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Biomedical subjects

H R Smith

Publications and source records attributed to H R Smith.

At least 19 recordsLinked to original sources
PubMed1995

Enteroaggregative strains of Escherichia coli belonging to serotypes O126:H27 and O44:H18 express antigenically similar 18 kDa outer membrane-associated proteins.

Outer membrane-associated proteins of 18 kDa were expressed by enteroaggregative Escherichia coli (EAggEC) belonging to serotypes O126:H27 and O44:H18, which hybridized with a probe derived from a plasmid necessary for enteroaggregative adhesion. The 18 kDa proteins expressed by strains of E. coli, belonging to these serotypes, were surface exposed and antigenically similar but not structurally identical.

Antigens, Bacterial↗
PubMed1995

Detection and differentiation of the gene for toxin co-regulated pili (tcpA) in Vibrio cholerae non-O1 using the polymerase chain reaction.

The polymerase chain reaction has been used to differentiate the gene which encodes the toxin co-regulated pili (tcpA) of the El Tor and classical biotypes of Vibrio cholerae O1. The same PCR primers were applied to strains belonging to non-O1 serogroups that produced cholera toxin. The size of fragment amplified was either identical to the tcpA of biotype El Tor (471 bp) or to the tcpA of biotype classical (617 bp). All strains belonging to the novel epidemic serogroup O139 generated a 471-bp fragment identical to El Tor tcpA. The present study suggests that there may be an association between non-O1 serogroup and tcpA type.

Bacterial Outer Membrane Proteins↗
PubMed1995

A family of murine NK cell receptors specific for target cell MHC class I molecules.

The Ly-49A molecule is an NK cell receptor specific for MHC class I molecules on target cells. When Ly-49A engages H-2Dd, Ly-49A+ NK cells become globally incapable of killing their targets in vitro. This interaction also occurs in vivo. Ly-49A belongs to a family of highly related molecules, including Ly-49C (5E6 antigen) and LGL-1 that also determine NK cell specificity. In the NK gene complex, the Ly-49 family is genetically linked to genes encoding NKR-P1 and CD69 that are structurally related and capable of activating NK cells. Finally, Ly-49 may be related to human molecules that are selectively expressed on NK cells and influence NK cell specificity. These findings highlight the emerging significance of the Ly-49 family in NK cell activity.

Animals↗
PubMed1995

Catabolite repression of the adhesion of Vero cytotoxin-producing Escherichia coli of serogroups O157 and O111.

The virulence traits that mediate Vero cytotoxin-producing E. coli (VTEC) adherence are unclear. Many VTEC strains possess the eaeA gene which is involved in the attaching and effacing effects of enteropathogenic E. coli (EPEC). Most eae-positive VTEC adhered to HEp-2 cells in a localized manner; however some strains did not adhere. Thus we investigated the adhesion of poorly adherent strains, especially those of serogroups O111 and O157. To establish a model, the adherence to HEp-2, INT407 and Caco-2 cells of 12 O157 VTEC and six O111 VTEC isolated from cases of human infection were studied after growth of the bacteria under different conditions. For adhesion tests mannose is usually added during prior broth culture of the bacteria, and during the period of attachment, so that any adhesion due to mannose-sensitive type 1 pili is inhibited. Bacteria cultured in peptone water in the absence of mannose adhered to all three lines; there were localized clusters of bacteria on 1%-82% cells, whether mannose was present during the attachment period or not. Bacteria grown in the presence of D-mannose, or any other sugar that was metabolized, showed little adherence (range 0-9%). alpha-Methyl-glucoside also caused marked inhibition of adhesion. It was concluded that inhibition of adhesion was due to catabolite repression.

Animals↗
PubMed1995

Do silicone breast implants cause autoimmune rheumatic diseases?

Current estimates are that up to a million women in the U.S. have breast implants with the predominant type being the silicone gel implant. Concerns have been raised regarding the safety of silicone gel breast implants with focus upon whether escaped gel might cause inflammatory and immune responses that subsequently lead to autoimmune rheumatic diseases such as progressive systemic sclerosis (scleroderma), systemic lupus erythematosus (SLE), Sjögren's syndrome or rheumatoid arthritis. A spectrum of illnesses ranging from local symptoms to systemic disease is seen in some patients with silicone breast implants, however, it remains to be determined whether such illnesses in these patients are coincidentally associated or are secondary to the implants. Our understanding of the relationship between the presence of autoimmune rheumatic diseases and silicone breast implants is limited. The available data indicate that silicone elicits a minimal immunological response as compared to conventional antigens. The histological, immunological and epidemiological experimental data derived from patients with silicone implants, as well as those from animal studies, are reviewed. These data do not convincingly demonstrate that there is a cause and effect relationship between silicone breast implants and autoimmune diseases. Further investigations are needed to clarify the interaction of silicone with the cellular and humoral immune systems, as well as with host and environmental factors.

