Changes in therapy and survival of metastatic renal cell carcinoma in Estonia.

BACKGROUND: Before the era of targeted therapies, cytokines were the main therapy for metastatic renal cell carcinoma (mRCC). Our aim was to analyze the changes in treatments and overall survival (OS) of all mRCC patients in Estonia in relation to the introduction of new medications. METHODS: All patients with mRCC who started medical therapy in Estonia during the years 2004-2012 were identified using the database of the Estonian Health Insurance Fund. Tumor and treatment data were gathered from medical records. Vital status data were obtained from the Estonian Population Registry. The only available therapy before 2008 was interferon alpha-2A (INFa2A), targeted agents added from 2008. For survival analysis, patients were divided into 2 groups: INFa therapy only (group 1) and INFa followed by targeted agents or targeted agents therapy only (group 2). RESULTS: Out of 416 identified patients, 380 were eligible for analysis. The most common 1st-line treatments were INFa (55%), sunitinib (32%) and INFa+bevacizumab (13%). 28% of patients received 2nd-line therapies and 15% 3rd-line treatments. Median survival of all patients was 13.7 months [95% confidence interval (CI) 11.3-16.2]; 7.6 months (CI 6.4-8.6) for group 1 and 19.8 months (CI 15.6-22.9) for group 2. In multivariate analysis, group 1 had nearly four times higher risk of dying than group 2 [hazard ration (HR) 3.88, 95% CI 2.64-5.72]. CONCLUSIONS: The implementation of targeted therapies significantly changed the outcomes of mRCC in Estonia: it prolonged median survival, reduced the risk of death and also enlarged the proportion of patients who received medical therapy.

Polygenic prediction of breast cancer: comparison of genetic predictors and implications for risk stratification.

BACKGROUND: Published genetic risk scores for breast cancer (BC) so far have been based on a relatively small number of markers and are not necessarily using the full potential of large-scale Genome-Wide Association Studies. This study aimed to identify an efficient polygenic predictor for BC based on best available evidence and to assess its potential for personalized risk prediction and screening strategies. METHODS: Four different genetic risk scores (two already published and two newly developed) and their combinations (metaGRS) were compared in the subsets of two population-based biobank cohorts: the UK Biobank (UKBB, 3157 BC cases, 43,827 controls) and Estonian Biobank (EstBB, 317 prevalent and 308 incident BC cases in 32,557 women). In addition, correlations between different genetic risk scores and their associations with BC risk factors were studied in both cohorts. RESULTS: The metaGRS that combines two genetic risk scores (metaGRS2 - based on 75 and 898 Single Nucleotide Polymorphisms, respectively) had the strongest association with prevalent BC status in both cohorts. One standard deviation difference in the metaGRS2 corresponded to an Odds Ratio = 1.6 (95% CI 1.54 to 1.66, p = 9.7*10- 135) in the UK Biobank and accounting for family history marginally attenuated the effect (Odds Ratio = 1.58, 95% CI 1.53 to 1.64, p = 7.8*10- 129). In the EstBB cohort, the hazard ratio of incident BC for the women in the top 5% of the metaGRS2 compared to women in the lowest 50% was 4.2 (95% CI 2.8 to 6.2, p = 8.1*10- 13). The different GRSs were only moderately correlated with each other and were associated with different known predictors of BC. The classification of genetic risk for the same individual varied considerably depending on the chosen GRS. CONCLUSIONS: We have shown that metaGRS2, that combined on the effects of more than 900 SNPs, provided best predictive ability for breast cancer in two different population-based cohorts. The strength of the effect of metaGRS2 indicates that the GRS could potentially be used to develop more efficient strategies for breast cancer screening for genotyped women.

Increasing kidney cancer incidence and survival in Estonia: role of age and stage.

