Construction of peptoliposomes for the incorporation of nutrient lipid supplements in insect cell culture media

Summary Nutrient liposomes that incorporate peptones instead of serum proteins can be used for the lipid supplementation of insect cells in lieu of or in addition to vertebrate serum to enhance the growth or differentiation of specific insect cell types in culture. The technique described here for the preparation of nutrient peptoliposomes involves the combination of both natural and synthetic phosphatidyl cholines (containing essential polyunsaturated fatty acids) with appropriate sterols. These are protected against premature oxidation by antioxidants (glutathione, alanine, protein hydrolysates) and antioxidant synergists (citrate, ascorbate, plus other amino acids) in the basal tissue culture medium as well as in the liposomal preparation itself (alphatocopherol acetate and protein hydrolysates). The lipid components are filter sterilized in chloroform and then dried before aqueous sonication, allowing the subsequent formation of a random array of unsized uni- and multilamellar liposomes, which are suitable as direct suppliers of lipid nutrients as well as nutrient carriers for lipid-soluble supplements.     

Transport of urea and amino acids through the isolated human placenta

By means of a method of two-way perfusion of the isolated human placenta the transport of urea from the fetal to the maternal placental circulation and the transport of amino acids in the opposite direction were studied. Experiments showed that the method provides for sufficiently complete perfusion of the intervillous space and creates suitable conditions for the study of placental transport. If the amino nitrogen concentrations in the two circulatory systems are equal, its concentration in the fetal circulation rises in the course of the experiment. On the addition of an amino acid to the maternal circulation, this increase develops to a greater degree. The results of these experiments confirm the view that amino acids are secreted by trophoblast cells into the fetal circulation.Laboratory of Biochemistry, Institute of Obstetrics and Gynecology, Academy of Medical Sciences of the USSR, Leningrad. (Presented by Academician of the Academy of Medical Sciences of the USSR M. A. Petrov-Maslakov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 81, No. 4, pp. 394–397, April, 1976     

Amino acid release from the cerebral cortex of the rabbit, its relationship with the electrocorticogram

The rate of release of five amino acids (alanine, glutamine, glutamic acid, glycine, GABA) from the surface of the sensorimotor cortex has been determined in rabbits under local anesthesia immobilized with gallamine. Concomitant recording of the electroencephalogram and of blood pressure has been carried out. The scope of the research was to determine if a difference in the release of these aminoacids could be correlated with variations in cerebral electrical activity. Two open-ended silver cylinders were adapted to the cortex and filled with saline. Every 10 min, the liquid was removed and assayed using a dansylating procedure followed by thin layer chromatographic separation and fluorimetric determination of the amino acids. The EEG of non treated animals, followed for a period of 100 min, remained synchronized for the majority of the recording; the biochemical determinations indicated a slight but constant fall in the rate of release of all the amino acids. A significant increase in the release accompanied the EEG desynchronisation induced either with eserine (0.2 mg/kg), amphetamine (2 mg/kg), or with electrical stimulation of the reticular formation.     

A way of preventing the free fatty acid concentration in the serum of rabbits from rising

A scheme of administration of nicotinic acid by mouth is suggested in order to prevent any increase in the free fatty acid concentration in the serum after intravenous injection of heparin or triiodothyronine, prolonged starvation, and thyroid feeding.Laboratory of Pathological Physiology, Institute of Experimental Endocrinology and Hormone Chemistry, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR N. A. Yudaev.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 78, No. 8, pp. 122–125, August, 1974.     

A Weaning Reaction to Microbiota Is Required for Resistance to Immunopathologies in the Adult

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A long term study of axonal transport in the central visual system following eye enucleation in the adult cat

Summary The effect of the enucleation of one eye on anterograde and retrograde labelling in geniculo-cortical, cortico-geniculate and commissural projections was investigated in adult cats by means of horseradish peroxidase (HRP) and tritiated aminoacids. It was found that in addition to the immediate decrease of retrograde labelling with HRP in the cortical projections from the deafferented A-laminae of the dorsal part of the lateral geniculate nucleus (Singer et al. 1977) there is a further reduction which lasts up to 75 days after enucleation. At 146 and 363 days after enucleation a slight increase in the number of labelled neurones was noted in the deafferented lamina. Qualitative assessment did not reveal any changes of anterograde labelling with tritiated amino acids in geniculo-cortical, cortico-geniculate and commissural axones. In addition, the retrograde labelling with HRP in cortico-geniculate and commissural projections seemed to be unaffected by eye enucleation.Deceased     

New high yield Cu(I) assisted 123I radioiodination of 15(p-I-phenyl)-9-methyl pentadecanoic acid,a potential myocardial tracer

A new high yield ¹²³I radioiodination (97%), based on the Cu(I) assisted isotopic exchange in an ethanol water mixture, of 15(p-I-phenyl)-9 methyl pentadecanoic acid, a potential myocardial tracer, is proposed. The method allows a true kit preparation of radioiodinated phenyl fatty acids for a substrate concentration of ±10^-6 moles. High specific activities (>0.1 mCi/g) can be obtained when coupling the labelling method to HPLC purification with an overall radiochemical yield of 75%.     

