Enhanced inhibition of RNA synthesis by amanitins in in vitro cultured cells.

The inhibition of RNA synthesis by alpha-amanitin on in vitro cultured cells is very slow. The action of various analogueoreover pretreatment of cell cultures with DEAE-dextran greatly enhanced the effect of beta-amanitin.     

Production of antibodies to amanitins as the basis for their radioimmunoassay.

The production of antibodies against amanitins is described. By means of these antibodies, a radioimmunoassay was developed which allows detection of as little as 0.5 ng of amanitins in 1 ml of serum. By this method, the clearance of alpha-amanitin from the blood of poisoned mice was measured.     

Influence of agents that act on DNA and RNA synthesis on the activity of poly(ADP-Rib) polymerase.

The activity of poly(ADP-Rib) polymerase is enhanced in the presence of spermine and spermidine. Among the adenosine-like antibiotics tested, only formycin B and showdomycin cause an inhibition of the enzyme, which is competitive to NAD. The activity of poly(ADP-Rib) polymerase is not reduced by rifamycin, alpha-amanitin and 2-phenylethanol.     

α-Amanitin: Inactivation by bovine lactoperoxidase

Summary The principle amatoxin, -amanitin, is found to be extremely sensitive toward lactoperoxidase catalyzed degradation, rather than iodination, of the indole nucleus. Extensive attenuation of inhibitor potency against eukaryotic DNA-dependent RNA polymerase II accompanies the treatment of -amanitin with lactoperoxidase, iodide and hydrogen peroxide.This work was supported in part by NIH grant 22147 and a grant from the University of Illinois.     

Fifty years of amanitin.

Pharmacokinetic studies have provided new insights into human Amanita poisoning, but it appears to be impossible to treat this intoxication by immunotherapy. New synthetic analogs have revealed structure-activity relationships that were unknown so far. The main toxin, alpha-amanitin, is in constant use as a tool in molecular biology and in biological research. First experiments have been reported in which amanitin bound to polymers could be internalized into tumor cells via a receptor-mediated endocytosis.     

Self-labeling of human polymorphonuclear leucocyte myeloperoxidase with 125iodine.

In order to obtain a radioimmunoassay (RIA) technique for the measurement of human plasma myeloperoxidase (MPO), we purified the enzyme from polymorphonuclear granulocytes (neutrophils), and compared three methods of labeling it with 125Iodine:chloramine T, lactoperoxidase, and an original technique of 'self labeling' based on the ability of the enzyme to oxidize and bind 125I in the presence of H2O2. The chloramine T technique produced a degraded protein, as well shown by a high non-specific binding of tracer to antibody. The lactoperoxidase technique did not succeed in labeling MPO with an adequate specific activity. In contrast, the self-labeling method gave a stable tracer with a specific activity of 23 microCi/micrograms MPO (85 MBq), a satisfactory level of immunoreactivity, and a low-specific binding (less than or equal to 3%). After labeling, purification of tracer was achieved by gel filtration chromatography in phosphate buffer (0.05 M; pH7) to which 0.1% poly-L-lysine was added. The labeled molecule remained stable for 40 days and could be used for RIA with a polyclonal antibody raised in rabbits.     

High salt effect on RNA synthesis in Krebs ascites tumor cells

The incubation of Krebs ascites tumor cells in medium with a high salt concentration resulted in a partial inhibition of nuclear RNA synthesis. The residual RNA polymerase activity in such nuclei was only slightly inhibited by low concentrations (50 nM) of alpha-amanitin. This finding suggested an inhibition of RNA polymerase II activity under conditions of medium hypertonicity. Indeed, RNA polymerase II, isolated from the nuclei of cells exposed to hypertonicity, revealed only half of the control activity. On the other hand, RNA polymerase I was not affected by hypertonicity. Moreover, chromatin fractions isolated from cells incubated in hypertonic or isotonic medium were equally template-active in RNA synthesis.     

