Homocysteine and folate levels in postmenopausal women.

OBJECTIVES: To assess total homocysteine (tHcy) and folate levels in postmenopausal women and investigate whether age, menopause duration, kind of menopause and tobacco use had an effect on these levels. METHODS: Total homocysteine and folate levels were measured in fasting blood samples of 200 postmenopausal women with normal thyroid and renal function tests. Patients were not receiving vitamins or hormone replacement therapy. RESULTS: Total homocysteine levels increased significantly after 60 years while folate levels showed a decrease trend after 65 years. Menopause duration had no effect on folate levels and increased significantly tHcy levels after >180 months duration. The kind of menopause did not influence tHcy and folate levels. Tobacco use reduced significantly folate levels. CONCLUSIONS: Age seems to be the principal factor influencing tHcy levels. We believe that decreased folate levels also reflect an age-associated inadequate dietary intake. Tobacco use did not alter tHcy levels; however, we found smoking to lower folate levels.

Mucin-like carcinoma-associated antigen (MCA) during normal pregnancy.

OBJECTIVE: To assess the usefulness of Mucin-like carcinoma-associated antigen (MCA) in monitoring pregnant patients with breast cancer. STUDY DESIGN: Maternal serum (MS) and amniotic fluid (AF) antigen values were measured by an enzyme immunoassay in 30 pregnant women during the second trimester, in 28 during the third and in 26 at parturition. Sera only from 26 women in the first trimester and from 26 healthy, non-pregnant women (controls) were also analyzed. RESULTS: Maternal serum MCA concentrations increased significantly with gestational age (p<0.0001). The frequency of elevated serum values was 5% in the first, 35% in the second and 100% in the third trimester and at parturition. Antigen values in AF were markedly higher than those in MS (p<0.0001) and increased also significantly with advancing gestation (p<0.0001). A strong correlation was observed between MS and AF antigen values (r=0.77, p<0.0001). Maternal serum values at parturition were dependent on the mode of delivery, being higher in the cases who delivered vaginally, compared to those delivered by elective caesarean section (p<0.006). CONCLUSION: Our data suggest that pregnancy affects significantly maternal serum MCA. Consequently, MCA seems to be a non-reliable marker in monitoring pregnant patients.

Hypothalamic-pituitary-thyroidal axis dysfunction and cortisol secretion in patients with nonclassical congenital adrenal hyperplasia.

OBJECTIVE: The purpose of this study was to evaluate thyroid function and TSH and cortisol (F) secretion in hyperandrogenemic women with nonclassical congenital adrenal hyperplasia (NC-CAH) due to 21-hydroxylase deficiency (Group A) when compared with women with hyperandrogenemic symptoms (menstrual irregularities, hirsutism, acne, seborrhea and sterility) of other etiologies (Group B). METHODS: Seventy-two women were subjected to stimulation of the adrenal cortex with i.v. ACTH administration in the early proliferative phase of the menstrual cycle. Basal plasma TSH, T3, T4, and FTI as well as basal and ACTH-stimulated plasma F and 17-hydroxyprogesterone levels were determined. RESULTS: According to internationally accepted criteria and HLA haplotyping, we diagnosed 28 NC-CAH patients as well as affected heterozygotes of the disease. No significant difference was found in the plasma T3, T4, or FTI or F concentrations between the women of the two groups. On the contrary, plasma TSH levels were significantly lower in patients with 21-hydroxylase deficiency when compared to the women with hyperandrogenemic symptoms of other etiologies. CONCLUSION: The results of this study support a dysfunction of the hypothalamic-pituitary-thyroidal axis due to altered ACTH secretion patterns.

Serum soluble E- and L-selectin in the very early neonatal period.

