Ethnic differences in HSV1 and HSV2 seroprevalence in Amsterdam, the Netherlands.

Herpes simplex virus type 1 (HSV1) and 2 (HSV2) infection can lead to significant morbidity, and HSV2 is considered a risk factor for HIV transmission. The majority of HSV-infected people are asymptomatic and unaware of their infection. We aimed to determine the HSV1 and HSV2 prevalence among various ethnic groups in a large urban area in the Netherlands. In 2004, serum samples from a population-based serum repository of 1,325 people over 18 years living in Amsterdam were tested for HSV1 and HSV2 antibodies in order to determine high-risk groups. Prevalence ratios were estimated and all analyses were weighted by sex, age, and ethnicity. In the general population of Amsterdam, 67% had HSV1 antibodies, 22% had HSV2 antibodies, 15% had HSV1 and HSV2 antibodies, and 26% had no indication of HSV infection. In multivariate analyses, HSV1 seroprevalence increased with age, and was higher among people of Turkish and Moroccan origin, homosexual men, and individuals with low educational level. HSV2 seroprevalence was associated with increasing age, Surinamese/Antillean background, and having a history of sexually transmitted infections (STI). These differences between ethnic groups in Amsterdam regarding the distribution of HSV1 and HSV2 infection emphasise the importance of an ethnic-specific approach of serological testing as well as campaigns aimed at behavioural change and counselling to raise awareness of the risk of HSV transmission.

Monitoring the potential introduction of the Swedish Chlamydia trachomatis variant (swCT) in the Netherlands.

This report describes the actions of public health experts in cooperation with specialists in sexually transmitted diseases (STD), epidemiologists and (molecular) microbiologists to investigate the possible introduction of the swCT variant in the Netherlands: 1. Investigating trends in CT epidemiology Result: STD surveillance and laboratory surveillance did not show any evidence of the introduction of the swCT variant in Holland. 2. Retesting samples by TaqMan PCR Result: Roche CT-negative samples suspected to be CT-positive on the basis of the clinical picture were retested by swCT TaqMan but did not harbour the swCT variant 3. Screening sample pools for the presence of the swCT variant Result: Four different sample pools covering a wide geographical range were tested by specific swCT Taqman assay, but the swCT variant was not detected in any of them. In conclusion, to date the swCT variant has not been found in the Netherlands. However, ongoing monitoring is needed until Roche and Abbott have adapted their CT nucleic acid amplification tests (NAATs) to detect the new variant.

Description of the ICTI consortium: an integrated approach to the study of Chlamydia trachomatis infection.

The use of an integrated approach to the study of Chlamydia trachomatis infection of the female genital tract, presented at the mini-symposium "Chlamydia trachomatis infections" and described in the thesis of Joseph M. Lyons, has resulted in the creation of the ICTI consortium. The ICTI consortium is based on strong interaction and collaboration between basic scientists, clinicians, epidemiologists, and health care policy makers. This translational approach will help to further the valuable insight into the immunopathogenesis of this sexually transmitted infection (STI) and the development of new intervention strategies, including the vaccines and screening programs necessary to effectively diagnose, treat and prevent C. trachomatis infection. A background of the need for this integrated approach is presented and the goals and participants of the consortium are described.

[Resistance of gonococci in the Netherlands; results of a survey of medical microbiology laboratories]. / Resistentie van gonokokken in Nederland; resultaten van een enquête bij medisch-microbiologische laboratoria.

OBJECTIVE: To collect information about the incidence ofgonorrhoea and gonococcal resistance in the Netherlands. METHOD: A questionnaire was sent to 39 medical microbiology laboratories to obtain information on current diagnostics and the susceptibility testing method, and on the number of positive results and the susceptibility pattern of gonococcal isolates in 2002 and 2003 (up to and including November). RESULTS: 32 laboratories participated in this survey. 13 laboratories used culture alone and 19 laboratories used culture and/or a molecular test. Gonorrhoea was diagnosed 2,666 times in 2002 and 2,190 times in 2003, with an incidence of 33.5 and 27.0 per 100,000 inhabitants, respectively. The rate of resistance to beta-lactam antibiotics (penicillin and amoxicillin) was 12.2% and 10.7% in 2002 and 2003, respectively, and the rates of resistance to tetracycline were 18.5% and 20.6%. An increase in the resistance to quinolones was observed from 6.6% in 2002 to 9.5% in 2003. Resistance to cephalosporins was low (0.5% in 2002 and 1.2% in 2003). Furthermore, regional differences in susceptibility were found within the Netherlands. CONCLUSION: The observed gonococcal incidence and resistance form the basis for a gonorrhoea prevention and treatment programme in the Netherlands.

