Biomedical subjects

C A Ison

Publications and source records attributed to C A Ison.

At least 55 records · Page 3Linked to original sources

PubMed1997

Induction of human endothelial tissue factor expression by Neisseria meningitidis: the influence of bacterial killing and adherence to the endothelium.

Tissue factor (TF), a small membrane bound high affinity receptor for factor VII, has an important procoagulant role in the haemostatic dysfunction associated with severe sepsis. Using an in vitro model of human endothelial TF expression, defined strains of Neisseria meningitidis were found to upregulate endothelial cell procoagulant activity (PCA) in a dose dependent manner. This TF response was detected with as little as 10(4) cfu/ml and reached similar levels to those seen with high concentrations of purified endotoxin (> 1 ng/ml). Treatment of N. meningitidis with either adult donor immune serum, penicillin or gentamicin failed to enhance this PCA. Limulus amoebocyte lysate assay of lipopolysaccharide in bacterial culture filtrates together with polymyxin B inhibition experiments suggest that endotoxin is largely responsible for endothelial TF induction by N. meningitidis. Incubation of endothelial cells with N. meningitidis B1940 and B1940 siaD- (an eight-fold more adherent unencapsulated isogenic strain), revealed a significantly greater TF response to B1940 siaD- (P < 0.01). In conclusion, bacterial adhesion to the vessel wall and therefore local levels of endotoxin may be important determinants of the endothelial procoagulant response to N. meningitidis and the consequent coagulopathy commonly associated with the disease.

Adult↗

PubMed1997

Plasmid analysis of Neisseria gonorrhoeae isolates and dissemination of tetM genes in southern Africa 1993-1995.

One group (145 isolates) of Neisseria gonorrhoeae was collected from municipal clinics in Bloemfontein in 1994 and a second group (65 isolates) in 1995. Penicillin and tetracycline MICs were determined and plasmid analysis performed to monitor antimicrobial susceptibilities in conjunction with the occurrence of plasmids in these isolates. The prevalence of penicillin resistance caused by beta-lactamase plasmids remained constant at 9% during the study period. Three high-level tetracycline-resistant strains (MICs 16 mg/L), the first to be detected in South Africa, were isolated in 1994. Although there was a reduction in the percentage of isolates harbouring 24.5 MDa conjugative plasmids (from 79% in 1994 to 46% in 1995), this was partially counteracted by an increase in TetM-encoding conjugative plasmids (25.2 MDa) from 2% to 18.5%. The tetM genes of 13 isolates shown to exhibit high-level tetracycline resistance were characterized as the American type. The American-type tetracycline resistance plasmid was demonstrated in 11 isolates. Digestion with Bg/l showed that two isolates harboured tetM-encoding plasmids that differed from the American- and Dutch-type plasmids described previously: one isolate contained a plasmid that produced two fragments of different sizes from those of the American-type plasmid and the second isolate possessed an American/Dutch hybrid plasmid. Auxotyping/serotyping and random amplified polymorphic DNA analysis revealed a predominant tetracycline-resistant family (NR/IA-6, genomic group I) in Bloemfontein. As there is a high incidence of chlamydial infections in southern Africa requiring tetracycline therapy, selective pressures exist in the environment for the maintenance and rapid spread of high-level tetracycline-resistant N. gonorrhoeae. It is possible that tetM genes may have emanated from Botswana and/or Namibia to Bloemfontein. The establishment of high-level tetracycline-resistant N. gonorrhoeae in Bloemfontein was seen to be complex as a related group of strains was identified, plasmid dissemination was evident and two new TetM-encoding plasmids were demonstrated. The appearance of these TetM-encoding plasmids indicates either that the American- and Dutch-type plasmids are continuing to evolve or that tetM genes are being introduced into different families of 24.5 MDa conjugative plasmids.

Cephalosporins↗

PubMed1997

Involvement of the gonococcal MtrE protein in the resistance of Neisseria gonorrhoeae to toxic hydrophobic agents.

