Production and Characterization of Antibodies against Each of the Three Subunits of the Bacillus cereus Nonhemolytic Enterotoxin Complex

The nonhemolytic enterotoxin (Nhe) is one of the two three-component enterotoxins which are responsible for diarrheal food poisoning syndrome caused by Bacillus cereus. To facilitate the detection of this toxin, consisting of the subunits NheA, NheB, and NheC, a complete set of high-affinity antibodies against each of the three components was established and characterized. A rabbit antiserum specific for the C-terminal part (15 amino acids) of NheC was produced using a respective synthetic peptide coupled to a protein carrier for immunization. Using purified B. cereus exoprotein preparations as immunogens, one monoclonal antibody against NheA and several antibodies against NheB were obtained. No cross-reactivity with other proteins produced by different strains of B. cereus was observed. Antibodies against the NheB component were able to neutralize the cytotoxic activity (up to 98%) of Nhe. Based on indirect enzyme immunoassays, the antibodies developed in this study were successfully used in the characterization of the enterotoxic activity of several B. cereus strains. For the first time, it could be shown that strains carrying the nhe genes usually express the complete set of the three components, including NheC. However, the amount of toxin produced varies considerably between the different strains.     

Retinal Oximetry with Scanning Laser Ophthalmoscope in Infants

Purpose

Dual wavelength retinal oximetry has been developed for adults, but is not available for infants. Retinal oximetry may provide insight into the pathophysiology of oxygen-mediated diseases like retinopathy of prematurity. More insight in the oxygen metabolism of the retina in infants may provide valuable clues for better understanding and subsequent prevention or treatment of the disease. The measurements of oxygen saturation are obtained with two fundus images simultaneously captured in two different wavelengths of light. The comparison in light absorption of oxygenated and deoxygenated hemoglobin can be used to estimate the oxygen saturation within the retinal vessels by means of a software algorithm. This study aims to make retinal oximetry available for neonates. The first step towards estimating retinal oxygen saturation is determining the optical density ratio. Therefore, the purpose of this study is to image healthy newborn infants with a scanning laser ophthalmoscope and determine the optical density ratio for retinal oximetry analysis.

Methods

Images of the retina of full-term healthy infants were obtained with an SLO, Optomap 200Tx (Optos), with two laser wavelengths (532nm and 633nm). The infant lay face down on the lower arm of the parent, while the parent supported the chest and chin with one hand, and stabilized the back with the other hand. No mydriatics or eyelid specula were used during this study. The images were analyzed with modified Oxymap Analyzer software for calculation of the Optical Density Ratio (ODR) and vessel width. The ODR is inversely and approximately linearly related to the oxygen saturation. Measurements were included from the superotemporal vessel pair. A paired t-test was used for statistical analysis.

Results

Fifty-nine infants, (58% female), were included with mean gestational age of 40 ± 1.3 weeks (mean ± SD) and mean post-natal age of 16 ± 4.8 days. A total of 28 images were selected for retinal oximetry analysis. The ODR was 0.256 ± 0.041 for the arterioles and 0.421 ± 0.089 for the venules (n = 28, p < 0.001). The measured vessel-width for the arterioles was 14.1 ± 2.7 pixels and for the venules 19.7 ± 3.7 pixels (n = 28, p < 0.001).

Conclusions

Retinal oximetry can be performed in newborn infants by combining an SLO and a dual-wavelength algorithm software. Sensitivity of the approach is indicated by the fact that the ODR measurements are significantly different between the arterioles and the venules. However, more variability in ODR is seen with the SLO approach in babies than is seen with conventional oximetry in adults. This approach is completely non-invasive, non-contact and even avoids the use of mydriatics or eyelid specula.     

Do foraging methods in winter affect morphology during growth in juvenile snow geese?