Antibodies, Antinuclear↗
PubMed1994

Ly-49 multigene family expressed by IL-2-activated NK cells.

The susceptibility of a target cell to killing by NK cells is inversely correlated with target cell expression of MHC class I molecules. We have recently demonstrated that a murine NK cell surface molecule, Ly-49, is a primary determinant of IL-2-activated NK cell specificity by apparently acting as an inhibitory receptor. Ly-49 seems to interact with polymorphic regions of certain MHC class I molecules on target cells, yet is expressed only by a subset (15 to 20%) of NK cells. If the interaction between Ly-49 and MHC class I represents a paradigm for understanding IL-2-activated NK cell specificity and putative inhibitory IL-2-activated NK cell receptors, these effector cells should express other molecules, perhaps related to Ly-49, that provide the inhibitory receptor function. We now report the isolation of cDNAs, encoding polypeptides related to Ly-49, from a CD3-, IL-2-activated NK cell library. Full length PCR products were also cloned and sequenced, confirming expression in IL-2-activated NK cells. These cDNAs were homologous to the Ly-49 cDNA and seem to be derived from distinct genes. The new cDNAs encode type II integral membrane proteins having lectin superfamily homology and 66 to 79% amino acid identity with Ly-49A. In contrast to previous reports, these cDNAs cannot be allelic variants because they were all isolated from C57BL/6-derived cells. On the basis of nucleotide homology, the cDNAs could be segregated into five groups; one with Ly-49 itself (now termed Ly-49A) and four others containing distinct sequences. One of the new groups comprised three cDNAs that shared identical sequences, but differed from one another by deletions or insertions of short DNA segments in the predicted extracellular region of the polypeptides. Thus, IL-2-activated NK cells express mRNAs for multiple, Ly-49-related molecules.

Animals↗
PubMed1994

Prospective study of verocytotoxin-producing, enteroaggregative and diffusely adherent Escherichia coli in different diarrhoeal states.

One hundred and eighty-one stool specimens from patients with various types of diarrhoea (135 patients) or from non-diarrhoeal controls (23 acute medical patients, 23 inflammatory bowel disease in remission) were investigated using a colony-blot DNA hybridization assay for the presence of Verocytotoxin-producing (VTEC), enteroaggregative (EAggEC) and diffusely adherent (DAEC) Escherichia coli. Twelve patients had probe-positive EAggEC in the stool and 8 of these had diarrhoea, 6 following recent travel. Eight patients had DAEC, 7 of whom had travellers' diarrhoea. Six of 10 (60%) travellers with gastroenteritis, but without a recognized enteric pathogen, were positive for EAggEC (4) or DAEC (2). Five of 10 (50%) travellers with gastroenteritis related to a recognized enteric pathogen also had DAEC identified in their stool. Of the 23 acute medical control patients 11 had been abroad, 4 of these were immigrants and had EAggEC. VTEC were not found and, with one exception, immunoassays for antibodies to E. coli O 157 and O 2 lipopolysaccharides were negative.

Adult↗
PubMed1994

Isolates of Escherichia coli O44:H18 of diverse origin are enteroaggregative.

One hundred thirteen strains of Escherichia coli O44:H18 isolated in several countries over 25 years were examined for adhesion to tissue culture cells and for hybridization with DNA probes. Fifty-nine strains were from sporadic cases of infection; 54 were from 12 outbreaks. Of the 113 strains, 85 showed aggregative adhesion to HEp-2 cells; 36 were from sporadic cases and 49 were from 9 outbreaks. All adhesive strains hybridized with the probe for enteroaggregative E. coli (EAggEC) and 1 nonadhesive strain was also positive. However, 80 of the 86 EAggEC probe-positive strains also hybridized with the probe for diffusely adherent E. coli (DAEC) derived from the daaC gene of strain F1845. The EAggEC and DAEC probes hybridized to different fragments of a large plasmid in the O44:H18 strains.

Aged↗
PubMed1994

The detection of Vero cytotoxin-producing Escherichia coli and Shigella dysenteriae type 1 in faecal specimens using polymerase chain reaction gene amplification.

Fifty consecutive faecal specimens received by the LEP were examined for the presence of Vero cytotoxin (VT) genes by polymerase chain reaction (PCR) gene amplification. Nineteen were positive by PCR and from 16 of these, VT positive Escherichia coli O157 were isolated. The remaining three samples were positive for VT genes by PCR but VTEC were not isolated. In a preliminary experiment, Shigella dysenteriae type 1 was isolated from a case of bloody diarrhoea following a positive amplification result.

