C5a induction of human interleukin 1. Synergistic effect with endotoxin or interferon-gamma

Interleukin-1 (IL-1) is a potent cytokine which possesses the ability to mediate systemic acute phase responses as well as local tissue inflammation. In these studies, we have examined the ability of C5a and C5a des Arg to induce IL-1 production in vitro. Human C5a and C5a des Arg were purified to homogeneity and were found to stimulate IL-1 release from freshly obtained human mononuclear cells into the extracellular medium. Only 2 hr of exposure to the purified complement components were necessary in order to stimulate IL-1 production. The minimal concentration of C5a required was 25 ng/ml, whereas 125 ng/ml of C5a des Arg induced comparable amounts of IL-1. This dose relationship was maintained at higher concentrations (150 ng/ml vs 750 ng/ml, respectively). That the effect was due to the anaphylatoxins themselves, and not endotoxin contamination, was shown by negative Limulus amebocyte lysate tests and employing preincubation of C5a/C5a des Arg with polymyxin B. The latter blocked a wide dose range of endotoxin-stimulated IL-1 production. However, when endotoxin was added to C5a or C5a des Arg, significant synergism in the stimulation of IL-1 production was observed, occurring at various concentrations of either agent. A similar synergism with C5a/C5a des Arg was seen with interferon-gamma. In these studies, IL-1 production was measured by bioassay employing cloned D . 10 . G4 . 1 murine T cells and by radioimmunoassay for human IL-1 beta; using C5a/C5a des Arg as stimulants, there was a high degree of correlation (r = 0.82) between the two assays. Since traumatic, infectious, and inflammatory diseases may result in the simultaneous appearance of these stimuli, the synergism described herein is likely to be clinically relevant.     

Sterol Composition and Ecdysteroid Content of Eggs of the Root-knot Nematodes Meloidogyne incognita and M. arenaria

Free and esterified sterols of eggs of the root-knot nematodes Meloidogyne incognita races 2 and 3 and M. arenaria race 1 were isolated and identified by gas-liquid chromatography-mass spectrometry. The major sterols of eggs of each race were 24-ethylcholesterol (33.4-38.8% of total sterol), 24-ethylcholestanol (18.3-25.3%), 24-methylcholesterol (8.6-11.7%), 24-methylcholestanol (7.7-12.5%), and cholesterol (4.6-11.6%). Consequently, the major metabolic transformation performed by Meloidogyne females or eggs upon host sterols appeared to be saturation of the sterol nucleus. The free and esterified sterols of the same race did not differ appreciably, except for a slight enrichment of the steryl esters in cholesterol. Although the sterol composition of Meloidogyne eggs differed from that of other life stages of other genera of plant-parasitic nematodes, the three Meloidogyne races could not be distinguished from each other by their egg sterols. Ecdysteroids, compounds with hormonal function in insects, were not detected by radioimmunoassay in the Meloidogyne eggs either as free ecdysteroids or as polar conjugates.     

ASSESSING NORTHERN ELEPHANT SEAL FEEDING HABITS BY STOMACH LAVAGE

Stomach lavaging was used to study the feeding habits of northern elephant seals ( Mirounga angustirostris ) found on San Miguel Island, California, during the spring of 1984. Fifty-nine elephant seals were chemically immobilized with an intramuscular injection of ketamine hydrochloride. Once immobilized, an animal's stomach was intubated, filled with 3–4 liters of water to create a slurry of the undigested food items, and evacuated into a collection device. The stomachs of 57 (96.6%) of the animals lavaged contained identifiable parts of prey. Twenty-nine different food items were identified, 12 of which have not been previously reported as prey of the northern elephant seal: two teleost fish, Coryphaenoides acrolepis (Pacific rattail) and another unidentified macrourid; two crustaceans, Pasiphaea pacifica (glass shrimp) and Euphausia sp.; six squid, Abraliopsis felis, Gonatus berryi, Histioteuthis dofleini, Cranchia scabra, Taonius pavo, and Galiteuthis sp. and two octopi, Octopus dofleini and Octopus rubescens.     

Chondroid bone arises from mesenchymal stem cells in organ culture of mandibular condyles

Mandibular condyles of fetal mice 19 to 20 days in utero comprising clean cartilage and its perichondrium were cultured for up to 14 days, and their capacity to develop osteoid and to mineralize in vitro was examined. After 3 days in culture the cartilage of the mandibular condyle appeared to have lost its inherent structural characteristics, including its various cell layers: chondroprogenitor, chondroblastic, and hypertrophic cells. At that time interval no chondroblasts could be seen; instead, most of the cartilage consisted of hypertrophic chondrocytes. By that time, the surrounding perichondrium, which contains pluripotential mesenchymal stem cells, revealed the first signs of extracellular matrix enclosing type I collagen, bone alkaline phosphatase, osteonection, fibronectin, and bone sialoprotein as demonstrated by immunofluorescent techniques. Electron microscopic examinations of the newly formed matrix revealed foci of mineralization within and along collagen fibers as is normally observed during bone development. The composition of the latter mineral deposits resembled calcium pyrophosphate crystals. Following 14 days in culture larger portions of the condyle revealed signs of osseous matrix, yet the tissue reacted positively for type II collagen. Hence, the condylar cartilage, a genuine representative of secondary-type cartilage, elaborated in vitro a unique type of bone that would be most appropriately defined as chondroid bone. Biochemical assays indicated that the de novo formation of chondroid bone was correlated with changes in alkaline phosphatase activity and 45Ca incorporation. The findings of the present study imply that mesenchymal stem cells that ordinarily differentiate into cartilage possess the capacity to differentiate into osteogenic cells and form chondroid bone.     