Self-labeling of human polymorphonuclear leucocyte myeloperoxidase with125iodine

In order to obtain a radioimmunoassay (RIA) technique for the measurement of human plasma myeloperoxidase (MPO), we purified the enzyme from polymorphonuclear granulocytes (neutrophils), and compared three methods of labeling it with¹²⁵Iodine: chloramine T, lactoperoxidase, and an original technique of self labeling based on the ability of the enzyme to oxidize and bind¹²⁵I in the presence of H₂O₂. The chloramine T technique produced a degraded protein, as well shown by a high non-specific binding of tracer to antibody. The lactoperoxidase technique did not succeed in labeling MPO with an adequate specific activity. In contrast, the self-labeling method gave a stable tracer with a specific activity of 23 Ci/gmg MPO (85 MBq), a satisfactory level of immunoreactivity, and a low-specific binding (3%). After labeling, purification of tracer was achieved by gel filtration chromatography in phosphate buffer (0.05 M; pH7) to which 0.1% poly-L-lysine was added. The labeled molecule remained stable for 40 days and could be used for RIA with a polyclonal antibody raised in rabbits.     

Melatonin,clock genes and mitochondria in sepsis

After the characterization of the central pacemaker in the suprachiasmatic nucleus, the expression of clock genes was identified in several peripheral tissues including the immune system. The hierarchical control from the central clock to peripheral clocks extends to other functions including endocrine, metabolic, immune, and mitochondrial responses. Increasing evidence links the disruption of the clock genes expression with multiple diseases and aging. Chronodisruption is associated with alterations of the immune system, immunosenescence, impairment of energy metabolism, and reduction of pineal and extrapineal melatonin production. Regarding sepsis, a condition coursing with an exaggerated response of innate immunity, experimental and clinical data showed an alteration of circadian rhythms that reflects the loss of the normal oscillation of the clock. Moreover, recent data point to that some mediators of the immune system affects the normal function of the clock. Under specific conditions, this control disappears reactivating the immune response. So, it seems that clock gene disruption favors the innate immune response, which in turn induces the expression of proinflammatory mediators, causing a further alteration of the clock. Here, the clock control of the mitochondrial function turns off, leading to a bioenergetic decay and formation of reactive oxygen species that, in turn, activate the inflammasome. This arm of the innate immunity is responsible for the huge increase of interleukin-1β and entrance into a vicious cycle that could lead to the death of the patient. The broken clock is recovered by melatonin administration, that is accompanied by the normalization of the innate immunity and mitochondrial homeostasis. Thus, this review emphasizes the connection between clock genes, innate immunity and mitochondria in health and sepsis, and the role of melatonin to maintain clock homeostasis.     

The Chemistry of the Universe: Historical Roots of Modern Cosmochemistry

During the second half of the twentieth century, the domain of geochemistry has greatly expanded and the field is today often seen as a branch of an extended chemistry of the Earth, called cosmochemistry. This paper is a historical introduction to cosmochemistry in which the wider cosmic aspects are surveyed up to about 1915, when nuclear physics changed the scene. These wider aspects or themes include, firstly, the attempts to determine the relative abundances of the elements, secondly, the extension of geochemistry to include physical geochemistry, thirdly, the study of meteorites and, fourthly, the spectroscopic study of the stars within the astrochemical tradition. Because of the lack of reliable data, a great deal of the protocosmochemistry described in the present paper was speculative. Nonetheless, by 1915 the contours of the cosmochemistry of the future were just visible and the developments here singled out can thus be seen as belonging to the prehistory of modern cosmochemistry.     

Aristotle, Copernicus, Bruno: centrality, the principle of movement and the extension of the Universe

This paper¹ studies the different conceptions of both centrality and the principle or starting point of motion in the Universe held by Aristotle and later on by Copernicanism until Kepler and Bruno. According to Aristotle, the true centre of the Universe is the sphere of the fixed stars. This is also the starting point of motion. From this point of view, the diurnal motion is the fundamental one. Our analysis gives pride of place to De caelo II, 10, a chapter of Aristotle’s text which curiously allows an ‘Alpetragian’ reading of the transmission of motion.In Copernicus and the Copernicans, natural centrality is identified with the geometrical centre and, therefore, the Sun is acknowledged as the body through which the Deity acts on the world and it also plays the role of the principle and starting point of cosmic motion. This motion, however, is no longer diurnal motion, but the annual periodical motion of the planets. Within this context, we pose the question of to what extent it is possible to think that, before Kepler, there is a tacit attribution of a dynamic or motive role to the Sun by Copernicus, Rheticus, and Digges.For Bruno, since the Universe is infinite and homogeneous and the relationship of the Deity with it is one of indifferent presence everywhere, the Universe has no absolute centre, for any point is a centre. By the same token, there is no place that enjoys the prerogative of being—as being the seat of God—the motionless principle and starting point of motion.     