Both E- and L-selectin are cell adhesion molecules. E-selectin is expressed by activated endothelial cells, whereas L-selectin by quiescent leukocytes and is rapidly cleaved off after activation. Both selectins take part in the first step of the 'adhesion cascade', the 'rolling of leukocytes', leading to the extravasation of the white cells to the sites of inflammation, infection or damage. For this reason their soluble forms (sE- and sL-selectin, respectively), are considered early and reliable markers of the immune activation and response. Moreover, sE-selectin has been reported to be a potent angiogenic factor and a reliable marker of infection and sepsis in neonates, as well as endothelial activation, while sL-selectin of the leukocyte function and maturity. Following informed maternal consent, we evaluated prospectively by ELISA, sE- and sL-selectin in the serum of 40 (19 females, 21 males), healthy, term, infection-free neonates, on the second and fifth day of life, and compared them with the respective values in 20 healthy adults (10 females, 10 males), with the purpose of examining the pattern of their values in the early postpartum days, and to establish reference values for both selectins. Values (mean+/-S.D.) of sE-selectin both on the second (139+/-48 ng/ml) and fifth day of life (111+/-35 ng/ml) were found to be highly increased, as compared with those in controls (48+/-13 ng/ml; P<4 x 10(-11) and P<4 x 10(-10), respectively), while sL-selectin values on both the second (674+/-223 ng/ml) and the fifth day of life (684+/-221 ng/ml), were significantly lower than those in controls (938+/-181 ng/ml); P<0.0001 and P<0.0003, respectively). A significant decrease was noted in sE-selectin values, from the second to the fifth day of life (P<10(-7)), while sL-selectin values showed no significant change in the same time interval. A strong correlation was found between values on the second and the fifth day of life of both sE- and sL-selectin (r(P)=0.885 and r(P)=0.813, respectively; P<0.00001). Neonatal values of both sE- and sL-selectin on the second or on the fifth day of life, did not depend on the perinatal factors, neonatal sex, or birth weight, mode of delivery, and maternal age or parity. In conclusion, in the very early neonatal period, our findings of highly increased sE-selectin, while low sL-selectin, suggest an immune and more specifically endothelial activation and an immature and decreased leukocyte function.

Tissue polypeptide specific antigen (TPS) throughout normal pregnancy.

AIM: TPS concentrations were measured throughout normal pregnancy in maternal serum (MS) and amniotic fluid (AF), in order to evaluate the usefulness of TPS in the follow-up of pregnancy breast cancer patients. PATIENTS AND METHODS: Following informed consent, 30 pregnant women during the 2nd trimester, 28 during the 3rd and 26 at parturition were included in the study. For comparison, 28 women in the 1st trimester and 28 healthy, non pregnant women (controls) were also studied. Both MS and AF antigen values were measured by an enzyme immunoassay (BEKI Diagnostics). RESULTS: Maternal serum TPS concentrations increased significantly with gestational age (p < 0.0001), being significantly higher in the 3rd trimester and during labor than those in the controls (p < 0.0001). Amniotic fluid TPS values were markedly elevated, compared with those in MS (p < 0.0001, paired-t-test), declining significantly from the 2nd to the 3rd trimester (p < 0.0015) and labor. Both MS and AF TPS values during labor depended on the mode of delivery, being higher in the cases terminated by vaginal delivery, compared to those by elective cesarean section. CONCLUSION: Maternal serum TPS values are influenced significantly by pregnancy, and thus, this antigen, as tumor marker seems to be reliable only during early pregnancy.

A comparative study of serum soluble vascular cell adhesion molecule-1 and soluble intercellular adhesion molecule-1 in preeclampsia.