Low-level resistance of Neisseria gonorrhoeae to toxic hydrophobic agents (HAs), including some antibiotics, is chromosomally mediated via the multiple transferable resistance (mtr) efflux system. The gene encoding the 48:3 kDa outer-membrane protein MtrE, which is associated with the mtr phenotype, was identified and is homologous to export-associated outer-membrane proteins, including the OprM (formerly OprK) lipoprotein of Pseudomonas aeruginosa. Insertional inactivation of the mtrE gene in N. gonorrhoeae strain FA19 resulted in the loss o the outer-membrane protein, with concomitant hypersusceptibility of the mutant strain to a range of HAs. The properties of this mutant confirmed the role of MtrE in multidrug resistance mediated by an active efflux mechanism. Secondary structure predictions for MtrE indicated a largely hydrophilic protein with a single alpha-helical transmembrane region. A transposon-like element, similar to that found downstream of the region containing the promoters for mtrR and mtrC in Neisseria meningitidis, was identified 63 bp downstream of the mtrE gene.

Amino Acid Sequence↗

PubMed1997

A longitudinal study of the vaginal flora over a menstrual cycle.

Healthy female volunteers participated in an anonymous study to monitor vaginal flora by taking daily vaginal samples and making a smear for later Gram-staining, as well as recording information on genital symptoms, sexual activity, contraceptive and bathing practices. A modification of Spiegel's criteria was used to categorize the Gram-stained smears, an intermediate category between normal flora and bacterial vaginosis (BV) being recognized. Of the 22 volunteers who completed the study, one was excluded because of pregnancy. Of the remaining 21 women, 10 (48%) had a normal flora throughout the study, 4 (19%) had an abnormal flora throughout and 7 (33%) had a basically normal flora which underwent a change to either an intermediate flora in 5 women or fully developed BV in 2 of them. In 5 (71%) of these women the change occurred within the first 9 days of the cycle. Transient changes in the vaginal microbial flora occurred predominantly in the first part of the menstrual cycle which suggests that in some women hormonal changes could have a role in the pathogenesis of bacterial vaginosis.

Candida↗

PubMed1996

Characteristics of gonococci isolated from men with urethritis in Dubai.

Neisseria gonorrhoeae were collected from men attending out-patient clinics in Dubai. The susceptibility to a range of therapeutic antibiotics and their auxotype and serotype was determined. The plasmid content of all penicillinase-producing strains was also analysed. Thirty-six strains of N. gonorrhoeae were isolated from specimens collected from 79 patients over a 24-day period. Of the 36 isolates, 9(25%) were penicillinase-producing N. gonorrhoeae (PPNG) and 15 (42%) were chromosomally resistant N. gonorrhoeae (CMRNG). CMRNG exhibited higher levels of resistance to cefuroxime, chloramphenicol, tetracycline and erythromycin than PPNG. All isolates were susceptible to ceftriaxone and spectinomycin. Three (8%) isolates showed reduced susceptibility (MIC, > or = 0.25 mg/l) to ciprofloxacin. Six isolates of PPNG carried the 4.4 MD and three the 3.2 MD penicillinase encoding plasmid. The total gonococcal population was phenotypically diverse, with 12 serovars, 6 auxotypes and 21 A/S classes. Gonorrhoea was found to be a major cause of urethritis in Dubai and the strains exhibited high levels of resistance to penicillin.

Adolescent↗

PubMed1996

The influence of capsulation and lipooligosaccharide structure on neutrophil adhesion molecule expression and endothelial injury by Neisseria meningitidis.

Host inflammatory response to meningococcal infection is believed to be a major determinant of disease severity. Isogenic mutants of Neisseria meningitidis serogroup B1940, which differ in expression of capsular polysaccharide and lipooligosaccharide (LOS), were used to examine host responses in a whole blood model of bacteremia and a model of endothelial injury. The parent organism caused significantly less neutrophil shedding of the adhesion molecule, L-selectin, than the three mutant organisms (P < .01) and was most resistant to the bactericidal activity of whole blood. Despite marked differences in bacterial adhesion to endothelial cells (P < .05), no damage was induced by organisms alone. Endothelial injury was observed when neutrophils were incubated with adherent, capsule-deficient organisms (P < .05). The degree of endothelial damage was related to the number of neutrophils adherent to the endothelium. Thus, bacterial capsulation and LOS structure can influence neutrophil activation and endothelial injury and, as such, may be important in the pathogenesis of meningococcal sepsis.

