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E3 ubiquitin ligases are of interest as drug targets due to their involvement in the regulation of the functions and interactions of several proteins. Various E3 ligase complexes are considered oncogenes or tumor suppres- sors associated with the development of melanoma. These proteins regulate the functions of various signaling pathways and proteins, such as p53 and Notch. The aim of the present study was to determine the expression levels of F-box and WD repeat domain-containing 7 (FBXW7), c-Myc, MDM2 and p53 proteins in samples from patients with dysplastic nevi or melanoma, and to evaluate their association with clinicopathological parameters and prognosis of the disease. Paraffin blocks with postoperative material from 100 patients diagnosed with dysplastic moles or melanoma were used in the present study. Tissue microarrays and immunohistochem- istry were used to examine FBXW7, c-Myc, MDM2 and p53 protein expression. The results revealed that there was significantly lower FBXW7 expression in advanced melanoma compared with dysplastic nevus, melanoma in situ and stage pT1 melanoma (P

Objectives: Sexual dysfunction was reported to compromise the quality of life in childhood cancer survivors. The aim of our study was to evaluate the reproductive health in long-term pediatric cancer survivors by conducting a crosscut survey. Material and methods: Childhood cancer survivors over 18 years of age, who were in remission for more than 5 years, were invited to complete a gender-specific questionnaire surveying on their reproductive health. Demographic and treatment data were retrieved from their medical records. Treatment modalities were reviewed for its potential gonadotoxicity. Results: 34 (17 males and 17 females, respectively) from 346 addressed survivors (9.8%) completed the questionnaire. Median age and follow-up after diagnosis was 27 (18–35) and 14 (3–25) years, respectively. Some respondents reported sexual concerns: 11.8% males experienced problems with penetration, two males (11.8%) who underwent semen analysis were found to be azoospermic. Similarly, 11.8% females reported delayed puberty, the average age of menarche was 14 (12–17) years, 29.4% females reported irregular menstrual cycles. Cyclophosphamide equivalent dose (CED) differed significantly between the patients treated for leukemia, lymphoma and solid tumors (3000 vs 4352 vs 6660 mg/m2, respectively, p = 0.014). Conclusions: Low prevalence of sexual dysfunction, fertility related disorders or delayed puberty in childhood cancer survivors was found. However, the results should be interpreted with caution taking into account a low response rate.

// Ausra Stumbryte 1, 2 , Zivile Gudleviciene 1 , Gabrielis Kundrotas 1 , Daiva Dabkeviciene 2 , Agne Kunickaite 3 and Saulius Cicenas 4, 5 1 Biobank, National Cancer Institute, LT-08660 Vilnius, Lithuania 2 Institute of Biosciences, Vilnius University Life Sciences Center, LT-10257 Vilnius, Lithuania 3 Department of Human and Medical Genetics, Faculty of Medicine, Vilnius University, LT-08661 Vilnius, Lithuania 4 Department of Thoracic Surgery and Oncology, National Cancer Institute, LT-08660 Vilnius, Lithuania 5 Faculty of Medicine, Vilnius University, LT-03101 Vilnius, Lithuania Correspondence to: Ausra Stumbryte, email: ausra.stumbryte@nvi.lt Keywords: lung cancer prognostic markers; SNPs; patient survival; mortality rate; HPV phylogenetic line Received: March 22, 2017 Accepted: November 01, 2017 Published: November 29, 2017 ABSTRACT Lung cancer (LC) is the second common and with the highest mortality oncological disease. Specific biomarkers for its diagnostics, treatment, and prognosis are still under the investigations. Aim of our study was to evaluate the relationship between the polymorphisms of TP53 pathway genes TP53 , MDM2 , MDM4 , the polymorphisms of HPV-associated genes MTHFR , CASP8 , CCR5 , and HPV infection with survival of LC patients. SNPs were genotyped using PCR-RFLP. qRT-PCR was used to detect, identify, and quantify HPV. No statistically significant differences were detected between individual SNPs and patient survival with stage I-IV LC. Cluster analysis of SNPs in genes MDM4 A/A, CCR5 wt/Δ32, MTHFR C/T, MDM2 T/T showed possible association with the worse survival. Patients who were diagnosed with C/T polymorphic variant of gene MTHFR tend not to survive stage III-IV LC ( P = .12). There is a tendency between MDM2 gene T/T variant and worse survival of patients diagnosed with late stage LC ( P = .11). HPV infection is very rear among LC patients (3 of 92). Overall, there is a link, although statistically insignificant, between specific SNPs and LC patient survival frequency and time, meanwhile the combination of specific SNPs showed a statistically significant measure. In conclusion, we determined statistically significant ( P = .04) link between the poor survival of LC patients after surgery and the combination of polymorphic variants C/T of the MTHFR and T/T of the MDM2 genes, whereas individually these SNPs do not show significant relationship with the survival of patients after surgery.

