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PubMed2004

Development of biomarkers based on diet-dependent metabolic serotypes: characteristics of component-based models of metabolic serotypes.

Our research seeks to identify a serum profile, or serotype, that reflects the systemic physiologic modifications resultant from dietary restriction (DR), in part such that this knowledge can be applied for biomarker studies. Direct comparison suggests that component-based classification algorithms consistently out-perform distance-based metrics for studies of nutritional modulation of metabolic serotype, but are subject to over-fitting concerns. Intercohort differences in the sera metabolome could partially obscure the effects of DR. Further analysis now shows that implementation of component-based approaches (also called projection methods) optimized for class separation and controlled for over-fitting have >97% accuracy for distinguishing sera from control or DR rats. DR's effect on the metabolome is shown to be robust across cohorts, but differs in males and females (although some metabolites are affected in both). We demonstrate the utility of projection-based methods for both sample and variable diagnostics, including identification of critical metabolites and samples that are atypical with respect to both class and variable models. Inclusion of non-statistically different variables enhances classification models. Variables that contribute to these models are sharply dependent on mathematical processing techniques; some variables that do not contribute under one paradigm are powerful under alternative mathematical paradigms. In practical terms, this information may find purpose in other endeavors, such as mechanistic studies of DR. Application of these approaches confirms the utility of megavariate data analysis techniques for optimal generation of biomarkers based on nutritional modulation of physiological processes.

Animals↗
PubMed2002

Characterization of diet-dependent metabolic serotypes: primary validation of male and female serotypes in independent cohorts of rats.

Our research seeks to identify serum profiles, or serotypes, that reflect substantial changes in food intake in both male and female rats. This report validates previously defined subsets of redox-active low-molecular-weight metabolites using independent cohorts of ad libitum consumption (AL) and energy or dietary restricted (DR) 6-mo-old male and female rats. In the male study, both hierarchical cluster analysis (HCA) and principal component analysis (PCA) distinguished the dietary groups of origin in the second male cohort with >85% accuracy using 56 analytically and biologically valid metabolites. Further analysis revealed that 29 metabolites (nine previously unidentified metabolites + 20 chosen from the 56 metabolites) enabled HCA to distinguish dietary groups at 100% efficacy. In the female study, the 63 previously identified serum metabolites were sufficiently robust to enable classification of the dietary intake of two female cohorts (cohorts 2 and 3) that were independent of the cohort on which these markers were initially identified (cohort 1). Classification accuracy was 94 and 100% using HCA and PCA, respectively, in the female cohort 2. HCA and PCA revealed that the 63-metabolite profile distinguished AL and DR samples at 91 and 100% accuracy in the female cohort 3, establishing the 63-metabolite dataset as our baseline profile. These studies used independent cohorts to validate and potentially improve upon previously defined metabolic serotype in male and female rats and set the stage for pattern recognition-based approaches to establish metabolome-based categorical separations.

Animals↗
PubMed1984

Purification, characterization and immunological properties of the serotype-specific capsular polysaccharide of Pasteurella haemolytica (serotype A1) organisms.

The serotype-specific capsular polysaccharide from two strains of Pasteurella haemolytica serotype A1 organisms was purified and characterized by chemical analysis and NMR spectroscopy. The polymer has the structure----3)-O-(2-acetamido-2-deoxy-4-O-acetyl-beta-D-mannopyranos yluronic acid)-(1----4)-O-(2-acetamido-2-deoxy-beta-D-mannopyranose)-(1----. The polysaccharide was immunogenic (able to evoke production of antibodies) for sheep but not for rabbits. Immuno electron-microscopy studies using the Protein A-gold technique showed the polysaccharide to be peripherally located on the bacterial surface. Reduction, oxidation and de-O-acetylation of the polymer did not appear to alter its immunological precipitability with specific antiserum, but all three treatments destroyed its ability to adhere to sheep erythrocytes at neutral pH. De-N-acetylation of the polymer destroyed both immunological precipitability and erythrocyte adherence.