Determination of the serological sex-specific (Sxs) antigen (“H-Y antigen”) in birds and mammals using high-titer antisera and a sensitive urease ELISA

Summary A rapid, sensitive, and reproducible enzyme-linked immunosorbent assay (ELISA), for the detection of the serological sex-specific (Sxs) antigen (formerly termed H-Y antigen; see Introduction), is described. This assay uses bovine lestes extract as the solid phase antigen, and high-titer anti-Sxs antisera and a urease-conjugated anti rat-IgG as the first and second antibody respectively. The urea containing substrate causes a pH shift in a positive reaction, which in turn is visualized by the use of bromocresol purple as a pH indicator. The method, and some representative applications of it, are described in detail.     

Fish cleaning behaviour of shrimp

Cleaning behaviour of five species of shrimp from three families was studied at three different geographic locations in an effort to gain a quantitative understanding of cleaning behaviour, and to compare a broad cross-section of cleaner shrimp species. Two shrimp from the genus Periclimenes , two from the genus Lysmata , and one from the genus Stenopus were used and 27 hours of recorded laboratory observations were made for each of the five shrimp species.
All shrimp species were inactive most of the observed time, and most spent less than 2% of the observed time cleaning fish hosts. Also, the shrimp spent more time cleaning the ventral rather than the dorsal surface of the fish because they were reluctant to board the fish. However, evenness in cleaning does not appear to be an indicator of overall excellence in cleaning because the two best cleaners (based on number and duration of cleaning bouts) were among the least even in their cleaning.
The fish cleaning behaviour of the shrimp appeared to be strictly stereotypic in form, but the stimulus-response and the total amount of cleaning differed greatly among the five species. A Cleaning Efficiency Index (CEI) was created in an attempt to incorporate significant aspects of the cleaning behaviour. According to this CEI, Lysmata grabhami was by far the most efficient (best) cleaner, CEI = 55–51, compared to the others; Stenopus hispidus , 33–78; Periclimenes pedersoni , 6–29; Periclimenes yucatunicus , 5–60; and Lysmata californien , 2–12.
The cleaners most widely distributed geographically have the highest CEI scores, while the most localized cleaners have the lowest CEIs. This relationship may allow the CEI score to be useful in determining a cleaner shrimp's potential geographical distribution, and may also serve as an indicator for the degree of phylogenetic relationship to other cleaner shrimps.     

A translocation associated, loss-of-function mutation in the nitrogen metabolite repression regulatory gene of Aspergillus nidulans can revert intracistronically

Summary The areA ^r -18 mutation is a loss-of-function mutation in areA, the positive acting regulatory gene mediating nitrogen metabolite repression in Aspergillus nidulans. It results from a reciprocal translocation which splits the coding region into 5 and 3 moieties. Surprisingly, we have selected rare intracistronic revertants of areA ^r -18. From crosses heterozygous for areA ^r -18 revertant alleles, duplication-deficiency progeny containing two copies of a substantial portion of chromosome IV but lacking part of chromosome III, including the 5 moiety of areA, have been obtained. For all four revertants analysed genetically, growth properties of these duplication-deficiency strains indicate that the reversion events involve the 3 portion of areA and that the 5 portion of areA is unnecessary for the revertant phenotype. This conclusion was directly confirmed for one revertant using Southern blotting. As all four reversion events involve additional chromosomal rearrangements, they probably fuse functional promoters, ribosome binding sites and in frame initiation codons to the 3 portion of the gene. In the course of characterisation of these mutations, new mapping data for a large region of chromosome IV have been generated, and a new reciprocal translocation activating the cryptic regulatory gene areB, whose product can substitute for that of areA, has been identified.     

Interactions between developmental processes,growth, and food selection in the larvae and juveniles of Rutilus rutilus (L.) (Cyprinidae)

Summary The aim of this study was to assess the effects of developmental events, occurring in fish during the first weeks after hatching, on the quantity and quality of the ingested food and on growth. The investigation was carried out with the larvae and juveniles of Rutilus rutilus, the single cyprinid species occurring in an oligotrophic subalpine lake in Tirol, Austria. Comparison between availability of prey in the water and gut contents suggests that the selection of food by the young fish is strongly influenced by developmental processes. For example, the prevalence of indigestible phytoplankton in the gut of young larvae can be taken as a sign of the not yet fully developed sensory and locomotory capacities of the young fish (El-Fiky et al. 1987). Furthermore, quantitative and qualitative changes in the gut contents correlate strongly with changes in the form and relative length of the gut, but reflect only weakly the availability of prey in the water. In the Seefelder See population of R. rutilus the switch from a phytoplankton to a cladoceran dominated diet is accompanied by an increase in relative growth rate by nearly one order of magnitude (Wieser et al. 1988).