BACKGROUND: Kidney cancer rates in Estonia are high. The study aimed to examine long-term trends in kidney cancer incidence, mortality and survival in Estonia, with special focus on age, birth cohorts, morphology and TNM stage. MATERIAL AND METHODS: Estonian Cancer Registry provided data on all incident cases of kidney cancer (ICD-10 C64), diagnosed in adults (age ≥15 years) in Estonia during 1995 - 2014. Relative survival ratios (RSR) were calculated and excess hazard ratios of dying were estimated with gender, age, period of diagnosis and TNM stage as independent variables. Joinpoint regression modeling was used to calculate estimated annual percentage change for incidence (1970-2014) and mortality (1995-2016) trends. Age-specific incidence rates were presented by birth cohort and period of diagnosis. RESULTS: Incidence increased significantly in both sexes, with the steepest rise seen for localized cancer. Cohort effects were pronounced particularly in men, while period effects were seen from the mid-1980s to mid-1990s in both sexes. Age-standardized five-year RSR for total kidney cancer increased by 13 percentage units (from 53% to 65%) over the study period; the increase was larger for renal cell carcinoma (from 63% to 78%). Survival increases of about five percentage units were seen for stages I/II and III. Age and gender were not associated with excess risk of dying from renal cell carcinoma after adjusting for stage. CONCLUSION: Estonia is currently among countries with the highest incidence of kidney cancer. The results suggest a combined effect of changing risk profiles in successive birth cohorts and increasing diagnostic activity around 1990. Large survival increase can mostly be attributed to earlier detection, but improved diagnosis and treatment have probably influenced stage-specific survival. High proportion of tumors with unspecified morphology and those with unknown stage among the elderly warrants further investigation of diagnostic and treatment practices.

A dual colour FISH method for routine validation of sexed Bos taurus semen.

BACKGROUND: Usage of sexed semen that allows to choose the gender of the calves, is commonly practiced in livestock industry as a profitable breeding alternative, especially in dairy farming. The flow cytometric cell sorting is the only commercially available method for bovine sperm sexing. For validation of the sexing procedure several methods have been developed including sperm fluorescence in situ hybridisation techniques. Latter usually include the use of pre-labelled nucleotides for probe synthesis which is relatively expensive approach compared to combined application of aminoallyl-dUTP and chemical binding of fluorescent dyes. Here a sex determining dual colour bovine sperm fluorescence in situ hybridisation method is presented which is considered more cost-effective technique than the previously reported approaches. RESULTS: The reliability of sex chromosome identifying probes, designed in silico, was proven on bovine metaphase plate chromosomes and through comparison with a commercially available standard method. In the dual colour FISH experiments of unsexed and sexed bovine sperm samples the hybridisation efficiency was at least 98%, whereas the determined sex ratios were not statistically different from the expected. Very few cells carried both of the sex chromosome-specific signals (less than 0.2%). CONCLUSIONS: A protocol for a dual colour bovine sperm FISH method is provided which is cost-effective, simple and fast for sex determination of spermatozoa in bull semen samples.

A case report and follow-up of the first live birth after heterotopic transplantation of cryopreserved ovarian tissue in Eastern Europe.

BACKGROUND: Ovarian insufficiency is a major concern for long-term cancer survivors. Although semen freezing is well established to preserve male fertility, the possibilities to secure post-cancer female fertility are mostly limited to oocyte or embryo freezing. These methods require time-consuming ovarian stimulation with or without in vitro fertilization (IVF) that evidently delays cancer therapy. Ovarian tissue cryopreservation and subsequent thawed tissue autotransplantation are considered the most promising alternative strategy for restoring the fertility of oncology patients, which has not yet received the full clinical acceptance. Therefore, all successful cases are needed to prove its reliability and safety. CASE PRESENTATION: Here we report a single case in Estonia, where a 28-year-old woman with malignant breast neoplasm had ovarian cortex cryopreserved before commencing gonadotoxic chemo- and radiotherapy. Two years after cancer therapy, the patient underwent heterotopic ovarian tissue transplantation into the lateral pelvic wall. The folliculogenesis was stimulated in the transplanted tissue by exogenous follicle-stimulating hormone and oocytes were collected under ultrasound guidance for IVF and embryo transfer. The healthy boy was born after full-term gestation in 2014, first in Eastern Europe. CONCLUSION: Despite many countries have reported the first implementation of the ovarian tissue freezing and transplantation protocols, the data is still limited on the effectiveness of heterotopic ovarian transplant techniques. Thus, all case reports of heterotopic ovarian tissue transplantation and long-term follow-ups to describe the children's health are valuable source of clinical experience.

