Enzyme-linked immunosorbent assays for detecting antibodies to Shiga-like toxin I,Shiga-like toxin II,andEscherichia coli O157:H7 lipopolysaccharide in human serum

Shiga-like toxin-producingEscherichia coli O157:H7 are important causes of bloody diarrhea and hemolytic uremic syndrome. To facilitate the epidemiologic study of these organisms, we developed enzyme-linked immunosorbent assays (ELISAs) for antibodies to Shiga-like toxin I (SLT I), Shiga-like toxin II (SLT II), andE. coli O157 lipopolysaccharide (LPS). We tested serum samples from 83 patients in two outbreaks ofE. coli O157:H7 diarrhea and from 66 well persons. Forty-three patients (52%) had at least one serum sample positive for anti-O157 LPS antibodies; among 26 culture-confirmed patients, 24 (92%) had at least one positive serum sample. Two (3%) of 66 control sera had positive anti-O157 LPS titers. ELISA results for SLT I and II were compared with those of HeLa cell cytotoxicity neutralization assays on both patient and control sera. Neutralization assays detected anti-SLT I antibodies in at least one serum sample from each of 17 (20%) patients and 7 (10.6%) controls, while 16 (19%) patients and 7 controls had positive titers by anti-SLT I ELISA. Although all serum samples, including control sera, showed nonspecific neutralization of SLT II, no antibody titers to SLT II were detected by either neutralization or ELISA. These results indicate that ELISAs for SLT I and SLT II antibodies are comparable to HeLa cell cytotoxicity neutralization assays. Both the ELISAs and neutralization assays are insensitive in detecting infected patients. However, the ELISA for antibodies toE. coli O157 LPS is both sensitive and specific, and may be more useful than assays for antitoxic antibodies in detecting persons withE. coli O157:H7 infection.     

Elemental and isotopic analyses of mammalian fauna from Southern Africa and their implications for paleodietary research

Elemental analyses of mammalian bone (e.g., strontium-calcium ratios, or Sr/Ca) distinguish between herbivores and carnivores; however, the relationships among herbivores are unclear. To study this question, a modern faunal sample from the Nagupande Tsetse Control Area (Zambezi drainage, Northwestern Zimbabwe) was used. This collection has the advantage of well-established geographical controls in addition to a varied fauna, which includes both bovids and suids. The grazing/browsing dietary status of each species was ascertained by means of isotopic analysis of carbon. Clear differences were seen in the δ¹³C of grazing and browsing animals; a specialized grazer was found to have significantly lower Sr/Ca than less specialized grazers and browsers. In this study it was also possible to examine differences in Sr/Ca by sex; female warthogs were found to have significantly lower Sr/Ca than males. The variation in certain animal groups was found to be abnormal. Implications for reconstruction of prehistoric human diets using trace-element techniques are discussed.     

Glycosaminoglycans vary in accumulation along the neuraxis during spinal neurulation in the mouse embryo

We have utilized the method of whole embryo culture for metabolic labeling of mouse embryos with [3H]glucosamine during closure of neural folds at the posterior neuropore (27- to 29-somite stage). Accumulations of newly synthesized glycopeptides, lactosaminoglycans, hyaluronate, and sulfated glycosaminoglycans (GAG) were assessed by ion-exchange chromatography of glycoconjugates isolated from labeled embryos. Accumulation of hyaluronate and sulfated GAG was greatest in the posterior neuropore and decreased progressively toward the hindbrain where neurulation was already complete. Hyaluronate comprised a progressively smaller proportion of total newly synthesized glycoconjugate from the posterior neuropore toward the cranial region and glycopeptides showed the opposite trend. Sulfated GAG and lactosaminoglycans showed no consistent differences in relative abundance along the neuraxis. Autoradiographic analysis of newly synthesized glycoconjugates revealed especially heavy incorporation into developing basement membranes, beneath the neuroepithelium and around the notochord, in the posterior neuropore and recently closed neural tube regions, but not at more cranial levels of the neuraxis. Predigestion of sections with a specific hyaluronidase showed a significant quantity of this glycoconjugate to be hyaluronate. These results are consistent with a role for neuroepithelial and notochordal basement membrane hyaluronate in spinal neurulation.     

