Importance of addition of HPV DNA testing to the cytology based cervical cancer screening and triage of findings with p16/Ki67 immunocytochemistry staining in 35 and 45 years old women LIBUSE trial data analysis.

OBJECTIVE: The study evaluates results of 2-years follow-up of patients in ages 35-36 and 45-46, who are participating in the project LIBUSE, that deals with efficacy of HPV DNA and Pap smear co-testing and p16/Ki67 dual staining in the Czech national cervical screening. DESIGN: Prospective observational study. SETTING: Department of Obstetrics and Gynecology, General University Hospital and 1st Medical Faculty, Charles University, Prague. MATERIALS AND METHODS: Out of all women enrolled in the project LIBUSE only those who were at the beginning of the study 35-36 and 45-46 years old were sellected. Conventional Pap smear and HPV DNA test (Cobas 4800, Roche Diagnostics) had been collected at the baseline. Women were stratified according to their results in the three risk groups: 1. low-risk, 2. high-risk and 3. intermediate risk, who subsequently underwent p16/Ki67 dual staining. All high-risk patients and those with positive result of dual staing were refered to the expert colposcopy. The cases with biopsy proven precancers or cancers were considered as „positive findings“. RESULTS: Altogether 352 women meet the age requirements. In 26 (7.6%) women had been proven HPV DNA positivity and out of the them 9 cases were HPV 16/18 positive. Severe cytological abnormality was found only in one patient (0.3%), who was simultaneously HPV positive. Ten women (2.8%) were classified as high-risk and directly refered to colposcopy. Another 18 patients underwent p16/Ki67 dual staining and 4 positive cases were refered to colposcopy too. After one year further 9 patients were classified as intermediate risk and 6 more were identified after two years of follow-up. Within two years 9 more patient were refered to colposcopy. After the entire period of follow-up in 10 patients biopsy confirmed precancer lesions, none of them had invasive cancer. CONCLUSIONS: Addition of HPV DNA testing with selective HPV 16/18 genotyping to the cytology based screening significantly increases sensitivity and safety of our cervical screening program.

Correlation between integration of high-risk HPV genome into human DNA detected by molecular combing and the severity of cervical lesions: first results of the EXPL-HPV-002 study.

OBJECTIVES: The aim of the EXPL-HPV-002 study is to evaluate the integration of 14 high-risk HPV as a biomarker of the severity and the progression of cervical lesions. Such a „triage biomarker“ would help to reduce the number of unnecessary colposcopies, to avoid over-treatment of lesions that spontaneously regress and to better target the lesions requiring treatment. DESIGN: EXPL-HPV-002 is a prospective, open-label, single arm, GCP study conducted at 2 clinical sites in the Czech Republic. SETTINGS: Investigations centers: Private Gynecology Center, Brno; Gynecological and Obstetrical Clinic, Brno; Genotyping central lab: NRL for Papillomaviruses and polyomaviruses, IHBT, Prague; Histology Central reading: Aeskulab Pathology, Prague; Molecular combing HPV test: Genomic Vision, Bagneux. METHODS: From June 2016 to May 2018, 688 patients aged 25-65, referred to colposcopy after an abnormal Pap-smear, were enrolled in the study. Among them 60% were found HPV high-risk. The study is divided in two phases: 1. a cross-sectional phase using data collected at first visit (colposcopy images ± histology, pap-smear for HPV genotyping and molecular combing) to study the association between HPV integration status versus colposcopy and histology grades; 2. a longitudinal phase using data collected in follow-up visits: cytology at 6, 18 and 30 months and colposcopy ± histology at 12, 24 and 36 months. A pap-smear collected at 12, 24 and 36 months allows to perform genotyping and molecular combing. HPV integration status is analyzed in comparison with the evolution of lesions, viral clearance and HPV genotype. HPV genotyping and molecular combing were performed on pap-smear samples in central laboratories. Histology data were reviewed by central reading. RESULTS: The transversal phase of the study is achieved and shows that the HPV integration into the human DNA, monitored by molecular combing, can significantly differentiate normal subjects from women with cervical lesions or cancer. CONCLUSION: HPV integration into the host genome, monitored by Genomic Visions technology, is a reliable diagnostic biomarker that will greatly help clinicians to improve their medical decision tree.

Effect of trapping method on species identification of phlebotomine sandflies by MALDI-TOF MS protein profiling.

