Laparoscopic removal of a giant paratubal cyst complicated by hydronephrosis.

Paratubal cysts represent approximately 10% of all adnexal masses. In most cases they are very small, but very few cases are reported in the literature where they exceed 15 cm of diameter. Furthermore, giant paratubal cysts complicated by bilateral hydronephrosis are unique. The Authors describe a case of a huge paratubal cyst (30 cm in diameter), in a 14 year old obese girl, treated by complete laparoscopic enucleation.

Beta-Thalassemia major and pregnancy.

OBJECTIVE: It was studied the clinical management and the medical outcomes of 6 pregnancies in 5 women affected by Beta Thalassemia major, based on last guidelines and pharmacological treatments. BACKGROUND: Paediatric Department and Department of Obstetrics and Gynaecology of the University of Catania. METHODS: These patients were taken among a group of 116 women affected by beta-thalassemia major divided into three subgroups, according to the characteristics of their menstrual cycle: 1) women with primitive amenorrhoea, 2) women with secondary amenorrhoea and 3) women with normal menstruation. Only one woman, affected by primitive amenorrhoea, needed the induction of ovulation. An accurate and detailed pre-pregnancy assessment was effected before each conception. This was constituted by a series of essays, including checks for diabetes and hypothyroidism, for B and C hepatitis and for blood group antibodies. Moreover were evaluated: cardiac function, rubella immunity and transaminases. Other pregnancy monitoring, and cares during labour and delivery were effected according to usual obstetrics practice. RESULTS: All the women were in labour when they were 38 week pregnant, and the outcome were six healthy babies born at term. There were no complications related to the pregnancy and to the immediate outcome after delivery. CONCLUSIONS: The improvements of current treatments, especially in the management of iron deposits, the prolongation of survival rate, will result in a continuous increase of pregnancies in thalassemic women. Pregnancy is now a real possibility for women affected by such disease. Although numerous complications can occur, vigilant monitoring by both experienced obstetricians and hematologists can lead to successful pregnancy outcomes (Tab. 1, Fig. 1, Ref. 16).

CD34 enrichment of umbilical cord blood.

OBJECTIVE: We describe the relation between the clamping time and blood volume collected, plus two enrichment systems for CD34+ stem cells from umbilical cord blood, to achieve an excellent recovery of cells with a high proliferative ability and bone marrow reconstitution. METHOD: After cord blood collection by an obstetrician, stem cell purification has been performed either with a combination of monoclonal antibodies, using the negative selection Stem Sep method, or with a positive cell selection based on their surface CD34 antigens, using the Mini Macs system. RESULTS: An excellent recovery of hematopoietic progenitors: Burst Forming Unit Erythroid; Colony Forming Unit Granulocyte and Macrophage; and Colony Forming Unit Granulocyte, Erythroid, Monocyte and Macrophage, inversely related to the rising of clamping time, has been achieved with the Mini Macs system (54 % of colonies, with 90 % purity). With the Stem Sep method, recovery of hematopoietic progenitors was 35 % (with 80 % purity). CONCLUSIONS: By an early clamping of umbilical cord blood, we obtained a higher number of CD34+ cells whose clonogenic activity increased with enrichment. This is particularly useful considering that the number of CD34+ stem cells contained in a unit of placental blood is enough for transplantation to a child, but not for an adult engraftment. Thus, using these methods, we can obtain a larger number of CD34+ stem cells, which increases the possibility of reducing graft-versus-host disease in adult patients, producing survival rates similar to those of transplantation of bone marrow from unrelated donors (Ref. 5).

Protection of ovarian tissue from radiotherapy.