OBJECTIVES: Maternal serum soluble vascular cell adhesion molecule-1 (sVCAM-1) and soluble intercellular adhesion molecule-1 (sICAM-1) were evaluated in preeclampsia to investigate whether these molecules could be helpful with regard to this pregnancy complication. STUDY DESIGN: The study population was composed of 30 preeclamptic patients with a mean gestational age of 35.5 +/- 4.6 weeks and 20 age-matched and gestational age-matched normotensive uncomplicated pregnancies (controls). Blood samples from 7 of the 30 preeclamptic patients and 15 of the 20 controls in the second trimester were also analyzed. Data were analyzed by parametric methods. RESULTS: Significantly higher maternal serum sVCAM-1 levels were found in both groups of preeclamptic patients with and without fetal growth restriction (981 +/- 145 ng/ml; n = 13; p < 0.0005 and 846 +/- 84 ng/ml; p < 0.02, respectively) compared with controls (668 +/- 186 ng/ml). In contrast, no significant difference was found in maternal serum sICAM-1 levels between preeclamptic and normotensive pregnancies, or in both adhesion molecules (1) in the controls between second and third trimester samples and (2) in the second trimester between pregnant women who developed preeclampsia later and gestational age-matched controls. CONCLUSION: These findings show a selective significant elevation of maternal serum sVCAM-1 in preeclampsia, with the highest values in cases complicated with fetal growth restriction, perhaps reflecting its angiogenic function. Hence, sVCAM-1 could be helpful in the diagnosis of this fetal complication in preeclampsia.

Serum leptin concentrations during the perinatal period.

We aimed to study maternal and infant serum leptin concentrations during the perinatal period and their relationship to the body weight of mothers and newborns. Serum leptin values were measured by enzyme-linked immunoadsorbent assay (ELISA) (R&D systems) in 26 healthy, term neonates during the first (N1) and fifth (N5) day after birth and were compared with serum leptin values in maternal blood (MS), amniotic fluid (AF), and umbilical cord (UC) at delivery. Twenty-five healthy, nonpregnant women, age and body weight-matched to the mothers, were used as controls (C). Infant serum leptin concentrations declined significantly after birth from UC to the N5 samples (p<0.003). MS leptin values were significantly higher than UC, N1, N5, and C values (p<0.001), while AF values were significantly lower than in controls (p<0.001). UC, but not MS leptin values correlated significantly with the birth weight of infants (r = 0.6; p<0.03). The elevated values of leptin in maternal serum and the regressing pattern of infant leptin values after birth suggest an additional, probably placental source of this protein during pregnancy, possibly contributing to the regulation of fetal body weight.

Soluble form of ICAM-1, VCAM-1, E- and L-selectin in human milk.

In breast milk and paired serum from 70 lactating women and 40 of their term, infection-free neonates, on the 2nd and 5th day postpartum slCAM-1, sVCAM-1, sE- and sL-selectin were measured by ELISA and compared with those in 26 healthy adults (controls). Seven infant formulas and fresh milk from five cows were also analyzed. Human colostrum values of slCAM-1, sVCAM-1 (similar to those in maternal and control serum), sE-selectin and sL-selectin (-10 and -100 times lower than in maternal and control serum) were significantly higher than those in milk, while they varied widely. None of the adhesion molecules was detected in fresh cow's milk or infant formulas. Exclusively breast-fed infants showed significantly higher values of slCAM-1 and sL-selectin on the 2nd day of life than those supplemented also with formula. Only slCAM-1 values correlated positively between colostrum and time-matched maternal serum. These findings show in human milk important amounts of slCAM-1 and sVCAM-1 but minimal amounts of sE- and sL-selectin, which could affect the immune system of the neonate.

Patterns of inflammatory cytokine serum concentrations during the perinatal period.

Inflammatory cytokines interleukin-1beta (IL-1beta), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) were measured in the serum of healthy, term neonates on the first (N1), fifth (N5) and 40th (N40) day after birth, compared with those in maternal serum (MS), umbilical cord (UC) and in adult controls. All three cytokines were significantly elevated in N1 and N5, compared with those in UC and adults (P < 0.0001). IL-1beta and IL-6 declined significantly from N1 to N40 (P < 0.0001), while TNF-alpha increased significantly from N1 to N5 and declined thereafter. TNF-alpha values in UC were significantly higher than in adults, but lower than in N40 (P < 0.0001), while IL-1beta and IL-6 values in UC did not differ from those in N40 and in adults. IL-1beta and IL-6, but not TNF-alpha values in MS were significantly higher than those in controls (P < 0.0001). IL-1beta values in MS were significantly higher than those in N1 (P < 0.0001), while those of IL-6 and TNF-alpha were significantly lower (P < 0.0001). Moreover, IL- 1beta values were dependent on the mode of delivery in N1 (P < 0.001), in MS (P < 0.02) and in UC (0.03), while IL-1beta and TNF-alpha values in N1 were strongly interrelated (r = 0.7; P < 0.01). In conclusion, the increased values of IL-1beta, IL-6 and TNF-alpha during the perinatal period might reflect a newborn immune response to the stress of delivery and to environmental changes after birth.