Trends in incidence and survival of cutaneous malignant melanoma in Estonia: a population-based study.

BACKGROUND: Previous studies have shown an increase in the incidence of cutaneous melanoma (CM) in Estonia, but also poor survival in international comparisons, with a significant survival gap between the sexes. The aim of this study was to analyze the time trends in CM incidence and relative survival by age, TNM stage and anatomical subsite among men and women in Estonia. MATERIALS AND METHODS: Data from the Estonian Cancer Registry were used to calculate age-standardized (World) and age-specific incidence of CM in 1995-2013, and five-year relative survival ratios (RSR) for cases diagnosed in 1995-2012 and followed through in 2014. Period hybrid analysis was used to calculate the most recent survival estimates for 2010-2014. RESULTS: Between 1995 and 2013, the age-standardized incidence of CM increased significantly in Estonia among both sexes, at a rate of around 4% per year. Among women, the proportion of trunk melanomas increased from 26% in 1995-1999 to 39% in 2010-2012 and became the most common site. The proportion of stage I cases and T1 tumors increased considerably. Women had more favorable stage distribution and thinner tumors than men. The age-adjusted five-year RSR increased significantly, from 64% in 1995-1999 to 81% in 2010-2014. The latest age-adjusted RSRs were 76% among men and 84% among women. Survival gains were the largest in patients below 50 years, those with head and neck or trunk melanomas, and stage III cancer. CONCLUSIONS: The proportion of stage I and T1 cases is lower in Estonia compared with the Scandinavian data and is likely a major contributor to the persisting overall survival deficit in Estonia. The apparent deficit in stage II survival also warrants further investigation. A public health program is necessary in Estonia to raise awareness of CM and to significantly increase early stage diagnosis.

Mammography use and mode of detection among breast cancer patients in Estonia.

The aim of this study was to examine past mammography use and mode of detection among breast cancer (BC) patients in Estonia, a country that has low screening coverage and high BC mortality. Women newly diagnosed with primary BC in Estonia in 2008-2010 were interviewed. Determinants of past mammography use and the detection of BC by mammography were studied using multivariate logistic regression. Among 977 participants, almost half reported no mammograms prior to the detection of BC. Overall, 22% of the cases were detected by mammography (16% by screening mammography). Detection by mammography was strongly related to age, past mammography use, and obesity. Among cases detected by mammography, 10% were stage III/IV at diagnosis (32% among cases detected by other modes). This study showed low mammography utilization and high rate of self-detection of BC in Estonia. Increased detection by mammography would help diagnose the disease at an earlier stage and consequently avoid premature BC deaths. Efforts should be undertaken to increase participation in screening and improve the availability of mammography among older and high-risk women. The results are likely to be relevant for other countries and population groups with low screening coverage.

Bovine sperm plasma membrane proteomics through biotinylation and subcellular enrichment.

A significant proportion of mammalian fertilization is mediated through the proteomic composition of the sperm surface. These protein constituents can present as biomarkers to control and regulate breeding of agricultural animals. Previous studies have addressed the bovine sperm cell apical plasma membrane (PM) proteome with nitrogen cavitation enrichment. Alternative workflows would enable to expand the compositional data more globally around the entire sperm's surface. We used a cell surface biotin-labeling in combination with differential centrifugation to enrich sperm surface proteins. Using nano-LC MS/MS, 338 proteins were confidently identified in the PM-enriched proteome. Functional categories of sperm-egg interaction, protein turnover, metabolism as well as molecular transport, spermatogenesis, and signal transduction were represented by proteins with high quantitative signal in our study. A highly significant degree of enrichment was found for transmembrane and PM-targeted proteins. Among them, we also report proteins previously not described on bovine sperm (CPQ, CD58, CKLF, CPVL, GLB1L3, and LPCAT2B) of which CPQ and CPVL cell surface localization was further validated. A descriptive overview of the bovine sperm PM integral and peripheral proteins is provided to complement future studies on animal reproduction and its relation to sperm cell surface. All MS data have been deposited in the ProteomeXchange with identifier PXD001096 (http://proteomecentral.proteomexchange.org/dataset/PXD001096).