Antigens, Bacterial↗

PubMed1996

Genes associated with meningococcal capsule complex are also found in Neisseria gonorrhoeae.

A homolog of the meningococcal cps locus region E has been identified in Neisseria gonorrhoeae immediately upstream of the gonococcal region D locus. Region E has no detectable function in capsule biosynthesis in Neisseria meningitidis or in lipopolysaccharide biosynthesis in either organism. The open reading frame is homologous to proteins of unknown function in Escherichia coli and Haemophilus influenzae. Further analysis of the N. meningitidis cps cluster has identified a second copy of region D encoding three additional open reading frames, including homologs of DNA methyltransferases. The organization of the region D and E genes in N. gonorrhoeae and N. meningitidis in relation to the cps genes provides some insight into the evolutionary origin of encapsulation in N. meningitidis.

Bacterial Capsules↗

PubMed1996

Antimicrobial agents and gonorrhoea: therapeutic choice, resistance and susceptibility testing.

INTRODUCTION: Neisseria gonorrhoeae, the causative agent of gonorrhoea is a particularly well adapted pathogen that has continued to evolve mechanisms to evade treatment with antimicrobial agents. THERAPEUTIC CHOICE: The choice of antibiotic for use in the first-line treatment of gonorrhoea should be made with knowledge of the susceptibility of the isolates of N gonorrhoeae to be encountered. RESISTANCE: High-level resistance to penicillin and tetracycline in N gonorrhoeae is plasmid-mediated and a major therapeutic problem. Penicillinase-producing N gonorrhoeae, first described in 1976, have now spread worldwide and tetracycline-resistant N gonorrhoeae, described in 1985, are becoming increasingly prevalent. Chromosomal resistance to penicillin is low-level and affects a range of antibiotics. High-level resistance to spectinomycin has been sporadic and has not limited its use whereas the emergence of resistance to ciprofloxacin will have a significant impact on its use for gonorrhoea. SUSCEPTIBILITY TESTING: A variety of methods are available including disc diffusion, breakpoint agar dilution technique, E-test and determination of the minimum inhibitory concentration (MIC). The choice of methodology will depend on the number and type of isolates and the facilities available for testing. DISCUSSION: Surveillance programmes to monitor levels of antibiotic resistant isolates are essential to ensure therapeutic success.

Anti-Bacterial Agents↗

PubMed1995

Specificity of the immune response to Haemophilus ducreyi.

The specificity of the antibody response to Haemophilus ducreyi in sera from patients attending a sexually transmitted disease clinic in South Africa has been studied using immunoblotting. Patients with chancroid were shown to have higher levels of IgG (mean 0.74, SD 0.34) to H. ducreyi than those with no history of chancroid (mean 0.34, SD 0.19). The pattern of the antibody specificity was highly variable between patients with culture proven chancroid but there was no observed strain specificity. In comparison, the patterns obtained using sera from patients without known exposure to H. ducreyi showed less variation between patients and were of less intensity at the dilution used. Sera from patients with chancroid recognised epitopes on proteins that varied in molecular weight between strains, particularly of 60-66kDa (10 of 36 patients) and 25-27kDa (8 of 36 patients). In addition epitopes were recognised on the GroEL and/or DnaK heat shock proteins in 13 of 36 sera tested. There was no apparent change in the epitopes recognised on proteins between the homologous and heterologous strains. Patterns of antibody specificity in sequential sera only varied in one of six patients tested.

Antibodies, Bacterial↗

PubMed1995

Whole blood model of meningococcal bacteraemia--a method for exploring host-bacterial interactions.