Breast cancer markers during normal pregnancy.

AIM: To measure MCA and CA153 concentrations in maternal serum (MS) and amniotic fluid (AF) paired samples during normal pregnancy, in order to evaluate the usefulness of these markers in monitoring pregnant patients with a history of breast cancer. PATIENTS AND METHODS: Serum and AF MCA and CA153 values were measured in 20 pregnant women during the 1st trimester, 29 cases in the 2nd, 26 in the 3rd and 20 at parturition and compared with those of 20 healthy, age-matched, non pregnant women (controls). RESULTS: MS values of MCA increased significantly with gestational age (p < 0.0001), being higher in the 3rd trimester and in labor than in control values (p < 0.0001). MCA values in AF were remarkably higher than those in MS and increased significantly with advancing gestation (p < 0.0001). In contrast, CA153 values in AF, which were marginally higher than in MS, did not differ significantly with the progression of pregnancy. CONCLUSIONS: Maternal serum MCA values are significantly influenced during pregnancy. Thus, this marker seems to be reliable only during early pregnancy. In contrast, CA153 remains a useful marker in monitoring pregnant breast cancer patients.

The incidence of 21 alpha-hydroxylase deficiency in Greek hyperandrogenic women: screening and diagnosis.

The purpose of this prospective study was to determine the incidence of any form of 21 alpha-hydroxylase deficiency among Greek women with hyperandrogenic symptoms, and to test the predictive value of basal serum 17-hydroxyprogesterone (17-OHP) in the early follicular phase as a screening index for patient preselection to adrenocorticotropic hormone (ACTH) testing. Eighty-eight unselected women with hyperandrogenic symptoms were examined in the Gynecological Endocrinology Unit of the Second Department of Obstetrics and Gynecology of Athens University. Using the ACTH-stimulated 17-OHP values at 60 minutes (17-OHP60) the study population was divided into four groups (A, B, C and D). Clinical and basal hormonal parameters as well as serum 17-OHP60 values and human leukocyte antigens were studied. Both clinical and basal hormonal parameters could be used to distinguish only patients with severe 21 alpha-hydroxylase deficiency (group A). In contrast, patients with moderate non-classical congenital adrenal hyperplasia (NC-CAH; group B), heterozygotes for NC-CAH (group C), and unaffected females (group D) can be diagnosed and classified only by serum 17-OHP60 values. In conclusion, the incidence of NC-CAH in Greek females with hyperandrogenic symptoms is 3.4%. The positive predictive value of basal 17-OHP is only 13% for this disease. Only 17-OHP60 helps to diagnose and classify moderate and mild forms of NC-CAH. Thus, it seems that ACTH testing is imperative in every subject suspected of this enzymatic disorder.

Soluble intercellular adhesion molecule-1 in newborn infants.

UNLABELLED: The aim of this study was to evaluate the effect of increasing postnatal age on soluble intercellular adhesion molecule-1 (sICAM-1), a very early and sensitive marker of immune activation and response in the serum of newborn infants. Serum sICAM-1 was measured by EIA (T Cell Diagnostics) in 20 healthy adults (controls) and in 43 (24 females/19 males) healthy neonates, of whom 28 were full term, and 15 were born at a gestational age between 35 and 38 weeks of pregnancy, on the 1st, 5th and 30th day of life. Neonatal serum sICAM-1 values showed a very significant increase (P < 0.01) from the 1st day (137.3+/-62.0 ng/ml) to the 5th day (259.3+/-124.0 ng/ml) and then to the 30th day of life (415.0+/-114.0 ng/ml), being significantly lower on the 1st day (P < 0.01), whereas significantly higher on the 30th day of life (P < 0.05), than those in healthy adults (305+/-195 ng/ml). Serum sICAM-1 values on the 1st day of life depended on both the mode of delivery (significantly higher in neonates born vaginally) and the gestational age at birth (significantly lower in those born at a gestational age over 38 weeks). A significant strong correlation was found in sICAM-1 values between the 1st and the 5th day following delivery (rp = 0.77, P < 0.009). CONCLUSION: The results of this study demonstrate a significant rise of serum sICAM-1 during the 1st month of life in healthy neonates suggesting a progressively increased activation of the neonatal immune system.

