Biomedical subjects

S Kadis

Publications and source records attributed to S Kadis.

11 recordsLinked to original sources

PubMed1992

Characterization of lipopolysaccharides from four Pasteurella haemolytica serotype strains: evidence for presence of sialic acid in serotypes 1 and 5.

Highly purified lipopolysaccharides (LPS) obtained from four strains of Pasteurella haemolytica representative of four different serotypes were studied to ascertain their overall structural elements and sugar and fatty acid compositions. SDS-PAGE analysis revealed that each LPS was of the smooth-type although they differed in migration patterns. Somewhat unusual features of these LPS included the presence of: (a) rhamnose in the core oligosaccharides of serotypes 2 and 3; and (b) sialic acid in the LPS of serotypes 1 and 5. The fatty acids, myristic, hydroxymyristic and palmitic occur in essentially equivalent amounts in each of these LPS. In addition, stearic acid was present in small amounts of serotypes 1 and 5.

Amino Sugars↗

PubMed1992

Protective efficacy of conjugate vaccines against experimental challenge with porcine Actinobacillus pleuropneumoniae.

In an attempt to protect pigs against swine pleuropneumonia induced by Actinobacillus pleuropneumoniae (SPAP) by neutralizing the effects of three virulence factors of A. pleuropneumoniae--the capsular polysaccharide (CP), the lipopolysaccharide (LPS), and the hemolysin protein (HP)--two subunit conjugate vaccines were prepared by covalently coupling the CP to the HP and the LPS to the HP. The CP, LPS, and HP were isolated from A. pleuropneumoniae, strain 4074, serotype 1, and the protective efficacy of the conjugate vaccines in swine experimentally infected with A. pleuropneumoniae was evaluated. Following a booster vaccination, a significant (P < 0.05) IgG antibody response to the CP, LPS, and HP was detected in the vaccinated pigs. The pigs vaccinated with the CP-HP and LPS-HP conjugates exhibited significantly less mortality (P < 0.05) and significantly greater weight gain (P < 0.001) than unvaccinated pigs. Vaccinated pigs exhibited significantly fewer and less extensive gross pulmonary lesions (P < 0.001) when compared with unvaccinated pigs. Thus, on the basis of mortality, weight gains, and pulmonary lesion formation, the two conjugate vaccines used in conjunction with one another provide noticeable protective efficacy against SPAP.

Actinobacillus Infections↗

PubMed1992

Effects of Actinobacillus pleuropneumoniae hemolysin on porcine neutrophil function.

In an attempt to gain insight into the events that take place during Actinobacillus pleuropneumoniae infection, the present study was designed to ascertain the effects of bacterial toxicity on porcine neutrophil functions and viability. Incubation of phagocytes (2 x 10(6)) with opsonized A. pleuropneumoniae 4074 (2 x 10(7) CFU) resulted in phagocytic uptake of less than or equal to 4%. At the same bacterium-to-phagocyte ratio, levels of lactate dehydrogenase activity of 74 and 81% were detected in the extracellular medium after 1.5 and 3 h of incubation, respectively. Furthermore, the ingested bacteria were not killed by the phagocytes. These effects were ascribed to hemolysin produced by the bacteria, because the presence of hemolysin-neutralizing antibody prevented overt cellular damage, significantly increased phagocytic uptake (P less than 0.001), and resulted in an approximately 10-fold decrease in the number of CFU of the ingested bacteria. Cytolytic doses of isolated hemolysin caused dose-related loss of cell viability, diminished bactericidal activity of toxin-treated phagocytes for Escherichia coli, and decreased the ability of the phagocytes to undergo a respiratory burst upon stimulation with phorbol myristic acetate. In contrast, sublytic doses of the hemolysin activated the phagocytes and caused them to respond to phorbol myristic acetate with increased generation of superoxide anion. Because heated (100 degrees C, 5 min) hemolysin preparations did not produce similar effects, we contend that the observed effects were not due to contaminating endotoxin. The data presented herein indicate that A. pleuropneumoniae hemolysin is a potent antiphagocytic virulence factor by virtue of its leukocidal activity. Sublytic doses of the toxin may have important effects on the oxidative metabolism of phagocytic cells.