Sandflies (Diptera: Psychodidae) (Newstead, 1911) are blood-feeding insects that transmit human pathogens including Leishmania (Trypanosomatida: Trypanosomatidae) parasites, causative agents of the leishmaniases. To elucidate Leishmania transmission cycles, conclusive identification of vector species is essential. Molecular approaches including matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) protein profiling have recently emerged to complement morphological identification. The aim of this study was to evaluate the effect of the trap type used to collect sandflies, specifically Centers for Disease Control (CDC) light or sticky traps, the two most commonly used in sandfly surveys, on subsequent MALDI-TOF MS protein profiling. Specimens of five species (Phlebotomus ariasi, Phlebotomus papatasi, Phlebotomus perniciosus, Phlebotomus sergenti, Sergentomyia minuta) collected in periurban and agricultural habitats in southeast Spain were subjected to protein profiling. Acquired protein spectra were queried against an in-house reference database and their quality assessed to evaluate the trap type effect. The results indicate that trap choice can substantially affect the quality of protein spectra in collected sandflies. Whereas specimens retrieved from light traps produced intense and reproducible spectra that allowed reliable species determination, profiles of specimens from sticky traps were compromised and often did not enable correct identification. Sticky traps should therefore not be used in surveys that deploy MALDI-TOF MS protein profiling for species identification.

[Nine-valent HPV vaccine - new generation of HPV vaccine]. / Nonavalentní HPV vakcína - nová generace ockování proti HPV.

OBJECTIVE: To evaluate current knowledge about new generation of HPV vaccine - nine-valent vaccine Gardasil9. DESIGN: Review article. RESULTS: The nine-valent vaccine against HPV 6/11/16/18/31/33/45/52/58 could improve efficacy of quadrivalent vaccine from 70 to 90 percent for cervical cancer. In addition this vaccine has covered around85-90% of HPV-related vulvar, vaginal and anal cancers. Efficacy and immunogenicity against HPV 6/11/16/18 is the same as for quadrivalent vaccine. Efficacy against HPV 31/33/45/52/58 associated lesions is 97%.

[Results of the Czech National Cervical Cancer screening programme]. / Výsledky Národního programu screeningu karcinomu delozního hrdla v Ceské republice.

INTRODUCTION: The nationwide Cervical Cancer Screening Programme in the Czech Republic was introduced in 2009. The aim of this article is to describe the results of this programme in the first few years after its transformation into a fully-fledged, organised programme. MATERIAL AND METHODS: During the first few years, a network of 37 accredited laboratories was stabilised, ensuring that examinations would be readily available across the Czech Republic. Although all women aged between 25 and 70 years fall within the target group of the current personalised invitation programme, women not in this age group are reimbursed for preventive screening examinations as well. The programme is equipped with an information support provided by the Institute of Biostatistics and Analyses of the Masaryk University, which runs the Cervical Cancer Screening Registry. RESULTS: Cervical cancer incidence and mortality rates in the Czech Republic have seen a significant decrease over the last decade. By the end of 2013, more than 11 million examinations were reported to the registry by the accredited laboratories, and more than 15,000 serious cervical lesions were detected. Analysis of the data from the registry makes it possible to monitor the quality of individual laboratories and the effectiveness of the entire programme. CONCLUSION: The promising start of the programme provides a strong argument for its continuation. The program--me has the potential to decrease cervical cancer incidence rates in the Czech Republic to values comparable with those reported by countries with advanced health care systems.

[Approach to population-based screening in the Czech Republic, methodology and first results of the personalised invitation of citizens to cancer screening programmes]. / Koncepce populacního screeningu v Ceské republice, metodika a první výsledky adresného zvaní obcanu do preventivních onkologických programu

In January 2014, a programme of personalised invitations was launched in the Czech Republic, with the objective of inviting insured persons to cancer screening programmes; namely breast cancer screening and cervical cancer screening in women, and colorectal cancer screening both in women and men. This programme aims at strengthening the current cancer prevention programmes, and to increase the currently inadequate participation of the target population in these programmes; therefore, personalised invitations are sent to citizens who have not participated in these programmes for several years and therefore at risk of developing a serious disease. The project is coordinated by the Czech Ministry of Health in cooperation with the expert medical societies involved (gynaecology, gastroenterology, gastrointestinal oncology, diagnostic radiology, general practice), representatives of health care payers, and other experts nominated by the Minister of Health. All health care payers invite their clients (insured persons) to preventive check-ups, covering all examinations needed. The project has been realised with the assistance of financial resources from EU funds. This article describes the methodology of personalised invitations which has been implemented nationwide, its data background, and the first results of the project in the first half of 2014, when almost 1.3 million Czech citizens were invited.