Advances researches in the diagnosis and treatment of childhood, adolescent and adult cancer have greatly increased the life expectancy of premenopausal women with cancer. However, one of the serious side effects of these treatments is the risk of damage to fertility. The ovaries are very sensitive to cytotoxic and radiotherapeutic treatment. The only established method of fertility preservation is embryo cryopreservation according to the Ethics Committee of the American Society for Reproductive Medicine (2005), but this option requires the patient to be of pubertal age, have a partner or use donor sperm, and be able to undergo a cycle of ovarian stimulation, which is not possible when the radiotherapy has to be initiated immediately or when stimulation is contraindicated according to the type of cancer. For patients who need immediate radiotherapy, cryopreservation of ovarian tissue is the only possible alternative. This manuscript reports the different techniques of cryopreservation and the results of transplantation of cryopreserved ovarian tissue. The current techniques allow cryopreservation of human ovarian fragments for a long time with good follicular survival rate after thawing. Numerous studies ultimately in this field have demonstrated to improve the survival rate of the oocytes and cryopreserved follicles. Moreover this manuscript includes a case of a 17-year-old girl who had to undergo pelvic irradiation for non-Hodgkin's lymphoma and the laparoscopic treatment to preserve the fertility (Fig. 2, Ref. 47).

[Innovative genetic, clinical and endocrinologic aspects in young females affected by premature ovarian insufficiency]. / Innovativi aspetti genetici, clinici ed endocrini in una giovane popolazione femminile affetta da insufficienza ovarica prematura.

AIM: Heterogeneity premature ovarian insufficiency (POI) is one of the reasons why there are different causes that contribute in determining this type of hormonal disorder. Although the causes have already been established for many types of premature ovarian failure, are still uncertain causes in most cases of idiopathic forms, despite the description of several candidate genes, including BMP-15 gene. The gene under study is precisely the BMP-15, which is part of the superfamily of Transforming Growth Factors-beta or the TGF-ß, which also belong to the growth differentiation factors (GDFs). METHODS: This study examined a sample of Sicilian women suffering from POI, carefully selected according to their age, since in these cases, the genetic factor probably has a greater impact. RESULTS AND CONCLUSION: Identify a mutant gene that causes ovarian failure may be important to make a diagnosis that can predict the possible future development of the disease. The outcome of the studies, however, has not found the gene in question, but it is hypothesized that this may be a direct consequence of the limited amount of women that was done the study, a case which may be rebutted by increasing the number of patients.

Evaluation of bone mass in young amenorrheic women with anorexia nervosa.

BACKGROUND: In the last 10 years an impressive increase in the frequency of food disorders (bulimia and anorexia) in teenagers has been recorded. Food disorders, present especially in girls, (part of more complex relational-familiar disorders) cause progressive repercussions on the general endocrine structure and in particular on the reproductive system of the subject. The most visible effect of this endocrine perturbation is the block of the ovarian function with secondary amenorrhoea and low levels of gonadotropins and estrogens due to the food disorder. In fact the qualitative and quantitative food deficit is responsible for the interruption of the normal hypothalamic and pituitary pulses regulating the correct monthly process of follicular growth. As in the physiological postmenopausal period, very low plasma levels of 17-b-estradiol (<30 pg/ml) can start the bone resorption with bone loss and/or osteoporosis related to the amount and the time of exposure to low levels of estrogens. OBJECTIVES: in our study we evaluate the influence of anorexia and amenorrhoea on bone mineral density. METHODS: We evaluate bone mineral density (BMD) with computerised bone mineralometry (MOC), a dual-energy-X-Ray (DEXA), in 18 women with anorexia nervosa and weight loss, 9-12 months amenorrhoea and with low plasma levels of 17-b-estradiol. The BMD was measured at the lumbar spine (L2-L4) and on total body in all subjects enrolled for this study. RESULTS: All the patients showed a decrease on the lumbar BMD with bone loss (49%) or osteoporosis (51%). On the other side the values of total body BMD were around the normal levels (86%) or showed bone loss (14%). CONCLUSIONS: Our data support the hypothesis that lower estrogenic levels associated with anorexia nervosa have an impact on BMD, especially at lumbar level, with early incidence for quantitative and qualitative lower food intake.

Uterine bleeding pattern during low dosage Noretisterone acetate and 17-b-Estradiol treatment in postmenopausal patients.

