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Tommaso Falcone

Publications and source records attributed to Tommaso Falcone.

At least 19 recordsLinked to original sources
PubMed2006

Cryopreservation of intact human ovary with its vascular pedicle.

BACKGROUND: The aim of this study was to assess the immediate post-thawing injury to the human ovary that was cryopreserved either as a whole with its vascular pedicle or as ovarian cortical strips. MATERIALS AND METHODS: Bilateral oophorectomy was performed in two women (46 and 44 years old) undergoing vaginal hysterectomy and laparoscopic hysterectomy, respectively. Both women agreed to donate their ovaries for experimental research. In both patients, one of the harvested ovaries was sectioned and cryopreserved (by slow freezing) as ovarian cortical strips of 1.0 x 1.0 x 5.0 mm(3) each. The other ovary was cryopreserved intact with its vascular pedicle. After thawing 7 days later, follicular viability, histology, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-digoxigenin nick-end labelling (TUNEL) assay (to detect apoptosis) and immunoperoxidase staining (to define Bcl-2 and p53 protein expression profiles) of the ovarian tissue were performed. Tissues from non-cryopreserved ovaries served as control specimens (two cases). RESULTS: The overall viability of the primordial follicles was 75 and 78% in intact cryopreserved-thawed (C-T) ovaries and 81 and 83% in ovarian cortical strips in the 46- and 44-year-old patients, respectively. Comparable primordial follicle counts, absence of features of necrosis, mean values of apoptosis and weak Bcl-2 and p53 protein expressions were observed both in the intact C-T ovary and in the C-T ovarian cortical strips. CONCLUSIONS: Cryoperfusion and cryopreservation of entire human ovary can be achieved with the maintenance of excellent viability of the superficial and the deeper tissues using a slow-freezing protocol. Cryopreservation injury is associated neither with significant alteration in the expression pattern of Bcl-2 and p53 proteins in the ovarian tissues nor with significant follicular damage.

Adult↗
PubMed2006

Tumour necrosis factor-alpha blockers: potential limitations in the management of advanced endometriosis? A case report.

Several studies have shown that tumour necrosis factor (TNF)-alpha levels are increased in the peritoneal fluid of women with endometriosis, with correlation between TNF-alpha concentrations and the degree of disease. It is also likely that elevation of peritoneal fluids' TNF-alpha levels may play a role in the pathogenesis of infertility associated with endometriosis. Use of drugs such as etanercept, a TNF-alpha receptor immunoglobulin fusion protein which inhibits TNF-alpha activity, showed in an animal study to reduce the severity of the disease, and the size of endometriotic foci. TNF-alpha blockers were recommended as a possible new line of therapy for endometriosis. Our case involved a 35-year-old Para 0, with rheumatic arthritis and stage 4 endometriosis. After 6 years of constant use of etanercept, she showed no improvement of endometriosis as demonstrated at laparoscopy. However, she underwent a successful IVF after the first attempt. TNF-alpha-blocker medications might not be beneficial for patients with advanced endometriosis. However, we cannot exclude the possible effect of these medications on early-stage endometriosis, and further study is required. Some of the immunologic abnormalities in the pelvis of patients with endometriosis could be the consequence of the disease and not the cause, and possibly suppression of immune cells and their products may not have a major effect on endometriotic lesions at an advanced stage. This also could explain why suppression of TNF-alpha showed no effect on infertility. However, use of TNF-alpha-blockers before IVF might increase the success rate in advanced endometriosis.

Adult↗
PubMed2006

Role of total antioxidant capacity in the differential growth of human embryos in vitro.