Additional file 5. Characterization of carboxylated QDs. (a) Image of QDs was made by Philips CM200 Field emission transmission electron microscope (TEM) equipped with light element EDX detector and Gatan Imaging Filter for PEELS and Energy Filtered TEM. Scale bar 20 nm. (b) 3D topography image of QDs was registered on mica surface with Innova atomic force microscope in the tapping mode using silicon nitride probes MPP12283. (c) Hydrodynamic size distribution of QDs was measured using a dynamic light scattering device Zeta Plus PALS. (d) The steady state absorption and photoluminescence spectra were recorded (λex = 625 nm) on Cary 50 UV–Vis spectrophotometer and Cary Eclipse fluorimeter, respectively. (e) PL decay curve of QDs was registered using FLS920 spectrometer equipped with 405 nm (66.9 ps) pulsed laser.

Background Human mesenchymal stem cells (MSCs) have drawn much attention in the field of regenerative medicine for their immunomodulatory and anti-inflammatory effects. MSCs possess specific tumor-oriented migration and incorporation highlighting the potential for MSCs to be used as an ideal carrier for anticancer agents. Bone marrow is the main source of MSCs for clinical applications. MSCs tracking in vivo is a critical component of the safety and efficacy evaluation of therapeutic cell products; therefore, cells must be labeled with contrast agents to enable visualization of the MSCs migration in vivo. Due to their unique properties, quantum dots (QDs) are emerging as optimal tools in long-term MSC optical imaging applications. The aim of this study was to investigate the uptake dynamics, cytotoxity, subcellular and extracellular distribution of non-targeted carboxylated quantum dots in human bone marrow MSCs at different cell growing densities. Results QDs had no negative impact on MSC viability throughout the experiment and accumulated in all observed cells efficiently; however, in some MSCs QDs induced formation of lipid droplets. At low cell growing densities QDs distribute within MSCs cytoplasm already after 1 h of incubation reaching saturation after 6 h. After 24 h QDs localize mainly in the perinuclear region of the cells in endosomes. Interestingly, in more confluent culture QDs localize mostly outside MSCs. QDs abundantly mark MSC long filopodia-like structures attaching neighboring cells. At high cell density cultivation, we for the first time demonstrated that carboxylated QDs localize in human bone marrow MSC extracellular matrix. Moreover, we observed that average photoluminescence lifetime of QDs distributed in extracellular matrix are longer than lifetimes of QDs entrapped in endocytic vesicles; thus, for the first time showing the possibility to identify and distinguish localization of QDs in various extracellular and intracellular structures using fluorescence-lifetime imaging microscopy without additional staining assays. Conclusion Carboxylated QDs can be used as nonspecific and effective dye for staining of human bone marrow MSCs and their specific extracellular structures. These results are promising in fundamental stem cell biology as well as in cellular therapy, anticancer drug delivery and tissue engineering. Electronic supplementary material The online version of this article (10.1186/s12951-019-0470-6) contains supplementary material, which is available to authorized users.

// Gabrielis Kundrotas 1,2 , Evelina Gasperskaja 1 , Grazina Slapsyte 1 , Zivile Gudleviciene 2 , Jan Krasko 3 , Ausra Stumbryte 2 and Regina Liudkeviciene 2 1 Department of Botany and Genetics, Faculty of Natural Sciences, Vilnius University, Vilnius, Lithuania 2 Biobank, National Cancer Institute, Vilnius, Lithuania 3 Laboratory of Immunology, National Cancer Institute, Vilnius, Lithuania Correspondence to: Gabrielis Kundrotas, email: // Keywords : human mesenchymal stem cells, genomic stability, long-term expansion, PCR arrays, senescence markers, Gerotarget Received : August 04, 2015 Accepted : February 05, 2016 Published : February 17, 2016 Abstract Human bone marrow mesenchymal stem cells (hBM-MSCs) hold promise for treating incurable diseases and repairing of damaged tissues. However, hBM-MSCs face the disadvantages of painful invasive isolation and limited cell numbers. In this study we assessed characteristics of MSCs isolated from residual human bone marrow transplantation material and expanded to clinically relevant numbers at passages 3-4 and 6-7. Results indicated that early passage hBM-MSCs are genomically stable and retain identity and high proliferation capacity. Despite the chromosomal stability, the cells became senescent at late passages, paralleling the slower proliferation, altered morphology and immunophenotype. By qRT-PCR array profiling, we revealed 13 genes and 33 miRNAs significantly differentially expressed in late passage cells, among which 8 genes and 30 miRNAs emerged as potential novel biomarkers of hBM-MSC aging. Functional analysis of genes with altered expression showed strong association with biological processes causing cellular senescence. Altogether, this study revives hBM as convenient source for cellular therapy. Potential novel markers provide new details for better understanding the hBM-MSC senescence mechanisms, contributing to basic science, facilitating the development of cellular therapy quality control, and providing new clues for human disease processes since senescence phenotype of the hematological patient hBM-MSCs only very recently has been revealed.

