Induction of ovulation in thalassemic patients.

The authors studied 19 thalassemic patients: 14 patients with secondary amenorrhea (group A), 5 patients with primary amenorrhea (group B) and 1 patient with anovulation (group C). All the patients underwent three cycles of induction of ovulation with FSH. The level of 17 beta-estradiol and the diameter of follicles were evaluated every three days. Normal gonadal function was maintained in three patients with positive response at Step 1 of the treatment.

Osteopenia in female beta-thalassemic patients.

Sixty-four women with thalassemia major aged 13-28 years were studied. 49 patients had primary amenorrhea and 15 had secondary amenorrhea. Bone density of the spine was performed using lunar DPX. The Z score was used to evaluate the degree of osteopenia. In 82% of the patients a Z score less than -2 was found. BMD correlated negatively with the duration of amenorrhea and age of thalassemic 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.

At term pregnancies in transfusion-dependent beta-thalassemic women.

The health background management and outcome of five pregnancies in women affected by Cooley Disease are described and the preconceptual guidance and care are considered. The patients were selected from a group of 103 thalassemic women divided into three subgroups according to their first and successive menstruation characteristics. Only one woman needed induction of ovulation. A precise and detailed pre-pregnancy assessment was done before each conception. All the women were in labour at 38 weeks' gestation, and five healthy babies were born at term, weighing between 2,600 and 3,200 g. The improvement in current treatments will result in a continuous increase in pregnancies in thalassemic women, making pregnancy a real eventuality for them. Furthermore, we are studying the possibility of collecting foetus umbilical cord blood after delivery, with the aim of obtaining complete marrow reconstitution in an attempt at heterologous transplantation to the mother.

Collection of fetal stem cells and newborn effects.

The aim of this work was to test fetal stem cells (FSC) number modification in relation to clamping time and newborn effect. The results show that a fast sample, between 20 and 40 seconds, from umbilical cord after fetus birth and before placental detachment assured a greater quantity of blood useful for the transplants; and that it was necessary to enrich the collected blood in CD34+ cells with specific clonogenic culture, as this is otherwise a small number for a donation to an adult. In the "new donors" the effects of the unconscious donation always depend on the clamping time, which should be the shortest possible to avoid blood overload, which is very dangerous in the presence of heart malformation.

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.

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.

Haematopoietic progenitors from umbilical cord blood.

BACKGROUND/AIM: The aim of this study is to improve the obstetrician-based cord blood collection system and an efficient recovery of CD34+ haematopoietic progenitor stem cells. METHODS: CD34+ cells were purified from total blood using a positive selection enrichment method, called Mini-Macs. RESULTS: The final yield of CD34+ cells we obtained was 10(4) cells/ml, with a CD34+ purity of 99%. CONCLUSION: Our results confirm that, by using this method, it is possible to get a significant stem cell number, thus improving transplanting both peripheral stem cells and umbilical cord ones.

Pregnancy outcome of a transfusion-dependent thalassemic woman.

A clinical case concerning a normal pregnancy outcome in a transfusion-dependent woman affected by homozygous beta thalassemia, whose partner was negative with regard to the "thalassemic trait", was reported. The patient showed no iron deposit problems, viral diseases that could have made the pregnancy management difficult or any complications during the gestation. Blood transfusion was not necessary during the following caesarean delivery. The outcome was a healthy female child, born at a gestational age of 38 weeks, showing neither malformations nor problems. This was possible due to a detailed preconceptual guidance and a pre-pregnancy assessment. The patient normally would have had a blood transfusion every 20 days and a strict desferrioxamine chelating therapy; however, this treatment was suspended during her pregnancy because of the well-recognised teratogenic effects of the drug. The average values of ferritin were just a little higher than before being pregnant. The foetus, due to her particular chelating activity, probably maintained these ferritin levels. A sample of 95 ml umbilical cord blood was taken during the delivery. It is well known that umbilical cord blood contains a good quantity of CD34+ stem cells, the haematopoietic progenitors. It was therefore collected for transplanting to the mother and for bone marrow reconstitution. Moreover, our experience suggests that desferrioxamine therapy during lactation does not alter iron excretion in breast milk. Therefore, women now affected by Cooley disease may possibly have a normal pregnancy without ovulation induction, intrauterine growth retardation, foetal loss and preterm labour.

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.

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.

Placing the newborn on the maternal abdomen increases the volume of umbilical cord blood collected.

Haematopoietic stem cell transplantation is an important therapy for certain haematological and malignant disorders. Umbilical cord blood contains a high proportion of potentially transplantable haematopoietic progenitor cells. However, the use of cord blood stem cell transplantation is limited by the low number of stem cells obtainable from a single cord blood donor. The aim of our study was to investigate the possibility that procedures during delivery might influence the number of umbilical cord blood haematopoietic progenitor cells available for transplantation. We assessed the effects of upper and lower positions of the newborn infant on the yield of cord blood stem cells in 51 vaginal deliveries. Neonates in the upper position group were placed by the midwife on the maternal abdomen immediately after birth, while those in the lower position group were placed on the delivery table, below the maternal introitus. The total volume of cord blood and the total number of CD34+ cells collected from babies in the upper position group were significantly higher than those from babies in the lower position group. There were no significant differences in cord blood haemoglobin levels and white blood cell counts between the two groups, nor were there any adverse effects in the newborn infants. The simple manoeuvre of placing the newborn on the maternal abdomen after delivery may thus increase the yield of transplantable haematopoietic progenitor cells in cord blood.

Early clamping of umbilical cord blood and foetal CD34 enrichment.

We describe the relation between clamping time and blood volume collected, and two enrichment systems of CD34+ stem cells from umbilical cord blood, in order to determine the criteria for an excellent recovery with high proliferative 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 owing to their surface CD34 antigens, using the Mini Macs system. An excellent recovery of haematopoietic progenitors--burst forming unit erythroid; colony-forming unit granulocyte and macrophage; and colony-forming unit granulocyte, erythroid, monocyte, and macrophage--inversely related to an increase in clamping time, was achieved using the Mini Macs system (54% of colonies with a 90% purity), while the recovery of haematopoietic progenitors was 35% (with a 80% purity) using the Stem Sep method. By clamping umbilical cord blood at an early stage, we obtained a greater number of CD34+ cells, 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 sufficient for transplanting into a child but not for an adult engraftment. Thus, using these methods, we obtain a larger number of CD34+ stem cells. This increases the possibility of reducing graft versus host disease in adult patients and produces survival rates similar to the ones we observed in transplantation of bone marrow from unrelated donors.

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.

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.