G. Mollica

Restless legs syndrome and pregnancy

Objective: To perform a large and detailed epidemiologic study on restless legs syndrome (RLS) during pregnancy and the puerperium. Methods: A structured clinical interview, assessing symptoms since the beginning of pregnancy, was performed to a population of 642 pregnant women at the time of delivery and at follow-up evaluation (1, 3, and 6 months after delivery). Main hematologic tests were also evaluated. A woman was considered affected if she met the International RLS Study Group criteria for RLS diagnosis. Results: Twenty-six percent of women were affected by RLS during their pregnancy. The disease was strongly related to the third trimester of pregnancy and tended to disappear reaching the time of delivery. Affected women presented lower values of hemoglobin and mean corpuscular volume compared with healthy subjects (both groups received the same supplemental iron and folate therapy). Conclusions: Pregnancy is associated with transient restless legs syndrome.

Relative influence of menopausal status, age, and body mass index on blood pressure.

The objective of this study was to assess the influence of menopausal status on blood pressure levels in a cross-sectional population study in the setting of a preventive health care program clinically conducted in Ferrara, Italy. The patients were 2397 healthy women 35 to 65 years old (489 in premenopause, 847 in perimenopause, 887 in spontaneous menopause, and 174 in surgical menopause); subgroups were also obtained with increasing duration of menopause (1 to > 5 years) matched with premenopausal and perimenopausal women by chronological age at onset of menopause. Measurements of sphygmomanometric blood pressure, age, age at menopause, duration of menopause, and body mass index were made. Postmenopausal women had higher blood pressure than premenopausal and perimenopausal subjects. After adjustment for body mass index, the blood pressure changes with menopausal status were still significant, but not after correction by age. The increased risk of hypertension of postmenopausal women also lost statistical significance after adjustment for age and body mass index. At multivariate analysis, blood pressure showed no significant relation with the duration of menopause, whereas age was a significant covariant for systolic blood pressure, and body mass index for systolic and diastolic blood pressure. A transient initial rise in blood pressure and body mass index was detected in surgical but not in spontaneous menopause. Blood pressure rises after menopause appear to be due more to increased body mass index and aging than ovarian failure per se.

Oxidative stress, body fat composition, and endocrine status in pre- and postmenopausal women.

Objective: To evaluate the role of menopause on the regional composition and distribution of fat in women and eventual correlations with the oxidative state. Design: In this observational clinical investigation, 90 women (classified for menopause status according to Stages of Reproductive Aging Workshop criteria) were evaluated for body mass composition and fat distribution by dual-energy x-ray absorptiometry and for oxidative status by determination of serum hydroperoxide levels and residual antioxidant activity. Results: Total body fat mass increases significantly in postmenopause (P < 0.05) by 22% in comparison with premenopause, with specific increases in fat deposition at the level of trunk (abdominal and visceral) (P < 0.001) and arms (P < 0.001). Concomitantly, the antioxidant status increases significantly (P < 0.001) by 17%. When data were adjusted for age by analysis of covariance, statistical significance disappeared for the increase in fat mass, but it was retained for antioxidant status (P < 0.05). Both antioxidant status and hydroperoxide level increased with trunk fat mass, as shown by linear correlation analysis (r = 0.46, P < 0.001 and r = 0.26, P < 0.05, respectively). Conclusions: The results of our investigation demonstrate that fat content increases in the upper part of the body (trunk and arms) in postmenopause and that age is the main determinant of this increase. During the comparison of premenopausal and postmenopausal women, we also detected a significant increase in antioxidant status. Apparently this change is mainly related to menopausal endocrine and fat changes.

Hormonal sensitivity of adenylate cyclase from human endometrium: Modulation by estradiol

In human endometria, a membrane-bound adenylate cyclase is present, which is recovered in high yield in a low-speed particulate fraction. Neither the specific activity of the enzyme nor the response to modifiers that act through the regulatory subunit of the complex, are modified during the proliferative or secretory phase of the cycle. Surprisingly, we found that in vitro treatment of secretory endometrial membranes with 17 beta-estradiol stimulates 3- to 4-fold the activity of adenylate cyclase. However this response does not occur on proliferative membranes. The activation by estradiol is independent of the presence of guanylylimidodiphosphate and is additive to that of the nucleotide. Possibly, as the consequence of the phenomenon, the concentration of cyclic AMP is significantly higher in curretage samples obtained from patients during the secretory rather than in the proliferative phase of the cycle. To our knowledge this is the first evidence of a target-cell membrane-directed effect of sex steroids in humans.