Bacterial Toxins↗
PubMed1994

Hybridization of strains of Escherichia coli O157 with probes derived from the eaeA gene of enteropathogenic E. coli and the eaeA homolog from a Vero cytotoxin-producing strain of E. coli O157.

A total of 375 Escherichia coli O157 strains were tested by colony hybridization with the eae probe from the central portion of the eaeA gene of the classical enteropathogenic E. coli strain E2348/69. They were also tested with a probe, eaeO157, from the C-terminal end of the eae gene homolog from a Vero cytotoxin (VT)-producing strain of E. coli (VTEC) of serotype O157:H7. Both probes hybridized with all 246 O157:H7 or H- VTEC strains tested. The majority were from human infections, and the remainder were from cattle. A further 10 strains (H7 or H-) hybridized with both eae and eaeO157 sequences but not with VT probes. They resembled O157 VTEC and were probably naturally occurring derivatives that had lost VT genes. The remaining 119 strains of O157 were from human, animal, and food sources and belonged to 16 H types other than H7 or were H-. They were VT negative and differed in their properties from O157 VTEC: generally they fermented sorbitol in 1 day, produced beta-glucuronidase, and could not be phage typed by the scheme for O157 VTEC. The eae probe but not the eaeO157 sequence hybridized with 18 H8 or H39 strains, predominantly from human diarrhea. The remaining 101 VT-negative strains hybridized with neither probe. However, 16 strains of O157:H45 hybridized with a probe for diffusely adherent E. coli and attached to HEp-2 cells in a diffuse pattern. Serogroup O157 comprises strains with heterogeneous properties. The eaeO157 probe is a valuable addition to the VT probes used to differentiate O157 strains.

Animals↗
PubMed1993

Use of digoxigenin-labelled oligonucleotide DNA probes for VT2 and VT2 human variant genes to differentiate Vero cytotoxin-producing Escherichia coli strains of serogroup O157.

Digoxigenin-labelled oligonucleotide DNA probes specific for B-subunit genes of Vero cytotoxin 2 (VT2) and a variant of VT2 (VT2vha) were used to differentiate 116 strains of Escherichia coli serogroup O157 belonging to phage types 1, 2, 4, 8, 14, and 49. Of these strains, 38% had sequences for both VT2 and VT2vha, 38% had sequences for VT2 only, and 24% had sequences for VT2vha only. Oligonucleotide probe hybridization subdivided strains of all of the phage types except phage type 1. The greatest variation in toxin gene pattern was observed with strains of phage type 14, for which there were six distinct patterns when the presence or absence of VT1 genes was also considered. Two strains from each phage type group were examined for bacteriophages encoding VT production. Two of the six VT2vha-producing strains carried phage from which DNA hybridized with the VT2vha-specific probe. Phages were not detected in the remaining four VT2vha strains, suggesting that genes may be chromosomally located or associated with a defective prophage. In contrast, seven of the eight VT2 strains carried phages from which DNA hybridized with the VT2-specific probe. Two strains of E32511 (O157:H-) were also investigated. One strain (E32511A) possessed gene sequences for both VT2 and VTvha and was shown to carry phage possessing gene sequences for VT2. With strain E32511B, however, phages were not detected and DNA hybridized only with the VT2vha probe. Analysis of total genomic DNA digested with restriction endonuclease EcoRI showed that polymorphisms were seen with VT2 strains and not with VT2vha strains.

Bacterial Toxins↗
PubMed1992

Vero cytotoxin-producing Escherichia coli O 157 in the United Kingdom.

A total of 226 cultures of Vero cytotoxin-producing Escherichia coli O 157 isolated from humans was received by the PHLS Laboratory of Enteric Pathogens during the six month period January to June 1992. A record monthly total of 122 isolates was received in June. Ten phage types (PT) were identified during this period; PT 2 (45%) and PT 49 (21%) predominated. In addition, 27 human sera with antibodies to E. coli O 157 lipopolysaccharide (LPS) were examined from other cases during this period, making a total of 253 cases of infection associated with E. coli O 157.

Bacterial Toxins↗
PubMed1992

Inflammatory cells and eicosanoid mediators in subjects with late asthmatic responses and increases in airway responsiveness.

