Mice were passively immunized with sera from blood donors active for rough lipopolysaccharides (LPS), the J5 (Rc chemotype) mutant of Escherichia coli O111:B4, and the Re595 (Re chemotype) mutant of Salmonella minnesota. All protected the mice against lethal challenge with smooth E. coli WF96 LPS, E. coli and Salmonella rough mutant LPS, or free lipid A. Epitopes recognized by monoclonal antibodies (MAbs) reacting with the LPS of S. minnesota Re595 or lipid A were localized in the 2-keto-3-deoxy-D-manno-octulosonic acid (KDO) region and on lipid A. Core-reactive MAbs reacted with their homologous Re LPS and with free lipid A. One, GL11, cross-reacted with the KDO alone. MAbs GL6, GL11, L.4, L.6, and L.8 protected the actinomycin D-sensitized mice against the lethal effects of LPS from E. coli WF96, Salmonella enteritidis, E. coli J5, S. minnesota Re595, and free lipid A. The GL11 antibody was also protective when injected after LPS challenge. These results indicate that antibodies directed against the core glycolipid of S. minnesota Re595 LPS may be useful as an additive form of therapy that may enable decreased mortality during gram-negative bacterial sepsis.
We prepared solutions of human IgM and IgG to various lipopolysaccharide (LPS) species. These were then tested, along with solutions of non-LPS specific human IgG or IgM, for their ability to confer passive immunity against experimental endotoxemia in two animal models. The immunoglobulins were first tested for an effect on the lethality induced by seven different LPSs in actinomycin-D sensitized mice, or by three different bacteria in normal mice. When the immunoglobulins were administered 1 h before challenge, a small protective effect was observed. This protection was dependent upon both the anti-LPS agent, the chemical composition of the LPS, or the strain of gram-negative bacteria used for injection. The anti-LPS IgM and IgG preparations reduced the mortality induced by Escherichia coli but not by Serratia marcescens or Klebsiella pneumoniae, indicating protection by strain-specific antibodies. When the antibodies were preincubated with LPS or bacteria for 30 min before administration, almost complete protection was seen. The influence of these immunoglobulin preparations or of human albumin (as a control) on the hypotensive and vascular-permeabilizing effects of LPS in rats was then studied. A dose-dependent inhibitory effect was observed with IgG preparations and albumin. At 200 mg/kg, anti-LPS IgG reduced the effects of LPS, while at 400 mg/kg, both anti-LPS and normal IgG preparations showed protection, as did human albumin used at the same dose. The IgM-enriched preparation worsened the initial hypotensive phase after LPS, whereas the anti-LPS IgM significantly reduced the second phase of the hypotension, but only at the largest dose of 400 mg/kg. In this second model using the rat, a clear difference between the activity of IgG and IgM was thus observed. We conclude that pretreatment with human immunoglobulins from large plasma pools modestly, but significantly, attenuated the effects of murine and rat Gram-negative sepsis, but that protection was incomplete. Our results suggest that single regimen intervention strategies may not be sufficient to influence the course of the disease.
ABSTRACT:Liège syrup is a Belgian traditional cooked fruit foodstuff, produced mainly from apples and pears. The process includes several hours of heating at high temperature during which complex chemical reactions occur, such as Maillard condensation between reducing sugars and amino acids. Aiming at understanding the modifications of the fruit juices during heating, different parameters were monitored throughout the process.It was shown that hydoxymethylfurfural was formed during the first step of concentration by heating. At the end of the process, hydroxymethylfurfural had totally disappeared and the deep brown color of the product suggested that this compound was transformed into melanoidins. A parallel increase in antioxidant capacity was also observed. To determine optimal conditions to reach high melanoidin content and high antioxidant capacity, different in vitro model systems were compared. It was shown that different combinations of an amino acid with glucose or fructose led to different levels of hydroxymethyfurfural, of melanoidins and antioxidant capacity. After heating of apple or pear puree, an increase of the antioxidant capacity and the hydroxymethylfurfural and melanoidin contents was observed when the heating time was doubled. An increase of the pH from 5 to 9 in apple marmalade's also induced an increase in antioxidant capacity and in hydroxymethylfurfural and melanoidins. However it was not the case in pear marmalade where only the increase in antioxidant capacity was observed. These results suggest that some parameters of the processing could be modified to improve the health-promoting effect of this traditional food (antioxidant properties and composition in hydroxymethylfurfural and melanoidins). The main factors affecting the quality of the final product were the cooking times, the temperature, the pH, the addition of reducing sugars or amino acids.
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