Tumour-associated antigens CEA, CA125, SCC and TPS in gynaecological cancer.

Pretreatment values of CEA, CA125, SCC and TPS were measured in 130 women with 1) ovarian carcinoma (n = 25), 2) breast cancer (n = 20), 3) endometrial cancer (n = 14), 4) cervical squamous cell carcinoma (n = 20), 5) cervical adenocarcinoma (n = 9) and 6) benign gynaecological diseases (n = 42) in order to evaluate the usefulness of multiple markers in diagnosing and monitoring patients with gynaecological cancer. Antigen values were significantly higher in the cancer groups than those in the benign one (p < 0.0001). CEA values were significantly elevated in the 2nd and 5th groups, CA125 in the 1st and 5th, SCC in the 4th and 5th, and TPS in the 1st, 2nd and 5th compared to the remaining groups (p < 0.04-p < 0.0001). In advanced stage diseases, significantly higher antigen values, except for SCC, than those in limited tumours were measured (p < 0.05-p < 0.0001). In conclusion, our results suggest that, multiple markers may be more efficient than the use of single markers in accurately identifying malignant from benign gynaecological diseases and in monitoring cancer patients.

Inflammatory cytokines in newborn infants.

Serum levels of IL-1beta, IL-6 and TNF-alpha were measured in 48 healthy, termed neonates on the 1st (N1), 5th (N5) and 40th (N40) day after birth, compared with those in maternal serum (MS), umbilical cord (UC) and adult controls. Cytokine values in N1 and N5 were significantly elevated, than those in UC and in controls (P<0.0001). IL-1beta and IL-6 declined significantly from N1 to N40 (P<0.0001), while TNF-alpha increased significantly from N1 to N5 and declined thereafter. MS infinity IL-1beta and IL-6, but not MS infinity TNF-alpha, were significantly higher than those of controls (P<0.0001). IL-1beta values depended on the mode of delivery. In conclusion, the increased concentrations of IL-1beta, IL-6 and TNF-alpha during the perinatal period might suggest their involvement in an inflammation-like process during normal parturition, and reflect also a newborn immune response to the stress of delivery and environmental changes.

Changes of angiogenin serum concentrations in the perinatal period.

The polypeptide angiogenin, a normal constituent of human plasma, might be involved in endothelium homeostasis, angiogenesis, and neovascularization accompanying various diseases. This study aimed at determining angiogenin serum concentrations in the perinatal period of healthy newborns and at forming a baseline for this protein, which in the future may serve as a diagnostic index in developmental errors of the placenta and/or newborn. One milliliter of blood was drawn on d 1 and 4 of life from 30 healthy full-term neonates, and angiogenin serum concentrations were measured by an enzyme immunoassay using a commercially available kit. In 10 cases angiogenin serum concentrations were also measured in the maternal serum before delivery and in the umbilical vein serum. Angiogenin serum concentrations (microgram/L) were significantly higher in maternal serum (225.7 +/- 49.6) compared with umbilical vein serum (119.0 +/- 34.2) (p < 0.0002), as well as that compared with day 1 (166.4 +/- 44.9) (p < 0.01) but not to d 4 neonatal serum (240.8 +/- 52.6). Angiogenin serum concentrations showed a statistically significant increase from d 1 to 4 (p < 10(-7)), as well as from umbilical cord serum to d 1 neonatal serum (p < 0.0002). A statistically significant correlation existed between values in umbilical cord serum and d 1 neonatal serum (r = 0.84, n = 10, p < 0.002) and between those in d 1 and 4 neonatal serum (r = 0.37, n = 30, p < 0.04). Sex, birth weight, or mode of delivery did not influence angiogenin serum concentrations. We conclude that a rapid increase of angiogenin serum concentrations to maternal levels takes place during the first four postnatal days in healthy full-term neonates.