OBJECTIVE: The objective of this study was to examine the relationship of early human embryonic development parameters with day 1 culture media total antioxidant levels (day 1 TAC). DESIGN: Prospective study. SETTING: Patients undergoing assisted reproduction (ART) in a teaching hospital. PATIENT(S): Patients undergoing conventional IVF (n = 153; 167 cycles) and intracytoplasmic sperm injection (ICSI; n = 105; 116 cycles) were included. Both fertilization and early cultures were performed in human tubal fluid (HTF) with 5% serum substitute supplement. INTERVENTION(S): Levels of total antioxidants in the central well (sample) and the outer well (control) of each embryo culture dish were measured. MAIN OUTCOME MEASURE(S): Fertilization rate and embryo quality at days 3 and 5 were recorded for each cycle. Age, parity, and demographic features were also compared. RESULT(S): After controlling for all demographic and clinical variables, day 1 TAC was related to fertilization rates in both groups of patients. Day 1 TAC was significantly related to high (>7) day 3 cell number, low (<10%) fragmentation rate, and blastocyst development rate in both conventional IVF and ICSI cycles. Day 1 TAC was related to pregnancy in ICSI but not conventional IVF cycles. CONCLUSION(S): Day1 TAC appears to be an important biochemical marker for early embryonic growth. Decreased embryonic fragmentation, enhanced cleavage rate, and increased blastocyst development rate may be partially related to day 1 TAC in the culture media. Whether this relationship is a cause or effect needs further assessment.

Adult↗
PubMed2006

Analysis of apoptotic cell death, Bcl-2, and p53 protein expression in freshly fixed and cryopreserved ovarian tissue after exposure to warm ischemia.

OBJECTIVE: To evaluate the effect of ischemia time on the expression of Bcl-2 and p53 proteins on freshly fixed and cryopreserved-thawed ovarian tissue. DESIGN: Experimental study using porcine animal model. SETTING: Biological Resources Unit, Cleveland Clinic Foundation. ANIMAL(S): Eight nonpregnant adult sows. INTERVENTION(S): Bilateral oophorectomy was performed in eight sows and the ovaries were subjected to time-dependent (1, 10, 20, and 30 min) warm ischemia (room temperature). Each specimen was divided into two parts, One was fixed as a fresh tissue (freshly fixed) and the other was subjected to cryopreservation, thawing, and fixation (cryopreserved). MAIN OUTCOME MEASURE(S): Apoptosis (TUNEL assay) and Bcl-2 and p53 protein expression (immunoperoxidase method) were assessed. RESULT(S): At 1, 10, 20, 30 min of warm ischemia the apoptotic indices were: 1) statistically significantly higher in the atretic (1.36 +/- 0.20, 1.59 +/- 0.20, 1.67 +/- 0.22, and 1.67 +/- 0.24, respectively) than in the nonatretic follicles (0.69 +/- 0.06, 0.69 +/- 0.06, 0.76 +/-, and 0.06, 0.71 +/- 0.06, respectively; P<.05); 2) not significantly different between freshly fixed and cryopreserved tissues; and 3) nonsignificantly higher with the increased duration of ischemia. Bcl-2 expression was seen in the granulosa but not in the theca cells of most of the healthy and occasional atretic follicles. p53 expression was seen only in few atretic follicles. Increased duration of ischemia was associated with insignificant incremental rise of the number of follicles with Bcl-2 expression (1.82 +/- 0.30, 2.01 +/- 0.44, 2.02 +/- and 0.35, 2.05 +/- 0.42 for healthy follicles at 1, 10, 20, and 30 min, respectively; P=.99). CONCLUSION(S): (1) Apoptosis is involved in follicular atresia; (2) Bcl-2 is induced by warm ischemia; and (3) cryopreservation insult does not alter the apoptotic signals with short tissue preparation time.

Animals↗
PubMed2006

Genetic polymorphism in the fibrinolytic system and endometriosis.

OBJECTIVE: Although most women experience retrograde menses during their reproductive life, endometriosis develops only in a small percentage. We hypothesized that persistence of a fibrin matrix in peritoneal pockets, as a result of hypofibrinolysis, could allow menstrually deposited endometrial fragments to initiate endometriosis. Fibrinolysis is modulated by several factors, and polymorphisms in the plasminogen activator inhibitor-1 (PAI-1) gene are considered to be one of the important determinants. The objective of this study was to evaluate PAI-1 genotypes in a group of women with or without endometriosis. METHODS: In 118 women (75 with laparoscopically confirmed endometriosis and 43 controls), genomic DNA was extracted from blood and the PAI-1 promoter genotype was determined by polymerase chain reaction amplification of DNA using specific primers for the 4G or 5G allele followed by gel electrophoresis. A portion of the polymerase chain reaction product was purified and sequenced to confirm the gel electrophoresis results. RESULTS: Endometriosis was more likely in patients with 4G/5G (odds ratio 38; 95% confidence interval [CI] 6-229) or 4G/4G (odds ratio 441; 95% CI 53-3,694) compared with 5G/5G PAI-1 genotype. Fifty-two of 75 women with endometriosis (69 %, 95% CI 58-79%) had the 4G/4G genotype compared with only 5 of 43 (12%; 95% CI 4-25%) controls. In contrast, the 5G/5G genotype associated with normal fibrinolysis was found in 2 of 75 (3%; 95% CI 0-9%) women with endometriosis compared with 24 of 43 (56%; 95% CI 40-71%) controls. CONCLUSION: Hypofibrinolysis, associated with the 4G allele of the PAI-1 gene, was found significantly more often in women with endometriosis compared with controls. Persistence of fibrin matrix could support the initiation of endometriotic lesions in the peritoneal cavity, explaining why some women with retrograde menstruation develop endometriosis while others do not. LEVEL OF EVIDENCE: II-2.

