Involvement of seminal leukocytes, reactive oxygen species, and sperm mitochondrial membrane potential in the DNA damage of the human spermatozoa.

Measurement of reactive oxygen species (ROS) producing leukocytes in semen has been a standard component of the semen analysis, but its true significance remains still unknown. In this study, we have correlated the number of seminal leukocytes to various semen parameters. We found a negative correlation between the leukocyte number and sperm concentration (rs  = -0.22; p = 0.01) and motility (rs  = -0.20; p = 0.02). In contrast, a positive correlation between the number of leukocytes and both seminal ROS (rs  = 0.70, p < 0.001; n = 125) and the number of spermatozoa with DNA fragmentation (rs  = 0.43, p = 0.032; n = 25) was found. However, only a trend of positive correlation between ROS and the number of spermatozoa with TUNEL-detected DNA fragmentation was observed. Moreover, this latter was not correlated with loss of sperm mitochondrial membrane potential (MMP) (10% vs 35%, rs  = 0.25, p = 0.08; n = 50). Overall these results indicate that the presence of high number of leukocytes in the ejaculate negatively affects key semen parameters, as sperm concentration and motility, associated with infertility conditions. Moreover, they suggest that leukocytes are the major source of the seminal ROS and cause of sperm DNA fragmentation. However, the absence of a clear correlation between ROS and sperm DNA fragmentation, and spermatozoa with damaged DNA and MMP loss, suggest that ROS produced by leukocytes might be not the only cause of DNA damage in spermatozoa and that intrinsic mitochondrial-dependent apoptotic pathways might not have a major impact on sperm DNA fragmentation.

Analysis of programmed cell death in mouse fetal oocytes.

We report a short-term culture system that allows to define novel characteristic of programmed cell death (PCD) in fetal oocytes and to underscore new aspects of this process. Mouse fetal oocytes cultured in conditions allowing meiotic prophase I progression underwent apoptotic degeneration waves as revealed by TUNEL staining. TEM observations revealed recurrent atypical apoptotic morphologies characterized by the absence of chromatin margination and nuclear fragmentation; oocytes with autophagic and necrotic features were also observed. Further characterization of oocyte death evidenced DNA ladder, Annexin V binding, PARP cleavage, and usually caspase activation (namely caspase-2). In the aim to modulate the oocyte death process, we found that the addition to the culture medium of the pan-caspase inhibitors Z-VAD or caspase-2-specific inhibitor Z-VDVAD resulted in a partial and transient prevention of this process. Oocyte death was significantly reduced by the antioxidant agent NAC and partly prevented by KL and IGF-I growth factors. Finally, oocyte apoptosis was reduced by calpain inhibitor I and increased by rapamycin after prolonged culture. These results support the notion that fetal oocytes undergo degeneration mostly by apoptosis. This process is, however, often morphologically atypical and encompasses other forms of cell death including caspase-independent apoptosis and autophagia. The observation that oocyte death occurs mainly at certain stages of meiosis and can only be attenuated by typical anti-apoptotic treatments favors the notion that it is controlled at least in part by stage-specific oocyte-autonomous meiotic checkpoints and when activated is little amenable to inhibition being the oocyte able to switch back and forth among different death pathways.

Influence of the hormonal status on somatic, psychopathological and mood symptoms in climacteric women.

OBJECTIVE: To evaluate the influence of hormonal status on somatic, psychopathological and mood symptoms in climacteric women. METHOD: 122 postmenopausal women have been evaluated by the PISA-system and P.O.M.S. (Profile of Mood States) to evaluate somatic, psychopathological and mood symptoms in a 3-intervention trial (perimenopausal women, postmenopausal women on replacement therapy, and postmenopausal women without any therapy). RESULT: We found no statistically significant difference among the three groups. A clear trend has however resulted: sex hormones seem to decrease the depressive mood, aggressiveness/anger and sexual dissatisfaction. CONCLUSION: Our results are inconclusive but they suggest that hormones influence some psychological and mood symptoms during the climacterium.

[Recruitment strategies for a randomized clinical study]. / Strategie di reclutamento per uno studio clinico randomizzato.

BACKGROUND: To value the level of acceptance of the Memphis Study, (a random clinical study suggested verifying the efficacy of a diet enriched with phytoestrogens to prevent menopausal problems), and underlining the reasons which induce the acceptance or refusal of the study. STUDY DESIGN: Prospective study. METHODS: Meeting groups have been held with 82 women needing Day Hospital treatment for Menopause at III Department of Obstetrics and Gynecology of the University of Bari. The features of women and the reasons that induce these++ acceptance or refusal of the study were valued by an anonymous questionnaire with a precoded reply. RESULTS: 92.9% of the women accepted participation in the study. The main reasons for acceptance were: 1) believing that the study was drawn up for women; 2) it was done by expert physicians; 3) it was not done for financial gain. The random standard was the main reason for refusal. CONCLUSIONS: With this method we had a very high acceptance, talking over the goals and problems that study aimed to address and to resolve. The chance offered to judge the reliability and competence of physicians is important.