Physical exertion during growth can affect ultimate size and density of skeletal structures. Such changes from different exercise regimes may explain morphological differences between groups, such as those exhibited by lesser snow geese (Chen caerulescens caerulescens; hereafter snow geese) foraging in southwest Louisiana. In rice‐prairie habitats (hereafter rice‐prairies), snow geese bite off or graze aboveground vegetation, whereas they dig or grub for subterranean plant parts in adjacent coastal marshes. Grubbing involves considerably more muscular exertion than does grazing. Thus, we hypothesized that rates of bone formation and growth would be lower for juveniles wintering in rice‐prairies than those in coastal marshes, resulting in smaller bill and skull features at adulthood. First, we tested this exertion hypothesis by measuring bills, skulls, and associated musculature from arrival to departure (November–February) in both habitats in southwest Louisiana, using both banded birds and collected specimens. Second, we used the morphological data to test an alternative hypothesis, which states that smaller bill dimensions in rice‐prairies evolved because of hybridization with Ross's geese (C. rossii). Under the exertion hypothesis, we predicted that bill and skull bones of juveniles would grow at different rates between habitats. However, we found that bill and skull bones of juveniles grew similarly between habitats, thus failing to support the exertion hypothesis. Morphometrics were more likely to differ by sex or change with sampling date than to differ by habitat. We predicted that significant, consistent skewness toward smaller birds could indicate hybridization with Ross's geese, but no skewness was observed in our morphological data, which fails to support the hybridization hypothesis. Further research is needed to clarify whether snow geese wintering in Louisiana represent a single polymorphic population that segregates into individually preferred habitats, which we believe at present to be more likely as an explanation than two ecologically and spatially distinct morphotypes.     

Winter diets of four seabird species in the Barents Sea after a crash in the capelin stock

Summary I describe the winter diets (early March) of four seabird species, black-legged kittiwake (Rissa tridactyla), northern fulmar (Fulmarus glacialis), glaucous gull (Larus hyperboreus) and Brüinnich's guillemot (Uria lomvia) collected in the south eastern Barents Sea, in open water approximately 70 km from the ice edge. All species preyed heavily on commercial fish species such as cod (Gadus morrhua), polar cod (Boreogadus saida) and redfish (Sebastes marinus/S. mentella). The median total length of fish eaten ranged from 51 mm in fulmars to 88 mm in Brünnich's guillemots and differed significantly among all species pairs except kittiwake and glaucous gull. Although the size of fish eaten differed among the four bird species they all fed upon much the same age categories of fish. Cod and redfish eaten were almost exclusively one year old, whereas polar cod was 1–4 years old. Other important food items were crustaceans (B unnich's guillemots), and squid Gonatus sp. (fulmars); the glaucous gull also preyed on other birds. Body weights and amounts of stored fat suggest that individuals of all species except Brüinnich's guillemots were in good physical condition. The diets described here differed substantially from those of birds caught mainly during the prelaying season near the colonies in this area. Some of these differences may be attributed to the very low density of capelin (Mallotus villosus) when this study was conducted.     

Long-term Anticoagulant Therapy after Myocardial Infarction in Women

A multicentre trial from five medical departments in Oslo has been carried out to determine the value in women patients of one year''s long-term anticoagulant therapy. Follow-up long-term laboratory control and anticoagulant dosage were performed at one centre (the Rikshospitalet). One hundred and fifty-nine patients were assigned randomly into two similar well-matched groups (control and treatment). Dosage was controlled by Thrombotest, aiming at 10–20% levels, and 50% of the tests were less than 14%. Compared with the control group, the treatment group showed a significant reduction in mortality and in reinfarction rate. No serious bleeding complications occurred. It is concluded that women benefit as much as men from long-term anticoagulant therapy.     

Mechanism of the Arginine Requirement for Adenovirus Synthesis: I. Synthesis of Structural Proteins

The mechanism of the arginine requirement for adenovirus was studied in cultures of KB cells infected with adenovirus type 2. Macromolecular synthesis was found to be severely impaired in uninfected cells under complete arginine deprivation, whereas an arginine concentration of 50 mum yielded a moderate and reversible inhibition of growth and nucleic acid synthesis. At this concentration, viral structural proteins were accumulated in excess although the virus yield was reduced more than 1,000-fold. The arginine-sensitive step appeared to occur early during the first 15 hr postinfection in the virus growth cycle. Virus-infected cells deprived of arginine to 50 mum showed, when reversed, a 4- to 5-hr lag period before the increase in virus growth was observed. Analysis of the radioactive pattern of labeled virions synthesized after reversion showed that all polypeptides were synthesized after addition of arginine to the medium, and none of the virion-polypeptides which are revealed by gel electrophoresis appeared to be preferentially synthesized after arginine reversion. The excess pool of structural proteins formed during depletion appeared to a large extent to be unavailable for virus assembly.     