Localization of vitamin D3-responsive alkaline phosphatase in cultured chondrocytes

Alkaline phosphatase activity appears to be altered when chondrocyte cultures are incubated with 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3). This study examined whether the hormone-responsive enzyme activity is associated with alkaline phosphatase-enriched extracellular membrane organelles called matrix vesicles. Confluent, third passage cultures of rat costochondral growth cartilage (GC) or resting zone chondrocytes (RC) were incubated with 1,25-(OH)2D3 or 24,25-dihydroxyvitamin D3 (24,25-(OH)2D3) and enzyme specific activity was assayed in the cell layer or in isolated matrix vesicle and plasma membrane fractions. Alkaline phosphatase-specific activity in the matrix vesicles was enriched at least 2-fold over that of the plasma membrane and 10-fold over that of the cell layer. Matrix vesicle alkaline phosphatase was stimulated by 1,25-(OH)2D3 in GC cultures and by 24,25-(OH)2D3 in RC cultures. The cell layer failed to reveal these subtle differences. 1,25-(OH)2D3 increased GC enzyme activity but the effect was one-half that observed in the matrix vesicles alone. No effect of 1,25-(OH)2D3 on enzyme activity of the RC cell layer or of 24,25-(OH)2D3 on either GC or RC cell layers was detected. Thus, response to the metabolites is dependent on chondrocytic differentiation and is site specific: the matrix vesicle fraction is targeted and not the cells per se.     

Effect of photoperiod and temperature on the life-cycle of different populations of the peacock butterfly Inachis io

The variation in response to photoperiod and temperature of different populations of the peacock butterfly, Inachis io (L.) (Lepidoptera: Nymphalidae), was investigated to test the extent to which species can adjust their response to the environment, and therefore maximise their reproductive potential. The photoperiodic (adult) diapause induction response varies between populations, and appears to be finely tuned to the local conditions. There is however variation within populations and the response can be adjusted in a population by selective breeding. The developmental rate is not significantly different between three latitudinally distinct populations, over the range of temperatures tested, and pupal weights are similar at given temperatures. However, pupal weights increase with decreasing development temperature. The implications of these findings are discussed with reference to modelling life history strategies.

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Résumé Inachis io (L.), observable dans une grande partie de l'Europe, y présente des différences quant au cycle biologique, au voltinisme et à la durée du développement. Trois populations provenant de la zone de transition entre monovoltinisme et bivoltinisme ont été échantillonnées pour examiner les différences de réponses à la photopériode et à la température, et pour évaluer l'étendue des possibilités d'adaptation de cette espèce à l'environnement et ainsi optimaliser son potentiel reproductif. L'induction photopériodique de la diapause est de type jour long pour toutes les souches examinées, mais la photopériode critique 50 (CPh50) varie suivant les populations et paraît étroitement ajustée aux conditions locales. Il y a cependant assez de variabilité à l'intérieur des populations pour que le seuil puisse être rapidement abaissé dans chaque population par des expériences d'élevage sélectif. Par contre, la vitesse de développement ne varie pas significativement entre les populations pour la gamme de température: 15–27°C. Les poids de chrysalides ne diffèrent pas suivant les populations, bien qu'ils augmentent quand la température de dévelppement diminue. On peut penser que des modèles, prédisant que la diminution du nombre de générations pendant une saison sera accompagnée d'une prolongation de la durée de développement et d'une augmentation de la taille, et que ceci est d'origine génétique et non le résultat seul du refroidissement de l'environnement, ne tiendront pas compte de l'absence de variation entre populations dans la relation entre température et développement.

     

Metabolic adaptations in goat mammary tissue during pregnancy and lactation

Metabolic adaptations of goat mammary tissue during pregnancy and lactation were monitored in serial biopsies of the tissue. Changes in the synthetic capacity of secretory cells were studied by combining measurements of enzyme activities with short-term culture of mammary explants to measure lactose, casein and total protein synthesis. By these criteria, the main phase of mammary differentiation began in late pregnancy and was essentially complete by Week 5 of lactation, coinciding with the achievement of peak milk yield. While milk yield declined after Week 5, the activities of key enzymes expressed per mg DNA and the rates of lactose and casein synthesis in mammary explants were maintained over a considerable period. The results suggest that changes in the synthetic capacity of epithelial cells may account for much of the rise in milk yield in early lactation, but are not responsible for the declining phase of milk production characteristic of lactation in ruminants.     

