Synchronous digestion of SV40 DNA by exonuclease III.

We have established an optimal condition for the synchronous digestion of SV40 DNA with Escherichia coli exonuclease III. Electron microscopy and polyacrylamide gel electrophoresis were used to obtain accurate measurements on the lengths of DNA before and after exonuclease III digestion. Based on this finding, a new method for determining the sequence of long duplex DNA can be realized. It involves (a) the synchronous digestion of the DNA from the 3' ends with exonuclease III, followed by (b) repair synthesis with labeled nucleotides and DNA polymerase, and (c) sequence analysis of the repaired DNA.     

Oviposition, fertility, and longevity and their relation to copulation in Antestiopsis lineaticollis (Heteroptera: Miridae)

Egg production, fertility, and longevity of solitary mated females and control pairs of males and females of Antestiopsis lineaticollis Stål. (=A. orbitalis bechuana Kirkaldy) were studied in the laboratory at 22–23° and 60–70% relative humidity. No significant differences were found in adult longevity, inter-oviposition period, number of eggs, post-reproductive period, and the number of eggs remaining in ovaries after the death of females. Control females had a significantly longer duration of oviposition than solitary ones. Thus, the physical presence of males or multiple mating by females has no influence on egg production in the Antestia bug.

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Zusammenfassung Eiproduktion, Fruchtbarkeit und Lebensdauer isolierter begatteter Weibchen sowie von Kontrollpaaren von Männchen und Weibchen von Antestiopsis lineaticollis Stål. (= A. orbitalis Kirkaldy) wurden im Labor bei 22–23° und 60–70% relativer Luftfeuchtigkeit untersucht. Es wurden keine signifikanten Unterschiede in der Lebensdauer der Adulten, den Perioden zwischen den Eischüben, der Anzahl der Eier, der Postreproduktionsperiode und der Anzahl der nach dem Tode der Weibchen in den Ovarien zurückgebliebenen Eier gefunden. Die Kontroll-Weibchen hatten eine signifikant längere Eiablagedauer als die isolierten. So hat also die physische Gegenwart der Männchen oder wiederholte Paarung der Weibchen bie der Antestia-Wanze keinen Einfluß auf die Eiproduktion.

     

Collaborative study of mutagenicity with Salmonella typhimurium TA102.

Thirty compounds of various chemical classes were investigated for mutagenicity in a collaborative study (3 laboratories) using Salmonella typhimurium TA102. With 5 compounds, namely hydrazine sulfate, phenylhydrazine, hydralazine, glutardialdehyde and glyoxal, mutagenicity was detected by all laboratories. Formaldehyde was assessed as weakly mutagenic in only 1 of 3 laboratories. The remaining 24 agents were uniformly described as non-genotoxic in TA102. In spite of the overall good qualitative agreement in the mutagenicity results between the 3 laboratories some quantitative discrepancies occurred in the dose response of the mutagenic compounds. Varying inter- and intra-laboratory differences in the spontaneous rate of revertants were obtained. The usefulness of the tester strain TA102 in routine mutagenicity testing is discussed.     

Impaired T cell functions during amphibian metamorphosis: IL-2 receptor expression and endogenous ligand production.

T cell functions are impaired during defined developmental stages of amphibian metamorphosis (Marx et al., 1987). Here we show, using a fluorescent anti-human IL-2 receptor antibody and flow cytometry, that during these stages, the splenocytes of Xenopus laevis, the South African clawed toad, have a progressively diminished capacity to express IL-2 receptors (IL-2R), after in vitro lectin stimulation. Preincubation with human rIL-2 specifically blocks binding of the anti-IL-2R antibody. Separation of an endogenous ligand bound to the IL-2R leads to a substantial increase in available epitope recognized by the anti-IL-2R antibody when pre- and postmetamorphic splenocytes are employed, but not when splenocytes of the prometamorphic stages are treated similarly. Thus, the cells from the prometamorphic stages are not producing significant quantities of the ligand. Finally, we demonstrate that human rIL-2 is not by itself mitogenic in the toad, but it can act as a co-stimulator of antigen-induced mitogenesis. Thus, an absence of an endogenous ligand (autologous IL-2?), coupled with a reduced capacity to express IL-2 receptors may be responsible for impaired T cell clonal expansion in metamorphosing Xenopus. Inhibition of T cell functions during this period is vital, since adult cells forming within the larval body bear surface proteins not found on larval cells (Flajnik et al., 1986).     