Comparison between steroid hormones and cortisol in serum and follicular fluid in stimulated and unstimulated cycles of in vitro fertilization patients.

The present study was undertaken to evaluate the effects of the concentrations of serum and follicular fluid steroids and cortisol levels on the establishment of pregnancies in in vitro fertilization (IVF). Our study group consisted of 42 women (group A) who received gonadotropins for induction of ovulation for IVF. The control group included 23 women (group B) who underwent in vitro fertilization without stimulation. Serum estradiol and progesterone levels were significantly higher in group A than in group B. Serum and follicular fluid cortisol levels were similar in both groups A and B. There was no significant difference in the fertilization rates of the stimulated or unstimulated cycles. However, there were no pregnancies in group B whereas there was a 28.5% pregnancy rate in group A. There were no correlations between the estradiol, progesterone and cortisol levels when compared to the oocyte maturity and the fertilization rates in both groups of patients.

Cyclic and individualized administration of gonadotropin-releasing hormone agonists plus progestogens: an alternative protocol for contraception.

Twenty-one women presenting with different diseases, with absolute or relative contraindications to hormonal contraception or the use of intrauterine devices, received 300-600 micrograms/day buserelin intranasally from the 1st to the 21st day, and 5 mg/day norethisterone acetate orally from the 16th to the 23rd day of the cycle for a total of 245 cycles. Serum luteinizing hormone (LH), follicle-stimulating hormone (FSH), estradiol and testosterone were determined on days 3-5 and 13-15 of the cycle, while progesterone determinations and ovarian sonography were performed during the second half of the cycle. According to progesterone values, 92.7% of the treatment cycles were anovulatory, while in one cycle pregnancy was detected (0.4%). Values of serum LH, FSH and estradiol were low, and in most of the cycles ovarian follicular development was limited to follicles < or = 11 mm. In 21 treatment cycles (9%), statistically significant increases in FSH (p < 0.0001) and LH (p < 0.02), as well as ovarian proliferation to preovulatory follicles or luteinized follicles, were found. It appears that in spite of the high cost of medication and monitoring of patients, this regimen could be useful as an alternative in cases where other forms of contraception are contraindicated or have failed.

Vaginal fluid and serum CEA, CA125 and SCC in normal conditions and in benign and malignant diseases of the genital tract.

Carcinoembryonic antigen (CEA), CA125, and squamous cell carcinoma antigen (SCC) were evaluated in paired vaginal fluid and serum samples from 69 women, mean age 40.6 (20-78) years. Fifteen of the subjects were normal females (controls), 12 were pregnant, 20 had benign gynecological diseases, 5 presented severe cervical dysplasias and 17 suffered from cancer of the genital tract. Highly elevated CEA, CA125 and SCC concentrations (median, range) were found in vaginal fluid: 186 ng/ml (12-5420); 890 U/ml (54-65000); 1600 ng/ml (27-13000) respectively, compared with those in the paired serum samples: 1 ng/ml (0.5-8.6); 12 U/ml (3.0-1590); 1 ng/ml (0.3-19). Vaginal fluid CEA, CA125 and SCC values were significantly different among the five studied groups (p < 0.0002; p < 0.02: p < 0.002 respectively), being significantly higher in the patients with benign gynecological diseases, compared with those in the patients with malignancies of the genital tract (p < 0.0001; p < 0.02; p < 0.005), and those in controls (p < 0.02; p < 0.007; p < 0.02 respectively). The results of this study suggest that: 1) CEA, CA125 and SCC seem to be normal constituents of vaginal fluid. 2) The distribution of CEA, CA125 and SCC between vaginal fluid and the circulation is affected by pregnancy, inflammation and cancer of the genital tract.