Sex differences in cancer survival in Estonia: a population-based study.

BACKGROUND: In Estonia, women have much longer life expectancy than men. The aim of this study was to examine sex differences in cancer survival in Estonia and to explore the role of age at diagnosis, stage at diagnosis and tumour subsite. METHODS: Using data from the population-based Estonian Cancer Registry, we examined the relative survival of adult patients diagnosed with nine common cancers in Estonia in 1995-2006 and followed up through 2011. Excess hazard ratios (EHR) of death associated with female gender adjusted for age, stage at diagnosis and tumour subsite were estimated. RESULTS: A total of 20 828 male and 13 166 female cases were analysed. The main data quality indicators were similar between men and women. Women had more cases with unknown extent of disease at diagnosis. Overall, the age-adjusted 5-year relative survival ratio was higher among women than men for all studied sites, but the difference was significant for cancers of mouth and pharynx (22% units), lung (5% units), skin melanoma (17% units) and kidney (8% units). The increase in survival over time was larger for women than men for cancers of mouth and pharynx, colon, rectum, kidney and skin melanoma. In multivariate analysis, women had a significantly lower EHR of death within five years after diagnosis for five of the nine cancers studied (cancers of mouth and pharynx, stomach, lung, skin melanoma and kidney). Adjustment for stage and subsite explained some, but not all of the women's advantage. CONCLUSIONS: We found a significant female survival advantage in Estonia for cancers of mouth and pharynx, stomach, lung, kidney and skin melanoma. The differences in favour of women tended to increase over time as from the 1990s to the 2000s, survival improved more among women than among men. A large part of the women's advantage is likely attributable to biological factors, but other factors, such as co-morbidities, treatment compliance or health behaviour, are also probable contributors to gender survival disparities in Estonia and merit further investigation. Our findings have implications for public health, early detection and cancer care in Estonia.

Time trends in population-based breast cancer survival in Estonia: analysis by age and stage.

BACKGROUND: Survival from breast cancer (BC) in Estonia has been consistently among the lowest in Europe. The aim of this study was to examine most recent trends in BC survival in Estonia by age and stage. The trends in overall BC incidence and mortality are also shown in the paper. MATERIAL AND METHODS: Estonian Cancer Registry data on all cases of BC, diagnosed in women in Estonia during 1995-2007 (n = 7424) and followed up for vital status through 2009, were used to estimate relative survival ratios (RSR). Period hybrid approach was used to obtain the most recent estimates (2005-2009). Stage was classified as localized, local/regional spread or distant. RESULTS: BC incidence continued to rise throughout the study period, but mortality has been in steady decline since 2000. The distribution of patients shifted towards older age and earlier stage at diagnosis. Overall age-standardized five-year RSR increased from 63% in 1995-1999 to 74% in 2005-2009. Younger age groups experienced a more rapid improvement compared to women over 60. Significant survival increase was observed for both localized and locally/regionally spread BC with five-year RSRs reaching 96% and 70% in 2005-2009, respectively; the latest five-year RSR for distant BC was 11%. Survival for T4 tumors was poor and large age difference was seen for locally/regionally spread BC. CONCLUSIONS: Considerable improvement in BC survival was observed over the study period. Women under 60 benefited most from both earlier diagnosis and treatment advances of locally/regionally spread cancers. However, the survival gap with more developed countries persists. Further increase in survival, but also decline in BC mortality in Estonia could be achieved by facilitating early diagnosis in all age groups, but particularly among women over 60. Investigations should continue to clarify the underlying mechanisms of the stage-specific survival deficit in Estonia.

Identifying women at risk for delayed presentation of breast cancer: a cross-sectional study in Estonia.