Amino Acids↗
PubMed1992

Nuclear-magnetic-resonance analysis of the capsular antigen of Actinobacillus pleuropneumoniae serotype 9. Its identity with the capsular antigen of Escherichia coli K62 (K2ab), Neisseria meningitidis serogroup H and Pasteurella haemolytica serotype T15.

The specific capsular antigen of Actinobacillus pleuropneumoniae serotype 9 was characterized by one-dimensional and two-dimensional high-field nuclear-magnetic-resonance methods, and by chemical analyses, as a teichoic-acid-type polymer of a repeating unit having the structure [formula: see text] The basic polymer structure is identical to capsular antigens of Neisseria meningitidis group H, Escherichia coli K62 (K2ab) and Pasteurella haemolytica serotype T15.

Actinobacillus↗
PubMed1999

Bacillus thuringiensis serotype H34 isolated from human and insecticidal strains serotypes 3a3b and H14 can lead to death of immunocompetent mice after pulmonary infection.

In 1995, we isolated a strain of Bacillus thuringiensis serotype H34 from severe human tissue necrosis. This bacterium was able to induce myonecrosis in immunosuppressed mice after cutaneous infection. Its potential pathogenicity for immunocompetent hosts was investigated in a mouse model of pulmonary infection. Mice infected intranasally by a suspension containing 10(8) spores died within 8 h in a clinical toxic-shock syndrome. In the same conditions, infection with a mutant without crystalline toxin, with the supernatant from a culture containing 10(8) bacteria ml(-1) and by the insecticidal strain serotypes 3a3b or H14 led to identical results. Lower inocula simply induced a local inflammatory reaction with bacterial persistence observed during the course of 10 days.

Animals↗
PubMed1997

Pseudomonas aeruginosa rfc genes of serotypes O2 and O5 could complement O-polymerase-deficient semi-rough mutants of either serotype.

Using a gene-replacement strategy and a mutated copy of the Pseudomonas aeruginosa O5 rfc gene, we were able to generate a rfc mutant in P. aeruginosa serotype O2. This mutant, which exhibits the semi-rough (SR) LPS phenotype, was used to isolate the O2 rfc gene. Mobilization of the O2 and O5 rfc genes into SR mutants of the heterologous serotype resulted in 'cross-polymerization' of O-repeat units, indicating that the genes are functionally exchangeable. Analysis of the nucleotide sequence of the rfc genes revealed that the two Rfc proteins are identical. The results of this study have enabled us to propose the linkage catalyzed by the O5 O-polymerase enzyme.

Base Sequence↗
PubMed1995

Pasteurella haemolytica serotype 2 contains the gene for a noncapsular serotype 1-specific antigen.

An ssa1-homologous genomic fragment cloned from Pasteurella haemolytica serotype 2 (ST2) enabled transformation of Escherichia coli DH5 alpha to a serotype 1 (ST1) phenotype through expression of the ST1-specific antigen (Ssa1). The Ssa1 protein expressed by ssa1-transformed E. coli was susceptible to heat and protease treatment and was distinct from P. haemolytica ST1-specific capsular polysaccharide. Electrophoretic analysis of in vitro-translated proteins, as well as the predicted amino acid sequence, demonstrated that Ssa1 proteins encoded from either ST1- or ST2-derived ssa1 genes were essentially identical. A comparison of the nucleotide sequences of ssa1 genes derived from P. haemolytica ST1 and ST2 revealed greater than 99% homology. Amino acid sequence homology of the predicted products of ST1- and ST2-derived ssa1 genes was greater than 98%. Northern (RNA) blot studies revealed that the presence of an increased level of ssa1 transcript in P. haemolytica ST1 grown as surface-adherent cultures on solid medium was correlated with a serologically detectable Ssa1 protein. Expression of the ssa1 transcript in ST1 was similarly upregulated by a high iron concentration in the growth medium.