An ex vivo whole blood model of meningococcal bacteraemia was developed to examine the total bactericidal activity of blood. Using a single defined donor and strains belonging to serogroups A, B and C and an unencapsulated strain, we demonstrated that the bactericidal mechanisms operating in whole blood varied with anticoagulant, serogroup and bacterial growth conditions. The choice of anticoagulant had a major effect on the survival of the serogroup A strain with 94% (SEM 7.6) survival in citrated blood compared to 19.7% (SEM 19.6) survival in heparinised blood after 60 min incubation. The serogroup C strain showed enhanced survival when grown in liquid medium compared to growth on solid medium (73.5%, SEM 7.5, and 8.2%, SEM 3.1, respectively, in citrated blood after 60 min). The pattern of survival of serogroup B and the unencapsulated strain were largely unaffected by these variables. Comparison with cell free conditions allowed the contribution of cellular components in meningococcal killing to be determined. Secreted levels of tumour necrosis factor and neutrophil elastase secreted during whole blood assays did not correlate with bacterial growth or viability indicating a lack of relationship between killing and activation of phagocytes.

Anticoagulants↗

PubMed1995

Opa-typing: a high-resolution tool for studying the epidemiology of gonorrhoea.

A single gonococcus possesses a family of 11 distinct and highly variable opa genes. The extensive variation and rapid evolution of the opa gene repertoire has been exploited to provide a high-resolution typing method for studies of the short-term transmission of gonorrhoea. The 11 opa genes are amplified with a single pair of primers by the polymerase chain reaction, digested with frequently-cutting restriction enzymes, and the fragments are fractionated on polyacrylamide to provide an opa-type. The method appeared to be highly discriminatory as the opa-types of gonococci, isolated world-wide over the last 30 years, were all different. Opa-typing discriminated between isolates of the same auxotype/serovar class. Similarly, there were 41 opa-types among 43 consecutive isolates from a sexually transmitted disease (STD) clinic. The two pairs of isolates from this clinic that gave the same opa-types were identical by other criteria and may have been from unsuspected sexual contacts. With one minor exception, identical opa-types were obtained from gonococci recovered from known sexual contacts. These results suggest that variation in the family of 11 opa genes evolves so rapidly that the opa-types of gonococci are distinguishable, unless the isolates are from sexual contacts or a short chain of disease transmission. The identification of gonococci with identical opa-types is therefore believed to be a good indicator that the individuals from which they were recovered were sexual partners, or part of a short chain of disease transmission.

Antigens, Bacterial↗

PubMed1995

A one-year survey of Neisseria gonorrhoeae isolated from patients attending an east London genitourinary medicine clinic: antibiotic susceptibility patterns and patients' characteristics.

OBJECTIVES: To collect epidemiological data on gonococcal infection in an east London genitourinary medicine (GUM) clinic; to perform antibiotic susceptibility testing on Neisseria gonorrhoeae isolates and relate results to patient data; to assess the efficacy of current first-line antibiotic therapy for treating gonorrhoea. METHODS: Gonococcal isolates were collected from 113 patients attending the GUM clinic at Newham General Hospital over a one year period. Isolates (104) were tested for susceptibility to various antibiotics. Plasmid profiles were obtained for penicillinase producing gonococci (PPNG) and isolates exhibiting high-level tetracycline resistance (TRNG). Epidemiological information was collected from clinic attenders by routine note-taking. RESULTS: PPNG (16) accounted for 15% of isolates tested, only three being acquired outside the United Kingdom (U.K.). Plasmid typing showed three types of beta-lactamase-encoding plasmids were represented (2.9 MDa, 3.2 MDa and 4.4 MDa). Amongst the non-PPNG isolates, high-level chromosomal resistance to penicillin (CMRNG) was found in 3.5%, intermediate resistance in 57.5% and full susceptibility in 39%. One isolate showed decreased susceptibility to ciprofloxacin (MIC = 0.06 mg/l). Three PPNG isolates also possessed a 25.2 MDa plasmid and expressed high-level tetracycline resistance encoded by tetM. All isolates were susceptible to cefixime, cefotaxime, azithromycin and spectinomycin. Most gonorrhoea (90%) was seen in local residents. The male:female case ratio was 2:1 with homosexually-acquired gonorrhoea accounting for only 3.5% of the total. Most patients (96%) had acquired gonorrhoea in the U.K.. A past history of gonorrhoea was more frequent in male patients. Concurrent chlamydial infection was seen in 31% females and 16% males. CONCLUSIONS: The high PPNG rate supports a previous decision to change first-line therapy from amoxycillin with probenecid to ciprofloxacin. There was no evidence of clinical treatment failure with ciprofloxacin. Cefixime, cefotaxime, azithromycin and spectinomycin all appear to be suitable alternative therapies. Acquisition of gonorrhoea abroad was associated with isolates exhibiting penicillin resistance but such isolates were also obtained from patients infected locally and without a history of foreign travel.