Actinobacillus pleuropneumoniae↗

PubMed1992

Preparation, characterization, and immunogenicity of conjugate vaccines directed against Actinobacillus pleuropneumoniae virulence determinants.

Conjugate vaccines were prepared in an attempt to protect pigs against swine pleuropneumonia induced by Actinobacillus pleuropneumoniae (SPAP). Two subunit conjugates were prepared by coupling the A. pleuropneumoniae 4074 serotype 1 capsular polysaccharide (CP) to the hemolysin protein (HP) and the lipopolysaccharide (LPS) to the HP. Adipic acid dihydrazide was used as a spacer to facilitate the conjugation in a carbodiimide-mediated reaction. The CP and the LPS were found to be covalently coupled to the HP in the conjugates as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and detergent gel chromatography analyses. Following a booster vaccination, pigs exhibited significantly high (P less than 0.05) immunoglobulin G antibodies against CP, LPS, and HP. The anti-CP and anti-LPS immunoglobulin G antibodies were found to function as opsonins in the phagocytosis of A. pleuropneumoniae by polymorphonuclear leukocytes, whereas antibodies to the HP neutralized the cytotoxic effect of the HP on polymorphonuclear leukocytes. No killing of A. pleuropneumoniae was observed when the effects of the antibodies were tested in the presence of complement. Thus, polysaccharide-protein A. pleuropneumoniae conjugates elicit significant antibody responses against each component of each conjugate, which could be instrumental in protecting swine against SPAP.

Actinobacillus pleuropneumoniae↗

PubMed1992

Inhibition of bactericidal activity of anticapsular antibody by nonspecific antibodies reactive with surface-exposed antigenic determinants on Actinobacillus pleuropneumoniae.

In an attempt to understand the mechanism of serum resistance in Actinobacillus pleuropneumoniae, in the present study we examined various interactions among the bacterial surface constituents, serum antibodies, and complement. Analysis of swine sera revealed the presence of anticapsular antibodies in convalescent-phase sera but not in preimmune sera. Both types of sera contained antibodies which reacted with each of 14 polypeptides present in saline extracts of the bacteria. Absorption of the preimmune sera with intact bacteria depleted antibodies to two of the polypeptides (27 and 32 kDa) and high-molecular-weight (greater than 97.4,000) components which did not stain with Coomassie blue. Data derived from complement consumption and C3-binding experiments indicated that the organism was capable of initiating complement activation and binding C3 during incubation in preimmune and immune sera. Experiments designed to evaluate the bactericidal effectiveness of anticapsular antibody revealed that the purified antibody was bactericidal only when preimmune sera absorbed with intact bacteria were used as a source of complement. The bactericidal effects of anticapsular antibody and absorbed preimmune sera were inhibited in a dose-dependent manner by heat-inactivated preimmune sera and immunoglobulin G derived from the sera. The inhibitory activity of the preimmune sera was neutralized by preincubating the sera with column fractions of the saline extract which contained either the 27- or the 32-kDa polypeptide. These results indicate that serum resistance in A. pleuropneumoniae 4074 could be related to inhibition of the bactericidal action of anticapsular antibody by nonspecific antibodies which recognize surface-exposed epitopes on the polypeptides.

Actinobacillus pleuropneumoniae↗

PubMed1979

Antibody formation in Corynebacterium renale-induced experimental pyelonephritis in the rat.

The immune defense system of the kidney was studied by inducing ascending pyelonephritis in rats with Corynebacterium renale. With the fluorescent antibody technique, C renale organisms were observed in the renal pelvis, but were not coated with antibody until they reached the medulla. Histopathologic evaluation of renal tissues collected serially after inoculation confirmed the presence of infection in the medulla when antibody coating occurred. Serum anti-C renale antibody concentrations increased after antibody-coated bacteria appeared in the urine and kidney. Free anti-C renale antibody was not detected in urine from infected rats, using the microagglutination assay. Antibody coating appears to occur only after C renale organisms invade the medulla during ascending pyelonephritis.

Animals↗

PubMed1978

Influence of iron on Corynebacterium renale-induced pyelonephritis in a rat experimental model.

