Production of anti-thymulin (FTS) monoclonal antibodies by immunization against human thymic epithelial cells

A monoclonal antibody specific for thymulin (FTS), a thymic hormone initially isolated from serum, was obtained by cell fusion using spleen cells from BALB/c mice immunized with cultured human thymic epithelial cells. Hybridomas were selected according to their capacity to produce antibodies binding specifically to thymic epithelial cells in culture (as assessed by indirect immunofluorescence) and their ability to absorb in vitro the biological activity of synthetic and natural hormone preparations and to induce in vivo the disappearance of endogenous circulating thymulin. In this way monoclonal antibodies were obtained that recognized a subpopulation of nonlymphoid cells on frozen sections of mouse and human thymuses. The epithelial nature of these cells was assessed using an antikeratin antiserum. The binding of the antibodies to thymic cells was completely abolished by its absorption with the synthetic hormone or normal (but not of thymectomized) mouse serum. The thymic specificity of the antibody was further confirmed by the complete absence of binding to sections of all the various lymphoid and epithelial organs examined (from both humans and mice). Double labeling experiments using the monoclonal antibody described above and a monoclonal antibody prepared by immunization with the synthetic peptide showed that the two antibodies bound to the same cell. These results provide further evidence for the exclusive presence of the thymic hormone thymulin in thymic epithelial cells.     

Extra-ribosomal spacer sequences in Triturus

We show that, in Triturus vulgaris meridionalis, sequences homologous to the rDNA "non-transcribed" spacer (NTS) are clustered at chromosomal loci where they are not associated with 18 S or 28 S rDNA genes: these sequences are referred to as the extra-ribosomal spacer sequences. Genomic clones containing such extra-ribosomal spacer sequences have been isolated. As shown by restriction mapping, these clones appear to consist mostly of repetitive BamHI fragments that are, in turn, internally repetitious and highly homologous to each other. The structure of the clones was confirmed by nucleotide sequence analysis, which also demonstrates the high degree of conservation between the BamHI elements and the homologous NTS sequences. An intriguing 12 base-pair homology between the extra-ribosomal spacer sequences and a Xenopus NTS enhancer sequence is reported. The possibility that a repetitive octanucleotide motif found within the BamHI elements could act as a recombination hotspot by virtue of its similarity with the Escherichia coli chi sequence is discussed.     

Effects of methyl palmoxirate on hypoxic rat atria.

Isolated rat atria in hypoxia released lactate into the bathing medium and underwent a decline of the contraction frequency which, in some cases led to a complete cessation of the pacemaker activity. A pronounced fall in the peak developed tension and a rise in the resting tension also appeared. The atria from 24 h fasted rats, which oxidize faster their reserve lipids than those from fed rats, exhibited greater functional disturbances during hypoxia, a lower lactate output and a smaller recovery of peak tension upon reoxygenation. Methyl palmoxirate, which is a selective inhibitor of carnitine palmitoyltransferase I, attenuated the decline of the beating rate and the rise of the resting tension in both groups of rats and the incidence of atrial arrest in the fasted rat group. The fall in the peak tension, lactate output and recovery upon reoxygenation were not altered by the inhibitor. These data indicate that methyl palmoxirate alleviates some of the hypoxic functional derangements. Hence, it may be inferred that inhibiting the oxidation of the fatty acid derived from the endogenous triacylglycerol is beneficial during oxygen-limited conditions and that these effects could not be ascribed to changes in the glycolytic flux.     

Vaccination in murine schistosomiasis with adult worm-derived antigens: variables influencing protection in outbred mice

Previous studies have shown that both permissive (mouse) and partially permissive (rabbit) hosts develop high levels of resistance against Schistosoma mansoni infection after vaccination with a multiple antigen extract (SE) obtained by incubation of living adult worms in saline, plus bacterial adjuvant. To investigate variables influencing SE-induced protection in murine schistosomiasis, a series of distinct vaccination protocols were performed focussing on the immunization dose, carrier systems, route, site and amplitude of challenge infection, and time between immunization and challenge. In addition, a new approach was adopted to evaluate SE protective activity, by means of population analysis of worm burden frequency distributions in a large scale study of vaccination in outbred Swiss mice. Distinct curves of frequency and a drastic difference in worm burden distribution of frequencies from SE-vaccinated x non-vaccinated mice were found. It was shown that SE could generate 75% mean protection in outbred mice even in the absence of adjuvant. In addition SE immunization was also able to induce full protection against lethal infection. SE-induced protection could be modulated by such parameters as dose of SE immunization/challenge interval, and route of cercariae injection. These data show that SE yields very high protective activity in outbred mice, and may provide a further insight for rational design of a vaccine in experimental schistosomiasis.     