Amino Acid Sequence↗
PubMed1991

The Lang strain of reovirus serotype 1 and the Dearing strain of reovirus serotype 3 differ in their sensitivities to beta interferon.

Replication of the Dearing strain of reovirus serotype 3 in mouse L cells was decreased 17- to 100-fold when a saturating dose of beta interferon (1,000 IU/ml) was used. Replication of the Lang strain of reovirus serotype 1 was inhibited only two- to threefold under similar conditions. It therefore appears that closely related strains of reovirus differ in their sensitivities to beta interferon treatment of mouse L cells.

Animals↗
PubMed1988

Use of monoclonal antibodies to serotype-specific antigens of Haemophilus pleuropneumoniae serotype 2 in passive immunization.

The prophylactic value of mouse monoclonal antibodies to the pig pathogen Haemophilus pleuropneumoniae was studied. Approximately 250 mg of purified mouse monoclonal antibody specific to capsular antigens of H pleuropneumoniae serotype 2 was given IV to five 9-week-old pigs. Five additional pigs from the same litter served as controls. On the following day, all pigs were given a lethal dose (5 x 10(9)) of H pleuropneumoniae serotype 2 into the trachea. Four controls and 1 pig that was given antibodies died within 24 hours. The surviving 5 pigs developed typical signs of pleuropneumonia. After 6 days, the pigs were euthanatized and their respiratory tracts were examined for pathologic changes. All 5 pigs had pathologic changes, but they were less severe in the 4 pigs that had been given antibodies, compared with those in the control pig.

Animals↗
PubMedDate not supplied

Difference between influences of homologous and heterologous maternal antibodies on response to serotype-2 and serotype-3 Marek's disease vaccines.

Marek's disease (MD) vaccines representing serotypes 2 (SB-1 strain of MD virus) and 3 (FC-126 strain of turkey herpesvirus) were administered to 1-day-old MD-susceptible chicks that either were free of antibodies or carried maternally derived antibodies against SB-1, FC-126, or a serotype-1 MD virus, CU-2 strain. Homologous antibodies delayed the development of vaccine virus viremias and inhibited vaccinal immunity, as judged by protection against challenge with the virulent JM-10 strain of MD virus 7 days postvaccination. Heterologous antibodies had little effect on vaccine responses. Antibodies were shown to interfere with both cell-associated and cell-free vaccine virus.

Animals↗
PubMed1985

The complete sequence of bluetongue virus serotype 10 segment 3 and its predicted VP3 polypeptide compared with those of BTV serotype 17.

The complete sequence of the large RNA segment 3 (L3) of bluetongue virus serotype 10 (BTV-10) has been determined from DNA copies of the viral RNA cloned in the E. coli plasmid pBR322. The L3 viral RNA is 2772 nucleotides long with a single open reading frame of 2706 nucleotides. The L3 predicted primary gene product (VP3) is 103 342 daltons and has a net charge at neutral pH of -5. The sequence of the L3 RNA species differs by 126 point mutations from that of BTV-17 (i.e., 95.5% homology; see M. Purdy, J. Petre and P. Roy, J. Virol. 51, 754-759, 1984). The predicted L3 primary gene products of the two viruses differ by 9 amino acids. These differences correspond to 9 point mutations and represent 0.15% of the sites where nucleotide substitution could cause an amino acid change. By contrast, another 114 point mutations in the genome correspond to 6.5% of the available sites where nucleotide substitutions could be silent (i.e., where a nucleotide substitution may not cause an amino acid change). Three point mutations are in the 3' non-coding region of the RNA species. The quantitative differences between the coding and silent mutations are interpreted as representing the result of gene product conservation.

Amino Acid Sequence↗
PubMed2005

Recombinant Mannheimia haemolytica serotype 1 outer membrane protein PlpE enhances commercial M. haemolytica vaccine-induced resistance against serotype 6 challenge.