Epigenetic alterations of human papillomavirus (HPV) genome play an important role in virus life cycle and carcinogenic progression. The aim of the current study was to investigate the correlation between the grade of cervical pathology and DNA methylation status within the L1 gene and the long control region (LCR) of HPV16, HPV18 and HPV51. HPV genomes were analyzed using bisulfite DNA modification procedure with the subsequent amplification of target DNA regions and sequencing. A collection of 202 cervical specimens was analyzed: 157 HPV16-positive specimens, 21 HPV18-positive specimens and 24 HPV51-positive specimens. This study revealed that methylation of CpG was significantly more prevalent in L1 gene as compared to LCR region of all three studied HPV types and the degree of DNA methylation level correlated with the severity of cervical neoplasia. An increased DNA methylation level of HPV16 promoter region in case of cervical cancer was determined.

Background. Mesenchymal stem cells (MSCs) are currently exploited in numerous clinical trials to investigate their potential in immune regulation, hematopoesis or tissue regeneration. The most common source of MSCs for clinical use is human bone marrow. To generate sufficient numbers of cells relevant to clinical use in most cases the high volumes (20-50 ml) of bone marrow aspirates are taken. Methods. In this pilot study, 8 healthy bone marrow donors were included. Two different MSC extraction methods were evaluated: MSCs extraction from 60 ml of bone marrow using density gradient and MSCs extraction from 6 ml using red blood cell (RBC) lysis. Results. Our results showed that after RBC lysis the efficient amount of human MSCs can be isolated from 10 times less bone marrow volume (6 ml). Moreover, using small volume of bone marrow the adequate therapeutical dose of MSCs could be achieved during similar period of time (3-4 weeks). In conclusion, we have shown that MSCs isolation using RBC lysis is an effective and more advantageous method in comparison to standard MSCs isolation using density-gradient. Using RBC lysis from small volume of bone marrow the same amount of MSCs were obtained as usually using large volume and density-gradient.

Human papillomavirus (HPV) is the main cause of cervical cancer. Therefore, the detection of oncogenic HPV types is important in predicting the risk of cervical cancer. The aim of the current study was to estimate the prevalence of 16 carcinogenic and potentially carcinogenic HPV types in the study group of Lithuanian women with various grades of cervical pathology in comparison to healthy women. A total of 824 cervical specimens were investigated for HPV DNA: 547 specimens of women with abnormal cytology and 277 specimens of healthy women. Cytological diagnosis was confirmed by histology. For the detection of HPV infection, HPV DNA was amplified by PCR using three different primer systems. HPV DNA was detected in 67.6% of specimens collected from women with abnormal cytology and 24.2% of specimens collected from healthy women. The frequency of HPV-positive specimens correlated with the severity of cervical pathology: it ranged from 50.0% in the subgroup of atypical squamous cells to 80.6% in cervical cancer. In cases confirmed by histology the frequency of HPV-positive specimens ranged from 68.6% in the subgroup of cervical intraepithelial neoplasia grade 1 to 89.2% in cervical intraepithelial neoplasia grade 3 or carcinoma in situ. HPV DNA-positive samples were further investigated for the presence of 16 HPV types by multiplex PCR. The most common HPV type was HPV 16 (detected in 42.3% of HPV-positive specimens) followed by HPV 31 (10.1%), HPV 33 (8.2%), and HPV 56 (5.7%). In contrast, the frequency of HPV 18 was lower as compared to other countries. J. Med. Virol. 87:461–471, 2015. © 2014 Wiley Periodicals, Inc.