Effect of 3-methyl-branching on the metabolism in rat hearts of radioiodinated iodovinyl long chain fatty acids

The metabolism of two new 3-methyl-branched iodovinyl fatty acids in rat hearts was evaluated by determining the subcellular and lipid pool distribution of these radiolabeled analogues after intravenous injection. Methyl branching had been introduced into the straight chain analogue, 19-iodo-18-nonadecenoic acid (IVN), to produce the monomethyl analogue, 19-iodo-3-(R,S)-methyl-18-nonadecenoic acid (BMIVN) and the dimethyl derivative, 19-iodo-3,3-dimethyl-18-nonadecenoic acid (DMIVN) in the hope of inhibiting oxidation. Since the presence of 3-methyl branching results in delayed myocardial clearance in rats, differences were sought in the lipid and subcellular distribution of these branched analogues that might correlate with the prolonged retention and reflect differences in metabolism. Hearts of rats injected intravenously with the radiolabeled fatty acids were removed and homogenized and the homogenates partitioned between the chloroform-methanol (organic) fraction and the aqueous fraction. Comparison of the distribution of radioactivity between the organic and aqueous fractions showed that most of the DMIVN and BMIVN activity was in the organic fraction with IVN activity initially divided equally between the two fractions. Identification of the lipid components of these organic fractions showed that there was slow incorporation of DMIVN into the triglyceride and polar lipid fractions with a slow loss from the free fatty acid fraction. With the straight chain IVN analogue which shows rapid washout from rat hearts, there was loss of activity from all 3 lipid components during the 60 min. The monomethyl branched BMIVN analogue demonstrated predominant storage in the polar lipid fraction with some incorporation into triglycerides. Subcellular distribution studies of the three analogues also showed differences that correlated with the observed differences in heart retention properties. With the unbranched IVN analogue, radioactivity was found primarily in the cytoplasmic fraction 30 min after injection, whereas the branched analogues demonstrated a much higher association with the microsomal and mitochondrial fractions of the heart. In rats fed prior to injection, these differences in the subcellular distribution profiles were minimized. The lipid and subcellular distribution patterns reported here for the methyl branched analogues as compared to those of the straight chain iodovinyl fatty acid may provide some understanding as to the mechanisms of retention in rat myocardium.Research supported by the Office of Health and Environmental Research, U.S. Department of Energy, under contract DE-AC0 5-840 R21400 with Martin Marietta Energy Systems, Inc.     

Antagonism of the hypermotility response induced by excitatory amino acids in the rat nucleus accumbens

Summary Several compounds have been shown to antagonize the excitation of single neurons produced by excitatory amino acids. This study was designed to determine the effectiveness of these compounds in antagonizing the hypermotility response to excitatory amino acids after intraaccumbens administration. Of the putative antagonists tested, D-aminoadipic acid, diaminopimelic acid and glutamic acid diethyl ester all showed significant inhibitory effects on excitatory amino acid-induced hypermotility while 2-amino-5-phosphonovaleric acid, -D-glutamylglycine, 2-amino-4-phosphonobutyric acid and cis-2,3-piperidine dicarboxylic acid were ineffective. D-Aminoadipic acid decreased N-methyl-aspartic acid-induced hypermotility while having no significant effect on the hypermotility responses induced by kainic or quisqualic acids. Diaminopimelic acid markedly decreased N-methyl-aspartic acid- and kainic acid-induced hypermotility but was totally ineffective on quisqualic acid-induced hypermotility. In contrast to D-aminoadipic acid, glutamic acid diethyl ester antagonized the increase in motility produced by kainic and quisqualic acids but not that produced by N-methyl-aspartic acid. The above data suggests that N-methyl-aspartic acid and quisqualic acid may produce their motor effects through the activation of two different receptors in the nucleus accumbens while kainic acid may mediate its hypermotility response through both N-methyl-aspartic acid and quisqualic acid receptors. However, a third receptor type activated solely by kainic acid cannot be excluded at this time.     

Prenatal Exposure to BPA: The Effects on Hepatic Lipid Metabolism in Male and Female Rat Fetuses

Bisphenol A (BPA) is an organic chemical compound widely used for manufacturing plastics. BPA exposure originates principally from the diet, but it can also originate from dermal contact. In over 90% of individuals, including pregnant women, BPA is detectable in several body fluids. The effects of this exposure on the fetus are under active investigation in several research laboratories. The aim of our work was to study the impact of prenatal exposure to BPA in the liver of rat fetuses from a sex-dependent point of view. We particularly investigated the effects of prenatal BPA exposure on hepatic lipids because of their crucial role, not only for the liver, but also for the whole-body functions. Our results demonstrate that the liver of rat fetuses, in utero exposed to a very low dose of BPA (2.5 µg/kg/day), displays significant modulations with regard to proteins involved in cholesterol and fatty acid biosynthesis and trafficking. Moreover, an impact on inflammatory process has been observed. All these effects are dependent on sex, being observable only in female rat fetuses. In conclusion, this work demonstrates that maternal exposure to BPA compromises hepatic lipid metabolism in female offspring, and it also reveals the perspective impact of BPA on human health at doses currently considered safe.