The course and prerequisites of Lys-plasminogen formation during fibrinolysis

Plasmin-catalyzed modification of the native plasma zymogen Glu1-plasminogen to its more reactive Lys78 form has been shown to be enhanced in the presence of fibrin. The aim of the present work has been to characterize the influence of fibrinopeptide release, fibrin polymerization, and plasmin cleavage of fibrin on the rate of Lys78-plasminogen formation. 125I-Labeled Glu1- to Lys78-plasminogen conversion was catalyzed by performed Lys78-plasmin, or by plasmin generated during plasminogen activation with tissue plasminogen activator or urokinase. The two forms of plasminogen were quantitated following separation by polyacrylamide gel electrophoresis in acetic acid/urea. Plasmin generated by plasminogen activator was monitored by a fixed-time amidolytic assay. The rate of Lys78-plasminogen formation was correlated, in separate experiments, to the simultaneous, plasmin-catalyzed cleavage of 125I-labeled fibrinogen or fibrin to fragments X, Y, and D. The radiolabeled components were quantitated after separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The results show that the formation of both bathroxobin-catalyzed des-A-fibrin and thrombin-catalyzed des-AB-fibrin leads to marked stimulation of Lys78-plasminogen formation, whereas inhibition of fibrin polymerization, with Gly-Pro-Arg-Pro, abolishes the stimulatory effect. The rate of Lys78-plasminogen formation varies markedly in the course of fibrinolysis. The apparent second-order rate constant of the reaction undergoes a transient increase upon transformation of fibrin to des-A(B) fragment X polymer and decreases about 10-fold to the level observed during fibrinogenolysis upon further degradation to soluble fragments Y and D.(ABSTRACT TRUNCATED AT 250 WORDS)     

Dispersal of the nuclear polyhedrosis virus of Neodiprion sertifer from soil to pine foliage with dust

Inclusion bodies (PIB) of nuclear polyhedrosis viruses (NPV) remain infectious for long periods in forest soils, but their transfer from soil to foliage is not well understood. Two instances of dispersal of the NPV of Neodiprion sertifer (Fourcroy) (Hymenoptera: Diprionidae) by dust are reported. The distributioo of diseased larval colonies in a young Scots pine stand indicated that dust caused by logging during winter could have started an epizootic. At another site there was a higher frequency of infected colonies near a forest road than further into the stand. The dust transmission hypothesis was tested by bioassays of road surface material and of samples of airborne dust collected on membrane filters. Both soil and air samples contained viable PIBs. Thus, virus dispersal with dust provided a likely explanation for the observed distributions of diseased larval colonies.

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Zusammenfassung Mortalität von N.sertifer Larven durch das wirtsspezifische Kernpolyedervirus ist zwischen den Massenvermehrungen der Sägewespe selten. Eine ständige Infektionskette bleibt nicht erhalten. Das einzige bekannte Reservoir von Virusinoculum zwischen den Massenvermehrungen ist der Waldboden. Die Übertragung von Virus vom Boden auf Kiefernnadeln wird als seltenes Ereignis betrachtet. Sie dürfte häufiger sein, wenn die bodenbedeckende Vegetation gestört ist. Es wird von zwei Studien berichtet, in denen offenbar NPV in windverfrachtetem Staub die Krankheit bei Larven verursachte.In einem jungen Bestand schottischer Kiefer, in dem keine neueren Meldungen von Virusauftreten vorlagen, wurden Proben genommen. Die Proben stammten aus verschiedenen Distanzen von einem ausgewachsenen, im vorherigen Winter gefällten Bestand. Die Befallsintensität sank hier scharf ab mit zunehmender Distanz vom Rand.Ferner wurde der Krankheitsbefall in Larvenkolonien an kleinen Bäumen nahe einer schmalen Forst-strasse und an Bäumen im Bestandesinnern untersucht: In Strassennähe war der Befall höher. Luftmuster wurden nahe der Strasse durch Membranfiltern gesaugt und in einem Biotest auf Virus untersucht. Wenn die Strassenoberfläche ungestört war, wurde kein Virus festgestellt. Wurde Staub aufgewirbelt, konnten infektiöse PIBs gefunden werden. Material von der Strassenoberfläche hatte eine hohe Virusaktivität.Die Übertragung des Virus vom Boden auf Kiefernnadeln dürfte demnach wichtig sein, um Krankheitsausbrüche auszulösen. Die beschriebenen Störungen waren vom Menschen verursacht. Ähnliches könnte sich unter andern Umständen ereignen, z.B. bei Windfall oder bei der Aktivität grösserer Tiere.