A role of thymidine phosphorylase and P53 tissue protein expression in biology of endometrial cancer.

Tumor suppressor gene p53, the most commonly mutated gene in human cancers has been shown to play an important role in the biology of gynaecological carcinomas. Thymidine phosphorylase is one of the most important angiogenic factors, which is connected with tumor expansion and invasiveness. The aim of the study was an evaluation of thymidine phosphorylase and p-53 tissue protein expression in human endometrial cancer cells by immunohistochemistry and comparison obtained data with clinicopathological factors as FIGO stage of disease and histopathologic grade. Endometrial cancer specimens were obtained from 55 postmenopausal patients (aged 52 to 74 years) operated by total abdominal hysterectomy with bilateral salpingo-oophorectomy. None of patients received preoperative pelvic irradiation. Histopathological typing and grading of the endometrial tumors (G-1, G-2, G-3) as well as myometrial invasion (<1/2, >1/2) were assessed using standard criteria, on hematoxylin-eosin sections. FIGO clinical stage of disease was determined. at the surgery. Thymidine phosphorylase overexpression was observed in 23 (41,8 %) cases. Although we found no statistically significant differences in TP expression between histopathologic grades, particular FIGO stages showed a significant trend of increase TP tumor overexpression. P53 protein overexpression was observed in tumor tissue in 21 cases (35,2%). It tended to be more frequent in cases of advanced disease as well as in low differentiated tumors. Although we found no statistically significant differences in p53 gene expression between groups of FIGO stage and histopathologic grade, we obtained a significant trend of increasing the P53 positive rate with both FIGO and tumour differentiation grading Joint assessment of thymidine phosphorylase and tumor suppressor genes expression may be of additional value in determining the biology of endometrial carcinoma. Key words: endometrial cancer, thymidine phosphorylase, p-53.

Evaluation of tumor angiogenesis and thymidine phosphorylase tissue expression in patients with endometrial cancer.

The formation of new blood vessels in endometrial cancer tissue is a main process, which leads to tumor progression, and is connected with tumor expansion and invasiveness. The aim of the study was evaluation of thymidine phosphorylase protein (TP) expression in human endometrial cancer cells by immunohistochemistry and comparison obtained data with intensity of angiogenesis process and clinicopathological factors as FIGO stage of disease and histopathologic grade. Endometrial cancer specimens were obtained from 55 postmenopausal patients (aged 52 to 74 years) underwent total abdominal hysterectomy with bilateral salpingo-oophorectomy. None of patients received preoperative pelvic irradiation. Histopathological typing and grading of the endometrial tumors (G-1, G-2, G-3) as well as myometrial invasion (<1/2, >1/2) were assessed using standard criteria, on hematoxylin-eosin sections. At the surgery, FIGO clinical stage of disease was determined. Thymidine phosphorylase overexpression was observed in 23 of 55 (41.8%) cases of endometrial cancer. Although we found no statistically significant differences in TP expression between histopathologic grades, particular FIGO stages showed a significant trend of increase TP tumor overexpression. Thymidine phosphorylase overexpression cases demonstrate higher intensity of angiogenesis in comparison to negative samples and results are statistically significant for t-test (p<0.0001). The most intensive new blood vessel formation was observed in G-2 of tumor differentiation grade (p=0.013 for ANOVA test) Mean angiogenic points density (APD) values in cases of G-1 histopathologic grade reached 135.7; values of G-2 and G-3 grades reached 213.8 and 162.8, respectively. Mean intensity of angiogenesis in the first FIGO stage of disease reached 160.0 APD, in stage II 205.6 APD, and in the third 286.9, respectively. Angiogenesis was more intensive in cases of advanced tumors - analysis of variance (ANOVA) confirmed statistically significant differences in APD values between FIGO stage groups (p=0.0007). In conclusion, thymidine phosphorylase expression correlates with increased microvessel density in endometrial cancer. The intensity of angiogenesis process increases according to FIGO stage of disease, which is connected with progressing of cancer disease. Thymidine phosphorylase can play an important role in endometrial cancer progression and could offer additional information about advance of disease.

Effect of calcium and calmodulin antagonists on contractile responses of the human uterine artery.