In some countries the cervical cancer incidence and mortality rates are much higher compared to the European average. The differences of HPV and its type prevalence between countries and regions influence cervical cancer incidence and mortality. Regarding the differences in cervical cancer incidence and mortality in Lithuania and Belarus, the aim of this study was to describe HPV infection level and HPVs type distribution among two study groups of patients with moderate or severe cervical intraepithelial neoplasia (CIN2-3) and cervical cancer. Our data shows that 74.2% [95% CI: 63.64÷84.76] of Lithuanian patients with cervical cancer and 85.6% [95% CI: 85.53÷92.85] of the study group with CIN2-3 were HPV positive, while in the study groups of Belarusian patients HPV infection was detected in 92.6% [95% CI, 74.25÷98.71] and 65.4% [95% CI, 44.36÷82.06] cases respectively. HPV 16 was the most prevalent type in Lithuanian as well as in Belarusian patients of the study groups. HPV 18 in Lithuanian patients of the study group with cervical cancer was identified in 10.2% [95% CI: 1.73÷18.67] and in the study group with CIN2-3 — in 2.6% [95% CI: 0.95÷6.15] of cases. HPV 18 was not detected in Belarusian patients of both groups.

Purpose: Matrix metalloproteinases (MMPs) are implicated in cancer cells invasion and metastasis processes and have been investigated as potential cancer biomarkers. In this study MMP-9 gene expression and MMP-9 -1562 C/T polymorphism in breast and non-small cell lung cancer patients’ blood and tumor samples and its correlation with clinicopathological parameters were investigated. Material/Methods: MMP-9 gene expression was assessed by reverse transcription – polymerase chain reaction method in 108 cancer patients’ blood and tumor samples. MMP-9 -1562 C/T polymorphism was determined by the polymerase chain reaction – based restriction fragment length polymorphism method. Results: Significant relationship of MMP-9 gene expression and tumor differentiation grade was found only between groups with G1 and G3 breast tumors. Low survival rates were identified among positive MMP-9 expression in blood and ductal carcinoma of the breast (p=0.01) and negative progesterone receptor reaction (p=0.04). Significant differences in the distribution among genotypes were found between groups with stage I and stages III/IV (p=0.005) as well as between groups with lymph node status N0 and N1 (p

Study Question The 16th European IVF-monitoring (EIM) report presents the data of the treatments involving assisted reproductive technology (ART) and intrauterine insemination (IUI) initiated in Europe during 2012: are there any changes compared with previous years? Summary Answer Despite some fluctuations in the number of countries reporting data, the overall number of ART cycles has continued to increase year by year, the pregnancy rates (PRs) in 2012 remained stable compared with those reported in 2011, and the number of transfers with multiple embryos (3+) and the multiple delivery rates were lower than ever before. What is Known Already Since 1997, ART data in Europe have been collected and re-ported in 15 manuscripts, published in Human Reproduction. Study Design, Size, Duration Retrospective data collection of European ART data by the EIM Consortium for the European Society of Human Reproduction and Embryology (ESHRE). Data for cycles between 1 January and 31 December 2012 were collected from National Registers, when existing, or on a voluntary basis by personal information. Participants/Materials, Setting, Methods From 34 countries (+1 compared with 2011), 1111 clinics reported 640 144 treatment cycles including 139 978 of IVF, 312 600 of ICSI, 139 558 of frozen embryo replacement (FER), 33 605 of egg donation (ED), 421 of in vitro maturation, 8433 of preimplantation genetic diagnosis/preimplantation genetic screening and 5549 of frozen oocyte replacements (FOR). European data on intrauterine insemination using husband/partner's semen (IUI-H) and donor semen (IUI-D) were reported from 1126 IUI labs in 24 countries. A total of 175 028 IUI-H and 43 497 IUI-D cycles were included. Main Results and the Role of Chance In 18 countries where all clinics reported to their ART register, a total of 369 081 ART cycles were performed in a population of around 295 million inhabitants, corresponding to 1252 cycles per million inhabitants (range 325-2732 cycles per million inhabitants). For all IVF cycles, the clinical PRs per aspiration and per transfer were stable with 29.4 (29.1% in 2011) and 33.8% (33.2% in 2011), respectively. For ICSI, the corresponding rates also were stable with 27.8 (27.9% in 2011) and 32.3% (31.8% in 2011). In FER cycles, the PR per thawing/warming increased to 23.1% (21.3% in 2011). In ED cycles, the PR per fresh transfer increased to 48.4% (45.8% in 2011) and to 35.9% (33.6% in 2011) per thawed transfer, while it was 45.1% for transfers after FOR. The delivery rate after IUI remained stable, at 8.5% (8.3% in 2011) after IUI-H and 12.0% (12.2% in 2011) after IUI-D. In IVF and ICSI cycles, 1, 2, 3 and 4+ embryos were transferred in 30.2, 55.4, 13.3 and 1.1% of the cycles, respectively. The proportions of singleton, twin and triplet deliveries after IVF and ICSI (added together) were 82.1, 17.3 and 0.6%, respectively, resulting in a total multiple delivery rate of 17.9% compared with 19.2% in 2011 and 20.6% in 2010. In FER cycles, the multiple delivery rate was 12.5% (12.2% twins and 0.3% triplets). Twin and triplet delivery rates associated with IUI cycles were 9.0%/0.4% and 7.2%/0.5%, following treatment with husband and donor semen, respectively. LIMITATIONS, REASONS FOR CAUTION The method of reporting varies among countries, and registers from a number of countries have been unable to provide some of the relevant data such as initiated cycles and deliveries. As long as data are incomplete and generated through different methods of collection, results should be interpreted with caution. Wider Implications of the Findings The 16th ESHRE report on ART shows a continuing expansion of the number of treatment cycles in Europe, with more than 640 000 cycles reported in 2012 with an increasing contribution to birthrate in many countries. However, the need to improve and standardize the national registries, and to establish validation methodologies remains manifest. STUDY FUNDING/COMPETING INTERESTS The study has no external funding; all costs are covered by ESHRE. There are no competing interests.

