ABSTRACT
Camelid-derived, single-domain antibodies (VHHs) have proven to be extremely powerful tools in defining the antigenic landscape of immunologically heterogeneous surface proteins. In this report, we generated a phage-displayed VHH library directed against the candidate Lyme disease vaccine antigen, outer surface protein A (OspA). Two alpacas were immunized with recombinant OspA serotype 1 from Borrelia burgdorferi sensu stricto strain B31, in combination with the canine vaccine RECOMBITEK Lyme containing lipidated OspA. The phage library was subjected to two rounds of affinity enrichment ("panning") against recombinant OspA, yielding 21 unique VHHs within two epitope bins, as determined through competition enzyme linked immunosorbent assays (ELISAs) with a panel of OspA-specific human monoclonal antibodies. Epitope refinement was conducted by hydrogen exchange-mass spectrometry. Six of the monovalent VHHs were expressed as human IgG1-Fc fusion proteins and shown to have functional properties associated with protective human monoclonal antibodies, including B. burgdorferi agglutination, outer membrane damage, and complement-dependent borreliacidal activity. The VHHs displayed unique reactivity profiles with the seven OspA serotypes associated with B. burgdorferi genospecies in the United States and Europe consistent with there being unique epitopes across OspA serotypes that should be considered when designing and evaluating multivalent Lyme disease vaccines.
Subject(s)
Lipoproteins , Lyme Disease , Single-Domain Antibodies , Animals , Dogs , Humans , Lyme Disease Vaccines , Epitopes , Antibodies, Bacterial , Bacterial Vaccines , Bacterial Outer Membrane Proteins , Lyme Disease/prevention & control , Antigens, Surface , Antibodies, MonoclonalABSTRACT
Visually perceived eye level (VPEL) and perceived pitch were measured while subjects viewed two sets of stimuli that were either upright or pitched top-toward or top-away from them. The first set of stimuli, a pair of vertical lines viewed at various angles of pitch, caused systematic changes in perceived pitch and upward and downward VPEL shifts for the top-toward and top-away pitches, respectively. Neither the perceived pitch nor the VPEL measures with these stimuli differed between monocular and binocular viewing. The second set of stimuli was constructed so that, when viewed at the appropriate pitch angle, the projected orientations of the lines in the retinal image of each stimulus were similar to those generated by a pair of vertical lines pitched by a lesser amount in the opposite direction. When viewed monocularly, these stimuli appeared pitched in the direction opposite their physical pitch, yet produced VPEL shifts consistent with the direction of their physical pitch. These results clearly demonstrate a dissociation between perceived pitch and VPEL. The same stimuli, when viewed binocularly, appeared pitched in the direction of their physical pitch and caused VPEL shifts indistinguishable from those obtained monocularly. The retinal image orientations of these stimuli, however, corresponded to those of vertical line stimuli pitched in the opposite direction. This finding is therefore consistent with the hypothesis that VPEL and perceived pitch are processed independently, but inconsistent with the specific version of this hypothesis which states that differences in VPEL are determined solely on the basis of the orientation of lines in the retinal image.
Subject(s)
Depth Perception/physiology , Models, Psychological , Adult , Cues , Female , Humans , Male , Middle Aged , Orientation , Photic Stimulation/methods , Retina/physiology , Retinaldehyde/physiology , Vision, Binocular/physiology , Vision, Monocular/physiologyABSTRACT
Obesity, and its associated complications, is one of the most costly diseases in modern civilisations. Dieting alone rarely gives good long-term results. The effect of the combination of nutritional education and moderately intensive physical exercise on the evolution of weight and Body composition has been analysed by bio-impedancemetry over a one-year period. Patients could be divided into four groups: patients lost after the 3-week nutritional course, patients neither dieting nor exercising, patients dieting and patients both dieting and exercising. The results for the four groups were the following: undeterminably, 5% loss compared to initial weight (and nearly 10% compared to reported maximum weight). 10% loss and 15% loss over one year. In the last group, Body composition showed a relative increase in muscle mass, which explains the lack of a drop in basal metabolic rates seen in the diet-alone group. This maintained metabolic rate probably prevented patients from weight cycling (yo-yo phenomenon). This result can be compared to other life-style changing studies or pharmacological treatments (Orlistat, sibutramine) of obesity, which resulted in an approx. 10% weight reduction.
Subject(s)
Body Composition , Body Weight , Obesity/therapy , Patient Compliance , Patient Education as Topic , Anti-Obesity Agents/therapeutic use , Cyclobutanes/therapeutic use , Diet, Reducing , Electric Impedance , Exercise , Female , Humans , Lactones/therapeutic use , Male , Middle Aged , Nutritional Physiological Phenomena , Obesity/physiopathology , Obesity/psychology , Orlistat , Retrospective Studies , Weight LossABSTRACT
The effects of selective inhibitions of both cyclo-oxygenase and lipoxygenase pathways were studied in the isolated, perfused and ventilated guinea-pig lungs. Leukotriene D4 (0.3 nmol) induced a significant bronchoconstriction. This effect was significantly inhibited by IPL 55712 (a SRS-A antagonist) and by Imidazole or Dazoxiben (specific thromboxane synthetase inhibitors), but aspirin and indomethacin were without significant effect on this broncho-constriction. Our results suggest that the principal component of leukotriene D4 induced bronchoconstriction in guinea-pig lungs is primary.
Subject(s)
Anti-Inflammatory Agents/pharmacology , Bronchi/drug effects , SRS-A/pharmacology , Animals , Cyclooxygenase Inhibitors , Guinea Pigs , In Vitro Techniques , Lipoxygenase Inhibitors , Lung/drug effects , Male , PerfusionABSTRACT
The effects of verapamil and propranolol on the LTD4-induced bronchoconstriction were studied in the isolated, perfused and ventilated guinea pig lungs. Verapamil (4.10(-5) et 1, 2.10(-4)M.1(-1], a calcium antagonist, produces a significative inhibition of LTD4-induced bronchoconstriction. On the other hand, propranolol (10(-7)M and 10(-4)M) does not increase the constrictor effects of LTD4 in guinea-pig isolated lung. These first results obtained in guinea-pig lung, confirmed the protective effect of verapamil, effect previously demonstrated in vivo or in vitro by using lung parenchymal strips. We have also confirmed the difference between the in vivo and in vitro effects of propranolol.
Subject(s)
Bronchi/drug effects , Lung/drug effects , Propranolol/pharmacology , SRS-A/pharmacology , Verapamil/pharmacology , Animals , Guinea Pigs , In Vitro Techniques , Male , Muscle Contraction/drug effects , Muscle, Smooth/drug effectsABSTRACT
The effects of selective inhibitors of both cyclo-oxygenase and lipoxygenase pathways were studied in the isolated, perfused and ventilated guinea-pig lungs. Leukotriene D4 (0,3 nmol) induced a significant bronchoconstriction. This effect was significantly inhibited by FPL 55712 (a SRS-A antagonist) and by Imidazole or Dazoxiben (specific thromboxane synthetase inhibitors), but aspirin and indomethacin were without significant effect on this bronchoconstriction. Our results suggests that the principal component of leukotriene D4 induced bronchoconstriction in guinea-pig lungs is primary.