BACKGROUND: Survival from breast cancer remains lower in Estonia than in most other European countries. More advanced stage and larger tumors that have impact on survival may be a result of delay in seeking help for breast cancer symptoms. The aim of this study was to identify determinants of delayed presentation among breast cancer patients in Estonia. METHODS: The study population included women with primary breast cancer diagnosed in Estonia in 2008-2010. All data were collected using structured personal interviews carried out by trained nurses in the hospital setting. Only patients with self-discovered symptoms were included in this analysis. Patient delay was measured as time elapsing from symptom self-discovery to first medical consultation. The effect of different factors on the likelihood of prolonged delay (>90 days) was evaluated using logistic regression. RESULTS: Among 703 eligible patients, median patient delay was 16 days (interquartile range 5-54 days). Seventeen percent of the patients had their first medical consultation more than three months after self-detection of symptoms. In multivariate analysis, the risk of prolonged delay was significantly associated with age 65 years and over (OR 2.27, 95% CI 1.23-4.20), current smoking (OR 2.09, 95% CI 1.21-3.61), symptoms other than painless breast lump or breast pain (OR 1.84, 95% CI 1.08-3.16), no history of mammograms (OR 1.83, 95% CI 1.13-2.95), having received no information on breast cancer during past year (OR 1.77, 95% CI 1.05-2.99), and previous benign breast problems (OR 1.65, 95% CI 1.01-2.67). Non-significant risk increase was seen with lower education. CONCLUSIONS: This study provides evidence that factors associated with delayed presentation of breast cancer in Eastern Europe are similar to those observed in Western countries. The results suggest that educational messages to general population should aim at increasing awareness of "non-lump" symptoms of breast cancer and encouraging women of all ages to present in a timely manner. Women at risk for delayed presentation such as smokers and women with no history of mammograms could be identified in the primary care setting.

Implementation of Risk-Stratified Breast Cancer Prevention With a Polygenic Risk Score Test in Clinical Practice.

Background: Breast cancer (BC) screening with mammography reduces mortality but considers currently only age as a risk factor. Personalized risk-based screening has been proposed as a more efficient alternative. For that, risk prediction tools are necessary. Genome-wide association studies have identified numerous genetic variants (single-nucleotide polymorphisms [SNPs]) associated with BC. The effects of SNPs are combined into a polygenic risk score (PRS) as a risk prediction tool. Objectives: We aimed to develop a clinical-grade PRS test suitable for BC risk-stratified screening with clinical recommendations and implementation in clinical practice. Design and methods: In the first phase of our study, we gathered previously published PRS models for predicting BC risk from the literature and validated them using the Estonian Biobank and UK Biobank data sets. We selected the best performing model based on prevalent data and independently validated it in both incident data sets. We then conducted absolute risk simulations, developed risk-based recommendations, and implemented the PRS test in clinical practice. In the second phase, we carried out a retrospective analysis of the PRS test's performance results in clinical practice. Results: The best performing PRS included 2803 SNPs. The C-index of the Cox regression model associating BC status with PRS was 0.656 (SE = 0.05) with a hazard ratio of 1.66. The PRS can stratify individuals with more than a 3-fold risk increase. A total of 2637 BC PRS tests have been performed for women between the ages 30 and 83. Results in clinical use overlap well with expected PRS performance with 5.7% of women with more than 2-fold and 1.4% with more than 3-fold higher risk than the population average. Conclusion: The PRS test separates different BC risk levels and is feasible to implement in clinical practice.

A Breast Cancer Polygenic Risk Score Is Feasible for Risk Stratification in the Norwegian Population.

BACKGROUND: Statistical associations of numerous single nucleotide polymorphisms with breast cancer (BC) have been identified in genome-wide association studies (GWAS). Recent evidence suggests that a Polygenic Risk Score (PRS) can be a useful risk stratification instrument for a BC screening strategy, and a PRS test has been developed for clinical use. The performance of the PRS is yet unknown in the Norwegian population. AIM: To evaluate the performance of PRS models for BC in a Norwegian dataset. METHODS: We investigated a sample of 1053 BC cases and 7094 controls from different regions of Norway. PRS values were calculated using four PRS models, and their performance was evaluated by the area under the curve (AUC) and the odds ratio (OR). The effect of the PRS on the age of onset of BC was determined by a Cox regression model, and the lifetime absolute risk of developing BC was calculated using the iCare tool. RESULTS: The best performing PRS model included 3820 SNPs, which yielded an AUC = 0.625 and an OR = 1.567 per one standard deviation increase. The PRS values of the samples correlate with an increased risk of BC, with a hazard ratio of 1.494 per one standard deviation increase (95% confidence interval of 1.406-1.588). The individuals in the highest decile of the PRS have at least twice the risk of developing BC compared to the individuals with a median PRS. The results in this study with Norwegian samples are coherent with the findings in the study conducted using Estonian and UK Biobank samples. CONCLUSION: The previously validated PRS models have a similar observed accuracy in the Norwegian data as in the UK and Estonian populations. A PRS provides a meaningful association with the age of onset of BC and lifetime risk. Therefore, as suggested in Estonia, a PRS may also be integrated into the screening strategy for BC in Norway.