Survival of patients with head and neck squamous cell carcinoma (HNSCC) is dependent on many factors — stage of the disease, treatment regimen, operation technique etc. Many authors discuss on association of survival with various biomarkers as HPV infection, p53 mutation and polymorphism or p16 expression. The objective of our study was to analyze the survival of HNSCC patients in association with HPV infection and p53 polymorphism.

Cervical cancer is caused by a persistent infection with high-risk human papillomavirus (hr-HPV). Accurate grading of premalignant precursor lesions, referred to as cervical intraepithelial neoplasia (CIN) is important for clinical management of patients. A promising candidate marker to identify high-grade CIN lesions is the cellular protein p16INK4a, an indirect indicator of cell cycle dysregulation commonly expressed in cervical dysplasias and carcinomas associated with hr-HPV. Like other cell cycle regulatory proteins, the diagnostic role of p16INK4A is still under consideration.The aim of the current study was to investigate the expression of p16INK4A mRNA by a quantitative reverse transcriptase PCR (qRT-PCR) in cervical specimens and evaluate the correlation between the p16INK4A expression levels and the grade of CIN as well as hr-HPV genotypes. The qRT-PCR assay for the detection of p16INK4A mRNA was developed and optimized. As an internal control, integrin gene was selected. The levels of p...

Background Cervical cancer is caused primarily by human papillomaviruses (HPV). The polymorphism rs1042522 at codon 72 of the TP53 tumour-suppressor gene has been investigated as a genetic cofactor. More than 80 studies were done between 1998 and 2006, after it was initially reported that women who are homozygous for the arginine allele had a risk for cervical cancer seven times higher than women who were heterozygous for the allele. However, results have been inconsistent. Here we analyse pooled data from 49 studies to determine whether there is an association between TP53 codon 72 polymorphism and cervical cancer.Methods Individual data on 7946 cases and 7888 controls from 49 different studies worldwide were reanalysed. Odds ratios (OR) were estimated using logistic regression, stratifying by study and ethnic origin. Subgroup analyses were done for infection with HPV, ethnic origin, Hardy-Weinberg equilibrium, study quality, and the material used to determine TP53 genotype.Findings The pooled estimates (OR) for invasive cervical cancer were 1.22 (95% CI 1.08-1-39) for arginine homozygotes compared with heterozygotes, and 1.13 (0.94-1.35) for arginine homozygotes versus proline homozygotes. Subgroup analyses showed significant excess risks only in studies where controls were not in Hardy-Weinberg equilibrium (1.71 [1.21-2.42] for arginine homozygotes compared with heterozygotes), in non-epidemiological studies (1.35 [1.15-1.58] for arginine homozygotes compared with heterozygotes), and in studies where TP53 genotype was determined from tumour tissue (1.39 [1.13-1.73] for arginine homozygotes compared with heterozygotes). Null results were noted in studies with sound epidemiological design and conduct (1.06 [0.87-1.29] for arginine homozygotes compared with heterozygotes), and studies in which TP53 genotype was determined from white blood cells (1.06 [0.87-1.29] for arginine homozygotes compared with heterozygotes).Interpretation Subgroup analyses indicated that excess risks were most likely not due to clinical or biological factors, but to errors in study methods. No association was found between cervical cancer and TP53 codon 72 polymorphism when the analysis was restricted to methodologically sound studies.Funding German Research Foundation (DFG).