BACKGROUND: Recent years have been characterized by progressive optimization of postmenopausal hormonal replacement therapy. More physiological therapeutic protocols have been, in fact, proposed to control the possible symptomatology and to prevent the associated risks, with estro-progestinic compounds characterized by lower effective dosages and suitable for the single patient need. However this therapy is not widely accepted by the women from our country for the fears and the inconvenience raised around such side effects as abnormal uterine bleeding and spotting. AIM: to obtain a good compliance and clinical benefits a continuous administration protocol of the hormonal replacement therapy, alternatively to the sequential one has been proposed. METHODS: Our research group has been observing a sample of 42 patients for 12 months, taking oral 17-b-estradiol 1 mg/noretisterone 0.5 mg in continuous administration. All of them were aged from 42 to 63 years and had been in symptomatic menopause for at least 3 months. The characteristics, the onset and the trend of vaginal bleeding were registered in appropriate monthly diaries. Endometrial thickness was evaluated by transvaginal sonography before starting the administration, not exceeding 4 mm in all the women considered. RESULTS: The incidence of bleeding (calculated as a percent of women who experienced a vaginal bleeding for al least a day during a menstrual cycle) was from 26% to 32% in the 1st trimester, reducing during the following months. At 6 months of therapy only 5% of women reported evident vaginal bleeding; at 12 months 90% of women complained with absence of bleeding or spotting. At 12 months no women showed an endometrial thickness over 6 mm. CONCLUSIONS: This observational study suggests that the majority of treated patients proved to be positively responsive to the treatment and that the 17-b-estradiol 1 mg/noretisterone 0.5 mg association reduces the incidence of bleeding and spotting with a sufficient endometrial protection from hyperplasia.

Comparison of dexa and ultrasonometry in the measurement of bone density.

The aim of our study was to evaluate the diagnostic agreement between quantitative sonography of the calcaneum (QUS) and dual energy X-ray absorptiometry (DEXA) of the spine and femur. 153 women enrolled in our study and were divided in three groups. Group A was composed of women aged between 45 and 55, Group B of women of 56-66 and Group C of women 67-77. Mean height cm 164+/-2.8; mean weight kg 68+/-3.2. The most concordant results were obtained in group B. This suggests that QUS screening for osteoporosis may be suitable for the "younger" perimenopausal patient.

Umbilical cord blood collection in Cesarean section: a comparison before and after placental delivery.

Fetal stem cells transplants depend on nucleated cells from fetal blood. This study was a prospective randomized trials to compare the collection of fetal blood by gravity into a bag containing anticoagulant, before and after delivery of the placenta. The obstetric and the newborn characteristics in the two group were not significantly different. The mean volume of fetal blood collected while the placental was still in utero was 74.93+/-7.1 ml as against 35.78+/-3.6 ml for collection of fetal blood after delivery of the placenta.

Influence of the kind of delivery on umbilical cord blood collection.

From December 1999 to February 2001, the Microbiological and Gynaecological Science Department of the University of Catania has been involved in the collection of umbilical cord blood samples. Eight hundred and sixty-three cord blood units were collected and sent to Sciacca's UCB bank. Among them, 429 were collected from newborns delivered vaginally, while the remaining were collected from Caesarean sections. The aim of this study was to evaluate the difference between umbilical cord blood samples collected during a vaginal delivery and those from a Caesarean section. In particular, the blood volume collected and cord blood CD34+ stem cell count were considered. The method of blood collection consisted of puncturing the umbilical cord vein with an 18-gauge needle and withdrawing the blood into a sterile bag immediately after clamping and newborn assistance. The blood was collected when the placenta was still in utero and the indication to the kind of delivery was, independently of the study, according to obstetrical good practice. The results of the analysis showed that the cord blood volume and the number of CD34+ cells collected were similar for the two groups. The higher median volume of blood collected from infants delivered by Caesarean section seems mainly due to the different clamping time, rather than to the kind of delivery.

Two CD34+ stem cells from umbilical cord blood enrichment methods.