Kinetic persistence of cruciform structures in reconstituted minichromosomes

Cruciforms persist in reconstituted minichromosomes, as revealed by cleavage with specific nucleases and hybridization with synthetic oligonucleotides. Relaxation by topoisomerase I suggests that cruciforms are located mainly on internucleosomal DNA and that their persistence on minichromosomes may be due to kinetic effects. The analysis of the kinetic behaviour of cruciforms in minichromosomes shows a definite velocity of reabsorption with respect to stable cruciforms in supercoiled naked DNA. An explanation based on suppression of the untwisting of linker DNA due to adjacent nucleosomes is proposed.     

Behavior of fish predators and their prey: habitat choice between open water and dense vegetation

Synopsis Behavior of largemouth bass, Micropterus salmoides, and northern pike, Esox lucius, foraging on fathead minnows, Pimephales promelas, or bluegills, Lepomis macrochirus, was quantified in pools with 50% cover (half the pool had artificial stems at a density of 1000 stems m⁻²). Both predators spent most of their time in the vegetation. Largemouth bass searched for bluegills and ambushed minnows, whereas the relatively immobile northern pike ambushed all prey. Minnows were closer to predators and were captured more frequently than bluegills. Even when minnows dispersed, they moved continually and eventually wandered within striking distance of a predator. Bluegills dispersed in the cover with predators. Bass captured the few bluegills that strayed into the open and pike captured those that approached too closely in the cover. The ability of predators to capture prey while residing in habitats containing patches of dense cover may explain their residence in areas often considered to be poor ones for foraging. The unit is sponsored jointly by the United States Fish and Wildlife Service, Ohio Department of NaturalResources, The Ohio State University, and the Wildlife Management Institute     

Neuroendocrine control of thymic hormonal production. II. Stimulatory effects of endogenous opioids on thymulin production by cultured human and murine thymic epithelial cells

Data have now accumulated to strongly demonstrate that several neuropeptides, including endogenous opioids, can have immunomodulatory functions. Most of the studies have so far focused on the direct action of these substances on lymphocytes. We decided to investigate whether thymic epithelial cells (TEC) - the major component of the thymic microenvironment - could also be modulated by endogenous opioids. Primary cultures of human and murine TEC were subjected to several opioids (alpha-beta- or gamma-endorphins, as well as met- or leuenkephalins) applied in concentrations ranging from 10(-6) to 10(-9) M. On the following days we measured the levels of thymulin (a chemically-defined thymic hormone known to stimulate some steps of T-cell differentiation) in the culture supernatants, as well as the numbers of thymulin containing cells, evaluated by immunofluorescence with an anti-thymulin monoclonal antibody. After treatment of TEC cultures with beta-endorphin or leu-enkephalin a significant increase in the levels of thymulin in the culture media was observed, paralleled by a rise in the percentage of thymulin containing cells. In addition, this stimulatory effect was dose-dependent. Preincubation of the opioids with the specific antibodies abrogated the opioid-induced stimulatory effect on TEC. Moreover, naloxone, an opioid receptor antagonist, blocked the effect of beta-endorphin on thymulin production, suggesting that the effect of this neuropeptide on epithelial cells was mediated by an opioid receptor. Importantly, no effect on thymulin production was observed with the other opioids used, whatever the dose. These results suggest that, at least in vitro, beta-endorphin and leu-enkephalin stimulate the hormonal function of the thymic epithelium. These findings lead to the general concept that the modulatory role of endogenous opioids on the immune system is not restricted to lymphocytes but can also take place at the level of cells belonging to T-cell differentiating microenvironments.