Objective: Head and neck squamous cell carcinoma (HNSCC) is a disease attributed to exposure to tobacco and alcohol. However, many epidemiological and molecular data suggest that human papillomavirus (HPV) infection may promote head and neck carcinogenesis. The objective of our study was to evaluate the prevalence of HPV and its various types in fresh HNSCC tumor samples using the polymerase chain reaction (PCR). Methods: 48 patients with primary diagnosed HNSCC carcinoma were investigated. HPV DNA was detected using PCR. HPV-positive samples were additionally tested for HPV16 and 18. Results: Of the 48 patients, 13 (27.1%) had detectable HPV. After HPV typing, HPV16 was found in only 4 cases (30.1%). Conclusions: Our study demonstrated a HPV detection rate of 27.1% in Lithuanian HNSCC patients; 30.1% of them harbored HPV16. There were no differences regarding the other risk factors (smoking, alcohol consumption and occupational history) between HPV-positive and HPV-negative patients with head and neck cancer in our study.

Cervical cancer remains an important cause of women morbidity and mortality. The progression of cervical pathology correlates with the HPV integration into the host genome. However, the data on the viral integration status in cervical dysplasias are controversial. The aim of the current study was to evaluate the status of HPV integration in two types of cervical pathology – invasive and non invasive cervical cancer (e.g. carcinoma in situ). 156 women were included in the study: 66 women were diagnosed with invasive cervical cancer (CC) and 90 with non invasive cervical cancer (carcinoma in situ, CIS). 74.2% [95% PI: 63.64÷84.76] of specimens collected from women with diagnosed CC and 85.6% [95% PI: 85.53÷92.85] of CIS specimens were positive for HPV. The most prevalent HPV genotype in both groups was HPV16. To evaluate HPV integration, three selected HPV16 E2 gene fragments were analyzed by PCR. In the majority of CC and CIS specimens the amplification of all three HPV16 E2 gene fragments was observed. The episomal HPV16 form was detected in the majority of CC and CIS specimens. The deletion of all three HPV16 E2 gene fragments was detected in 9.4% of CC specimens and 2.2% of CIS specimens. Finally, integration status could not be used as diagnostical additional test to distinguish between invasive and non invasive cervical cancer.

Human papillomavirus (HPV) is the main cause of cervical cancer. Therefore, the detection of oncogenic HPV types is important in predicting the risk of cervical cancer. The aim of the current study was to estimate the prevalence of 16 carcinogenic and potentially carcinogenic HPV types in the study group of Lithuanian women with various grades of cervical pathology in comparison to healthy women. A total of 824 cervical specimens were investigated for HPV DNA: 547 specimens of women with abnormal cytology and 277 specimens of healthy women. Cytological diagnosis was confirmed by histology. For the detection of HPV infection, HPV DNA was amplified by PCR using three different primer systems. HPV DNA was detected in 67.6% of specimens collected from women with abnormal cytology and 24.2% of specimens collected from healthy women. The frequency of HPV-positive specimens correlated with the severity of cervical pathology: it ranged from 50.0% in the subgroup of atypical squamous cells to 80.6% in cervical cancer. In cases confirmed by histology the frequency of HPV-positive specimens ranged from 68.6% in the subgroup of cervical intraepithelial neoplasia grade 1 to 89.2% in cervical intraepithelial neoplasia grade 3 or carcinoma in situ. HPV DNA-positive samples were further investigated for the presence of 16 HPV types by multiplex PCR. The most common HPV type was HPV 16 (detected in 42.3% of HPV-positive specimens) followed by HPV 31 (10.1%), HPV 33 (8.2%), and HPV 56 (5.7%). In contrast, the frequency of HPV 18 was lower as compared to other countries.

Human carbonic anhydrase XII (CA XII) is a single-pass transmembrane protein with an extracellular catalytic domain. This enzyme is being recognized as a potential biomarker for different tumours. The current study was aimed to generate monoclonal antibodies (MAbs) neutralizing the enzymatic activity of CA XII. Bioinformatics analysis of CA XII structure revealed surface-exposed sequences located in a proximity of its catalytic centre. Two MAbs against the selected antigenic peptide spanning 167-180 aa sequence of CA XII were generated. The MAbs were reactive with recombinant catalytic domain of CA XII expressed either in E. coli or mammalian cells. Inhibitory activity of the MAbs was demonstrated by a stopped flow CO2 hydration assay. The study provides new data on the surface-exposed linear CA XII epitope that may serve as a target for inhibitory antibodies with a potential immunotherapeutic application.