Adult↗

PubMed1994

Naturally occurring PIA/PIB hybrids of Neisseria gonorrhoeae.

The major outer membrane protein of Neisseria gonorrhoeae, Por, functions as a porin and is thought to occur as one gene with two alleles. The immunologically distinct epitopes of the two subclasses, IA and IB, have allowed the development of serotyping schemes. Clinical isolates of N. gonorrhoeae are believed to express only a single type of Por, either IA or IB. We have encountered two clusters of isolates that react with antibodies to both IA and IB Por. Isolates within the clusters are indistinguishable by the phenotypic characteristics tested. In addition, the amplification of the por gene in representative isolates showed that the por gene of the hybrids gave similar restriction digest patterns within but not between the clusters.

Base Sequence↗

PubMed1994

A longitudinal study of bacterial vaginosis during pregnancy.

OBJECTIVE: To determine the longitudinal changes in the incidence of vaginosis in pregnancy. DESIGN: A prospective study of women during pregnancy. SETTING: A District General Hospital in North-West London. SUBJECTS: Seven hundred and eighteen pregnant women attending antenatal clinics. At their first attendance and subsequently, Gram-stained vaginal smears were examined and Mycoplasma hominis and Gardnerella vaginalis were sought by culture. RESULTS: Initially, 87 (12%) women had bacterial vaginosis diagnosed on Gram-stained reading of the vaginal smears. Examination of further smears, obtained from 176 women at 36 weeks of gestation, showed that those whose vaginal flora was normal initially, and who went to term, rarely developed vaginosis (three of 127, 2.4%). Samples were obtained at 36 weeks gestation from 32 women who had bacterial vaginosis initially, and went to term. In almost 50% (15 of 32) of these a normal lactobacillus-dominated flora had regenerated. Thirty-five women (5%) had initial vaginal smears graded as intermediate. From this group, six of the 17 (35%) women from whom samples were obtained at 36 weeks gestation still had flora of an intermediate pattern; 10(59%) now had normal flora and only one (6%) had developed bacterial vaginosis. Women with bacterial vaginosis were more likely to be culture-positive for M. hominis than those with normal flora (34/78 versus 10/563, odds ratio 42.73 (18.9 to 102.3) P < 0.001), or to be culture-positive for G. vaginalis than those with normal flora (35/78 versus 21/563, odds ratio 21.0 (10.75 to 41.2) P < 0.001). CONCLUSION: Pregnant women do not commonly develop bacterial vaginosis after 16 weeks gestation, and if present, it remits spontaneously in approximately half of those who reach term. As bacterial vaginosis is associated with increased rates of second trimester miscarriage and preterm delivery, any treatment aimed at its eradication in pregnancy should be given no later than the beginning of the second trimester of pregnancy.

Adult↗

PubMed1993

Antibodies directed against Haemophilus ducreyi heat shock proteins.

The response to heat shock in Haemophilus ducreyi has been investigated by labelling newly synthesized proteins with a pulse of 35S-methionine at a range of temperatures between 30-40 degrees C. Proteins were then separated by SDS-PAGE and analysed by autoradiography of the resultant profiles. Four heat shock proteins (HSP) were identified of apparent molecular weight < 14, 58.5, 74 and 78 kDa. The three larger proteins appeared to be independently controlled and inducible in a range of H. ducreyi strains. The protein profiles were also probed by Western blot with serum from an immunized rabbit, serum from a rabbit infected intradermally with H. ducreyi, mouse monoclonal antibodies (MAb) and a pool of human sera from patients with chancroid. The rabbit sera, the human serum pool and MAb (BB11) reacted with epitopes on the HSP of 58.5 kDa, whereas only the immunized rabbit serum and another MAb (CC11) reacted with epitopes on the HSP of 78 kDa. The HSP of 58.5 kDa appears to be immunogenic and may play a part in the host response to H. ducreyi and the pathogenesis of chancroid.