Galanin – 25 years with a multitalented neuropeptide

The skin, the largest organ of the body, functions as a barrier between the body proper and the external environment, as it is constantly exposed to noxious stressors. During the last few years, the concept of an interactive network involving cutaneous nerves, the neuroendocrine axis, and the immune system has emerged. The neuroendocrine system of the skin is composed of locally produced neuroendocrine mediators that interact with specific receptors. Among these mediators are neuropeptides, including members of the galanin peptide family--galanin, galanin-message-associated peptide, galanin-like peptide, and alarin--which are produced in neuronal as well as nonneuronal cells in the skin. Here we review the expression of the galanin peptides and their receptors in the skin, and the known functions of galanin peptides in different compartments of the skin. We discuss these data in light of the role of the galanin peptide family in inflammation and cell proliferation.     

Crustal layering, simplicity, and the oil industry: The alteration of an epistemic paradigm by a commercial environment

This paper proposes that the gradual alteration of the predominant epistemic paradigm in crustal seismology in the interwar period—namely, simplicity—came about because of the strong influence of a particular commercial environment, i.e. the oil industry. I begin by demonstrating the interwar predominance of Jeffreys’ ‘simplicity postulate’ and his probabilistic epistemology, highlighting the espousal by several seismologists (Bullen, Stoneley, Byerly), whose crustal models drew on mathematical idealisations. Next, I demonstrate that the renunciation of simplicity in the 1930s came about too quickly, and, above all, too heterodoxically to have been the result of new geological evidence. Rather, I argue, the paradigm shift among seismologists was a result of the significant rise in seismic exploration generated by the oil industry. Driven by market demands, American petroleum companies pioneered new technologies, organised research initiatives, and trained young geophysicists who, through the fusion of experimentalism and field experience, brought about fundamental progress in earthquake seismology. Remarkably, historians of science have almost entirely failed to recognise the interwar primacy of the simplicity paradigm as well as its subsequent renunciation. More importantly, they have failed to acknowledge the role the oil industry played in contributing to this renunciation and to the development of new paradigms in seismology.     

Differential diurnal variations of anandamide and 2-arachidonoyl-glycerol levels in rat brain

The endogenous ligands of cannabinoid receptors, also known as endocannabinoids, have been implicated in many physiological and pathological processes of the central nervous system. Here we show that the levels of the two major endocannabinoids, anandamide and 2-arachidonoyl-glycerol (2-AG), in four areas of the rat brain, change dramatically between the light and dark phases of the day. While anandamide levels in the nucleus accumbens, pre-frontal cortex, striatum and hippocampus were significantly higher in the dark phase, the opposite was observed with 2-AG, whose levels were significantly higher during the light phase in all four regions. We found that the activity of the fatty acid amide hydrolase, which catalyzes the metabolism of anandamide, was significantly lower during the dark phase, thus providing a possible explaination for the increase in anandamide levels. However, the activities of monoacylglycerol lipase and diacylglycerol lipase, two of the possible enzymes catalyzing the degradation and biosynthesis of 2-AG, respectively, changed significantly only in the striatum. These data suggest that the levels of the two major endocannabinoids might be under the control of endogenous factors known to undergo diurnal variations, and underscore the different roles, suggested by previous studies, of anandamide and 2-AG in neurophysiological processes.Received 9 December 2003; received after revision 15 January 2004; accepted 20 January 2004     