Influence of spontaneous and surgical menopause on atherogenic metabolic risk

We evaluated serum total-, LDL-, HDL-, cholesterol and triglycerides in 858 women. Sixty-seven were in premenopause, 307 in perimenopause, 326 in spontaneous menopause, and 158 women underwent hysterectomy. Of these, 101 had bilateral ovariectomy, 24 with unilateral ovariectomy and 33 with retention of both ovaries. After correcting for chronological age and body mass index by variance analysis, it was found that total-, LDL- and HDL-cholesterol significantly increased during menopause without changes in the total cholesterol/HDL-cholesterol, LDL-cholesterol/HDL-cholesterol and triglycerides/HDL-cholesterol ratio. The changes gradually occurred during the months preceding spontaneous cessation of menses. Atherogenic metabolic risk (as relative risk of high total and LDL-cholesterol, estimated by logistic regression analysis) significantly increased in all groups of women in reference to premenopause. The risk observed in surgical menopause with bilateral ovariectomy was higher than in spontaneous menopause, and conservation of ovaries in hysterectomized women seems to protect only partially against such an increase.

Human vascular kinin receptors of the B2 type characterized by radioligand binding

1 The human umbilical vein responds to bradykinin (BK) with contractions that are mediated by B2 receptors. In the present study, the corresponding vascular smooth muscle B2 binding sites have been investigated. 2 [3H]‐BK, a full agonist labelled ligand, was used to demonstrate a single binding site giving a Kd value of 0.51±0.02 nM and a Bmax of 24±1 fmol mg−1 protein. Scatchard plots were linear (r=0.98) in the 0.05–5 nM range of concentrations. Non‐specific binding was found to be 30% of total binding. 3 Competition binding curves gave the following order of potency for various B2 receptor agonists: BK‐[Hyp3]‐BKLys‐BK>>[Aib7]‐BK>>>[desArg9]‐BK, which is typical of B2 receptors. There was no binding to B1 receptors since the selective B1 receptor ligand, Lys‐[desArg9]BK was inactive up to 10 μM (n=4). 4 Characterization of the binding site with antagonists, performed with three chemically distinct series of peptide and non‐peptide compounds, revealed a high affinity of Hoe 140 (D‐Arg‐[Hyp3,Thi5,D‐Tic7,Oic8]‐BK) (Ki 0.17 nM; n=4) which was more potent that FR 173657 ([(E)‐3‐(6‐acetamido‐3‐pyridyl)‐N‐[N‐[2,4‐dichloro‐3‐[(2‐methyl‐8‐quinolinyl)oxymethyl]phenyl]‐N‐methylaminocarbonylmethyl] acrylamide]) (Ki 1.94 nM; n=4), D‐Arg‐[Hyp3,D‐Phe7,Leu8]‐BK (Ki 256 nM; n=4) and Win 64338 (phosphonium, [[4[[2[[bis(cyclohexylamino)methylene]amino]‐3‐(2‐naphthalenyl)‐1‐oxopropyl]amino]phenyl]methyl]tributyl, chloride, monohydrochloride) (Ki 1,450 nM; n=4). 5 The present study describes and characterises B2 receptor binding sites in the vascular smooth muscle of the human umbilical vein. The binding assay appears to be suitable for studying new agonists or antagonists designed to activate or block the B2 receptor class that mediate the majority of the physiopathological effects of kinins in man.

Oophorectomy and Spine Bone Density: Evidence of a Higher Rate of Bone Loss in Surgical Compared with Spontaneous Menopause