Mannheimia haemolytica outer membrane protein PlpE, a major immunogenic outer membrane lipoprotein has identical sequences in serotypes 1 (S1) and S6. Recombinant outer membrane lipoprotein PlpE (rPLpE) from M. haemolytica S1 was added to commercial M. haemolytica S1 vaccines to determine if it would enhance vaccine-induced immunity against heterotypic M. haemolytica S6 challenge. Serum antibody responses to M. haemolytica whole cells, leukotoxin and rPlpE were measured. Experiment 1 consisted of four vaccine groups: controls, 100 microg rPlpE, M. haemolytica Bacterin-Toxoid (One Shot) and M. haemolytica Bacterin-Toxoid + 100 microg rPlpE. Vaccines were given on day 0. On day 21, calves were challenged transthoracically with M. haemolytica S6. Lung lesion scores and percentage lesion reduction were 6.3 +/- 2.0 for controls, 3.6 +/- 2.4 for rPlpE vaccinates (42.9% reduction), 3.4 +/- 1.5 for One Shot-vaccinates (46.0% reduction), and 2.4 +/- 1.4 for One Shot/rPlpE vaccinates (61.9% reduction). Experiment 2 consisted of four vaccine groups: controls, 100 microg rPlpE, M. haemolytica toxoid (Presponse), and M. haemolytica toxoid+100 microg rPlpE. On day 28, calves were challenged transthoracically with M. haemolytica S6. Lung lesion scores and percentage lesion reduction were 8.1 +/- 2.2 for controls, 4.4 +/- 4.7 for the rPlpE vaccinates (45.7% reduction), 4.8 +/- 2.2 for Presponse-vaccinates (40.7% reduction), and 2.0 +/- 1.2 for Presponse/rPlpE vaccinates (75.3% reduction). These results indicate that addition of rPlpE from M. haemolytica S1 can enhance commercial M. haemolytica vaccine-induced resistance against experimental challenge with M. haemolytica S6.

Amino Acid Sequence↗
PubMed1974

Serotypes of trachoma agent isolated in the Gambia: with an observation on the relation between serotype and morphology.

Of 60 TRIC agents isolated from Gambian children with trachoma, 25 were serotype 1 and the remainder type 2. There was a pronounced difference in the proportions of these types in the two villages studied. In the village with a predominance of type 2 strains, TRIC agents remained confined to 2 adjacent compounds over a 14 month observation period. All 19 type 1 strains examined were characterized by the appearance in yolk sac smears of compact aggregates of elementary bodies; such aggregates were seen in only 2 of 35 type 2 strains, and may reflect a chemical difference in the surface of the elementary bodies or in a substance elaborated during their replication.

Animals↗
PubMed2001

The enterohaemorrhagic Escherichia coli (serotype O157:H7) Tir molecule is not functionally interchangeable for its enteropathogenic E. coli (serotype O127:H6) homologue.

A major virulence determinant of enteropathogenic Escherichia coli (EPEC) is the Tir molecule that is translocated into the plasma membrane where it orchestrates cytoskeletal rearrangements. Tir undergoes several phosphorylation events within host cells, with modification on a tyrosine essential for its actin-nucleating function. The EHEC (serotype O157:H7) Tir homologue is not tyrosine phosphorylated implying that it uses an alternative mechanism to nucleate actin. This is supported in this study by the demonstration that EHEC Tir is unable to functionally substitute for its EPEC homologue. Like EPEC, the EHEC Tir molecule is phosphorylated within host cells, with the actin-nucleating dysfunction correlated to an altered modification profile. In contrast to EHEC Tir, the EPEC Tir molecule mediated actin nucleation whether delivered into host cells by either strain. Thus, it would appear that EHEC encodes specific factor(s) that facilitate the correct modification of its Tir molecule within host cells. Domain-swapping experiments revealed that the N-terminal, alpha-actinin binding, Tir domains were functionally interchangeable, with both the actin-nucleating dysfunction and altered modification profiles linked to the EHEC C-terminal Tir domain. This tyrosine-independent modification process presumably confers an advantage to EHEC O157:H7 and may contribute to the prevalence of this strain in EHEC disease. The presented data are also consistent with EPEC and EHEC sharing non-phosphotyrosine phosphorylation event(s), with an important role for such modifications in Tir function. An EHEC-induced phosphotyrosine dephosphorylation activity is also identified.