Chemistry and Chemists at the London Institution 1807-1912

The London Institution, established in the City of London in 1807, was devoted, as its full title proclaimed, to the 'advancement of Literature and the Diffusion of Useful Knowledge'. With its extensive lecture programme, splendid reference library, reading rooms, laboratory and other amenities, it provided for its members a scientific and cultural centre, modelled on the highly successful and fashionable Royal Institution in London's West End. Among its scientific activities, chemistry long maintained a leading role, in terms of both the sheer volume and variety of its presentations, and the high standing of its lecturers; they included Faraday, Playfair, Hofmann, Roscoe, Odling, Norman Lockyer, Meldola, and Sir William Ramsay, as well as other visiting lecturers, specially selected for their ability to present their subject in an interesting and attractive fashion to a wider lay public. The laboratory of the Institution, although limited in size and facilities, was the scene of instruction in practical chemistry, and between 1863 and 1884 attained the reputation of a significant centre of chemical research during the successive tenure of the professorship in chemistry by J. A. Wanklyn and H. E. Armstrong. Their publications, appearing under the device 'From the Laboratory of the London Institution', were a frequent feature of the leading chemical periodicals. Thus, within its many-sided activities, the Institution promoted significantly the public appreciation of the function of chemistry, as a contributor both to pure knowledge, and to technical and economic progress. It achieved this in an environment of influential City merchants, manufacturers and financiers and doubtless led to beneficient, if unrecorded, consequences. It was only towards the close of the nineteenth century, when the universities had become increasingly concerned with the systematic study of the discipline, that chemistry lost its direct impact in the London Institution, but continued to maintain a presence within its cultural framework.     

Fred Neufeld and pneumococcal serotypes: foundations for the discovery of the transforming principle

During the first decade of the twentieth century, the German bacteriologist Fred Neufeld, later Director of the Robert Koch-Institute in Berlin, first described the differentiation of pneumococci into serotypes on the basis of type-specific antisera. This finding was essential for subsequent research at the Rockefeller Institute of Medical Research (RIMR) in New York, and elsewhere, aiming for the conquest of human pneumococcal pneumonia, including antiserum therapy, the discovery that the type-specific antigens were carbohydrates, and the development of effective multivalent pneumococcal polysaccharide vaccines. Moreover, on the basis of pneumococcal serotypes Fred Griffith, in 1928 in London, discovered pneumococcal transformation, and Oswald T. Avery and coworkers, in 1944 at RIMR, identified DNA as the transforming substance. This sequence of events, leading to today’s knowledge that genes consist of DNA, was initiated by a farsighted move of Simon Flexner, first Director of the RIMR, who asked Neufeld to send his pneumococcal typing strains, thus setting the stage for pneumococcal research at RIMR. Here, we describe Fred Neufeld’s contributions in this development, which have remained largely unknown.     

Leibniz on the requisites of an exact arithmetical quadrature

In this paper we will try to explain how Leibniz justified the idea of an exact arithmetical quadrature. We will do this by comparing Leibniz's exposition with that of John Wallis. In short, we will show that the idea of exactitude in matters of quadratures relies on two fundamental requisites that, according to Leibniz, the infinite series have, namely, that of regularity and that of completeness. In the first part of this paper, we will go deeper into three main features of Leibniz's method, that is: it is an infinitesimal method, it looks for an arithmetical quadrature and it proposes a result that is not approximate, but exact. After that, we will deal with the requisite of the regularity of the series, pointing out that, unlike the inductive method proposed by Wallis, Leibniz propounded some sort of intellectual recognition of what is invariant in the series. Finally, we will consider the requisite of completeness of the series. We will see that, although both Wallis and Leibniz introduced the supposition of completeness, the German thinker went beyond the English mathematician, since he recognized that it is not necessary to look for a number for the quadrature of the circle, given that we have a series that is equal to the area of that curvilinear figure.     

The ‘Darwin-Marx correspondence’: a correction and revision

The logical links between the Judaeo-Christian doctrine of creation and the practice of natural philosophy on the one hand, and the rejection of belief in demonic agency on the other, were made explicit in the seventeenth century by, among others, Balthasar Bekker (1634–98), whose ideas I argue to have been not without influence. In section 1, I present the accounts of three historians of the opposition to belief in witchcraft and of the decline of the witch-persecution, Hugh Trevor-Roper, Keith Thomas, and Brian Easlea. In section 2, I maintain that Bekker has been underestimated both by Trevor-Roper and by Easlea. In section 3, I investigate more generally some of the connections between the new natural philosophy and belief in supernatural interventions, cast doubt on the view that rejection of belief in witchcraft and the devil requires rejection of belief in creation, and thus supplement or qualify the accounts of Trevor-Roper, Thomas, and Easlea of why belief in witchcraft faded away.