[Inguinal lymphogranuloma venereum in a man having sex with men: perhaps an example of the missing link to explain the transmission of the recently identified anorectal epidemic]. / Inguïnale lymphogranuloma venereum bij een man die seks heeft met mannen; wellicht een voorbeeld van de ontbrekende schakel ter verklaring van de transmissie bij de recent vastgestelde anorectale epidemie.

A 38-year-old man who had sex with men, presented at the outpatient department for Sexually Transmitted Diseases in Amsterdam with a painful, red, fluctuating swelling in the left groin and general discomfort. He had been sexually active in the population of men who have sex with men, in which an anorectal lymphogranuloma venereum (LGV) epidemic has recently been discovered. Unlike other cases where there was anorectal involvement, this patient was the first case of LGV with the classical inguinal presentation although he had not visited the tropics where the inguinal form of LGV occurs as an STD. Routine investigation using PCR on material from urethra and rectum and from the urine, repeatedly failed to detect LGV. However, PCR on pus aspirated from the enlarged lymph node demonstrated Chlamydia trachomatis serovar type L2. Treatment with doxycycline 100 mg twice daily was started. This case illustrates that routine analysis from urethra and rectum and of urine may fail to detect LGV. Furthermore, this case of a patient who probably had LGV initially in the urethra may be the missing link in explaining the route of transmission of the anorectal LGV epidemic.

[Syphilis epidemic and an increase of the number of HIV infections among homosexual men attending the Amsterdam venereal disease clinic]. / Syfilisepidemie en stijging van het aantal HIV-infecties onder homoseksuele mannen op de Amsterdamse SOA-polikliniek.

The registered number of cases of early infectious syphilis and of (ano)genital gonorrhoea among the attendees of the outpatient clinic for sexually transmitted diseases of the Amsterdam municipal health service shows a strong increase for both diagnoses in the period 1990-2001, notably in the last few years. Nearly all of this increase is accounted for by homosexual men. Syphilis increased mostly among men aged 35 years and over, gonorrhoea mostly among younger men. The population of older men also showed a distinct increase since 1997 in HIV incidence.

[Increased Neisseria gonorrhoeae quinolone resistance in Amsterdam]. / Toename van chinolonresistentie bij Neisseria gonorrhoeae in Amsterdam.

In addition to a rise in the number of cases of gonorrhoea, the susceptibility of Neisseria gonorrhoeae to antibiotics is also a cause for concern. After a period of high resistance rates to penicillin and tetracycline between 1985 and 1995, resistance rates have dropped considerably in recent years, probably due to changes in treatment regimens. However, recently we have seen an increasing number of quinolone-resistant N. gonorrhoeae isolates in Amsterdam, the Netherlands, a development that has previously been reported in other parts of the world. Some form of national resistance monitoring for gonococci is therefore urgently required to allow timely detection of changes in N. gonorrhoeae resistance.

Amplified fragment length polymorphism fingerprinting for identification of a core group of Neisseria gonorrhoeae transmitters in the population attending a clinic for treatment of sexually transmitted diseases in Amsterdam, The Netherlands.

Amplified fragment length polymorphism analysis seems well suited for studying the epidemiology of isolates of Neisseria gonorrhoeae obtained from patients attending the Sexually Transmitted Disease Outpatient Clinic in Amsterdam, The Netherlands. It shows potential to identify the core group of transmitters.

Prevalence and risk factors of HSV-1 and HSV-2 antibodies in European HIV infected women.

OBJECTIVES: To investigate the prevalence and risk factors of HSV-1 and HSV-2 antibodies in HIV infected women and the association between recurrent genital ulcerations and HIV disease progression in HSV-2 positive women. METHODS: The presence of HSV antibodies was tested in 276 of the 487 women participating in a European cohort study of HIV infected women. Prevalence rate ratios described the association between HSV infection and its risk factors, using log binomial regression. Generalised estimating equations (GEE) analysis was performed to determine the impact of markers of HIV disease progression on recurrent genital ulcerations. RESULTS: The prevalence of HSV-1 and HSV-2 antibodies was 76% (95% confidence interval (95% CI): 71-81) and 42% (95% CI: 36-50); 30% (95% CI: 24-35) of the women had antibodies against both HSV-1 and HSV-2. The prevalence of HSV-1 was 86% (95% CI: 80-92) in southern Europe compared with 69% (95% CI: 57-79) and 67% (95% CI: 55-77) in central and northern Europe (p=0.002). This geographical variation remained after adjustment for other risk factors. An increasing number of years of sexual activity (p=0.0002) and a history of prostitution (p=0.0001) were independently associated with HSV-2 prevalence. In HSV-2 positive women, symptomatic cases of HSV infection were minimal, but increased with decreasing CD4 count. CONCLUSION: In HIV infected women, the prevalence of HSV antibodies is high and symptomatic cases of HSV infection are minimal, but increase with decreasing CD4 count. HSV-2 but not HSV-1 was related to sexual behaviour (that is, a history of prostitution and the number of sexually active years) in this group of HIV infected women.

