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C A Ison

Publications and source records attributed to C A Ison.

At least 19 recordsLinked to original sources

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

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

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

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

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

Concordance of auxotype/serovar classes of Neisseria gonorrhoeae between sexual contacts.

One hundred and three known sexual-contact pairs of patients with culture-proven gonorrhoea who attended St Mary's Hospital, London between May 1989 and February 1991 were identified. All isolates from these patients were serotyped and auxotyped and compared for type concordance within sexual-contact pairs. Serotype was concordant in 80 (78%) of 103 sexual-contact pairs, auxotype in 88 (85%) and auxotype/serovar (A/S) class in 66 (64%) on the first screening. All pairs of isolates showed concordance in both serotype and auxotype when typing was repeated using a single set of serotyping reagents and of auxotyping media. Seventeen serovars, 9 auxotypes and 36 A/S classes were found in this population. Our results suggest that both serotyping and auxotyping may be used as markers to allow tracing of sexual-contact pairs, but that a single set of reagents should be used to ensure maximum reliability.

Contact Tracing

A longitudinal study of pelvic inflammatory disease.

OBJECTIVE: To study the microbiology and long term prognosis of pelvic inflammatory disease (PID). DESIGN: A prospective study of women with laparoscopically confirmed PID. SETTING: Teaching hospital in central London. SUBJECTS: 23 women with PID. OUTCOME MEASURES: Microbiological investigations at the time of diagnosis and at follow up; subsequent fertility and the occurrence of pelvic pain. RESULTS: PID diagnosed by laparoscopy was regarded as moderate to severe in 15 cases. Neisseria gonorrhoeae, Chlamydia trachomatis, Mycoplasma hominis and Ureaplasma urealyticum or a combination of these micro-organisms were detected most frequently in the cervix, less often in the endometrium and least in the tubes, C. trachomatis being the micro-organism found most commonly in the tubes. There was serological evidence of acute chlamydial infection in 13 of 20 cases in which paired sera were available and a serological response to M. hominis in 9 of 12 cases in which it was possible to evaluate the results. On the basis of microbiological and serological results, C. trachomatis appeared to be the most important aetiological agent in 10 cases, N. gonorrhoeae in four cases, M. hominis in three cases and U. urealyticum in none; in five cases, two of these micro-organisms appeared to be of equal importance. After 1 to 3 years, 33% of the women were having difficulty conceiving and 56% continued to complain of pelvic pain. CONCLUSION: It is difficult to be certain of the infectious cause of PID in any given case. However, the evidence that N. gonorrhoeae and C. trachomatis are pathogens is very strong. M. hominis may be responsible for a few cases on its own or together with other micro-organisms.

Cervix Uteri

The dominance of a multiresistant strain of Neisseria gonorrhoeae among prostitutes and STD patients in The Gambia.

OBJECTIVE: To study the epidemiology of antibiotic resistant strains of Neisseria gonorrhoeae from sexually transmitted disease clinics in The Gambia. MATERIALS AND METHODS: One hundred and sixty five strains of N gonorrhoeae were tested for their antibiotic susceptibility, auxotype, serotype, and plasmid content. RESULTS: Of the total population 84 (51%) were non-penicillinase producing (nonPPNG) and 81 (49%) penicillinase-producing N gonorrhoeae (PPNG). There were 16 serovars, five auxotypes and 33 auxotype/serovar (A/S) classes in the total population and the nonPPNG. Among PPNG only five serovars, two auxotypes and nine A/S classes were found. One A/S class predominated, NR/IB-7 (86 isolates), of which 66 (77%) were PPNG and the remainder were chromosomally-mediated resistant N gonorrhoeae (CMRNG). These strains also showed reduced susceptibility to ciprofloxacin, ceftriaxone and tetracycline and were evenly distributed among patient groups. CONCLUSION: We have identified a relatively homogeneous gonococcal population with a core group of isolates exhibiting high levels of antibiotic resistance.

Drug Resistance, Microbial

Epidemiology of penicillin resistant Neisseria gonorrhoeae.