Thymus-replacing activity from the metamorphic spleen of Xenopus laevis

Anuran metamorphosis offers an interesting vertebrate immunological paradigm, for adult cells that arise within the immunocompetent larval body are MHC Class I disparate from those of the larva. The animals, in order to avoid immune self-destruction during this transition period, are made unresponsive to these modified-self cells by an impairment of T-cell functions. However, it remains to be discovered how an animal with compromised T-cell functions can protect itself from those environmental pathogens protection from which is thymus dependent. During metamorphosis, larval and adult immunocytes capable of reacting to each other coexist within the animal. Their interaction might stimulate the secretion of a cytokine capable of circumventing T-cell functions by acting directly on B cells. Here, we report that such an activity is released in vitro by metamorphic and not by adult splenocyte suspensions. This activity will amplify in vitro anti-hapten responses by immunized, but not carrier-primed, adult splenocytes. The activity is unaffected by dexamethasone and, since it will amplify anti-hapten responses in T-cell-depleted immunized adult splenocyte suspensions, the antibody-producing (B) cell population may be affected directly. Two radiolabeled protein peaks of 65 kD and 40 kD were obtained by SDS-PAGE analysis from secreting, metamorphosing, but not from adult, splenocytes.     

Should I stay or should I go? Mycorrhizal plants are more likely to invest in long‐distance seed dispersal than non‐mycorrhizal plants

Seed dispersal and mycorrhizal associations are key mutualisms for the functioning and regeneration of plant communities; however, these processes have seldom been explored together. We hypothesised that obligatory mycorrhizal plants will be less likely to have long‐distance dispersal (LDD) syndromes since the probability of finding suitable mycorrhizal partners is likely to decrease with distance to the mother plant. We contrasted the mycorrhizal status and LDD syndromes for 1960 European plant species, using phylogenetically corrected log‐linear models. Contrary to our expectation, having specialised structures for LDD is more frequent in obligate mycorrhizal plants than in non‐mycorrhizal plants, revealing that lack of compatible mutualists does not constrain investment in LDD structures in the European Flora. Ectomycorrhizal plants associated with wind‐dispersing fungi are also more likely to have specialised structures for wind dispersal. Habitat specificity and narrower niche of non‐mycorrhizal plants might explain the smaller investment in specialised structures for seed dispersal.     

Morphology and surface ultrastructure of <Emphasis Type="Italic">Dadaytrema oxycephala</Emphasis> (Digenea: Cladorchiidae) with a new host record from Peruvian Amazon floodplain

In this study, a new host record for Dadaytrema oxycephala (Digenea: Cladorchiidae) is described from an important Amazonian ornamental fish. The parasite was identified based in the general morphology and surface ultrastructure of the young and mature trematodes using scanning electron microscopy. The digenetic trematodes were found in the intestine of 15 of 80 specimens (18.7%) of Brochis multiradiatus captured from two floodplain lakes of the Amazonas River, Department of Loreto, Peru and no clinical signs were observed in the parasitized organ. This is the first report of D. oxycephala parasitizing fish of the genus Brochis in the Amazon basin and the first to report this parasite infection in an ornamental fish from Peru. Given that international trade of ornamental fish is a major factor for transboundary spreading of fish parasites and taking account that Peru is the second largest exporter of the Amazonian countries, there is a fundamental need of constant monitoring for diagnosis and timely control of infections by digeneans in order to better prevent introduction in new environments, dissemination and future disease outbreaks.     

A Living Biobank of Breast Cancer Organoids Captures Disease Heterogeneity

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Modulation of the Aplysia gill withdrawal reflex by dopamine

The ability of dopamine to modulate gill contractions was tested in Aplysia. When dopamine was perfused through the gill vasculature, gill contractions caused by siphon stimulation (gill withdrawal reflex) and by depolarization of the gill motor neuron L7 were increased in amplitude, as compared with those evoked during seawater perfusion. Habituation of gill movements, brought about by repetitive stimulation of the siphon or of L7, was prevented by dopamine. Despite the absence of reflex habituation, the number of action potentials in central gill motor neurons, evoked by siphon stimulation, showed normal decrement. Dopamine's effects were blocked when the ctenidial nerve was cut or when L7 hyperpolarized. These data suggest that dopamine acts peripherally to increase the efficacy of L7's synaptic transmission onto gill muscle or elements of the gill neural plexus.     

An immunocytotoxicity assay for neural cells in monolayer cultures

The immunological analysis of cell surface constituents which may characterize neuronal and glial populations, though still in its infancy, will greatly facilitate the investigation of several important problems in neurobiology. One critical component of such analyses is the way by which a given antiserum can be shown to be active on, and possibly selective for neurons and glial cells from normal neural tissues. This report describes the use of monolayer cultures of normal neural cells for recognition and quantitative titration of antisera directed against them. Sera were collected from rabbits immunized with chick embryo spinal cord cell susptnsions, and found to be reactive to the same cells in the initial cell dissociate as well as in subsequent monolayer cultures of different in vitro ages. A monolayer assay procedure was developed, which (i) uses small numbers of cells and small volumes of immune reagents, with the possibility of further scaling down; (ii) applies equally to cultures using different substrata; (iii) permits differential counts of morphologically different cultured cells; (iv) allows to recognize cytological damage imposed by the immune serum in the presence, though not in the absence, of complement; and (v) quantitatively titrates the immune activity with 10- to 20-fold higher sensitivity than other titration procedures. While the study was not intended to investigate the possible specificities of the new antisera, it provided the unexpected observation that non-neuronal cells in these spinal cell cultures were considerably less sensitive than neurons to the complement-dependent action of the antisera.