Ph. D. thesis of Punya Boonyaviroj.Established Investigator of the Chief Scientist's Bureau, Israeli Ministry of Health.     

Influence of heavy water (D2O) on the multiplication of Adeno and Mengo virus

Zusammenfassung Der Einfluss von Deuteriumoxyd (50%) auf die Vermehrung von Mengo-Virus (kleiner RNS-Virus) und Adeno-7-Virus (ein DNS-Virus) in L-Zellen wurde untersucht. Nach 8 h im H₂O-Medium ist der Titer vom Mengo-Virus signifikant erhöht. 24 h nach Infektion sind die Titer von Mengo-Virus in H₂O und D₂O enthaltenden Medien identisch. Der Adenovirustiter in H₂O enthaltendem Medium ist 24 und 48 h nach Infektion signifikant höher als im Medium mit D₂O.     

New C26 δ-lactones from the Dufour's gland of the urticating antTetramorium aculeatum

(6R^*)-{(2S^*)-2-hydroxyheneicos-12-enyl}-5,6-dihydro-2H-pyran-2-one (1)^o is the major constituent of the secretion of freshly dissected Dufour's gland of the urticating antTetramorium aculeatum. In solution, compound 1 is slowly transformed into (1S^*, 5R^*, 7S^*)-7-(nonadec-10-enyl)-2,6-dioxabicyclo[3.3.1]nonan-3-one (2)^o on standing. The structures of compounds 1 and 2 have been established on the basis of their spectral and chemical properties. Compound 1 could be responsible for the urticating properties of the ant.^o IUPAC numbering.     

Elemental sulphur (S<Subscript>8</Subscript>) in higher plants — biogenic or anthropogenic origin

In the course of investigating lipophilic air pollutants in the epicuticular wax ofPinus sylvestris L. needles, elemental sulphur, S₈, was found in all samples. An investigation was conducted to determined the origin of this substance. No correlation between the level of S₈ in the needles and human activities in the sampling area could be found, contrary to what would have been expected of an anthropogenic compound. The internal lipids ofP. sylvestris as well as the epicuticular wax of historical herbarium material and seedlings grown in clean, filtered air, and the epicuticular wax of several other species, both gymnosperms and angiosperms, also contained S₈. Quantitation of S₈ inP. sylvestris gave levels of 7.2±2.9 μg/g wax, 3.8±1.9 μg/g internal lipid and 0.43±0.17 μg/g total needle dry weight. Almost 0.1% of the total sulphur in pine needles is S₈, and approximately half of the total S₈ is found in the wax. The results suggest that S₈ is endogenous in many higher plants. A function for S₈ as part of an antifungal defence system is possible.