Adult↗
PubMed2005

Peritoneal fluid leptin is associated with chronic pelvic pain but not infertility in endometriosis patients.

BACKGROUND: Leptin influences the proinflammatory immune responses and has angiogenic activity in vitro and in vivo. The objective of this study was to evaluate the peritoneal fluid levels of leptin in patients with endometriosis and idiopathic infertility and compare them with a control group of tubal ligation/reanastomosis patients. METHODS: In this observational, prospective controlled study, peritoneal fluid from 108 women was obtained while they underwent laparoscopy for pelvic pain, infertility, tubal ligation or sterilization reversal. We measured the concentration of leptin in the peritoneal fluid and compared the levels among women who were divided into groups according to their post-surgical diagnosis. Sixty patients were diagnosed with endometriosis, 10 with idiopathic infertility and 38 had undergone tubal ligation or reanastomosis (control group). RESULTS: Peritoneal fluid leptin was significantly higher in endometriosis 14.62+/-9.79 (mean+/-SD) ng/ml compared to idiopathic infertility [0.92+/-1.57 ng/ml (P=0.0007)] and to controls [0.78+/-1.94 ng/ml (P<0.0001)]. Leptin levels were positively correlated with the stage of endometriosis (r=0.45; P=0.03), and with pelvic pain in endometriosis patients (r=0.49; P=0.001). Peritoneal fluid leptin levels in patients with idiopathic infertility were comparable to controls. CONCLUSIONS: Higher levels of leptin were observed in peritoneal fluid of patients with endometriosis compared to those without the disease. These data suggest that the proinflammatory and neoangiogenic action of leptin may contribute to the pathogenesis of endometriosis. Moreover, leptin may play a role in endometriosis-associated pain.

Ascitic Fluid↗
PubMed2005

Relationship between cytokines and the embryotoxicity of hydrosalpingeal fluid.

OBJECTIVE: The exact chemical composition of hydrosalpingeal fluid is unknown. The objective of this study was to characterize cytokines in hydrosalpingeal fluid (HSF) and examine their possible role in the embryo development. STUDY DESIGN: HSF was aspirated at laparoscopic salpingectomy in eight infertile women. Levels of IL-1beta, IL-13, IL-8, IL-6 and TNF-alpha in the HSF were determined by quantitative immunoassay kits. Two-cell mouse embryos were incubated with 0, 25, 50 and 75% concentrations of HSF. The blastocyst development rate (BDR) of mouse embryos was measured at each HSF concentration. RESULT(S): The embryotoxicity of HSF was concentration dependent. An increase in the HSF concentration resulted in significant decrease in % BDR (p < 0.01). IL-1beta was present in six of the eight HSF samples with a mean (+/-SD) concentration of 0.9 +/- 0.8 pg/mL. IL-13 was not detected in any of the HSF samples. IL-8, IL-6 and TNF-alpha were detected in all samples with a mean (+/-SD) concentration of 4741.2 +/- 6554.4 pg/mL, 204.8+/-132.8 pg/ml and 12 +/- 12.8 pg/mL respectively. IL-6 was positively correlated with BDR (r = 0.53; p < 0.04). CONCLUSION(S): We demonstrated for the first time the presence of IL-1beta, IL-8, IL-6 and TNF-alpha and the absence of IL-13 in human hydrosalpingeal fluid. IL-6 was positively related to the BDR.