The authors describe the relation between clamping time and blood volume collected, and two enrichment systems of CD34+ stem cells from umbilical cord blood, to determine an excellent recovery with high proliferate ability and bone marrow reconstitution. After an obstetrician-based cord blood collection, the purification of stem cells was performed either with a combination of monoclonal antibodies (negative selections) using the Stem Sep method, or with a positive cells selection thanks to their surface CD34 antigens, using the Mini Macs system. An excellent recovery of hematopoietic progenitors, burst-forming unit erythroid, colony-forming unit granulocyte and macrophage, and colony-forming unit granulocyte, erythroid, monocyte, and macrophage, inversely related to the rising of clamping time, was performed with the Mini Macs system (54% of colonies, with a 90% purity), while with Stem Sep method, hematopoietic progenitor recovery was 35% (with an 80% purity). By applying early clamping of the umbilical cord blood a greater number of CD34+ cells was obtained and their clonogenic activity increased with enrichment. This is particularly useful, considering that the number of CD34+ stem cells contained in a unit of placental blood is enough for transplanting to a child, but not for an adult engraftment. Thus, using this method, a larger number of CD34+ stem cells can be obtained, which increases the possibility to reduce graft versus host disease also in adult patients, producing survival rates similar to the ones obtained with transplantation of bone marrow from unrelated donors.

Influence of sport and low animal saturated fats intake diet on lipid dismetabolisms in postmenopausal women.

BACKGROUND: An important aetiopathogenetic factor leading to menopausal age pathologies (such as hypertension, diabetes, cardiovascular diseases and so on) is certainly represented by the worsening of lipid dismetabolism. This condition is generally characterized by an increase in total mean cholesterol levels, LDL-cholesterol (low density lipoprotein-cholesterol), VLDL-cholesterol (very low density lipoprotein-cholesterol), triglycerides, with reduction of HDL-cholesterol (high density lipoprotein-cholesterol) mean levels. All these metabolic modifications are triggered or increased by the hypoestrogenemia typical of postmenopausal age. The aim of our study was to evaluate the influence on postmenopausal women's lipid metabolism of a low contribution of animal saturated fat diets, associated with physical training. METHODS: We enrolled in our study 36 mild hypercholesterolemic postmenopausal women (non smokers and not complaining of hypertension and diabetes) and we put them through aerobic training, consisting of 1 hour of a light continuous run, repeated 4 times per week. The final control of their metabolic status and of their lipid order was executed after 6 months, but during this period, all the women were observed bi-weekly or monthly, to assure a high uniform compliance in the group. None of the women enrolled took specific drugs for the lipid metabolism during the study. RESULTS: After 6 months from the beginning of the diet, with associated physical training, an important weight loss was observed. Increases of HDL-cholesterol mean levels (p<0.05) associated with a decrease of total cholesterol mean levels (p<0.05) were reported in all the subject examined. A statistically non-significant decrease in LDL and VLDL cholesterol mean levels was also noticed. CONCLUSIONS: A mild lipid dysmetabolism in postmenopause may be corrected advantageously by an opportune diet associated with moderate, but constant, physical training.

VLA-2 and VLA-5 cell adhesion molecules expression in CD34+ cells from umbilical cord blood and from bone marrow.

BACKGROUND/AIMS: Umbilical cord blood contains a large number of early hematopoietic cells with high proliferating capacity, that has been used as an alternative to bone marrow transplantation. The aim of this study is to investigate the number of two cell adhesion molecules in cord blood and in bone marrow. METHODS: We investigated two integrins, named VLA-2 and VLA-5 (Very Late Appearing Antigen), expressed in the surface of CD34+ cells. The CD34+ cells, isolated with MACS CD34+ isolation kit, were labelled with the appropriate monoclonal antibodies. RESULTS: Cell adhesion molecules showed highly expressed in both cord blood and bone marrow CD34+ cells. CONCLUSION: There are no significant differences between the two sources of CD34+ populations.

Differences in umbilical cord blood units collected during cesarean section, before or after the delivery of the placenta.