[Territorial research on the availability of phytoestrogen rich foods]. / Indagine territoriale sulla disponibilità di alimenti ricchi di fitoestrogeni.

BACKGROUND: To verify the availability of foods containing phytoestrogens, the quality of available products, the degree of knowledge of these foods by dealers, the willingness of dealers to give cooperation in preparation of the MENFIS study, a study assessing the efficacy of a phytoestrogen rich diet on the long term effects of menopause. METHODS: Perspective research by questionnaires carried out in the herbalist's shops of Bari. RESULTS: For the aims of the study the herbalist's shops resulted in being the most suitable, because they were the most supplied with these products and with natural foods, with a high degree of knowledge of these foods and a helpfulness towards the users, perhaps because of the strong motivation towards this type of diet. CONCLUSIONS: The easy availability of these products, the good preparation of managers of the herbalist's shops and their good experience is a sound basis for those intending to start dietary programs with phytoestrogen rich foods.

[Severe preeclampsia. Appropriate management and socio-cultural factors]. / La preeclampsia severa. Adeguato management e fattori socioculturali.

BACKGROUND: Preeclampsia in its severe form is characterized by a high incidence of both maternal and fetal morbidity and mortality. Its prevention is therefore one of the fundamental goals of modern obstetrics. METHODS: Twenty-two women with severe preeclampsia have been perspectively examined and compared with 66 women without preeclampsia with the aim to evaluate the effect of the prenatal care and the influence of sociocultural variables. RESULTS: Differences between the two groups weren't observed with regard to sociocultural features, but the women with severe preeclampsia were treated later and less controlled during pregnancy. CONCLUSIONS: Preeclampsia requires a careful surveillance and an intense care, above all in its severe form, to avoid the high tribute of maternal and fetal complications that this syndrome still causes today.

[Phytoestrogen-containing food and prevention of postmenopausal osteoporosis and cardiovascular diseases]. / Fitoestrogeni alimentari e prevenzione dell'osteoporosi e della malattia cardiovascolare postmenopausale.

Food phytestrogens and prevention of postmenopausal osteoporotic and cardiovascular disease. Phytestrogens are diphenolic compounds, widely found in plants and foods, with structural and biological estrogen-like similarities. Their anti-estrogenic effects are well known and studied due to the possibility to prevent some tumors such as breast and prostate cancer. In menopause they have an estrogenic-like action on lipidic and bone metabolism. Phytestrogens rich foods can positively affect the postmenopausal osteoporotic and cardiovascular pathology.

Postmenopausal users of long-term hormonal replacement therapy: social-cultural features.

OBJECTIVE: To study of influence of social-cultural factors on the effects of long-term hormonal replacement therapy (HRT) in preventing cardiovascular disease and menopausal osteoporosis. METHOD: We examined, perspectively the social-cultural features of 394 postmenopausal women who attended the menopausal out-patient department and were eligible for long term HRT. RESULTS: The HRT users were thinner and younger, with earlier menopause, physically more active, healthier, with more oophorectomies and with more professional work. Data shows that, as a whole, a greater number of healthy women are selected or self-selected for HRT. CONCLUSION: Evaluating the results of long-term HRT in preventing cardiovascular disease and menopausal osteoporosis, the social-cultural features of HRT users must be carefully considered before any mass preventive interventions.

Spontaneous and induced release of prostaglandins, interleukin (IL)-1beta, IL-6, and tumor necrosis factor-alpha by placental tissue from normal and preeclamptic pregnancies.

PROBLEM: The objective of this study was to clarify the role of the main proinflammatory cytokines (interleukin [IL]-1, IL-6, tumor necrosis factor [TNF]-alpha) in the pathogenesis of preeclampsia and how these cytokines affect one another and the production of prostaglandins (PGs). METHOD OF STUDY: The concentrations of cytokines and PGs in supernatants of placental tissue from preeclamptic and normal women were determined by enzyme-linked immunosorbent assay. RESULTS: The concentrations of the PGs from unstimulated preeclamptic placental tissue were significantly higher compared to the concentrations of PGs from normal unstimulated placental tissue. Significant levels of IL-1beta were observed only in the supernatants of preeclamptic placental tissue. Of interest, an increase in TNF-alpha production was detected in the supernatants of IL-1-stimulated preeclamptic placental tissue. CONCLUSIONS: The overproduction of TNF-alpha may be related not only to the effect of a stimulant like IL-1beta, but mainly to the lack of mechanisms down-regulating the production of TNF-alpha.

