European seroepidemiology network 2: Standardisation of assays for seroepidemiology of varicella zoster virus.

BACKGROUND: The aim of the European Sero-Epidemiology Network (ESEN2) is to harmonise the serological surveillance of vaccine-preventable diseases in Europe. OBJECTIVE: To allow comparison of antibody prevalence in different countries by standardising results into common units. STUDY DESIGN: For varicella zoster virus (VZV), a reference laboratory established a panel of 148 samples, characterised by indirect enzyme-immunoassay (ELISA), indirect immunofluorescence, and complement fixation test. Fifty-seven samples were also studied by the fluorescence antibody to membrane antigen test. The geometric mean of the antibody activity (GMAA) obtained from four ELISA determinations was used to characterise each sample of the panel as positive (GMAA: >100 mIU/ml), equivocal (GMAA: 50-100 mIU/ml) or negative (GMAA: <50 mIU/ml) for antibody to VZV (anti-VZV). Thirteen laboratories, using five different ELISA tests, tested the panel. RESULTS: Agreement with the reference laboratory was above 85% in all cases, and the R(2) values obtained from regression analysis of the quantitative results were always higher than 0.87. Finally, the regression equations could be used to convert national values into a common unitage. CONCLUSION: This study confirmed that results for anti-VZV obtained by different ELISA methods can be converted into common units, enabling the comparison of the seroprevalence profiles obtained in the participant countries.     

Acute Methanol Poisoning: Prevalence and Predisposing Factors of Haemorrhagic and Non‐Haemorrhagic Brain Lesions

The purpose was to study the prevalence and predisposing factors of brain lesions in survivors of acute methanol poisoning. Clinical data on 106 patients with methanol poisoning were collected during the Czech mass poisoning outbreak. Of 83 survivors, in 46 (55%) patients, follow‐up examinations including magnetic resonance imaging of brain (MR) were performed 3–8 and 24–28 months after discharge from the hospital. Of 46 patients with a median age of 49 (interquartile range, 35–57) years, 24 (52%) patients had a total of 40 abnormal brain findings with haemorrhagic lesions detected in 15 (33%) and non‐haemorrhagic lesions found in 9 (19%) patients. The patients with haemorrhagic brain lesions were more acidemic (lower arterial blood pH, higher base deficit) and had higher glycaemia and lactacidaemia on admission than those without haemorrhages (all p < 0.05). Thirteen of 32 (41%) of patients with systemic anticoagulation and 2 of 14 (14%) of patients without it had haemorrhagic lesions (p = 0.080). Bleeding complications during the treatment occurred in 4 of 15 (27%) patients, and 5 of 15 (33%) patients had conditions predisposing to haemorrhage in the group with haemorrhagic lesions. In three cases with a series of computer tomography (CT)/MR performed during hospitalization, the necrotic lesions in the brain remained non‐haemorrhagic during hospitalization and haemorrhagic lesions were detected on the follow‐up MR examinations only. No association between brain haemorrhages and systemic anticoagulation during dialysis was found: brain haemorrhages might occur in severely poisoned patients treated without systemic anticoagulation, whereas treatment with high doses of heparin might not lead to brain haemorrhages.     

Proliferative potential and phenotypic analysis of long-term cultivated human granulosa cells initiated by addition of follicular fluid

Purpose

The aim of this study was to develop and optimize a strategy for long-term cultivation of luteinizing human granulosa cells (GCs).

Methods

GCs were cultivated in DMEM/F12 medium supplemented with 2% fetal calf serum. In vitro proliferation of GCs was supported by follicular fluid as well as FSH and growth factors.

Results

The cultured GCs were maintained for 45 days with a doubling time of 159 ± 24 h. GCs initiated by the addition of follicular fluid and cultivated under low serum conditions reached 10 ± 0.7 population doublings. GCs maintain the typical phenotypic expression and the telomere length according to specific culture conditions.

Conclusion

Our present study has demonstrated that GCs can be maintained in vitro for at least 45 days and this cell model can be beneficial when studying hormonal regulation associated with follicular maturation and preparation of oocytes for fertilization.     