[Performance indicators in screening programmes]. / Indikátory kvality screeningových programu

Breast, colorectal and cervical cancer screening programmes make it possible to decrease the population mortality rates of these diseases. How-ever, complex standards of the quality of care must be introduced and followed in order to maintain a favourable ratio between the benefits and risks arising from population-wide screening programmes. Such programmes should be organized and population-based, ensuring that quality control is performed at all levels. This review introduces the system of quality control in the Czech cancer screening programmes, and provides specific examples of performance indicators that are usable and/ or being used in these programmes. Cancer screening programmes in the Czech Republic are equipped with a comprehensive information background which involves monitoring of the cancer burden in the population, monitoring of the screening process based on clinical data, and monitoring of the screening process based on administrative data. In particular, the specific performance indicators describe the success rate of take up of the target population, ability of the screening test to reveal (sensitivity) or to exclude (specificity) the screened condition, correct employment of subsequent diagnostic methods or treatment of detected cancers or precancerous lesions where applicable. In the Czech breast cancer screening programme, these indicators are routinely used in order to monitor the individual centres; in both colorectal and cervical cancer screening programmes, these indicators are used to monitor the entire programme, whereas the system of quality control for individual centres is under continuous development. A project of personalized invitations was launched in 2014, and its results are regularly evaluated in cooperation with the Czech National Reference Centre and the Ministry of Health of the Czech Republic.

[Prebioptic methods in the cervical cancer screening]. / Prebioptické metody pri screeningu karcinomu delozního hrdla.

OBJECTIVE: To evaluate the agreement between Pap smears, colposcopic findings and definitive excisional specimen in patients with abnormal Pap smears. DESIGN: Retrospective study. SETTING: Center of outpatient gynecology and primary care, Brno; Department of Obstetrics and Gynecology, University Hospital, Olomouc; Department of Medical Biophysics, Palacky University, Olomouc.Subjects and methods of the study: This retrospective study assessed the correlation between colposcopy and histopathology of woman who had abnormal Pap smears. Colposcopic chart review included participants from 2008 to 2012 who attended colposcopic clinic, center of outpatient gynecology care, Brno. RESULTS: One thousand nine hundred and twenty five patients screened by cytology, submitted to colposcopy and subjected to cone biopsy were selected. Cytopathological results were compared with colposcopic findings and results obtained on the basis of histological analyses of cone biopsy specimen. Agreement of cytology and histopathological diagnosis was in 1199 patients (62.3%). Agreement of colposcopic diagnosis and cervical pathology was matched in 1492 (93.5%). Agreement of colposcopy and cytology was found in 1022 pacientek (64.1%). False negatives of cytology in high grade lesions were in 36.4%. CONCLUSION: Strength of agreement between colposcopic diagnosis and cervical pathology was found to be very good while for cytology high percentage of false negative results was seen. Expert colposcopists and high quality standard cytopathologic and bioptic laboratories are necessary for nationwide cervical screening programmes. KEYWORDS: cervical cancer, colposcopy, oncological cytology, histology, CIN, cone biopsy.

[Role of prebioptic and bioptic methodsin the screening and diagnosis of cervical cancer]. / Prebioptické a bioptické metody pri screeningu a diagnostice karcinomu delozního hrdla.

OBJECTIVE: To describe the available prebioptical and bioptical methods used in screening and diagnosis of cervical cancer. DESIGN: Review article. SETTING: Center of outpatient gynecology and primary care, Brno; Department of Obstetrics and Gynecology, University Hospital, Olomouc.Subjects and methods of the study: In a review article there are outlines of the prebioptical investigative techniques and related diagnostics aim to detect, verificate and treat cervical lesions. It describes and compares various methods, including their mutual correlation. CONCLUSION: The oncological cytology, colposcopy and HPV testing have an irreplaceable role in diagnosis of cervical cancer. Appropriate combination gives us a chance to achieve high sensitivity in detection of severe lesions of the cervix.

The role of indigenous phlebotomine sandflies and mammals in the spreading of leishmaniasis agents in the Mediterranean region.

An updated view of the establishment and spread of the leishmaniases in Europe is presented, mostly with respect to newly emerging and re-emerging foci and the incrimination of neglected as well as new reservoir hosts. At the same time, a concept of specific versus permissive vectors reassesses the potential role of various sandfly species in Leishmania transmission and considers the risk of introduction of exotic Leishmania species in Europe. The leishmaniases are dynamic diseases and the circumstances of transmission are continually changing in relation to environmental, demographic and human behavioural factors. Changes in the habitat of the natural hosts and vectors, immunosuppressive conditions (like infection with human immunodeficiency virus (HIV) or organ transplantation-associated therapies in humans) and the consequences of war, all contribute to the transformation of the epidemiology of leishmaniasis. Such changes should be considered when studying the spread of the disease throughout Europe for targeted control measures to safeguard public health.

