PTEN mutations as predictive marker for the high-grade endometrial cancer development in slovak women.

Endometrial carcinoma (ECa) is one of the most common neoplasia of the female genital tract. The phosphatase and tensin (PTEN) homolog is the most frequently mutated tumor suppressor gene in endometrial carcinoma. PTEN encodes a phosphatase, a key regulatory enzyme involved in a signal transduction pathway that regulates cell growth, migration and apoptosis. The study evaluates an association between the morphological appearance of endometrial hyperplasia and ECa, and the presence of PTEN variations, PTEN protein´s level and intracellular localization. A total of 67 archived formalin-fixed and paraffin-embedded human biopsy tissue specimens with normal proliferative and secretory endometrium, endometrial hyperplasia without atypia and endometrial atypical hyperplasia, endometrioid the grade G1 and G3 and serous subtype of ECa were evaluated by sequencing for the presence of mutations in coding regions of PTEN gene of endometrial epithelial cells. The PTEN gene expression and intercellular localization of PTEN protein were evaluated immunohistochemically by immunoreactive score (IRS). PTEN mutation spectrum in endometrial carcinoma was identified for Slovak population. 28 non-silent mutations were identified in PTEN, twelve of them were novel, not annotated in Catalogue of Somatic Mutations in Cancer. Higher frequency of PTEN mutations was observed in serous carcinoma compared to global average. No correlation was observed between samples´ IRS, PTEN cellular localization and identified mutations. PTEN sequencing can be beneficial for patients considering prognosis of disease and sensitivity to treatment.

Blood testosterone in middle aged males heavily exposed to endocrine disruptors is decreasing more with HCB and p,p'-DDE related to BMI and lipids, but not with Σ15PCBs.

OBJECTIVE: This work was aimed to evaluate the fundamental relations between the blood levels of testosterone (TEST) and persistent organochlorinated pollutants (POPs) related to body mass index (BMI) and blood lipids in a cohort of heavily exposed males from Eastern Slovakia. METHODS: In 429 middle aged (41-55 years) males heavily exposed to POPs the levels of 15 polychlorinated biphenyl congeners (Σ15PCBs), hexachlorobenzene (HCB), and dichlorodiethyl-dichloroethylene (p,p'-DDE) were measured by gas chromatography/mass spectrometry and the total testosterone (TEST) by electrochemiluminiscent immunoassay. RESULTS: After classifying the values of BMI, TEST, HCB, p,p'-DDE, and Σ15PCBs in quintiles and evaluating mutual interrelations of individual quintile counts in pairs of variables with chi-square, statistically significant interrelation was found for BMI/TEST (<0.0001) and HCB/TEST (p<0.001), but not for p,p'-DDE/TEST (p<0.6036) and Σ15PCBs/TEST (p<0.3246). Moreover, highly significant negative correlation was found between HCB and TEST by means of both Pearson (p<0.01) and Spearman rank correlations (p<0.0001). However, similar correlations performed between p,p'-DDE and Σ15PCBs did not reveal statistical significance. Finally, highly significant positive correlations were found between HCB and BMI, age, total lipids, and triglycerides. However, these correlations were less significant for p,p'-DDE and not significant or even negligibly negative for Σ15PCBs. In contrast, correlations of TEST with BMI and lipid fractions were significantly negative. CONCLUSION: It appears that HCB might play a role in a decrease of TEST in males with relatively narrow age range of males highly exposed to POPs. Highly significant positive correlation of HCB with BMI and blood lipids points out the role of BMI as an imaginary compartment closely related to the total body fat mass and representing a depot of POPs which is closely related to the level of POPs and lipids in blood. However, the differences in the affinity of individual POPs to BMI and blood lipids as well as the mechanism of their different relation to blood TEST levels remain to be still explained.

Alpha-fetoprotein, carcinoembryonic antigen and beta2-microglobulin in adult population highly exposed to organochlorinated pollutants (PCB, DDE and HCB).