Umbilical cord blood is largely employed as an alternative source of stem cells in the treatment of hemato-oncological diseases. Current results show that the success rate of purified umbilical cord blood engraftment is comparable to that obtained using bone marrow, and it is directly related to the number of pluripotent stem cells transplanted. The technique of fetal blood collection varies among different umbilical cord blood banks. Many authors collect umbilical cord blood during vaginal delivery, after placental detachment, while others collect it while the placenta is still within the uterus. In a previous randomized trial, we showed a greater collection of umbilical cord blood before placental detachment during vaginal delivery. The present study was performed to determine whether umbilical cord blood collection before placental detachment (group A) during cesarean section is superior to that after placental delivery (group B) puncturing the umbilical vein once and using a closed bag system. To accomplish this, 47 pregnant women subjected to cesarean section were enrolled in the study. Twenty-one of them were allocated to group A, while the remaining 26 formed group B. The volume of umbilical cord blood collected from the patients of group A was greater than that collected from patients of group B. The cord blood volume collected was 90.7 +/- 6.0 versus 60.9 +/- 13.7 ml; the cord blood nucleated cell number was 10.1 +/- 1.2 x 10(8) vs. 7.1 +/- 0.8 x 10(8); and the mean cord blood CD34+ cell number was 20.0 +/- 6.0 x 10(5) vs. 16.4 +/- 2.4 x 10(5), respectively.

Two enrichment methods to obtain CD34+ stem cells from umbilical cord blood.

We describe the relation between umbilical cord clamping time and two different enrichment system of CD34+ stem cells from umbilical cord blood with the proliferative ability and bone marrow reconstitution of the stem cells obtained. After an obstetrician performed the cord blood collection, the purification of stem cells was performed either with a combination of monoclonal antibodies (negative selections) using the Stem Sep method, or with a positive cells selection based on their surface CD34 antigens using the Mini Macs system. An excellent recovery of haematopoietic progenitors (Burst Forming Unit Erythroids (BFUE); Colony Forming Unit Granulocytes and Macrophages (CFU-GM); and Colony Forming Unit Granulocytes, Erythroids, Monocytes and Macrophages (CFU-GME)), inversely related to the increase in clamping time, was performed with the Mini Macs system (54% of colonies, with a 90% purity). With Stem Sep method, haematopoietic progenitors recovery was 35% (with a 80% purity). By applying early clamping of umbilical cord blood we obtained a greater number of CD34+ cells and their clonogenic activity was increased with enrichment. This is a useful technique considering that the number of CD34+ stem cells usually contained from a unit of placental blood is enough for the transplant to a child, but not for an adult. Thus, using these methods, we can get a larger number of CD34+ stem cells which reduces the risk of Graft versus Host Disease also in adult patients, producing survival rates similar to the those obtained with transplantation of bone marrow from unrelated donors.

Divergent effects of corticotropin releasing hormone on endothelial cell nitric oxide synthase are associated with different expression of CRH type 1 and 2 receptors.

1. Endothelium is a target for an array of factors involved in inflammation. Endothelial cells express receptors for CRH, a neuropeptide produced during inflammation. We report both the concentration-dependent inhibitory effect of CRH upon cytokine-stimulated nitrite release by H5V murine endothelioma cells, and its stimulatory one in HUVEC cells. 2. Western blot analysis showed that CRH inhibits cytokine-stimulated iNOS protein in H5V cells, and, instead, potentiated it in HUVEC cells. 3. H5V cells expressed both CRH receptors (CRH-R1 and R2) mRNAs, whereas HUVEC cells expressed the CRH-R2 mRNA solely. 4. CRH increased medium nitrites and iNOS protein expression in H5V cells pretreated with the selective CRH-R1 antagonist CP 154,526. However, the selective CRH-R2 antagonist anti-Svg-30 failed to produce similar effects. In fact, anti-Svg-30 inhibited CRH-induced increase of nitrite release and iNOS expression in HUVEC cells. 5. Our results confirm the activating role of CRH on endothelial cells, although it suggests its possible inhibitory role in the late phase of the inflammatory response. NO-mediated effects of CRH on endothelial cells could be exploited in therapeutic strategies related to inflammatory and/or degenerative diseases.