OBJECTIVE: To study the epidemiology of Neisseria gonorrhoeae that exhibit both chromosomal and plasmid-mediated resistance to penicillin. MATERIALS AND METHODS: A total of 1589 strains of N gonorrhoeae isolated from patients attending St Mary's Hospital, London were tested for both their susceptibility to penicillin and for their auxotype and serotype. RESULTS: Of the 940 non-penicillinase producing N gonorrhoeae, 840 were considered penicillin sensitive (MIC less than or equal to 0.5 mg/l) and 100 were chromosomally-mediated resistant N gonorrhoeae (CMRNG), (MIC greater than or equal to 1.0 mg/l). Of the 649 penicillinase producing N gonorrhoeae (PPNG), 429 carried the 4.4 megadalton (MDa) penicillinase encoding plasmid and 220 carried the 3.2 MDa plasmid. CMRNG were predominantly serogroup IB (90%). PPNG with 3.2 MDa plasmid were the only group more often serogroup IA (58%) than IB (42%). Serovar IA-1/2 and requirement for arginine, hypoxanthine and uracil (AHU) were associated with increased susceptibility to penicillin whereas serovar IB-5/7 was associated with decreased susceptibility in nonPPNG. There was a significant difference in the distribution of the IA and IB serovars between PPNG carrying either the 4.4 MDa or 3.2 MDa plasmid. AHU and PAOU requiring strains were not found among PPNG and were uncommon among CMRNG. CONCLUSION: Some clear associations have been found but the pattern among PPNG appears more complex and in most instances could be related to clusters of a single strain over a short time span.

Chromosomes

Current status of serotyping of Neisseria gonorrhoeae.

Protein I has become the basis of serotyping of N. gonorrhoeae. A panel of 12 anti-protein I monoclonal antibodies is supplied by Syva Company, Palo Alto, CA, through distributors for research purposes only. These distributors are responsible for storing the monoclonal antibodies, supplying research groups and training new users, particularly in the interpretation of coagglutination reactions. A European Workshop was held in Heidelberg, Federal Republic of Germany in July 1988 to discuss serotyping with these antibodies. The current uses of serotyping of N. gonorrhoeae include epidemiological studies, clinical purposes and surveillance of antibiotic resistance and plasmid carriage. Predominant serovars may be subtyped either by additional antibodies or with the use of another technique, such as auxotyping, determination of antibiotic sensitivities, plasmid analysis or genetic fingerprinting. Conversely, there is growing evidence that it may be appropriate to group certain serovars together. Data collected during prevalence studies could be combined with clinical information and an international data bank set up. This supposes a close future collaboration of all groups involved in the epidemiology of N. gonorrhoeae.

Antibodies, Monoclonal

Transfer of beta-lactamase plasmids by conjugation in Neisseria gonorrhoeae.

Conjugation experiments in vitro have been used in an attempt to explain certain epidemiological phenomena seen amongst penicillinase producing Neisseria gonorrhoeae (PPNG) isolated at St Mary's Hospital, London. These include changes in plasmid profiles, a lack of beta-lactamase encoding plasmids in PAOU-requiring strains and a difference in the serological classification of strains of PPNG and non-PPNG isolated from the same clinic. It was shown that acquisition of the conjugative plasmid by beta-lactamase producing transconjugants does vary, but is not related to the auxotype/serovar (A/S) class of, or type of beta-lactamase plasmid carried by the donor. It was not possible to transfer either the 3.2MDa or 4.4MDa plasmids to PAOU-requiring strains of N gonorrhoeae. The conjugation frequency from a single donor to different recipients was shown to be variable, but was not influenced by the serovar of the recipients. It was shown that the transfer and acceptance of beta-lactamase encoding plasmids is variable but it was not possible to identify the factors responsible.

Conjugation, Genetic

Penicillin and cephalosporin resistance in gonococci.