Time-gated luminescence assay using nonmetal probes for determination of protein kinase activity-based disease markers.

A novel nonmetal optical probe ARC-1063 whose long-lifetime luminescence is induced by association with the target protein kinase is used for the measurement of the concentration of catalytic subunit of protein kinase A (PKAc) in complicated biological solutions. High affinity (K(D) = 10 pM toward PKAc) and unique optical properties of the probe enable its application for the measurement of picomolar concentrations of PKAc in the presence of high concentrations of other proteins. The described assay is applicable in the high-throughput format with the instrument setups designed for lanthanide-based time-gated (time-resolved) luminescence methods. The assay is used for demonstration that extracellular PKAc (ECPKA) is present in plasma samples of all healthy persons and cancer patients but great care must be taken for procedures of treatment of blood samples to avoid disruption, damage, or activation of platelets in the course of plasma (or serum) preparation and conservation.

Precise, Genotype-First Breast Cancer Prevention: Experience With Transferring Monogenic Findings From a Population Biobank to the Clinical Setting.

Although hereditary breast cancer screening and management are well accepted and established in clinical settings, these efforts result in the detection of only a fraction of genetic predisposition at the population level. Here, we describe our experience from a national pilot study (2018-2021) in which 180 female participants of Estonian biobank (of >150,000 participants in total) were re-contacted to discuss personalized clinical prevention measures based on their genetic predisposition defined by 11 breast cancer-related genes. Our results show that genetic risk variants are relatively common in the average-risk Estonian population. Seventy-five percent of breast cancer cases in at-risk subjects occurred before the age of 50 years. Only one-third of subjects would have been eligible for clinical screening according to the current criteria. The participants perceived the receipt of genetic risk information as valuable. Fluent cooperation of project teams supported by state-of-art data management, quality control, and secure transfer can enable the integration of research results to everyday medical practice in a highly efficient, timely, and well-accepted manner. The positive experience in this genotype-first breast cancer study confirms the value of using existing basic genomic data from population biobanks for precise prevention.

Identification of Bull Semen Microbiome by 16S Sequencing and Possible Relationships with Fertility.

Reports on the use of 16S sequencing for the identification of bacteria in healthy animals are lacking. Bacterial contamination of bull semen can have a negative effect on the sperm quality. The aims of this study were threefold: to identify bacteria in the semen of healthy bulls using 16S sequencing; to investigate the differences in the bacterial community between individual bulls; and to establish if there was a relationship between the bacteria isolated and bull fertility. Semen from 18 bulls of known fertility was used for the DNA extraction and 16S sequencing; 107 bacterial genera were identified. The differences in the amplicon sequence variants (ASVs) and the numbers of genera between bulls were noted. Negative correlations (p < 0.05) between several bacterial genera with Curvibacter, Rikenellaceae RC9-gut-group and Dyella spp. were seen. Other negatively correlated bacteria were Cutibacterium, Ruminococcaceae UCG-005, Ruminococcaceae UCG-010 and Staphylococcus, all within the top 20 genera. Two genera, W5053 and Lawsonella, were enriched in bulls of low fertility; this is the first time that these bacteria have been reported in bull semen samples. The majority of the bacteria were environmental organisms or were species originating from the mucous membranes of animals and humans. The results of this study indicate that differences in the seminal microbiota of healthy bulls occur and might be correlated with fertility.

Genotype-first approach to the detection of hereditary breast and ovarian cancer risk, and effects of risk disclosure to biobank participants.