Study question The 13th European in vitro fertilization (IVF)-monitoring (EIM) report presents the results of treatments involving assisted reproductive technology (ART) initiated in Europe during 2009: are there any changes in the trends compared with previous years? Summary answer Despite some fluctuations in the number of countries reporting data, the overall number of ART cycles has continued to increase year by year and, while pregnancy rates in 2009 remained similar to those reported in 2008, the number of transfers with multiple embryos (3+) and the multiple delivery rates declined. What is known already Since 1997, ART data in Europe have been collected and reported in 12 manuscripts, published in Human Reproduction. Study design, size, duration Retrospective data collection of European ART data by the EIM Consortium for the European Society of Human Reproduction and Embryology (ESHRE); cycles started between 1st January and 31st December are collected on a yearly basis; the data are collected by the National Registers, when existing, or on a voluntary basis. Participants/materials setting, methods From 34 countries (-2 compared with 2008), 1005 clinics reported 537 463 treatment cycles including: IVF (135 621), intracytoplasmic sperm injection (ICSI, 266 084), frozen embryo replacement (FER, 104 153), egg donation (ED, 21 604), in vitro maturation (IVM, 1334), preimplantation genetic diagnosis/screening (PGD/PGS, 4389) and frozen oocyte replacements (FOR, 4278). European data on intrauterine insemination using husband/partner's semen (IUI-H) and donor (IUI-D) semen were reported from 21 and 18 countries, respectively. A total of 162 843 IUI-H (+12.7%) and 29 235 IUI-D (+17.3%) cycles were included. Data available from each country are presented in the tables; total values (as numbers and percentages) refer to those countries where all data have been reported. Main results and the role of chance In 21 countries where all clinics reported to the ART register, a total of 399 020 ART cycles were performed in a population of 373.8 million, corresponding to 1067 cycles per million inhabitants. For IVF, the clinical pregnancy rates per aspiration and per transfer were 28.9 and 32.9%, respectively and for ICSI, the corresponding rates were 28.7 and 32.0%. In FER cycles, the pregnancy rate per thawing was 20.9%; in ED cycles, the pregnancy rate per transfer was 42.3%. The delivery rate after IUI-H was 8.3 and 13.4% after IUI-D. In IVF and ICSI cycles, 1, 2, 3 and 4+ embryos were transferred in 24.2, 57.7, 16.9 and 1.2%, respectively. The proportions of singleton, twin and triplet deliveries after IVF and ICSI (combined) were 79.8, 19.4 and 0.8%, respectively, resulting in a total multiple delivery rate of 20.2%, compared with 21.7% in 2008, 22.3% in 2007, 20.8% in 2006 and 21.8% in 2005. In FER cycles, the multiple delivery rate was 13.0% (12.7% twins and 0.3% triplets). Twin and triplet delivery rates associated with IUI cycles were 10.4/0.7% and 10.3/0.5%, following treatment with husband and donor semen, respectively. Limitations, reasons for caution The method of reporting varies among countries, and registers from a number of countries have been unable to provide some of the relevant data such as initiated cycles and deliveries. As long as data are incomplete and generated through different methods of collection, results should be interpreted with caution. Wider implications of the findings The 13th ESHRE report on ART shows a continuing expansion of the number of treatment cycles in Europe, with more than half a million of cycles reported in 2009. The use of ICSI has reached a plateau. Pregnancy and delivery rates after IVF and ICSI remained relatively stable compared with 2008 and 2007. The number of multiple embryo transfers (3+ embryos) and the multiple delivery rate have shown a clear decline.

Overall, head and neck sqamous cell carcinoma accounts for more than 550,000 cases annually worldwide. It is well known that human papillomavirus (HPV) is the main risk factor for cervical cancer development. As the incidence and the mortality of cervical cancer are closely related to the HPV prevalence, we hypothesized that there is the same association between HPV prevalence and head and neck squamous cell carcinoma. Therefore we performed the study aiming to compare the level of HPV infection and HPV type distribution between two groups of Lithuanian and Belarusian patients with head and neck sqamous cell carcinoma. One hundred ninety head and neck sqamous cell carcinoma patients were included in the study, 75 from Lithuania and 115 from Belarus. PCR was used for HPV detection and typing. The distribution of HPV infection among head and neck sqamous cell carcinoma patients was similar in the Lithuanian (20.0%) and Belarusian (18.3%) patient groups, however differences were found in the distribution of HPV types.