To determine the relationship of inflammatory cells and eicosanoid mediators to the pathogenesis of the late asthmatic response (LAR) and increases in nonspecific airway responsiveness, we studied bronchoalveolar lavage (BAL) cells and fluid in 27 subjects 12 hours after inhaled antigen challenge. Methacholine challenge was performed before antigen challenge and 24 hours later (12 hours after BAL). Eight subjects had no LAR (-LAR, less than or equal to 10% fall in FEV1), nine subjects had an equivocal LAR (+/- LAR, 11% 25% fall in FEV1), and 10 subjects had a definite LAR (+LAR, greater than 25% fall in FEV1). Subjects developing +LAR had increased airway responsiveness at baseline compared with that of subjects developing an +/- LAR, but not with subjects having -LAR. If airway responsiveness was markedly increased at baseline, further increases after antigen challenge were often not observed. We found that both percent neutrophils and eosinophils increased in BAL as the severity of the LAR increased, but significant differences between the groups with -LAR and +LAR were only observed when both cell types were considered together. In addition, there was a significant correlation between the combined cell percentages and the severity of the LAR as determined by fall in FEV1. Likewise, increases in airway responsiveness were associated with significant increases in both neutrophil and eosinophil numbers, but only neutrophils correlated with the change in airway responsiveness after antigen challenge. However, despite the significant physiologic and cellular differences that we found between our groups, no significant differences could be found in BAL eicosanoid-mediator concentrations.(ABSTRACT TRUNCATED AT 250 WORDS)

Airway Resistance↗
PubMed1992

Aerosolized antigen exposure without adjuvant causes increased IgE production and increased airway responsiveness in the mouse.

Inhalation of an antigen, ovalbumin (OVA), in the absence of adjuvant has been demonstrated to induce an immune response that is associated with increased airway responsiveness. Determination of OVA-specific serum IgE and IgG antibody responses revealed an early increase in antibody titers that were initially restricted to the IgE class. Subsequently, IgG antibody titers increased and IgE antibody plateaued. Furthermore, we observed a tenfold increase in the number of lymphocytes caused by a predominant expansion of CD3+ T cells in the peribronchial-associated lymph modes (PBLNs) of sensitized animals compared with the numbers of cells in control animals or in the gut-associated lymphoid tissue. The sensitized animals demonstrated an increase in airway responsiveness to intravenous methacholine challenge. Analysis of in vitro immunoglobulin production by spleen mononuclear cells revealed increased spontaneous IgE production that was more than fourfold enhanced in the presence of OVA, but IgG production was not increased. Spleen and PBLN lymphocytes, but not lymphocytes from gut-draining lymph nodes, demonstrated a proliferative response to OVA. Control animals exhibited no proliferative response to OVA. Histopathologic examination of the sensitized lung revealed an absence of acute inflammatory cells (e.g., neutrophils and macrophages), lymphocytes, or monocytes at the time of the increased airway hyperresponsiveness. These data indicate that, after sensitization of mice by inhalation of antigen, the animals develop a specific IgE antibody response, expansion of PBLN lymphocyte numbers, and increased airway hyperresponsiveness in the absence of signs of airway inflammation.

Adjuvants, Immunologic↗
PubMed1992

Properties of Vero cytotoxin-producing Escherichia coli of human origin of O serogroups other than O157.

The 48 Vero cytotoxin-producing Escherichia coli (VTEC) examined for properties associated with virulence were of human origin and represented 17 O serogroups other than O157 and O26. Only Vero cytotoxin production was common to all the strains. About 60% produced enterohemolysin and hybridized with the CVD419 probe derived from plasmid sequences of E. coli O157. Thirteen strains gave localized adherence (LA) to HEp-2 cells. All of these hybridized with the E. coli attaching and effacing (eae) gene probe and were positive in the fluorescence actin staining test, properties characteristic of strains that efface intestinal microvilli. A further 5 strains were eae probe-positive but did not give LA. None of the VTEC hybridized with a probe specific for the enteropathogenic E. coli adherence factor. Seven strains adhered to HEp-2 cells in a diffuse or aggregative pattern but did not hybridize with probes for these phenotypes. Non-O157 E. coli strains are diverse in their properties, although some may share virulence mechanisms with other diarrheogenic E. coli.

Animals↗
PubMed1992

Evaluation of a Salmonella-specific DNA probe by colony hybridization using non-isotopic and isotopic labeling.

A 2.3 kilobase (kb) Salmonella probe, JEO402-1, and two subfragments, F1214 (1.3 kb) and F1217 (0.8 kb), have been evaluated by colony hybridization using pure cultures of Salmonella serovars and non-salmonella bacteria. JEO402-1, and its subfragments, F1214 and F1217, hybridized to all of 156 different Salmonella serovars tested, while there was no reaction to 112 non-salmonella strains belonging to 19 genera and 37 species of Enterobacteriaceae. Together with previously published results, the JEO402-1 probe has now been shown to detect a total of 396 Salmonella strains belonging to 214 serovars of Salmonella subspecies I-VI. A total of 178 non-salmonella strains representing 23 genera and 51 species of Enterobacteriaceae have all tested negative with JEO402-1. The hybridization results obtained using a digoxigenin-labeled probe were similar to those obtained with 35S isotopic labeling when complete colony lysis was ensured.

Bacterial Typing Techniques↗