1. The dependence on extracellular calcium of contractile responses of intramyometrial arteries (0.5-2 mm diameter), as well as the effects of various types of calcium antagonists on these responses, were studied. Contractions were induced by K-depolarization (K) and noradrenaline (NA). 2. Whereas the K response was completely abolished in a calcium-free medium containing 2 mM LaCl3, the NA response was substantially maintained. 3. Nimodipine strongly inhibited the K response but had a relatively weak effect on the NA response; the IC50 values for the K and NA responses being 2 nM and 6 microM, respectively. Corresponding values for verapamil were about 0.7 and 10 microM. 4. Calmodulin antagonists, particularly trifluoperazine and flunarizine, caused a greater inhibition of the NA than of the K response. 5. These results indicate that besides the extracellular calcium which appears to be the major source of activator calcium, there is an intracellular pool of calcium which can be utilized to activate, albeit to a limited extent, drug-induced contractile responses.

Corticotrophin-releasing hormone and ACTH levels in maternal and fetal blood during spontaneous and oxytocin-induced labour.

The changes in corticotrophin-releasing hormone (CRH), ACTH and dehydroepiandrosterone (DHEA) in maternal and fetal plasma were estimated in women undergoing spontaneous and oxytocin-induced labour to correlate hormone changes with the mode of parturition. Blood was sampled from a maternal peripheral vein 2 days before labour, during the second stage of labour and on the second postnatal day, and also from umbilical vessels just after delivery. Hormone concentrations were measured by RIA and ELSA methods. The maternal plasma CRH concentration before labour was significantly higher in the group of women delivered spontaneously and declined during the labour through to the second postnatal day. Measured in umbilical vessels, CRH as well as ACTH concentrations were higher in the umbilical vein than artery. The mean maternal plasma ACTH was similar in both groups before delivery, then increased significantly in both groups during the labour, decreasing on the second day after delivery. There were no changes in DHEA concentrations among the groups and at all time points of collection. No correlations between CRH and ACTH or DHEA were observed. Our results suggest that the maternal pituitary can respond to stress factors during delivery but peripheral CRH, probably mainly of placental origin, is not a major modulator of pituitary action.

Treatment of cancer patients with a low-density-lipoprotein delivery vehicle containing a cytotoxic drug.

A cytotoxic drug (vincristine, VC) was incorporated into low-density lipoprotein (LDL) and given to cancer patients for the first time by repeated intravenous injection. Individuals presenting with ovarian or endometrial cancer received four or five weekly doses of 1.4 mg/m2 LDL/VC. The uptake of LDL/VC by the adrenal cortex and the liver was minimised by concurrent administration of prednisolone and chenodeoxycholic acid. No febrile, allergic or other reaction attributable to the LDL occurred, and no side effect on haemopoietic, adrenal or liver functions was observed. The neurotoxic side effects commonly seen during VC therapy appeared to be reduced. These results suggest that directed cytotoxic therapy might be achieved in humans through the use of LDL as a carrier. Thus, dose-range and comparative studies using LDL/VC vs VCSO4 are warranted in malignancies in which treatment with the latter drug has been established.

Effect of ovarian steroids and diethylstilbestrol on the contractile responses of the human myometrium and intramyometrial arteries.

Estriol, estradiol, progesterone and diethylstilbestrol in the concentration range of 0.2-40 microM inhibited the spontaneous contractions of the myometrium in a dose-dependent manner but the differences in IC50 values obtained with different hormones were not statistically significant. All these hormones caused a concentration-dependent inhibition of the K(+)-induced contraction. The IC50 values were lowest for diethylstilbestrol and highest for estriol. Vasopressin at concentrations of 1.5 x 10(-6) - 1.8 x 10(-3) U/ml stimulated myometrial contractions. These responses were also inhibited by ovarian steroids and diethylstilbestrol. The IC50 values for estriol and progesterone were significantly higher than for estradiol or diethylstilbestrol. The values for estriol and progesterone did not differ significantly. In the uterine arteries, which lacked spontaneous activity, ovarian steroids and diethylstilbestrol inhibited contractions induced by K+ depolarization. As with myometrium, the lowest effect was observed with estriol and the highest with diethylstilbestrol. A dose-dependent inhibition by all four hormones (0.2-40 microM) of vasopressin-induced contractile responses of the uterine arteries was observed. With the lowest concentration of progesterone, however, the arterial response to vasopressin was enhanced. The increases by progesterone (0.02 and 0.2 microM) of responses induced by vasopressin were statistically significant (P < 0.05). The present data strongly suggest that, in human myometrium and uterine arteries, ovarian steroids and diethylstilbestrol cause a more pronounced inhibition of receptor-mediated than of voltage-dependent Ca2+ channels. The increase by a very low (physiological) concentration of progesterone of vasopressin-induced responses in both myometrium and arteries may be of significance in the pathophysiology of dysmenorrhea.