Adenomyosis and uterus rupture during labor.

A spontaneous uterus rupture occurred during the labour of a 37-week-pregnant woman showing a precocious rupture of membranes. It appeared enlarged and the cut surface of the myometrium showed coarse trabeculations. The histological examination showed a hypertrophic gravidic myometrium with heterogeneous areas of fibrosis and adenomyosis, necrotic decidual foci and hyperplastic cervical canal glands. Our experience suggests that a silent and spontaneous uterine rupture, is possible even in the absence of the principal risk factors.

High sperm aneuploidy rate in unselected infertile patients and its relationship with intracytoplasmic sperm injection outcome.

BACKGROUND: Men with oligoasthenoteratozoospermia (OAT) frequently undergo intracytoplasmic sperm injection (ICSI) as a treatment for their infertility. However, there is an increased risk of transmitting chromosomal abnormalities to the offspring given that natural selection is bypassed by the use of this technique and patients have an increased rate of sperm aneuploidy which, in addition, may negatively affect ICSI outcome. For this reason, the rate of sperm aneuploidy in unselected patients undergoing ICSI and its impact on ICSI performance have been evaluated. METHODS: Aneuploidy and diploidy were evaluated in spermatozoa separated by swim-up for oocyte injection, using DNA probes for chromosomes 8, 12, 18, X and Y. RESULTS: ICSI patients had sperm aneuploidy and diploidy rates significantly higher than those of 13 normozoospermic men who served as controls. Although the total aneuploidy rate varied considerably between the 18 patients, 15 of them (83%) had values above the upper range of the control group. Eighteen ICSI cycles were performed with an overall fertilization rate of 95% and a pregnancy rate of 39%. The aneuploidy rate of the 11 patients whose wives did not achieve pregnancy was slightly higher than that of pregnant couples, but the difference did not reach statistical significance. However, 10 patients in this group (91%) had a sperm aneuploidy rate well above the upper limit of the controls as compared with two patients in the "pregnant" group (29%). CONCLUSIONS: This study has shown that unselected patients undergoing ICSI had an elevated sperm aneuploidy rate. Lack of pregnancy was associated with a tendency towards an increased aneuploidy rate; however, pregnancy occurred even in the presence of an elevated sperm aneuploidy rate.

Endocrine and metabolic effects of insulin sensitizers in the treatment of patients with polycystic ovary syndrome and hyperinsulinaemia.

We evaluated 16 women with polycystic ovary syndrome (PCOS) and 67 controls, aged 18-34 years. All subjects had an oral glucose tolerance test (OGTT), showing a normal response. They were subdivided into 2 groups (normoinsulinaemic, hyperinsulinaemic) undergoing 7 days of octreotide treatment. This caused modifications in body mass index, waist-hip ratio and blood pressure. No modifications were observed in basal luteinizing hormone (LH), follicle-stimulating hormone, androgens, 17alpha-hydroxyprogesterone, oestradiol, sex-hormone-binding globulin (SHBG), prolactin and cortisol serum concentrations in normoinsulinaemic patients. A significant decrease in the concentration of LH, androgens, and a significant increase in SHBG were observed in the hyperinsulinaemic patients. In hyperinsulinaemic patients, the treatment restored a regular insulinaemic response to an OGTT. No modifications of glycaemic response were detected in normoinsulinaemic ones. In hyperinsulinaemic patients, a decompensation of the glycaemic response was observed. Several agents (insulin sensitizers) decrease insulin secretion and androgen concentration. They could be considered an approachable long-term therapy for hyperinsulinaemic hyperandrogenic patients. In the next few years the pharmacotherapy for PCOS could be greatly expanded, allowing some patients, such as hyperinsulinaemic ones, to be treated with insulin sensitizers.