Animals↗

PubMed1993

Non-radioactive ribotyping of Haemophilus ducreyi using a digoxigenin labelled cDNA probe.

Haemophilus ducreyi, the causal organism of chancroid, has increased in significance recently due to its association with HIV transmission. Most previous typing systems have exploited phenotypic characteristics. Detection of ribosomal RNA cistrons, ribotyping, was successfully developed to examine H. ducreyi, but required the use of 32P. We have used digoxigenin to define ribotypes from 30 strains of H. ducreyi from diverse geographical locations. This was achieved by agarose gel electrophoresis of restriction enzyme (RE) digested DNA extracts. These extracts were vacublotted onto nylon membrane and probed using digoxigenin labelled complementary DNA probe, prepared from Escherichia coli 16S and 23S ribosomal RNA. From 19 REs tested, Ava II, Hinc II, Bgl II and BstE II gave clear ribotypes. The ribotypes of BstE II and Bgl II used together gave the highest index of discrimination (D = 0.95), 16 types, and showed good reproducibility. This non-radioactive method demonstrates the three important features of a typing system; discrimination, typability and reproducibility.

Bacterial Typing Techniques↗

PubMed1993

Antimicrobial susceptibilities and serotyping of Neisseria gonorrhoeae in southern Africa: influence of geographical source of infection.

One hundred and ninety-two strains of Neisseria gonorrhoeae isolated from migrant mine-workers were tested for their susceptibility to antibiotics, auxotyped and serotyped. Of the total, 93 (48%) were acquired locally and 64 (33%) from different geographical locations. Plasmid-mediated resistance to penicillin was found in 28 (14.6%) strains and was associated predominantly with the presence of 5.0 kb penicillinase encoding plasmid (18/28, 64%). Chromosomal resistance to penicillin (MIC > or = 1 mg/l) was detected in 14 (7.3%) strains. Resistance to tetracycline was chromosomally and not plasmid-mediated. Antibiotic resistance was encountered most commonly among strains acquired in Natal. The overall gonococcal population was sensitive to ceftriaxone, ciprofloxacin, spectinomycin and azithromycin. Nine auxotype/serovar (A/S) classes were encountered among penicillinase-producing N. gonorrhoeae (PPNG) compared to 24 A/S classes among non-PPNG strains. The most common A/S class was NR/IA-6 which accounted for 38% of PPNG and 15% of non-PPNG.

Africa, Southern↗

PubMed1993

Neisseria gonorrhoeae isolated at St. Mary's Hospital London, 1980-91.

OBJECTIVE: To describe and discuss the trends in the isolation of Neisseria gonorrhoeae from patients attending the Genitourinary Medicine Clinic at St. Mary's Hospital, Paddington, London between 1980 and 1991. DESIGN: A retrospective study of the total number of gonococci isolated over an eleven year period was performed. In addition, for the years 1988-1991 the number of isolates from homosexual men was analysed by age of the patient, site of infection and HIV antibody status of the patient. RESULTS: The total number of N. gonorrhoeae isolates identified declined markedly between 1980 and 1989 from 3670 to 750 isolates. Over the same time period the number of specimens screened for N. gonorrhoeae fell by 50%. In 1990 there was an increase in N. gonorrhoeae isolates but this was not maintained, and in 1991 the number of N. gonorrhoeae fell to its lowest level of 638 isolates. The decrease since 1980 occurred in both men and women although the number of rectal isolates from men showed a steeper decline reaching its lowest level of 24 isolates in 1988. The number of rectal isolates from homosexual men has since increased with a peak in 1990. Many of the infections among homosexual men occurred in older men and included insignificant number of patients who were HIV positive. CONCLUSION: Gonorrhoea among attenders at St. Mary's Hospital has declined dramatically since 1980 following trends reported from much of Europe. The increase in gonococcal isolates since 1989 and the peak in 1990 are unexplained but are coincident with a higher number of isolates from homosexual men.

Adult↗