Clinical tumour markers in ovarian cancer.

Within past few years, the measurement of serological, histochemical and molecular genetic markers has had an increasing influence on clinical decisions about initial treatment and follow-up. This review presents data concerning the most studied and interesting markers in ovarian cancer. CA 125, CA 19.9, TATI, CASA, CEA, TPA, TPS and CYFRA21-1 are now the most widely used serological tumour markers for management of ovarian cancer patients. Ras oncogenes, C-erb2 proto-oncogene, p53 suppressor gene and Bcl-2 oncogene are examples of currently used molecular genetic markers. As histochemical markers-proliferation markers, flow cytometric analysis, thymidine labelling index, Ki-67 nuclear antigen or differentiation markers are nowadays the ones most often determined. Some of these markers might be useful adjuncts for monitoring response to therapy, including early detection of tumour reactivation to allow curative therapy and rapid detection of treatment failure. The study of these markers may also lead to a better understanding of the biological characteristics of ovarian cancer. Numerous tumour markers characterized in this paper have been recognized as promising prognostic factors. The information derived from studies of these markers also represents the most promising avenue towards new treatment strategies; nevertheless to validate these factors, prospective studies of a large patient population are needed.

Uterine effects of N-alpha-triglycyl-(8-lysine)-vasopressin and 8-lysine-vasopressin in the first trimester of pregnancy.

Myometrial effects of a vasopressin hormonogen (N-alpha-triglycyl-(8-lysine)-vasopressin, TGLVP) and of 8-lysine-vasopressin (LVP) were studied in early pregnancy. Intrauterine pressure was recorded in 28 women, who were to have a therapeutic abortion at 8-12 weeks of gestation, and intravenous injection of TGLVP (0.5 and 1 mg) or LVP (0.1, 0.2 and 0.4 IU) were given, or control recordings without drug injection were obtained. TGLVP generally caused a biphasic increase in uterine activity with a rise in uterine tone as the most conspicuous initial effect followed by an increase in amplitude and duration of contractions. In the higher dose group the uterine activity remained significantly higher than in the controls during the rest of the recording period of 4-7 h. The magnitude of effect with the two doses of TGLVP did not differ significantly. The initial myometrial response to LVP resembled that to TGLVP but the effects disappeared within 10-45 min. In the controls the uterine activity did not change. It is suggested that vasopressin analogues could have a therapeutic value in the induction of abortion, but further basic studies are required to define this.

Effect of an oral contraceptive in primary dysmenorrhea--changes in uterine activity and reactivity to agonists.

Mechanisms for the therapeutic effect of oral contraceptives in dysmenorrhea were studied by recording intrauterine pressure on the first day of menstrual bleeding in women with moderate to severe symptoms and after three weeks of oral contraceptive therapy (150 micrograms levonorgestrel + 30 micrograms ethinyl estradiol, daily). Spontaneous uterine activity and reactivity to intravenous injections of vasopressin (6 pmol/kg body weight; n = 8) or prostaglandin F2 alpha (12 nmol/kg body weight; n = 9) at the two sessions were compared. During the first recording when all women had dysmenorrhea, the uterine activity and reactivity to both agonists were pronounced. After therapy, when the women felt essentially no pain, a statistically significant decrease in spontaneous uterine activity in terms of total pressure area, frequency and amplitude of contractions was observed. The agonist injections induced less pain at the second recording, although the magnitude of responses, superimposed on the much smaller uterine activity at this time, were not significantly different from those at the first recording during dysmenorrhea. The mechanism of pain relief by oral contraception in dysmenorrhea could be a lesser impact of the decreased contractile activity on uterine blood flow, abolishing the local ischemia. A reduced uterine reactivity to agonists might also to some extent contribute to the therapeutic effect.