To compare the influence of spontaneous and surgical menopause on bone loss, we measured with dual x-ray absorptiometry (DXA) the spinal bone mineral density (BMD) in 513 women recruited at the Menopause Clinic at Ferrara University Hospital. One hundred one women were premenopausal with regular menstrual cycles; 185 women were perimenopausal with irregular periods or with absence of menstruation for <11 months; 160 women had spontaneous menopause with at least 12 months of amenorrhea; 67 women had a surgical menopause (hysterectomy with bilateral oophorectomy) prior to which they had regular menstruation. To minimize the age bias on BMD, all postmenopausal patients were selected to have the age range at menopause corresponding with the chronological age range (45–53 years) of premenopausal women used as reference. Moreover, to evaluate the influence of time since menopause on BMD, all postmenopausal women were stratified in five categories according to time lapsed since their last menses or oophorectomy. BMD values of spontaneous and surgical menopause do not appear to differ significantly (0.908 4pM 0.146 and 0.885 ± 0.129 g/cm2, mean ± SD). However, the difference between the menopausal groups becomes evident when BMD results take into account the interval since menopause. After 61–144 months of amenorrhea, women who had undergone spontaneous menopause had a cumulative bone loss of 21.8% in comparison with premenopausal BMD, whereas women who had undergone surgical menopause had a bone loss of 25.8%. The yearly percentage of bone loss values of surgical menopause (ranging from 3.72 to 7.93) settled to ∼1% per year after 5 years from oophorectomy, whereas the percentage values of spontaneous menopause (ranging from 1.75 to 4.65) settled to 1% per year after 3 years since the last menses. The difference between bone loss rates of spontaneous and surgical menopause, evaluated by comparison of regression coefficients (— 0.027 and —0.051, respectively) of linear regressions of BMD values on time since menopause, was statistically significant (p ≤ 0.001). Odds ratio (OR) of osteopenia (as Tscore, ≥ — 1) was significantly higher in surgical menopause (OR, 10.36; CI, 24.69–4.34) compared with spontaneous menopause (OR, 7.11; CI, 14.73–3.43). Our data support the evidence that women undergoing bilateral oophorectomy while still menstruating are at a higher risk of osteopenia than women undergoing menopause spontaneously.

Papilloma and Polyoma DNA Tumor Virus Sequences in Female Genital Tumors

Human papillomaviruses (HPVs) and BKV, JCV, and SV40 polyomaviruses (PYVs) are oncogenic viruses associated with different human tumors. Our aim was to determine if PYV and HPV sequences could be detected in human genital tumors. HPV types 6b, 11, 16, and 18 and PYV were investigated in 22 genital tumor samples and the corresponding adjacent normal tissues, by PCR and filter hybridization. HPV and PYV sequences were also searched in six sperm fluid and four peripheral blood cell (PBC) samples. HPV-16 sequences were revealed in 7 of 14 cervical tumors and 1 of 1 vaginal adenocarcinoma, whereas 1 of 14 cervical carcinoma tested positive for HPV-18. Interestingly, each normal cervical tissue surrounding the neoplasm obtained from the same patient was positive for HPV type-16 and -18 with the same prevalence detected in tumors. BKV sequences were found in 9 of 14 cervical tumors, 1 of 7 vulvar tumors, and 1 of 1 adenocarcinoma, but also in normal tissues from cervix (13 of 14), vulva (6 of 7), sperm fluid (5 of 6) and PBC (3 of 4) samples. SV40 sequences were detected in 1 of 14 normal cervical tissue, 2 of 6 sperm fluids and 1 of 4 PBCs. None of the samples were JCV positive. To our knowledge, this is the first investigation reporting on the simultaneous association of both HPV and PYV with human genital tumors. These results suggest that PYV, together with HPV, may be involved as a cofactor in the onset/progression of human genital tumors, and raise the possibility that PYV act synergistically with HPV to enhance their pathogenicity in vivo. In addition, HPV and PYV may complement each other in infecting human genital tissues.

Sex Steroid Hormones Influence the cAMP Content in Human Endometrium during the Menstrual Cycle

The measurement of the tissue concentrations of cyclic adenosine monophosphate (cAMP) in human endometrium shows that the levels of the nucleotide vary during the menstrual cycle, being 11.4 +/- (SE) 2.5 and 37.7 +/- (SE) 10.1 pmol/mg protein in the proliferative and secretory phase, respectively. The individual determinations of cAMP are significantly correlated to the estradiol/progesterone blood concentration ratio: by this means we obtained a superimposable distribution with the results of histologic examination suggesting that the hormones have a direct action in determining the cAMP levels in this tissue. The relevance of these observations for the physiology of the endometrium is discussed.

Short-Term Effect of Oophorectomy on Lipoprotein Metabolism

The influence of steroid hormones on lipoprotein metabolism has been investigated in menstruating women undergoing oophorectomy; indeed, by this procedure, it is possible to alter selectively the secretion of the hormones and thus to determine the influence of their withdrawal on blood lipids. Within 3 months of surgery we observed significant rapid changes in cholesterol levels of total and HDL-bound components (early decrease and subsequent increase) without alteration of the triglyceride component of the lipoproteins. Similar biphasic changes were observed for the apolipoproteins A and B, determined by immunological methods. These results are suggestive of an action of sex hormones on the lipoprotein metabolism, both on the lipid and the protein moieties. They can be explained by a primary action of the hormones on the apolipoprotein metabolism and a secondary apoprotein-mediated effect on the lipid component of the lipoproteins.