Chromosomal localization of zein genes by in situ hybridization in Zea mays

Summary In order to localize the genes coding for zein, the major storage protein of maize endosperm, zein ¹²⁵I-mRNA and ³H-cDNA labelled at high specific activity were used for in situ hybridization on heterozygous interchanges and paracentric inversions of the KYS strain of Zea mays. The analysis of the diplotene-metaphase I microsporocytes indicated the presence of zein structural genes on the long arm of chromosomes 4 and 5, the short arm of chromosome 7 and the distal segment of the long arm of chromosome 10. The two hybridization sites on chromosomes 7 and 10 are found near opaque-2 and opaque-7 loci which are known to regulate zein synthesis. The present data are discussed in relation to results obtained by other authors using genetical mapping of zein genes.     

Delayed-type cutaneous hypersensitivity to melanoma antigens and its implication in active specific immunotherapy in cancer

Summary In this study, we explored whether soluble tumor-cell surface-associated antigens (TAA) might be derived from autochthonous as well as allogeneic sources as immunogens for active specific immunotherapy. Using two popular cell membrane-bound antigen extraction techniques (3 M KCl and isotonic-hypotonic NaCl), we examined the immunogenic potential of such TAA and the specificity of immunologic host reactivity through a delayed-type cutaneous hypersensitivity reaction (DTH) as a guideline for their immunogenic potential in a human malignant melanoma model system. We found that either extraction technique could provide soluble TAA from both autochthonous and allogeneic sources capable of eliciting DTH. While evidence of positive DTH with autochthonous TAA reaffirms the immunogenicity of such TAA, the specificity of host reactivity against TAA derived from allogeneic sources is extremely difficult to establish, even with TAA partially purified by column chromatography in Sephadex G-200. Patients exhibited reactivity to other TAA derived from tumors of different histologies and often to more than one component isolated by column chromatography. Furthermore, when a group of melanoma patients was tested against a panel of melanoma antigens in any random combination, DTH to allogeneic TAA was seen in an unpredictable order and with inconsistent frequency. We conclude, therefore, that while autochthonous antigen immunizations may be justified, more careful studies will be necessary to define the antigenic profile of a given tumor (individual specificity vs shared specificity), establish specificity of alloantigens, and devise suitable methods for testing immunologic specificity for alloantigens, before rational immunotherapy with allogeneic tumor antigens will be feasible.     

Gas chromatographic method for the simultaneous determination of hydralazine and its acetylated metabolite in serum using a nitrogen-selective detector

A relatively simple gas chromatographic method has been developed for the quantitative determination of hydralazine simultaneously with its acetylated metabolite, 3-methyl-s-triazolo[3,4-α]phthalazine (MTP). The proteins were removed by means of sulfosalicylic acid and Sure-Sep®. On treatment with formic acid, hydralazine and its internal standard were converted into their formylated derivatives. These derivatives, MTP and its internal standard were extracted with toluene and determined by gas chromatography with a nitrogen-selective detector. The lower limits of detection for hydralazine and MTP were 0.13 and 0.27 μmol/l, respectively.     

Amino acid transport in pig lymphocytes Enhanced activity of transport system asc following mitogenic stimulation

Changes in neutral amino acid transport activity caused by addition of phytohaemagglutinin-P to quiescent peripheral pig lymphocytes have been evaluated by measurements of ¹⁴C-labelled neutral and analogue amino acids under conditions approaching initial entry rates. Utilizing methylaminoisobutyric acid, the best model substrate of System A, we confirmed our previous report (Borghetti, A.F., Kay, J.E. and Wheeler, K.P. (1979) Biochem. J. 182, 27–32) on the absence of this transport system in quiescent cells and its emergence following stimulation. Furthermore, we demonstrated the presence in quiescent cells of an Na⁺-dependent transport system for neutral amino acids that has been characterized as System ASC by several criteria including intolerance to methylaminoisobutyric acid, strict Na⁺-dependence, the property of transtimulation and specificity for pertinent substrates such as alanine, serine, cysteine and threonine. Analysis of the relationship between influx and substrate concentration revealed that two independent saturable components contribute to entry of alanine in quiescent cells: a low affinity ($\text{K}{}_{\mathrm{<mtext>m</mtext>}}\text{= ≈4}\text{mM}$) and a high affinity ($\text{K}{}_{\mathrm{<mtext>m</mtext>}}\text{= ≈0.2}\text{mM}$) component. The high affinity component could be inhibited in a competitive way by serine, cysteine and threonine, but methylaminoisobutyric acid did not change appreciably its constants. The enhanced activity of alanine transport through the ASC system observed in activated cells resulted from a large increase in the capacity ($\text{V}$) of the high affinity component without any substantial change in the apparent affinity constant ($\text{K}{}_{\mathrm{<mtext>m</mtext>}}$).     

Ovipositional deterrents in Dacus oleae

The deterrent substances diverting D. oleae females from ovipositing on already attacked olives are contained, at least partly, in the juice which trickles from the oviposition wounds. Surprisingly, the water fraction of the olive juice had limited deterrent activity. The principal deterrent stimuli are present in the oil fraction. Acetophenone and benzaldehyde are likely to be involved.Some liposoluble volatile substances can be deterrent to the females as vapours. The deterrent power of olives containing D. oleae larvae is probably linked to these substances.Most deterrent hydro-soluble substances had two contiguous hydroxyls, characterized by a comparative acidity and located in the ortho position in diphenols and somewhat analogously in glycolic acid.Amongst orthodiphenols, Pyrocatechol-the simplest compound-was the most active substance.