Genotype-first approach allows to systematically identify carriers of pathogenic variants in BRCA1/2 genes conferring a high risk of familial breast and ovarian cancer. Participants of the Estonian biobank have expressed support for the disclosure of clinically significant findings. With an Estonian biobank cohort, we applied a genotype-first approach, contacted carriers, and offered return of results with genetic counseling. We evaluated participants' responses to and the clinical utility of the reporting of actionable genetic findings. Twenty-two of 40 contacted carriers of 17 pathogenic BRCA1/2 variants responded and chose to receive results. Eight of these 22 participants qualified for high-risk assessment based on National Comprehensive Cancer Network criteria. Twenty of 21 counseled participants appreciated being contacted. Relatives of 10 participants underwent cascade screening. Five of 16 eligible female BRCA1/2 variant carriers chose to undergo risk-reducing surgery, and 10 adhered to surveillance recommendations over the 30-month follow-up period. We recommend the return of results to population-based biobank participants; this approach could be viewed as a model for population-wide genetic testing. The genotype-first approach permits the identification of individuals at high risk who would not be identified by application of an approach based on personal and family histories only.

An anaplastic lymphoma kinase (ALK) fusion oncogene positive metastatic sarcomatoid carcinoma of the lung with good response to crizotinib.

Primary sarcomatoid carcinoma (SC) of the lung is a rare tumor that accounts for less than 1% of all lung cancers and compared to other non-small cell lung cancers (NSCLC) they appear more aggressive with poorer prognosis and response to treatment. Carcinosarcoma is one of the subtypes of SC. We report a case of carcinosarcoma with ALK-EML4 fusion gene in a 50-year-old male patient with a good response to therapy with crizotinib. An ALK rearrangement is a rare finding in SC, but as this case demonstrates, it may occur and it is necessary to perform the ALK testing in these tumors to find possible targeted treatments for better outcomes.

Changes in the natural history of renal cancer: comparison of Estonian data from the periods 1986-1988 and 1996-1998.

To explain considerable increase in survival of renal cancer patients in Estonia during last decades, we compared the stage distribution, diagnostic and treatment methods for the patient groups diagnosed in the periods 1986-1988 and 1996-1998. A significant difference in stage distribution was detected with an increase for stages I-II and a decrease for stages III-IV in 1996-1998. There was a shift in primary diagnostic methods from intravenous urography and angiography to ultrasonography and computed tomography. In multivariate analysis the independent prognostic factors for overall survival were age, stage, and operation status. In conclusion, the increase in the survival of renal cancer patients has been due to the larger number of cases with the earlier stage, which is associated with the application of ultrasonography and computed tomography. Another factor for better survival was the higher operation rate among patients with stage IV disease, a possible factor was the change in operation techniques.

The effects of ovarian function on estrus synchronization with PGF in dairy cows.

Milk progesterone concentration (P4), milk yield, milk composition, ovarian structures and pregnancy status were studied in 108 cows treated with two doses of PGF 14 days apart and inseminated at fixed time (TAI) 80-82 h later. The synchronization protocol was started at 70+/-1.4 days after parturition. Milk P4 profiles revealed that anestrus, failure of luteolysis following treatment with PGF and failure to ovulate following luteolysis were the main reasons for low pregnancy rate with TAI. Anestrous cows had a higher percentage of milk fat (P<0.05) and higher fat to protein ratio (P<0.01), and cows that did not undergo luteolysis had higher milk yield (P<0.05) and lower percentage of milk protein (P<0.05) than cows that responded to PGF treatment. Cows that did not undergo luteolysis and cows that did not ovulate following luteolysis had lower milk P4 during the luteal phase preceding the second PGF injection (P<0.01 and P<0.05, respectively). Pregnancy rates 24 and 47 days after TAI in cows that responded as expected to the synchronization treatment were 62% and 54%, respectively. Pregnancy was precluded in non-responsive cows. The largest follicle at the time of TAI in cows experiencing late embryonic mortality was smaller (P=0.02) than in cows that successfully maintained pregnancy. Results suggest that a primary reason for low pregnancy rate in dairy cows after administration of PGF and TAI is inappropriate ovarian function prior to, or following treatment.