[Breast cancer in pregnancy. Resistance to screening]. / Il carcinoma mammario in gravidanza. Resistenze allo screening.

BACKGROUND: An increasing number of women decide to become pregnant after 30 years of age, an age which begins to superimpose that of greater incidence of breast cancer, and therefore an ever-increasing number of women are destined to become pregnant during the growth of their cancers. Pregnancy associated breast cancer has a worse prognosis, but this prognosis is caused mainly from a delay in the diagnosis; for this reason the aims of this study have been to ascertain the reasons for this delay. METHODS: A sample of 500 pregnant women at term at the age of over 30 years spontaneously applied to the II Dept of Obstetrics and Gynecology at the University of Bari, were perspectively examined through a questionnaire about non-instrumental breast checks carried out during the pregnancy. RESULTS: Only the 7.4% of the women have been subjected to breast examination during pregnancy, most (67.6%) without troubles. The habit of self-examination has fallen from 28.4% to 6.2%. A greater care for cervical carcinoma has been underlined. CONCLUSIONS: Pregnancy is a moment of less attention for the physician and patient about breast cancer, and this causes a delay in the diagnosis with consequent prognostic worsening. For this reason there must be a greater interest to this problem and a different diagnostic approach.

[The multipara today: a socio-biological analysis]. / La multipara oggi. Analisi socio-biologica.

OBJECTIVE: To evaluate if and in which way work affects the choice of pregnancy and what is the outcome of pregnancy in multipara. METHODS: A retrospective study of 820 women who delivered at the Dept. of Obstetrics and Gynecology II at the University of Bari, in the period 1993-1996 was performed. RESULTS AND CONCLUSIONS: The results show that work and its problems influence the choice of having more children and that multiparity is related to a greater incidence of premature deliveries.

Smoking and menopause.

A retrospective study on 293 women who attended the Physiopathology of Human Reproduction menopausal out-patient clinic at Bari University Hospital throughout 1995, has been carried out to evaluate correlations between smoking and climacteric syndrome. From the data we have discovered an earlier onset of menopause in the smokers. Fewer smokers were married and they worked more, drank more, and were more educated. The incidence of climacteric syndrome did not differ between smokers and non-smokers.

Sexuality in the climacterium.

A retrospective study on 421 women who spontaneously attended the Menopausal Center of Physiopathology of Human Reproduction at Bari University has been carried out to evaluate the incidence of the decrease of libido in climacterium and factors that affect it. The results show a high incidence of decreased libido (46%) and a correlation with age, dyspareunia and housewife status.

[Pregnancy in women over 35 years of age. What risk?]. / La gravidanza nelle ultratrentacinquenni. Quale rischio?

A retrospective study of women who are delivered at the Dept. of Obstetrics and Gynecology at the University of Bari, in the period 1991-1994, was performed to evaluate if over 20 week pregnancies in women aged 35 years and over were at higher risk for maternal and neonatal complications than pregnancy in women aged 20 to 29 years. The results of these 4 years show that pregnancy in women aged 35 and over is burdened with a greater incidence of maternal complications and cesarean sections, but age is not a risk for neonatal outcomes. Moreover, most working women were obliged to delay the first birth child, but work does not increase the number of complications.

[Incidence of climacteric syndrome]. / Incidenza della sindrome climaterica.

A retrospective study on 300 women who attended the menopausal out-patient clinic of Physiopathology of Human Reproduction at Bari University Hospital from 1994 to 1995, was carried out to evaluate the incidence of climacteric syndrome in premenopause together with its correlations with the social and cultural environment. The results don't show differences between spontaneous and surgical menopause, and minor differences with premenopause. No relationship has been discovered between the onset of the menarche and the onset of spontaneous menopause. The hot flushes are more frequent in women with low level of learning.

Ultrasound diagnosis of heterotopic pregnancy with viable fetuses.

Combined pregnancy is an under-estimated and increasing clinical condition. Its recognition is usually made post-operatively. Ultrasound findings are not specific. Diagnosis is possible only when the cardiac activity of the extrauterine and intrauterine fetus can be detected. In the literature, 18 pre-operative sonographic demonstrations of such cases are reported. A correct echographic assessment can show a definite diagnosis or indicate a diagnostic laparoscopy minimizing maternal morbidity and mortality and improving the prognosis of the intrauterine fetus. In this report, ultrasound diagnostic aspects are discussed.

Respiratory complications during artificial barbiturate coma.

Artificial barbiturate coma was induced in 13 patients with disorders of consciousness from traumatic or spontaneous diseases. Early respiratory complications consisting with Adult Respiratory Distress Syndrome (ARDS) were found in 10 out of these 13 patients; on the contrary, only 2 out of 13 patients, treated with routine intensive care therapy without barbiturate, showed analogous respiratory complications. The literature on the subject and the possible pathophysiological mechanism of respiratory distress are discussed.