Angiogenesis and Bcl-2 protein expression in patients with endometrial carcinoma.

Considering the particular importance of angiogenesis and tumor suppressor genes expression in solid tumors, angiogenesis and Bcl-2 protein expression were evaluated in order to specify their role in the biology of endometrial carcinoma. Clinical material comprised 66 patients (postmenopausal, aged 52 to 76 years) with endometrial adenocarcinoma. For evaluation of angiogenesis immunohistochemical method was applied using DAKO EPOS Anti-Human Von Willebrand Factor/HRP antibodies. Morphometric method was applied to count angiogenic points (microvessels + single endothelial cells), using a light microscope with morphometric appliance. Angiogenic points density (APD) was defined as the density of AP per square mm. Immunohistochemical staining for Bcl-2 cytosomic protein expression was performed using MoAb124 (dilution 1:80, Dako A/S, Denmark) monoclonal antibodies. The percentage of 10% positive cells was considered as Bcl-2 positive tissue expression. Positive cytoplasmic reaction of Bcl-2 in 51.3% of patients with Stage I endometrial cancer, and in 23.8% and 0% of patients with II and III FIGO stage, respectively, was observed. No relationship between Bcl-2 tissue cytoplasmatic expression and tumor grade was found. However, an inverse correlation between cytoplasmatic expression of Bcl-2 and FIGO stage was observed. The APD (angiogenic points density) was increasing with the clinical (FIGO) stage of endometrial cancer, but it was not observed in the case of tumor histologic grade. Bcl-2 expression and angiogenesis may be a useful parameter in evaluation of the biology of endometrial adenocarcinoma as the study conducted showed the influence of Bcl-2 protein expression upon angiogenesis.

Evaluation of angiogenesis, p-53 tissue protein expression and serum VEGF in patients with endometrial cancer.

Endometrial carcinoma occurs mostly in post-menopausal women. Classical methods of prognostication, as FIGO stage and histopathologic grade, could be improved by applying additional techniques, utilizing molecular biology and immunochemistry. p-53 tumor suppressor gene, the most commonly mutated gene in human cancers has been shown to play an important role in the biology of gynecologic carcinomas. Angiogenesis, a process of formation of new vessels, being connected to tumors progression and metastatic potential was shown to be linked with tumor suppressor genes expression. The aim of the study was to evaluate relationships between intensity of tumor angiogenesis, serum levels of Vascular Endothelial Growth Factor (VEGF) and tissue p-53 protein expression in endometrial adenocarcinoma. Angiogenic Point's Density (APD) was calculated in hot spots areas using the morphometric appliance. For detection of p53 protein in tumor samples, LSAB + Kit Alkaline Phosphatase (DAKO) was used. VEGF levels were assessed in patient's blood sampled before the operation. Overexpression of p53 protein was found in tumor tissue in 35.2% of cases and mean angiogenic points density was greater in p53 positive cases. Serum levels of VEGF were above the cut off level in 54.5% of patients, in those cases angiogenesis was also elevated. In cases of p53 overexpression, VEGF levels tended to be greater as compared with p53 negative cases. In conclusion, our study demonstrated that angiogenesis was more intensive in p53 positive cases, confirming the hypothesis of tumor suppressor-gene regulation of the process of neovascularization. Serum levels of VEGF were borderline-significantly higher in cases of p53 overexpression, they were also correlated to the angiogenesis. Joint assessment of angiogenesis and tumor suppressor genes expression may contribute to reliable evaluation of the biology of endometrial carcinoma.

The influence of lead ions on uterine activity in the rat.