[Systemic enzyme therapy in the treatment of recurrent vulvovaginal candidiasis]. / Systémová enzymoterapie v lécbe recidivující vulvovaginální kandidózy.

OBJECTIVE: Phase I: A prospective evaluation of long-term systemic enzyme therapy (Wobenzym; WE) effects on the frequency of recurrent vulvovaginal candidiasis (RVVC) episodes. Phase II: A retrospective analysis of possible positive effects remaining in the next 3 years. TRIAL DESIGN: Original work - pilot project consisting of prospective phase I and retrospective phase II trials. METHODS: Composition of animal and plant proteo-lytic enzymes (WE) is a common component in the treatment of chronic or recurrent inflammatory diseases and has been also shown to have immunomodulatory effects. Project involving 7 gynecology practices has been started in 2005 - 2007 to evaluate the effectiveness of WE in the complex treatment of RVVC. The trial involved 62 women with at least 4 microscopically confirmed episodes of RVVC in the last 12 months (year 1; 4-9 episodes, mean 4.4 episodes per patient-year). From the beginning of the trial, participants took WE in the dose of 2× 8 tbl/day for 10 weeks and were monitored for 12 months (year +1). All infections of RVVC were treated according to usual practice of the particular gynecologist. The number of RVVC episodes during the year -1 was compared to the number of RVVC during the year +1. To evaluate possible long-term effects of the WE treatment, retrospective analysis of the data from 3 years following the phase I (year +2, +3, +4) was performed. Complete data for 54 women were collected (87.1% of the former group of patients). All data were processed with regular statistics methods. RESULTS: Mean number of RVVC in the year +1 has decreased from 4.4 to 0.5 per patient-year (i.e. by 88.5%; p < 0.001). All women experienced an improvement, 63% of them experienced no recurrence. The lower incidence of RVVC remained also for the phase II (year +2: 0.91; year +3: 0.57; year +4: 0.52 episodes of RVVC per patient-year). The difference, as compared to the mean incidence of RVVC before the treatment (year -1), remains significant (p < 0.001) although women, who became pregnant during the trial, were not excluded from the observed population. If only non-pregnant women were analyzed (41 women), the mean incidence of RVVC was even lower (year +2: 0.69; year +3: 0.39; year +4: 0.44 episodes of RVVC per patient-year). CONCLUSION: 10 weeks of systemic enzyme therapy (WE) in women with RVVC significantly reduced recurrence of this disease not only for the first year following the treatment, but also for the next 3 years. An explanation of the basis for this effect needs further research.

[Epidemiology of genital warts in female population of Czech Republic]. / Epidemiologie genitálních bradavic mezi zenami v Ceské republice.

OBJECTIVE: The aim of study was to evaluate prevalence of genital warts in Czech Republic. SUBJECT: Multicentric prospective observation study. SETTING: HPV College. METHODS: During 6 month (February 2010 - July 2010) 20 private gynaecological centers in all Czech Republic were counting up the number of genital warts cases. Risk factors, therapy and knowledges about genital warts were evaluated. RESULTS: There were 637 patients with genital warts in cohort of 70 980 patients. The prevalence of genital warts was 0.89%. The most frequent risk factor was cigarette smoking in 37%. Main strategy for treatment were podophyllin local application and cold knife excision. CONCLUSION: The prevalence of genital warts in our study has shown importance for its prevention by rules of safety sex and HPV vaccination against HPV type 6 and 11.

Phlebotomus sergenti (Parrot, 1917) identified as Leishmania killicki host in Ghardaïa, south Algeria.

Since 2005, an outbreak of human cutaneous leishmaniasis (CL) in Ghardaïa, south Algeria, was studied and one output of these investigations was the identification of two Leishmania species, Leishmania major and Leishmania killicki, as the CL causative agents. In the present study, we were curious to focus on sand fly fauna present in this area and detection of Leishmania-positive sand fly females. Sand flies (3717) were collected during two seasons using sticky papers and CDC light traps in urban, rural and sylvatic sites. Twelve Phlebotomus species were identified. Phlebotomus papatasi was dominant in the urban site while Phlebotomus sergenti and Phlebotomus riouxi/chabaudi were dominant in the sylvatic site. Out of 74 P. sergenti females captured by CDC light traps in the sylvatic site populated by Ghardaïas' Gundi (Massoutiera mzabi), three ones were hosting Leishmania promastigotes. PCR-RFLP and sequencing of seven single-copy coding DNA sequences identified the promastigotes as L. killicki. Furthermore, laboratory experiments revealed that L. killicki isolate sampled from a CL patient inhabiting the studied region develop well in P. sergenti females. Our findings strongly suggest that the human cutaneous leishmaniases caused by L. killicki is a zoonotic disease with P. sergenti sand flies acting as hosts and vectors and gundi rodents as reservoirs.