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Résumé Les jus d'olive qui sourt des blessures dues à la ponte, contient des substances inhibitrices qui empêchent les attaques ultérieures de la mouche de l'olivier.Une telle activité était attribuée à des substances hydrosolubles et, en particuliet, au dihydroxyphenil ethanol.On a testé l'activité inhibitrice des fractions huileuse et acqueuse obtenues par centrifugation de la pulpe des olives, rendue homogène.De facon inattendue, la fraction huileuse s'estavérée posséder une activité inhibitrice environ 20 fois supérieure à celle de la fraction acqueuse.Les différences d'activités inhibitrices d'huile d'olives mûres ou non, ou d'huiles commerciales, permet de penser que l'activité inhibitrice principale est liée à des substances liposolubles présentes dans l'huile.En outre, on a constaté que la pulpe d'olives, broyée, présente une action inhibitrice en l'absence de contact direct avec les femelles. Des substances inhibitrices volatiles sont aussi présentes dans l'huile.Des inhibiteurs volatiles qui se libèrent des tissus lacérés par les larves, peuvent expliquer l'activité inhibitrice des olives attaquées durant la période de développement des larves. Les substances inhibitrices dont la femelle couvre la surface cireuse des fruits, sont, en effet, facilement éliminées par la pluie.Parmi les substances liposolubles dont on connaît la présence dans l'huile, l'aceto-phénone et le benzaldehyde, possèdent un pouvoir inhibiteur prononcé.Les substances hydrolsolubles jouent un rôle mineur dans l'ensemble des stimuli inhibiteurs. Le pouvoir inhibiteur relatif est lié à deux hydroxyles contigus à une fonction acide présente en position ortho dans les diphénols, ceci par analogie avec l'acide glycolique qui possède un hydroxyle en caractérisé par une certaine acidité.Le plus actif orthodiphénol s'est avéré être la pyrocatéchine. Aussi une part des inhibiteurs liposolubles sont des substances liées au biochimisme des phénols.

     

Lipid peroxidation induced by cercosporin as a possible determinant of its toxicity

The photodynamic action of cercosporin was assayed in various kinds of natural and artificial membranes. Cerosporin induces lipoperoxidation of liposomes, rat liver and pea internode mitochondria and microsomes, estimated both as malondialdehyde (MDA) formation and O2 consumption. Cercosporin-induced lipoperoxidation is inhibited by either singlet oxygen quenchers, free radical trapping agents or EDTA. Superoxide anion (O2-), hydrogen peroxide and hydroxyl radicals (.OH) are not involved in the activity of cercosporin. In addition cercosporin, by chelating iron, lowers the lipoperoxidation induced by such a metal. Therefore cercosporin stimulates, through singlet oxygen production, the hydroperoxide formation but, at the same time, it inhibits the continuation of the iron-mediated free radical chain. The present results suggest that cellular lipid peroxidation has a certain relevance to toxic activity of cercosporin.     

Effects of diffusible products of peroxidation of rat liver microsomal lipids

The effects on cellular structures of products of peroxidation of rat liver microsomal lipids were investigated. A system containing actively peroxidizing liver microsomal fraction was separated from a revealing or target system by a dialysis membrane. The target system, contained in the dialysis tube, consisted of either intact cells (erythrocytes) or subcellular fractions (liver microsomal fraction). When liver microsomal fractions were incubated with NADPH (or an NADPH-generating system), lipid peroxidation, as measured by the amount of malonaldehyde formed, occurred very rapidly. The malon-aldehyde concentration tended to equilibrate across the dialysis membrane. When the target system consisted of erythrocytes, haemolysis occurred abruptly after a lag phase. The lysis was greatly accelerated when erythrocytes from vitamin E-deficient rats were used, but no haemolysis was observed when erythrocytes from vitamin E-treated rats were used. When, in the same system, freshly prepared liver microsomal fractions were exposed to diffusible factors produced by lipid peroxidation, the glucose 6-phosphatase activity markedly decreased. A similar decrease in glucose 6-phosphatase activity, as well as a smaller but significant decrease in cytochrome P-450, was observed when the target microsomal fractions were exposed to diffusible factors derived from the peroxidation of liver microsomal lipids in a separate preincubation step. These and additional experiments indicated that the toxicological activity is relatively stable. Experiments in which the hepatic microsomal fractions destined for lipid peroxidation contained radioactively labelled arachidonic acid, previously incorporated into the membranes, showed that part of the radioactivity released from the microsomal fraction into the incubation medium entered the dialysis tube and was recovered bound to the constituents of the microsomal fractions of the target system. These results indicate that during the course of the peroxidation of liver microsomal lipids toxic products are formed that are able to induce pathological effects at distant loci.     

Manipulating Representations

The present paper proposes a definition for the complex polysemic concepts of consciousness and awareness (in humans as well as in other species), and puts forward the idea of a progressive ontological development of consciousness from a state of ‘childhood’ awareness, in order to explain that humans are not only able to manipulate objects, but also their mental representations. The paper builds on the idea of qualia intended as entities posing regular invariant requests to neural processes, trough the permanence of different properties. The concept of semantic differential introduces the properties of metaphorical qualia as an exclusively human ability. Furthermore this paper proposes a classification of qualia, according to the models–with different levels of abstraction–they are implied in, in a taxonomic perspective. This, in turn, becomes a source of categorization of divergent representations, sign systems, and forms of intentionality, relying always on biological criteria. New emerging image-of-the-world-devices are proposed, whose qualia are likely to be only accessible to humans: emotional qualia, where emotion accounts for the invariant and dominant property; and the qualic self where continuity, combined with the oneness of the self, accounts for the invariant and dominant property. The concept of congruence between different domains in a metaphor introduces the possibility of a general evaluation of truth and falsity of all kinds of metaphorical constructs, while the work of Matte Blanco enables us to classify conscious versus unconscious metaphors, both in individuals and in social organizations.