Spontaneous uterine activity and reactivity to oxytocin, naproxen and PGF2 alpha were studied in vitro in 80 rats, which for 2-8 weeks had been exposed to different concentrations and combinations of Pb2+, Zn2+ and Cu2+ in their water supply or had been given clean water for control. In rats given only Pb2+ in concentrations of 1000 and 500 ppm for 6 weeks the uterine activity was significantly increased, whereas in groups given the other ions alone, or Pb2+ for 6 weeks followed by 2 weeks of clean water or Zn2+ or Cu2+ no change was observed. The responses to the oxytocin, naproxen and PGF2 alpha did not differ. These results suggest that contamination with lead ions might be one of the etiological factors involved in conditions with increased uterine activity.

The relationship between cadmium and lead burdens and preterm labor.

OBJECTIVE: To study the possible involvement of lead and cadmium in mechanisms of preterm labor. METHOD: Estimations of lead and cadmium in blood, myometrium and placenta of Swedish and Polish women in preterm labor (n = 17) or delivered at term (n = 13). RESULT: There was a positive correlation between pregnancy week and content of lead in blood. The mean blood concentration of cadmium in preterm delivered women, 2.0 micrograms/l, was significantly higher than that in term delivered patients, 0.6 microgram/l. In Polish patients the concentration of lead in blood, 37.9 micrograms/l, was higher than that in Swedish women of 11.2 micrograms/l. Cadmium concentrations in myometrium and placenta of Polish women, 0.2 and 0.3 microgram/g, respectively, were also significantly higher than the corresponding concentrations in Swedish patients, 0.04 and 0.1 microgram/g and this difference remained when only nonsmokers were compared. Smokers and nonsmokers did not have significantly different cadmium levels. CONCLUSION: These results do not support the concept of lead causing increased myometrial activity in preterm labor, but indicate a higher uptake of lead and cadmium of Polish than of Swedish women. Further investigations are warranted to delineate the importance of the elevated concentration of cadmium in the blood of women with preterm labor.

Influence of high lead and cadmium soil content on human reproductive outcome.

The influence of lead and cadmium on human reproductive outcome was studied in areas of Eastern Poland with high specific soil contamination with these heavy metals. By personal interviews, information on socioeconomic background factors as well as obstetrical and gynecological histories were obtained from 136 women living in villages with high amounts of lead and cadmium and from 269 women of nearby villages with no increased content of these metals. Physical examination and estimations of blood concentrations of lead and cadmium were also performed in 89 and 175 women from these areas, respectively. In the contaminated areas, there were significantly fewer women with three or more pregnancies (39 vs. 52%) and deliveries at full term (35 vs. 44%) than in nearby villages with normal concentrations of these heavy metals, and the weight of preterm newborns was lower (1930 vs. 2225 g). Furthermore, hypertension was more common (15 vs. 22%) and of higher magnitude (mean blood pressure 140 vs. 129 mmHg) in the study group than in the control women. The mean blood concentrations of lead in the two groups. 675 and 621 micrograms/dl did not differ statistically, whereas the concentrations of cadmium of 29 and 25 micrograms/dl were significantly higher in the study than in the control group. There was a significant correlation between cadmium levels and number of preterm deliveries. These results suggest that cadmium and lead could be factors of importance in reproductive outcome.

[Birth environmental modification: needs of mother and child in modern obstetrics]. / Modyfikacja srodowiska narodzin--potrzeby matki i dziecka a potrzeby nowoczesnego poloznictwa.

This paper evaluates changes which all ready has been done and still are taking place in obstetrics according to World Health Organisation recommendations in relation to women's increasing expectation from obstetric hospitals and maternity units. The needs of parturients and theirs children are not in contrary to the needs of modern obstetrics. An attempt has been done to disclose concurrent points to work up the best obstetrics health care model after 2000 year. The author emphasizes that progressive technicalization in obstetrics could not indicate its dehumanization.

[Evidence-based medicine and medical databases in obstetrics and gynecology]. / Medycyna oparta na dowodach i medyczne bazy danych w poloznictwie i ginekologii.