OBJECTIVE: It is aimed to obtain some general information about the prevalence of certain biomarkers in highly exposed population and on the interrelations between their serum level as related to that of some major organochlorines (OCs). METHODS: The level of alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA) and beta2-microglobulin (beta2-MG) as well as that of polychlorinated biphenyls (Σ15PCBs), dichlorodiphenyl-dichloroethylene (DDE) and hexachlorobenzene (HCB) was estimated in 2046 adults (834 males and 1212 females) from highly polluted Eastern Slovakia. RESULTS: Great majority of blood levels was lower than two specific units used for individual markers, while the prevalence of values higher than two specific units of appropriate markers. At the same time, the prevalence of all markers level higher than 2 specific units was highly significantly increasing with of stratified PCBs level quintiles which were also positively related to these of DDE and HCB. Some significant correlations between biomarkers level and age were also observed. CONCLUSIONS: Although from the data obtained within this multipurpose field survey any notable interrelations between AFP, CEA and beta2-MG and some specific diseases and/or malignant processes could not be retrospectively specified, from the data obtained it appears that some of such interrelations cannot be definitely excluded.

Polychlorinated biphenyls and testosterone: age and congener related correlation approach in heavily exposed males.

OBJECTIVE: It was aimed to evaluate some fundamental correlations of 15 individual PCB congeners and their sum with serum testosterone level in highly and long-term exposed males with special respect to minimize the interfering effect of age. METHODS: A total of 834 males from eastern Slovakia (age range of 21-78 years; median, 75th and 90th percentile of 48, 54 and 58 years, respectively) were examined consisting of 432 males from highly polluted area and 402 males from the area of background pollution. In all of them the serum level of 15 polychlorinated biphenyl congeners (PCBs), hexachlorobenzene (HCB) and dichlorodiethyl-dichloroethylene (DDE) was measured by gas chomatography/mass spectrometry and total testosterone in serum was measured with the aid of electrochemiluminiscent immunoassay. Pearson's correlation coefficients for each individual PCB congener as well as for Sigma15PCBs with testosterone were assessed in the cohort of all 834 males and also in the cohort of 444 males with age range of 41-55 years in which any significant negative influence of age on testosterone level has not been found and thus the interfering effect of aging on that level was apparently minimized. RESULTS: In the cohort of 834 males with high level of Sigma15PCBs (median = 885; range = 211-77,084; 5% - 95% = 377 - 4051 ng/g lipid) and highly significant negative correlation with age (r= 0.303; p<0.000) a significant negative correlation (p<0.05) with testosterone has been observed only for two mono-ortho-congeners (CB-105 and -118). However, in the cohort of 444 males aged 41-55 years any significant correlation for individual PCB congeners and for Sigma15PCBs with testosterone did not appear. CONCLUSION: In a large cohort of highly exposed males with minimized interfering effect of age any significant correlations between 15 PCB congeners analyzed and total testosterone were not found.

Thyrotropin versus thyroid hormone in regulating bone density and turnover in premenopausal women.