Non-penicillinase producing Neisseria gonorrhoeae isolated at St Mary's Hospital, London were examined for the prevalence of resistance to penicillin and for decreased susceptibility to cefuroxime. Of the 941 non-PPNG tested 100 (10.6%) were resistant to penicillin (minimum inhibitory concentration, MIC, greater than or equal to 1 mg/l) and were considered to be chromosomally-resistant N gonorrhoeae (CMRNG). Decreased susceptibility to cefuroxime (MIC, greater than or equal to 0.5 mg/l) was detected in 79% of the CMRNG. The CMRNG were also more often prototrophic and of serogroup IB than the remaining non-PPNG. The correlation coefficient for resistance to penicillin and cefuroxime was high, 0.79. Transformation experiments with both genetically-defined strains and transformants obtained using DNA from clinical isolates, showed that increased resistance to cephalosporins was acquired in three steps in close association with penicillin. We think this suggests that the loci controlling resistance to the cephalosporins are identical or closely linked to those controlling penicillin resistance.

Cefuroxime

Laboratory methods in genitourinary medicine. Methods of diagnosing gonorrhoea.

Gonorrhoea is normally diagnosed presumptively by the presence of intracellular Gram-negative cocci on a Gram stain and confirmed by culture of the causative organism, Neisseria gonorrhoeae. Alternative methods have been evaluated extensively for the detection of gonococci in clinical specimens including immunological techniques such as ELISA and immunofluorescence, DNA probes, genetic transformation and the limulus lysate assay. Some of these tests have proved as sensitive and specific for the detection of gonorrhoea in symptomatic men as the Gram stain but offer no advantage in time or cost. In women, no test has been found that shows a sensitivity and specificity sufficiently adequate for clinical use. Culture in men and women remains the method of choice for diagnosis. In addition the need to obtain the infecting organism for antibiotic susceptibility testing has not been overcome. In contrast, the rapid identification of N gonorrhoeae can be achieved within four hours using either monoclonal antibodies or by the detection of preformed enzymes. New methods for both the detection and identification of N gonorrhoeae should be carefully evaluated particularly for use in cases of child and sexual abuse where medico-legal problems may arise.

Adult

Sampling methods for monitoring changes in gonococcal populations.

A total of 160 consecutive isolates of Neisseria gonorrhoeae was collected over a 3-month period. They were tested for their susceptibility to penicillin, erythromycin and spectinomycin and the auxotype and the serotype determined. We have evaluated two sampling methods, the collection of every fifth isolate and the first 20 isolates (10 male and 10 female) each month, to determine whether either is representative of the total population. There was no significant difference between either method of sampling and the total for detecting the predominant auxotypes and serovars or the distributions in antibiotic susceptibility. It is possible to monitor major changes in a gonococcal population, particularly susceptibility to antibiotics, using a sample of the total population.

Bacterial Typing Techniques

Characterisation of monoclonal antibodies for detection of Mobiluncus spp. in genital specimens.

Monoclonal antibodies were raised to the anaerobic curved rods, Mobiluncus curtisii subsp. curtisii NCTC 11656, M. curtisii subsp. holmesii NCTC 11657 and M. mulieris NCTC 11658. Three antibodies reacted with the two subspecies of M. curtisii and, when used in combination against clinical isolates, showed 100% sensitivity and specificity in immunofluorescence studies. Immunoblotting showed that two of these antibodies reacted with an epitope on a protein which had an electrophoretic mobility corresponding to a Mr of 75 Kda in the absence of a reducing agent and 82 Kda in its presence in both type strains and in clinical isolates. The third antibody reacted with an epitope in type strains which had a mobility corresponding to 75 Kda and was unaffected by a reducing agent. However, in clinical isolates the epitope was present on a protein of 75 Kda or 71 Kda, or on both. A fourth antibody showed reactivity with M. mulieris NCTC 11658 alone, but only 6 (24%) of 25 clinical isolates gave positive results by immunofluorescence. The epitope is believed to be present on a protein of greater than 90 Kda. All four antibodies were shown by immunogold staining to be directed against epitopes exposed on the cell surface.

Antibodies, Bacterial