Cervical cancer is one of the most common cancers in Lithuania. It is the fourth cancer in women according to the morbidity. The incidence rate of cervical cancer has been increasing in Lithuania since 1992. One of the important risk factor for cervical cancer is human papillomavirus (HPV) infection. The aim of this study is to compare the prevalence of human papillomavirus among cervical cancer patients and women with cervical intraepithelial lesions and healthy women, to determine its types and HPV 16 variants. Polymerase chain reaction and DNA sequencing were used for analysis of specimens taken from cervix uteri. Human papillomavirus was detected in 92.7% of cervical cancer patients and in 26.7% of control women (p0.0001). It is important to notice that prevalence of HPV in women with high-grade squamous intraepithelial lesions was higher than in the control group. HPV 16 (55.8% in squamous cell carcinoma patients and 35.3% in cervical adenocarcinoma patients) and its European HPV 16 L83V variant were detected more frequently in cervical cancer patients. Studies on HPV, prevalence of its types and variants are very important for HPV based preventive strategies, especially for the vaccination.

Cervical cancer is the fourth most common cancer for women in Lithuania. One of the important cervical cancer risk factors is human papillomavirus (HPV) infection. Recent literature has considered p53 allelic polymorphism to be a putative predisposing factor for cervical carcinoma development.The aim of this study was to elucidate the prevalence of HPV, especially HPV 16, in cervical cancer patients and in healthy women, to investigate the distribution of p53 gene 72 codon polymorphism and to correlate these to cervical cancer risk in Lithuanian women.588 women were included in the study: 212 women with primary diagnosed cervical cancer (case group) and 376 healthy volunteers (control group).A high prevalence of HPV DNA was detected in cervical cancer patients, 92.0%, and in control women, 23.6% (P0.0001). HPV 16 is the most frequent HPV type in cervical cancer patients. In the case of squamous cell carcinoma, this type was detected in 55.8%, in adenocarcinoma - 35.3%. In the control group, this type was detected in 19.0%. A statistically significant difference in the distribution of p53 alleles between the case and the control groups was found.Cervical cancer risk in Lithuanian patients is associated with HPV infection (OR = 75.39; 95% CI 33.61-192.98), especially HPV 16 type (OR = 100.3; 95% CI 46.05-238.59) and p53 homozygous Arg/Arg allele (OR = 2.10; 95% CI 1.10-4.19).

Background. Since the implementation of the cervical cancer screening program in Lithuania in 2004, cervical cancer incidence rates have stabilized during a 4-year period: in 2006 and 2007, 508 and 485 new cases, respectively, were diagnosed. Human papillomavirus (HPV) infection is one of the main risk factors for cervical cancer and development of intraepithelial lesions. However, not only HPV, but also HPV type, is a very important factor for malignant transformation. Cervical intraepithelial lesions with HPV 16 and 18 more frequently progress to cancer. To date, in Lithuania, studies only on HPV prevalence and risk factors have been carried out, and less attention has been paid to the identification of HPV types. The aim of this study was to identify the most common HPV types in women with various cytological lesions. Material and methods. A total of 246 women with various cytological lesions (atypical squamous cells of undetermined significance [ASCUS], low-grade squamous intraepithelial lesion [LSIL], and high-grade squamous intraepithelial lesion [HSIL]) were included into the study. All the women were screened for HPV infections followed by HPV typing for types 6, 11, 16, 18, 31, 33, 45, and 59. Polymerase chain reaction was used. Results. Less than half (45.5%) of women with cytological lesions were infected with HPV. The highest prevalence of HPV was detected in women with HSILs (62.1%) and CIN2 (86.7%). HPV typing revealed that the most frequent type was HPV 16 (64.3%), HPV 18 and HPV 33 accounted for 5.4% and 4.5% of cases, respectively. Based on cytologic diagnosis, HPV 16 was more frequently found in women with HSILs than women with ASCUS (77.8% vs. 50.0%). Conclusions. The prevalence of HPV infection in women with cytological lesions was 45.5%. The highest prevalence of HPV was detected in women with HSILs (cytologic investigation) and CIN2 (histologic investigation). HPV 16 is the most common type in women with various cervical intraepithelial lesions.