Actins↗
PubMed1988

Molecular divergence of the serotype-specific plasmid (pSLT) among strains of Salmonella typhimurium of human and veterinary origin and comparison of pSLT with the serotype specific plasmids of S. enteritidis and S. dublin.

Molecular variants of the serotype-specific plasmid (SSP) of Salmonella typhimurium (pSLT) were recognised in clinical and veterinary isolates by restriction enzyme fingerprinting. Three had undergone minor DNA rearrangements, whereas two had acquired resistance determinants to a wide range of antimicrobial agents including gentamicin, trimethoprim, tetracycline, streptomycin, ampicillin (Ap) and kanamycin (Km). One of the latter was the result of co-integrate formation with an IncX, conjugative R-plasmid that specified ApKm resistance. The co-integrate plasmid (pOG669) was incompatible with, and displaced, pSLT and its molecular variants. The restriction fingerprints of SSPs of S. enteritidis and S. dublin were compared with pSLT. All were related at the 35% level on the basis of a Dice coefficient of similarity. The SSPs of S. enteritidis and S. dublin were incompatible with the co-integrate plasmid pOG669. Whereas in S. enteritidis this resulted from incompatibility with the pSLT component (the SSP was compatible with the IncX component), the converse was found with S. dublin.

Animals↗
PubMed1989

Discovery of new morganocin types of Morganella morganii in strains of diverse serotype and the apparent independence of bacteriocin type from serotype of strains.

A collection of 45 serologically distinct strains of Morganella morganii from several European countries was typed by morganocin production (p) and sensitivity (s). This permitted their differentiation into 33 morganocin p/s types; 15 new types of morganocin and one new morganocin sensitivity type were found and are defined. Morganocin production and sensitivity characteristics appeared to be unrelated to the O and H antigens of the strain. The finest strain recognition in M. morganii might be achieved by a combination of bacteriocin and serotyping methods.

Bacterial Typing Techniques↗
PubMed1991

Fine serotyping of human immunodeficiency virus serotype 1 (HIV-1) and HIV-2 infections by using synthetic oligopeptides representing an immunodominant domain of HIV-1 and HIV-2/simian immunodeficiency virus.

In this study, enzyme immunoassays for detection of type-specific antibodies to human immunodeficiency viruses (HIV) were developed by using short peptides corresponding to sequences located within the immunodominant domain of the transmembrane glycoproteins of both HIV-1 and HIV-2-simian immunodeficiency virus (SIV). The assays were highly sensitive with currently available sera from various geographical areas. Furthermore, they appeared to be more specific in HIV serotyping than the Western blot (immunoblot) assay, since all of the sera were clearly discriminated as one or the other type. It was also shown that in contrast to HIV-1, the C-terminal cysteine residue (amino acid 620, SIV from captive macaques, Mm142 strain) of the HIV-2-SIV peptide is not necessary for recognition of the peptide by antibody to HIV-2.

Amino Acid Sequence↗
PubMed1993

Analysis of serotype-specific antibodies to Trichosporon cutaneum types I and II in patients with summer-type hypersensitivity pneumonitis with monoclonal antibodies to serotype-related polysaccharide antigens.

Summer-type hypersensitivity pneumonitis is the most prevalent type of hypersensitivity pneumonitis in Japan. We constructed a sandwich enzyme-linked immunosorbent assay system for diagnosis of summer-type hypersensitivity pneumonitis in which monoclonal antibodies were used to bind serotype-related polysaccharides to plastic plates, and this system was proven to have sufficient sensitivity and specificity.

Adolescent↗