Antimicrobial resistance of Neisseria gonorrhoeae and emerging ciprofloxacin resistance in the Netherlands, 1991 to 1998.

Surveillance of antibiotic resistance in Neisseria gonorrhoeae showed a decrease in the percentage of beta-lactamase-producing isolates but an increase in intermediately penicillin-resistant strains and strains resistant to a high level of tetracycline. MICs for the ciprofloxacin-resistant isolates that emerged increased, and these isolates had mutations in gyrA and parC similar to those observed in the Far East.

Biofilm production by Staphylococcus epidermidis isolates associated with catheter related bacteremia.

The mean biofilm production of 22 Staphylococcus epidermidis isolates associated with catheter related bacteremia was significantly higher than that of 32 nose isolates from healthy individuals. This difference was due to seven catheter related isolates. These findings do not show a clear association between biofilm production and virulence.

Effectiveness of resins in neutralizing antibiotic activities in bactec plus Aerobic/F culture medium.

Incorporating resins in blood culture media can effectively reduce the activities of several antibiotics. It was shown that the activities of some generally used antibiotics decreased by 80 to 90% within 2 h in Bactec Plus Aerobic/F resin-containing culture medium. Bactec vials containing resins were still found to be positive for bacteria when antibiotics were present. The addition of beta-lactamase shortened the detection time irrespective of the presence of resins.

Binding of blood coagulation factor VIII and its light chain to phosphatidylserine/phosphatidylcholine bilayers as measured by ellipsometry.

Factor VIII is a plasma protein which plays an essential role in the coagulation system. When assembled with the enzyme Factor IXa on a phospholipid membrane, it functions as a cofactor in the enzyme complex that cleaves the zymogen Factor X to Factor Xa. We studied the binding of both Factor VIII and the Factor VIII light chain to planar phospholipid bilayers consisting of 25% dioleoylphosphatidylserine and 75% dioleoylphosphatidylcholine (PSPC) by ellipsometry. Equilibrium-binding studies revealed that both Factor VIII and its light chain bind with high affinity to PSPC bilayers. The binding affinity of Factor VIII, with a dissociation constant Kd of 0.24 nM, was comparable with that of the Factor VIII light chain (Kd 0.49 nM). Maximal binding was 2.3 mmol of protein per mol of PSPC for Factor VIII and 7.1 mmol of protein per mol of PSPC for the Factor VIII light chain. Adsorption kinetics of both Factor VIII and its light chain conformed to the classical Langmuir adsorption model yielding dissociation constants calculated from the rates of adsorption that were similar to those obtained by equilibrium-binding studies. In contrast, measurements of rates of desorption revealed a deviation from those expected for a single class of binding sites. The desorption rate of Factor VIII increased with increasing residence time on the lipid membrane. This indicates transition of Factor VIII to a configuration with a lower binding affinity. As this time-dependent change in affinity could affect the validity of the measurement of binding parameters, in particular equilibrium-binding determinations carried out on a long timescale, binding affinity was also estimated from adsorption kinetics at half-maximal surface coverage, a relatively rapid procedure for the determination of the affinity. A Kd of 0.087 nM was obtained under these conditions. Measurement of equilibrium binding to small PSPC vesicles, a system in which equilibrium is rapidly attained, resulted in similar binding parameters (Kd = 0.13 nM and a maximal binding of 2.8 mmol of protein per mol of PSPC). These data confirm the results of equilibrium binding to planar bilayers. Taken together, our results indicate that Factor VIII, by means of its 80 kDa light chain, binds to PSPC bilayers with a dissociation constant below the concentration of Factor VIII in plasma and therefore may readily bind to exposed phospholipid membranes under physiological conditions.

Treatment with monoclonal anti-CD3 antibody protects against lethal Sendai virus infection by induction of natural killer cells.