Adult↗
PubMed2005

Effect of pentoxifylline in reducing oxidative stress-induced embryotoxicity.

PURPOSE: To 1) evaluate the embryotoxic effects of hydrogen peroxide on mouse embryo development and 2) examine if pentoxifylline can reverse hydrogen peroxide induced embryotoxicity. METHODS: Prospective in vitro study examining the effects of varying concentrations of hydrogen peroxide and pentoxifylline on the blastocyst development rate alone as well as in combination. RESULTS: A dose-dependent decrease in % BDR was seen with increasing concentrations of H2O2. High concentrations of hydrogen peroxide (> 60 microM) were embryotoxic. Pentoxifylline (500 microM) was able to reduce the embryotoxic effect of hydrogen peroxide. Percent blastocyst development rate increased from 44% in hydrogen peroxide alone to 85% in hydrogen peroxide and pentoxifylline coincubation. CONCLUSIONS: Pentoxifylline may be beneficial in reducing hydrogen peroxide induced embryo damage and improve IVF outcome. Patients with endometriosis-associated infertility may benefit from the use of pentoxifylline without significantly affecting embryo development.

Animals↗
PubMed2005

Infliximab may reverse the toxic effects induced by tumor necrosis factor alpha in human spermatozoa: an in vitro model.

OBJECTIVE: To examine the toxic effects of tumor necrosis factor alpha (TNF-alpha) on ejaculated spermatozoa and evaluate the ability of infliximab to reverse these effects. DESIGN: Prospective controlled study. SETTING: Cleveland Clinic Foundation, Cleveland, Ohio. PATIENT(S): Thirty-one healthy sperm donors. INTERVENTION(S): Exposure of human spermatozoa to varying concentrations of TNF-alpha (100, 300, 400, 500 pg/mL, and 2.5 microg/mL) and infliximab (400 microg/mL). MAIN OUTCOME MEASURE(S): Sperm motility, functional integrity of plasma membrane, and DNA fragmentation. RESULT(S): Spermatozoa quality declined following incubation with TNF-alpha in a dose-dependent and time-dependent manner. Sperm motility and membrane integrity were higher in the samples incubated with TNF-alpha plus infliximab than in the samples treated with TNF-alpha only. These parameters improved significantly and were comparable with both controls and sperm incubated with infliximab alone. Similarly, the percentage of spermatozoa with DNA fragmentation improved significantly following incubation with TNF-alpha plus infliximab and again was comparable with both controls and sperm incubated with infliximab alone. CONCLUSION(S): Spermatozoa may be exposed to abnormal levels of TNF-alpha in the male reproductive tract or during their passage into the female reproductive tract (in cases of endometriosis). Exposing spermatozoa to pathological concentrations of TNF-alpha can result in significant loss of their functional and genomic integrity. Infliximab could potentially be used to help treat female infertility caused by endometriosis in those with elevated levels of TNF-alpha in their peritoneal fluid.

Analysis of Variance↗
PubMed2005

Robotics in reproductive medicine.

OBJECTIVE: To review the history, development, current applications, and future of robotic technology. DESIGN: The MEDLINE database was reviewed for all publications on robotic technology in medicine, surgery, reproductive endocrinology, its role in surgical education, and telepresence surgery. SETTING: University medical center. CONCLUSION(S): Robotic-assisted surgery is an emerging technology, which provides an alternative to traditional surgical techniques in reproductive medicine and may have a role in surgical education and telepresence surgery.

Animals↗
PubMed2005

Effects of peritoneal fluid on preimplantation mouse embryo development and apoptosis in vitro.

To evaluate the effect of peritoneal fluid (PF) on preimplantation mouse embryo development and apoptosis, PF was obtained from women presenting with (n = 7) and without endometriosis (n = 7). Mouse embryos were cultured to the blastocyst stage in human tubal fluid medium alone (control) or 10% PF. Embryo development was assessed and the total cell number per embryo was determined by Hoechst 33258 staining. Allocation of inner cell mass and trophectoderm in blastocysts, and incidence of apoptosis were determined using confocal microscopy. The blastocyst development rate was significantly lower in the presence of 10% PF (P < 0.01). Total cell number and trophectoderm in blastocysts cultured in culture media alone was significantly higher than in the presence of PF (P = 0.034 and P = 0.01 respectively). A higher incidence of apoptosis was seen in blastocysts cultured in culture media alone, compared with those cultured in the presence of PF (P = 0.04). PF decreases the development of early mouse embryos to the blastocyst stage, as well as the incidence of apoptosis in the resulting mouse blastocysts. No difference was seen in the effect of PF from women with and without endometriosis on mouse embryo development and apoptosis.