Interactions amongst species in a guild of subtidal benthic herbivores

Summary The subtidal coralline flats of northeastern New Zealand support a characteristic guild of grazing herbivores. The most important members of this guild are an echinometrid echinoid, patellid, turbinid and trochid gastropods. Densities of these herbivores fluctuate through time. Interactions within and among the different species of echinoids and gastropods were investigated experimentally. Different combinations of species were caged at densities up to 5 times that of ambient for a 24 week period in an experiment designed to differentiate between intra- and interspecific competition.The echinoidEvechinus chloroticus and the turbinid gastropodCookia sulcata exhibited reduced mean dry weight with increasing intraspecific densities. There was little evidence of density-related mortality in these species. The limpetCellana stellifera showed comparatively large losses of weight and enhanced mortalities in intraspecific experimental treatments but this was not related to density.Investigation of interspecific interactions amongstEvechinus andCookia revealed no evidence of a negative influence of one species on the other. In terms of dry weight,Cookia was indifferent to the presence ofEvechinus, andEvechinus benefited in the reciprocal interaction.Cookia also enjoyed an enhanced mean dry weight when in the presence ofCellana compared to the equivalent intraspecific treatments. There were no coherent trends in proportional mortality in any treatments with enhanced interspecific densities. Cellana, in the presence ofCookia, exhibited a dramatic decrease in mortality rate and increase in mean dry weight. The presence of the turbinid gastropod was clearly beneficial to the limpet when compared to the intraspecific treatments with enhanced intraspecific densities and the control cages containingCellana at ambient density. We suggest that subtidal areas constitute poor habitats for limpets in the absence of agents such asCookia which may provide or maintain suitable sites for attachment and grazing.For the combinations of densities and species investigated there was a consistent trend towards positive interspecific interactions. It seems unlikely that at the sites investigated interspecific competition could act to restrict distributions, or limit abundances of species.     

Critical nuclear DNA size and distribution associated with S phase initiation

Using HeLa S-3 cells synchronized by selective detachment, in this paper we report a parallel study of nuclear morphology and autoradiography grain patterns between middle G₁ and middle S phases: Our results show two distinct [³H]-thymidine labeling patterns. The first “peripheral” labeling pattern has a characteristic nuclear size distribution, in contrast to the heterogeneous and varying size distributions of Feulgen-stained nuclei, and apparently is characteristic of very early S phase. The sizes of the second labeling pattern—homogeneous or inhomogeneous grain distribution throughout the nucleus—are equal or larger than the first and vary with S phase progression. Together, the corresponding nuclear sizes of the labeled nuclei represent the larger extreme of nuclear areas, and the labeling index closely parallels the fraction of nuclei with areas larger than the minimum size of the labeled nuclei. These results suggest a characteristic nuclear size (reflecting unique intranuclear DNA distribution) as a necessary, if not sufficient, requirement for S phase initiation. Parallel experimentation with rat liver cells—synchronized in vivo by partial hepatectomy and analyzed by thin section autoradiography—confirms the existence of a peripheral labeling pattern in both the very early part and the very late part of S phase, which reconciles our data with previous results and points to the fact that both initiation and termination sites for DNA replication are near the nuclear periphery.     

Partial trisomy 11p with interatrial septal aneurysm. Case report and literature review

A newborn female infant presented with hypotonia, joint hyperextensibility, cardiac murmur, macroglossia, and hepatosplenomegaly. Karyotype of the child revealed partial trisomy of chromosome 11p derived from a paternal balanced translocation. Echocardiogram obtained in the newborn period suggested interatrial aneurysm, which was confirmed on post-mortem examination. Interatrial septal aneurysm is a rare abnormality not previously described in partial trisomy 11p.     

Effect of various excitatory agonists on the secretion of 5-hydroxytryptamine from permeabilised human platelets induced by Ca2+ in the presence or absence of GTP

Addition of GTP markedly enhances the ability of thrombin to cause a leftward shift in the Ca2+ dose/response curve for 5-hydroxytryptamine secretion from permeabilised human platelets. Little effect is observed on addition of GTP in the absence of thrombin. Neither ADP nor adrenaline, in the presence or absence of GTP, causes such a shift, whereas 5-hydroxytryptamine does so to a small extent but only in the presence of GTP. The leftward shift in the Ca2+ dose/response curve induced by 12-O-tetradecanoyl-phorbol-13-acetate or 1-oleyl-2-acetylglycerol is not enhanced by addition of GTP. The thrombin concentration required for half-maximal enhancement of the response to Ca2+ is markedly reduced by addition of GTP. The results support the postulate that the effects of excitatory agonists in this system correlate with their ability to activate phospholipase C and provide further evidence for a role for GTP in signal transduction between the receptor and phospholipase C.