Evidence-based medicine is a new trend in both teaching medicine and supporting the clinical decisive process, answering the clinical questions. The basis of the evidence-based medicine comprises of analysing and interpreting current and reliable medical publications concerning certain subject. The important condition, which has to be fulfilled is a possibility of an easy access to current medical information--via internet, medical publications' databases and publications themselves. It is crucial to use the proper hardware system (local net) and proper software system--tools for using evidence-based medicine. Regarding the medical research, nowadays, database systems became unavoidable. By using such a programs process of data collection, control and analysis became both easier and more reliable thanks to eliminating many "human based" errors. In following paper own experience in introducing evidence-based medicine and medical databases in the field of obstetrics and gynaecology will be presented.

[Iron deficiency as a risk factor during the perinatal period]. / Niedobór zelaza jako czynnik zagrozenia w okresie perinatalnym.

Iron deficiency as a risk factor in the perinatal period--the review of the literature. The presence of iron is necessary for the metabolic processes, erythropoiesis, oxygen transport and respiration, thermoregulation and humoral and cellular immunity. Iron deficiency decreases the amplitude of adaptation of the pregnant women. It might be responsible for abortion or preterm labour as well as disarrangement of myometrial activity during labour and puerperium. Congenital fetal malformations and intrauterine growth retardation might be also stimulated by iron deficiency.

[The management of labor overweight women with intrapartum fetal oxygen saturation monitoring]. / Sródporodowy nadzór z wykorzystaniem pulsoksymetrii plodowej u rodzacych z nadwaga.

OBJECTIVE: Overweight is one of the most common problems that occurs in pregnancy. The aim of this study was to evaluate the usefulness of fetal pulse oximetry during labour and to compare fetal oxygen saturation between cases with pregravid overweight and normal weight. MATERIAL AND METHODS: Fetal oxygen saturation was continuously recorded with use of Nellcor N-400 fetal pulse oximeter in 20 cases of pregravid overweight and 30 control cases of normal weight. Distribution of fetal oxygen saturation values during 4 periods of labour was analyzed and compared between the examined groups together with neonatal umbilical artery pH values, Apgar score, birth weight and percentage of ceasarian sections performed. RESULTS: We noticed statistically important differences in fetal oxygen saturation between analyzed groups. Mean oxygen saturation value was lower in the overweight group, at the end of first stages of the labour (47% vs 52%) and at the second stages (42% vs 46%) We noticed differences in birth weight too. No significant differences in neonatal umbilical artery pH, ceasarian sections, newborns mean 1 minute Apgar score were observed between analyzed groups. CONCLUSION: Fetal pulse oximetry is a a useful method for intensive surveillance of the fetus at risk of hypoxemia during the labour. A lower fetal oxygen saturation value during labor by pregravid overweight were observed.

[Acid-base balance in capillary blood of newborns with perinatal hypoxemia symptoms]. / Równowaga kwasowo-zasadowa krwi wlosniczkowej noworodków z objawami niedotlenienia okoloporodowego.

The purpose of the study was to compare acid-base parameters in capillary blood of newborns from groups with hypoxaemia symptoms and from the control group (physiological deliveries). 111 newborns were divided into 8 groups. Group I consisted of 33 newborns from uncomplicated labour while groups II-VIII involved newborns from complicated labor with symptoms of hypoxaemia. Capillary blood was taken in the 10th and 60th minute and after 24 hours of newborn's life. In newborns with pathological CTG, green amniotic fluid and prolonged first stage of labour--higher duration of acidosis was observed.

[Trial for assessment of values: normal, threatening, and pathologic parameters of acid-base equilibrium in umbilical vessel blood and capillary blood in newborns]. / Próba okreslenia zakresów wartosci: prawidlowych, zagrozenia i patologicznych parametrów równowagi kwasowo-zasadowej krwi naczyn pepowinowych plodów i krwi wlosniczkowej noworodków.

A prospective study in 33 newborns infants born spontaneously at term was carried out. First stage labour duration was up to 10 hours while the second one was no longer than 60 minutes. The key criterion for newborns selection from physiological deliveries was a normal heart rate baseline throughout the whole period of labour. Acid-base parameters in umbilical blood vessels and capillary blood were performed on an Blood Analyzer Corning 168. The results allowed to define normal and pathological ranges of the analysed indicators. They may be used for evaluation of newborn's postnatal conditions.