Intraspecific variability of natural populations of Phlebotomus sergenti, the main vector of Leishmania tropica.

An intraspecific study of Phlebotomus sergenti was performed on populations from Turkey, Syria, Israel, and Uzbekistan by four different approaches: geometric morphometrics, RAPD analysis, internal transcribed spacer 2 (ITS2) sequencing (nuclear marker), and cytochrome B sequencing (mitochondrial marker). In RAPD analysis, distinct clades were formed in accordance with the geographical origin of the specimens. There was no distinct grouping according to place of origin within the Turkish samples from various localities in south-eastern Anatolia, which suggests a gene flow between populations separated spatially by the Amanos mountains, a mountain range of a considerable altitude. The results of ITS2 rDNA sequencing complied with the previously published intraspecific division of P. sergenti into two branches, northeastern and southwestern. However, mtDNA haplotypes formed three lineages with specimens from Turkey and Israel, sharing a common clade. A previously postulated hypothesis about a complex of sibling species within P. sergenti is therefore questionable. Cytochrome B seems to be a more discriminative marker for intraspecific variability assessment.

[Genital warts and HPV vaccination]. / Genitální bradavice a HPV vakcinace.

OBJECTIVE: To present and overview of incidence of, and cost of care for, genital warts. SUBJECT: Review. SETTING: Department of Obstetrics and Gynecology, Palacky University and Faculty University, Olomouc; Office gynecology and primary care centre, Brno; Department of Obstetrics and Gynecology, Charles university in Prague-First Faculty of Medicine and General Faculty Hospital, Prague. METHOD: Literature review of incidence of, and cost of care for, genital warts in some european countries, North America and Australia. CONCLUSION: Genital warts exert a considerable impact on health services, a large proportion of which could be prevented through immunisation using the quadrivalent human papillomavirus vaccine.

Czech National Cancer Screening Programmes in 2010.

BACKGROUNDS: All three cancer screening programmes recommended by the Council of the EU are available to defined target age groups in the Czech Republic. Organized programmes for screening of breast, colorectal and cervical cancer have been initiated in the last decade. MATERIALS AND METHODS: A system for information support, as an essential component of organized screening programmes, has been implemented in all screening programmes. It comprises the Czech National Cancer Registry to monitor the cancer burden and population impact of the programmes, the National Reference Centre as a provider of nationwide insurance claims data, and the specialised databases of all three programmes, which collect information on screening, diagnostics and final diagnoses. RESULTS: Early diagnostics of malignant neoplasms and progress in therapy have helped to stabilize mortality, even in diagnoses with increasing incidence. The coverage of the Czech screening programmes has constantly been rising; however, it is still insufficient: 51.2%, 17.9% and 48.4% of the target population was covered at the end of 2008 in breast, colorectal and cervical screening programmes, respectively. In 2008, a total of 468,419 women underwent screening mammography and 2,128 tumours were detected (4.5 per 1,000 screened). According to the screening colonoscopy registry, more than 13,000 men and women underwent preventive colonoscopy in 2009, 4,085 patients were diagnosed with adenoma and 619 with colorectal cancer, mostly in the early stages. The information system for cervical screening was implemented in 2009 and has been running in pilot mode; the first results are expected at the end of 2010. CONCLUSION: The system for information support within organised cancer screening programmes enables monitoring of the performance of screening and diagnostic centres and thus helps to maintain continuous quality improvements, which are a necessary presumption for replicating the promising results of clinical trials. To achieve a substantial impact on population incidence and mortality, a large increase in test coverage in target populations will be necessary. The programmes should be transformed to a population-based form, which involves inviting all people in the target population to be screened.

[Screening of cervical cancer--theoretical background]. / Screening karcinomu delozního hrdla--teoretické predpoklady.

OBJECTIVE: To show the basic rules for creation of creening programme in case of cervical cancer's screening. DESIGN: Review article. SETTING: Centrum of gyneco-oncology prevention, Havírov. METHODS: Study of the current literature. CONCLUSION: Screening programme should be set up economic possibilities of state. It must cover all risk group. It should have high specificity and sensitivity. This test should be connected with diagnostic methods and appropriate therapy.