OBJECTIVE: This cross-sectional study aimed to evaluate the interrelations between endogenous TSH level on one side and the status of bone mineral density (BMD) and bone metabolic turnover (BMT) on the other in pooled four groups of premenopausal women either without or with a long-term L-thyroxine treatment. METHODS: Serum levels of free thyroxine (FT4), thyrotropin (TSH), calcium (Ca), alkaline phosphatase (ALP), osteocalcin OC) and cross linked N-telopeptide of type 1 collagen (NTx) as well as urinary calcium (U-Ca/24h), bone mineral density of lumbar spine L 1-4 (BMD-L) and femoral hip (BMD-F) were estimated in a cohort of 151 premenopausal women (median 36 years) consisting of four groups: Group 1, 40 healthy untreated women, while three other groups consisted of patients previously treated for about 5 years; Group 2, 41 patients with genuine hypothyroidism treated by L-thyroxine (50-100 microg daily); Group 3, 40 patients with genuine hyperthyroidism treated by Carbimazol (10-15 mg daily); Group 4, 30 patients treated by suppressive doses of L-thyroxine (100-150 microg daily) after thyroidectomy for thyroid cancer (n=10) or because of progressively growing benign goitre (n=20). RESULTS: When using multiple correlation analysis (Pearson's r) in pooled 151 women, TSH showed significant positive correlation with BMD-L (p<0.01) and BMD-F (p<0.001) and, at the same time, significant negative correlation with serum level of BMT markers such as ALP (p<0.05), OC (p<0.05) and NTx (p<0.01), while the correlation of FT4 with BMD-L, BMD-F was significantly negative (p<0.001 for both) and that with all BMT markers was significantly positive (p<0.05 to <0.001). Thus, it appeared that higher TSH level was associated with increased bone mineral density and, at the same, with decreased bone metabolic turnover. These interrelations were further supported by the findings of significantly lower BMD-F (p<0.01), BMD-L (p<0.001) and significantly higher ALP, OC and NTX (all at p<0.001) in the group of 36 women with TSH level<0.3 mU/l as compared to the group of 115 women with TSH level range of 0.35-6.3 mU/l). CONCLUSIONS: Irrespectively of thyroid diagnosis and/or previous long term thyroxine treatment in some groups, this cross sectional study showed that, after the pooled group of 151 women has been redistributed according to the actual TSH level, the bone mineral density and the level of bone turnover markers was significantly more favorable in 115 subjects with TSH level range of 0.35-6.3 mU/l than these in 36 women with TSH<0.3 mU/l.

High prevalence of prediabetes and diabetes in a population exposed to high levels of an organochlorine cocktail.

AIMS/HYPOTHESIS: A heavily polluted area of Eastern Slovakia was targeted by the PCBRISK cross-sectional survey to search for possible links between environmental pollution and both prediabetes and diabetes. METHODS: Associations of serum levels of five persistent organic pollutants (POPs), namely polychlorinated biphenyls (PCBs), 2,2'-bis(4-chlorophenyl)-1,1-dichloroethylene (p,p'-DDE), 2,2'-bis(4-chlorophenyl)-1,1,1-trichloro-ethane (p,p'-DDT), hexachlorobenzene (HCB) and beta-hexachlorocyclohexane (beta-HCH), with prediabetes and diabetes were investigated in 2,047 adults. Diabetes and prediabetes were diagnosed by fasting plasma glucose in all participants and by OGTT in 1,220 compliant participants. RESULTS: Our population was stratified in terms of individual POPs quintiles and associations between environmental pollution, prediabetes and diabetes were investigated. Prevalence of prediabetes and diabetes increased in a dose-dependent manner, with individuals in upper quintiles of individual POPs showing striking increases in prevalence of prediabetes as shown by OR and 95% CI for PCBs (2.74; 1.92-3.90), DDE (1.86; 1.17-2.95), DDT (2.48; 1.77-3.48), HCB (1.86; 1.7-2.95) and beta-HCH (1.97; 1.28-3.04). Interestingly, unlike PCBs, DDT and DDE, increased levels of HCB and beta-HCH seemed not to be associated with increased prevalence of diabetes. Nevertheless, individuals in the 5th quintile of the variable expressing the cumulative effect of all five POPs (sum of orders) had a more than tripled prevalence of prediabetes and more than six times higher prevalence of diabetes when compared with the 1st referent quintile. CONCLUSIONS/INTERPRETATION: Increasing serum concentrations of individual POPs considerably increased prevalence of prediabetes and diabetes in a dose-dependent manner. Interaction of industrial and agricultural pollutants in increasing prevalence of prediabetes or diabetes is likely.

The level of TSH appeared favourable in maintaining bone mineral density in postmenopausal women.