HIV infection and high risk behavior among patients attending an STD referral clinic in Prague, Czech Republic

OBJECTIVE: This survey was conducted to determine the prevalence of HIV infection and associated risk behaviour in a high risk population of clients attending an STD clinic in Prague, Czech Republic. METHODS: Between September 1994 and January 1995, clients entering the Apolinar STD clinic in Prague, Czech Republic, were enrolled in a blinded, unlinked HIV antibody seroprevalence study. Non-identifying demographic characteristics, STD diagnoses, HIV risk exposures, and voluntary HIV testing experience were extracted from medical charts. RESULTS: Of 1394 patients for whom serum was available for testing, one was positive for HIV (HIV prevalence 0.07%, 95% confidence interval (CI) 0.01, 0.41%). This person was one of 28 men having sex with men (MSWM) (HIV prevalence among MSWM 3.6%, 95% CI 0.6, 17.7%). Among the 775 male clients, 75.5% had heterosexual unprotected sex, 11.1% had sex with high risk partners or prostitutes, 3.6% were MSWM, 1.0% were injecting drug users (IDUs), 0.7% were both MSWM and IDUs, and 6.8% and 1.8% had other or no recognized risk for HIV/STDs, respectively. Among the 619 female clients, 74.5% had heterosexual unprotected sex, 11.6% were prostitutes, 7.8% had sex with high risk partners, 1.1% were IDUs, and 3.9% and 2.3% had other or no recognised risk, respectively. The 304 adolescent patients (age 11-19 years) differed significantly (p < 0.05) in risk behaviour and STD diagnoses from the 1090 patients who were 20 years and older. Adolescents were significantly more likely to be female (58.6% v 40.5%, OR = 2.1), IDUs (3.6% v 0.4%, OR = 10.2), prostitutes (8.9% v 4.7%, OR = 2.0), and have sex partners with STDs (7.6% v 4.4%, OR = 1.8). The adolescent patients were also significantly more likely to be diagnosed with gonorrhoea (21.1% v 12.3%, OR = 1.9) and non-gonococcal urethritis (27.6% v 17.2%, OR = 1.8), and significantly less likely to have been tested previously for HIV (19.1% v 31.9%, OR = 0.5). CONCLUSIONS: HIV infection is currently uncommon in this population. However, the high rates of unprotected sex, prostitution, injecting drug use, and STDs, especially among adolescents, provide the basis for an epidemic in this population. Aggressive prevention education should be started before adolescence.


     

Standardization of measles, mumps and rubella assays to enable comparisons of seroprevalence data across 21 European countries and Australia

The aim of the European Sero-Epidemiology Network is to establish comparability of the serological surveillance of vaccine-preventable diseases in Europe. The designated reference laboratory (RL) for measles, mumps, rubella (MMR) prepared and tested a panel of 151 sera by the reference enzyme immunoassay (rEIA). Laboratories in 21 countries tested the panel for antibodies against MMR using their usual assay (a total of 16 different EIAs) and the results were plotted against the reference results in order to obtain equations for the standardization of national serum surveys. The RL also tested the panel by the plaque neutralization test (PNT). Large differences in qualitative results were found compared to the RL. Well-fitting standardization equations with R2> or =0.8 were obtained for almost all laboratories through regression of the quantitative results against those of the RL. When compared to PNT, the rEIA had a sensitivity of 95.3%, 92.8% and 100% and a specificity of 100%, 87.1% and 92.8% for measles, mumps and rubella, respectively. The need for standardization was highlighted by substantial inter-country differences. Standardization was successful and the selected standardization equations allowed the conversion of local serological results into common units and enabled direct comparison of seroprevalence data of the participating countries.     

Short report: HIV infection among commercial sex workers and injecting drug users in the Czech Republic

Since the first HIV case was diagnosed in the Czech Republic in 1985, there is a lack of information regarding the epidemiology of HIV infection in most high-risk groups. To determine the prevalence of, and risk factors for, HIV among female and male commercial sex workers (FCSW and MCSW, respectively) and injecting drug users (IDUs), cross-sectional studies were conducted in the cities of Cheb, Usti nad Labem, Ostrava, and Prague of the Czech Republic. A total of 1,277 subjects were enrolled, which included 585 FCSWs, 230 MCSWs, and 462 IDUs. The HIV prevalences were 0.7% (95% CI: 0.2-1.7%), 0.9% (95% CI: 0.1-3.1%), and 0.2% (95% CI: 0.005-1.2%) among FCSWs, MCSWs, and IDUs, respectively. Although low HIV prevalences were found, ongoing sentinel surveillance studies, which address modifiable behavioral and biologic risk factors among high-risk groups, are necessary to guide strategies to stem the tide of the epidemic in this country.