C57BL/6 mice are protected from a lethal pneumonia caused by Sendai virus when treated with low doses of mAb directed to the CD3 Ag. The protective mechanism is not due to an accelerated Sendai virus-specific Th cell, CTL, or antibody response but to a strong NK cell response via the in vivo induction of lymphokines. Antibodies directed against the NK1.1 and asialo GM1 marker totally reversed the protective effect of anti-CD3 treatment. In vivo treatment with rIL-2 also induced NK activity and induced antiviral protection. Treatment with anti-CD3 protects when given in a narrow time window (1 day before until 1 day after Sendai virus inoculation), indicating that NK activity is protective in the early phase of virus infection.

The platelet alloantigen Zwa or PlA1 is expressed by cultured endothelial cells.

Recently, the synthesis by cultured human endothelial cells of a membrane protein complex immunologically related to platelet glycoprotein (GP) IIb/IIIa complex was demonstrated. Since platelet GP IIIa is known to carry the platelet alloantigen Zwa or PlA1, studies were performed to establish whether this antigen is also expressed on endothelial cells. The present report describes the results of these studies, which provide evidence for the presence of the Zwa or PlA1 antigen on the surface of cultured human endothelial cells. This evidence is based on the following observations: (1) cultured endothelial cells react with anti-Zwa (PlA1) antibodies as shown by indirect immunofluorescence; (2) two proteins are precipitated by anti-Zwa (PlA1) antibodies from lysates of 125I-labelled endothelial cells with an electrophoretic mobility corresponding with that of GP IIb and IIIa; (3) anti-Zwa (PlA1) reacts specifically, as shown by immunoblotting of sodium-dodecylsulphate polyacrylamide gels of solubilized endothelial cells, with a protein with a mobility similar to that of platelet GP IIIa.

Cultured human endothelial cells synthesize a plasma membrane protein complex immunologically related to the platelet glycoprotein IIb/IIIa complex.

We have previously demonstrated that endothelial cells synthesize a plasma membrane protein indistinguishable from platelet glycoprotein (GP) IIa. The present study provides evidence for a further analogy between the platelet and the endothelial cell membrane by showing that cultured endothelial cells also synthesize a membrane protein complex immunologically related to the platelet GP IIb/GP IIIa complex. This evidence is based on the following observations: (1) C17, a murine monoclonal antiplatelet GP IIIa antibody, consistently precipitates two proteins, apparent molecular weights, respectively, 115,000 and 125,000 reduced and 95,000 and 135,000 nonreduced, from metabolically (35S-methionine) as well as surface 125I-labeled cultured human endothelial cells; (2) upon crossed immunoelectrophoresis of solubilized endothelial cells against a polyclonal rabbit antiplatelet antiserum and 125I-labeled C17 IgG, a single precipitate of the protein(s) recognized by C17 is observed. As judged by their mobility in 9% polyacrylamide gels, both endothelial proteins appear to have a somewhat larger molecular weight than their platelet counterparts. Patterns obtained by crossed immunoelectrophoresis are also indicative of a difference in electrophoretic behavior of the platelet GP IIb/IIIa complex and the endothelial cell protein complex.

Vascular endothelial cells synthesize a plasma membrane protein indistinguishable from the platelet membrane glycoprotein IIa.

To define the role of membrane components that function in endothelial cell physiology and to characterize them biochemically, we have attempted to prepare monoclonal antibodies specific for endothelial cells. Several clones were obtained producing antibodies which bound to endothelial cells and also to platelets. The antibody of one of these clones, CLB-HEC 75, was studied in more detail. This antibody is directed against a single protein which is synthesized constitutively by endothelial cells and is expressed on the surface of both endothelial cells and platelets. The CLB-HEC 75 antigen was isolated from Nonidet P-40-solubilized endothelial cells and platelets by immunoprecipitation and exhibited an apparent molecular weight by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of approximately 145,000 in the presence of 2-mercaptoethanol. Two-dimensional polyacrylamide gel electrophoresis and crossed immunoelectrophoresis revealed that the mobility of the CLB-HEC 75 antigen relative to platelet glycoproteins Ib, IIa, IIb, and IIIa fits previously defined criteria for platelet membrane glycoprotein IIa. The CLB-HEC 75 antigen isolated from endothelial cells co-migrated under all conditions tested with the antigen from platelets. These results indicate that endothelial cells share a plasma membrane protein indistinguishable from platelet membrane glycoprotein IIa. This protein may be a component involved in the interaction of endothelial cells with their environment including coagulation factors, platelets, and the subendothelial matrix. CLB-HEC 75 may serve as a useful tool for studying these processes.