OBJECTIVE: Since the positive role of thyrotropin (TSH) in bone remodeling has been recently emphasized, this cross-section study is aimed to evaluate the association of bone status with the level of TSH and free thyroxine (FT4) in the cohort of postmenopausal women after long-term treatment of thyroid disorders and age matched controls. METHODS: Urinary calcium (dUCa) and serum level of TSH, FT4 and of bone turnover markers (BTMs) such as alkaline phosphatase (ALP), osteocalcin (OC), cross linked N-telopeptide of type 1 collagen (NTx) as well as lumbar spine L 1-4 (BMD-L) and femoral hip (BMD-F) mineral density were determined in 113 postmenopausal women consisting of 42 patients with Graves disease treated by carbimazole, 32 patients with thyroid cancer treated with L-thyroxine and 39 age matched women without any thyroid and osteological disorders. For statistical evaluation t-test, Pearson's correlation coefficient and linear multiple regression were used. RESULTS: To compare the association of TSH versus FT4 with BMD and BTMs the pooled cohort of all 113 women was divided in two groups in terms of TSH level: 1. 34 women with low TSH (>or=0.50 mU/l); 2. 79 women with normal TSH (0.51-4.3 mU/l). In spite of significantly higher FT4 level, the Group 2 with normal TSH level had significantly higher BMD-L and BMD-F (p<0.001) and, in contrast, significantly lower urinary dUCa, ALP, OC (all at p<0.001) and NTx (p<0.01) as compared to the Group 1 with low TSH level. Linear multiple regression showed highly significant influence of TSH on BMD-L and BMD-F0 (p<0.001) independent of age, FT4 and body mass index, while that of FT4 was not significant. The strength of linear interrelation between all variables used was finally tested by Pearson's correlation coefficient (Table 3) which was highly positive for TSH with BMD-F and BMD-L, but highly negative for TSH with serum NTx, OC, ALP) and urinary calcium (dUCa). In contrast, no significant correlation was found between the level of FT4 and BMD. CONCLUSIONS: Irrespectively of FT4 level, postmenopausal women with normal TSH level showed a favorable bone status as compared to these with low level of TSH which is consistent with the view that TSH itself possibly participates in playing a favorable role in influencing the bone mineral density in adult women.

Increased prevalence and coincidence of antinuclear and antithyroid antibodies in the population exposed to high levels of polychlorinated pollutants cocktail.

OBJECTIVES: Because of well known association between the exposure to persistent organochlorinated pollutants (POPs) and impaired immune system, it was attempted to check possible coincidence of nuclear and thyroperoxidase antibodies with the levels of major POPs. METHODS: Antinuclear antibodies. (ANA) were estimated by indirect immunofluorescence test using Hep2- cells and thyroperoxidase antibodies (TPOab) by electrochemiluminiscent immunoassay in the cohort of 253 adults (82 males and 171 females) aged 21-75 years, among them 144 (46 males and 98 females) from the area polluted (POLL) by polychlorinated biphenyls (PCB) and 109 (36 males and 73 females) from the area of background pollutrion (BCGR). In the same cohort fifteen congeners of PCB and also total DDE (2,2'-bis(4-chlorophenyl)-1,1-dichloroethylene) and hexachlorobenzene (HCB) were estimated by high resolution gas chromatography/mass spectrometry. RESULTS: Prevalence of ANA only was significantly higher in POLL than in BCGR in males (p < 0.001) and females (p < 0.001) and the same was true for the prevalence of TPOab in males (p < 0.05) and females (p < 0.01) from POLL. In addition, also the prevalence of coincident ANA+TPOab in males (p < 0.001) and females (p < 0.05) was significantly higher in POLL. In a total of 253 pooled males and females from both areas and stratified in terms of PCB level quintiles. The prevalence of ANA in the 4th and 5th quintile of each among three pollutants (PCB, DDE and HCB) was significantly higher (p < 0.01 or < 0.001) and showed the parallel increase with the level of all pollutants. CONCLUSIONS: Significantly increased prevalence of ANA either only or in coincidence with TPOab was found related to increasing level of PCB, DDE and HCB.