Growth of Corynebacterium renale in vitro on low-iron medium (1.34 micron) was only slightly less than that on high-iron media (7.16 and 9.85 micron). However, studies on C. renale-induced pyelonephritis using the rat as an experimental model revealed that C. renale cultivated in high-iron media was capable of producing pyelonephritis, but when grown on low-iron medium, these bacteria were noninfective. This apparent avirulence of the bacteria cultivated on low levels of iron could be reversed by injecting the rats intramuscularly with ferric ammonium citrate.

Animals↗

PubMed1978

Minimal nitrogen requirements of Corynebacterium renal strains.

Corynebacterium renale strain 10849 was grown in a chemically defined medium containing glucosamine, ammonium sulfate, and 5 amino acids as possible nitrogen sources. Although glucosamine was slightly stimulatory, its omission from the medium had a minimal effect on growth, and washed cells introduced into glucosamine-free medium grew readily through 10 serial transfers, demonstrating that this compound was not required for growth. Individual omissions of isoleucine, valine, methionine, and glutamine resulted in lengthened lag periods and reduced growth rates in initial transfers, but recovery occurred in subsequent serial transfers so that by the 3rd or 4th transfer, growth rates and cell crops were only slightly less than in control cultures in complete medium. Omission of cystine resulted in a permanently low growth rate and reduced cell crop, but this was remedied by substituting various nonnitrogenous compounds containing reduced sulfur. Strain 10849 and 6 additional strains were then serially cultured in a minimal defined medium in which sodium thioglycolate provided reduced sulfur and ammonium sulfate served as sole nitrogen source. Since only ammonium ion was required as the nitrogen source, it could be concluded that C renale, which rapidly hydrolyzes urea, should find an adequate source of nitrogen for growth in the urinary tract of animals.

Ammonia↗

PubMed1977

Experimental Rat model for Corynebacterium renale-induced pyelonephritis.

The laboratory rat was able to serve as a model for ascending pyelonephritis after implantation of a zinc disk coated with Corynebacterium renale into the urinary bladder because it satisfied three different criteria for infection. The production of an alkaline urine and the presence of significant numbers of C. renale in the kidneys, as well as distinct pyelonephritic lesions as revealed by histological examination, were observed in all rats infected with C. renale. Control rats that harbored sterile disks in their urinary bladders exhibited none of the above effects.

Animals↗

PubMed1977

Influence of acetohydroxamic acid on experimental Corynebacterium renale pyelonephritis.

The role of Corynebacterium renale urease in the establishment of pyelonephritis was studied by the oral administration of acetohydroxamic acid (AHA), a urease inhibitor, to experimentally infected rats. The bacteria were introduced by surgical insertion of a zinc disc containing 1 X 10(6) colony-forming units of C-renale into the urinary bladder whereas sterile discs were implanted in the bladders of the control animals. Daily administration of AHA via the drinking water did not halt the development of pyelonephritis. Larger doses, given by gavage, did accomplish this goal; that is, the pH of the urine was lowered, the number of colony-forming units of C. renale in the kidney was reduced drastically, and pyelonephritic lesions were observed in the kidney by light-microscopic examination. All experimental rats developed cystitis in varying degrees of severity. About 70% of the intact AHA given by gavage was excreted in the urine 24 h after administration of this compound. Rats implanted with a urease-negative mutant of C. renale displayed no signs of pyelonephritis but did develop cystitis.

Animals↗

PubMed1976

Effect of hydroxamic acids on growth and urease activity in Corynebacterium renale.

Studies were conducted on the effect of four different hydroxamic acids (HA), hydroxyurea, acetohydroxamic acid, p-flurobenzoylhydroxamic acid and sorbylhydroxamic acid, on the growth and urease activity of Corynebacterium renale. The addition of each of these HA, at concentrations ranging form 10(-3) to 10(-5) M, to medium containing urea as the sole nitrogen source resulted in a lengthened lag period of growth the extent of which depended upon the concentration of each HA tested as well as the structure of the compound; that is, the size and (or) complexity of the side chain attached to the common terminal group of the molecule. However, the maximal growth levels achieved following conclusion of the exponential phase were not affected by the HA. Investigations on the effect of these HA on the urease activity of intact cells as well as cell-free extracts revealed that in each case the enzymatic activity was inhibited by each of the HA tested. The extent of inhibition with the intact cells was aobut one-half of that observed with cell-free extracts. Direct incubation of cell-free extracts as well as intact cells with each of the HA tested was required for maximal inhibition.

Cell-Free System↗