Animals↗
PubMed2005

Fertility preservation and pregnancy outcome after malignancy.

PURPOSE OF REVIEW: The overall survival and cure rates of patients with childhood and adult malignancies have improved dramatically, but cancer treatment can be associated with diminished reproductive potential. However, research on the preservation of fertility in these patients has given patients new options. This article discusses the mechanisms of reproductive failure after cancer therapy and the currently available fertility preservation strategies. RECENT FINDINGS: Ovarian transposition is still a viable option if radiotherapy is to be used alone. Modifications in assisted reproductive technology that decrease peak estradiol levels are ideal for breast cancer survivors. Embryo freezing technology offers excellent pregnancy rates. Oocyte freezing is available for women without a partner, but there is more limited experience with this technique. Understanding the concepts of graft function after the autotransplantation of frozen-thawed ovarian tissue has resulted in great strides in the technical requirements for success. SUMMARY: Gonadotropin-releasing hormone analogues are the only available medical protection means for gonadotoxic chemotherapy. Assisted reproductive technology offers excellent results, but the protocols require a delay in implementing chemotherapy. Despite recent reports of embryo development after the transplantation of cryopreserved-thawed ovarian tissue, clinical experience is limited and the technique remains experimental.

Antineoplastic Agents↗
PubMed2005

Protein supplementation and the incidence of apoptosis and oxidative stress in mouse embryos.

OBJECTIVE: To estimate the effect of protein supplementation of culture media on reactive oxygen species production and incidence of apoptosis in preimplantation mouse embryos. METHODS: A total of 72 two-cell mouse embryos were cultured in human tubal fluid (HTF) alone (HTF-alone, control) and 71 embryos in HTF with protein supplementation (10% serum substitute supplement; HTF-SSS) for 72 hours. Total cell number per embryo was determined by staining with Hoechst 33258. Allocation of inner cell mass and trophectoderm in blastocysts and incidence of apoptosis were determined by confocal microscopy. Levels of reactive oxygen species in culture media were measured by chemiluminescence assay using luminol as probe. RESULTS: Blastocyst development, total cell number, and the inner cell mass/trophectoderm ratio were similar between the 2 groups. The blastocyst hatching rate was significantly higher in the HTF-SSS group than in the HTF-alone group (20% compared with 4%, P = .007). Level of reactive oxygen species was significantly higher in HTF-alone compared with HTF-SSS at 24 hours (median and interquartile range 28 [13, 43] compared with 0 [0, 1], P = .02), 48 hours (24 [21, 26] compared with 2 [1, 2], P = .02), and 72 hours (26 [9, 32] compared with 2 [2, 3], P = .03). The incidence of apoptosis in blastocysts cultured in HTF-SSS was significantly lower than those in HTF-alone group (mean +/- standard deviation 2.38 +/- 0.68 and 5.81 +/- 1.11, respectively, P = .001). CONCLUSION: Protein supplementation of culture media improves the hatching rate and reduces reactive oxygen species levels and the incidence of apoptosis in mouse preimplantation embryos.

Animals↗
PubMed2005

Ovarian hyperstimulation syndrome.

OBJECTIVE: Ovarian hyperstimulation syndrome (OHSS) is a rare iatrogenic complication of ovarian stimulation usually occurring during the luteal phase or during the early part of pregnancy. OHSS is a potential complication of ovarian induction by almost every agent used for ovarian stimulation. Today, due to aggressive treatment protocols including the development of in vitro fertilization and cryopreservation with the goal of obtaining sufficient numbers of oocytes and embryos, an increased risk of developing OHSS is present. OHSS is now becoming increasingly more recognized due to the higher number of women undergoing assisted reproductive techniques. DESIGN: Review of the literature regarding ovarian hyperstimulation syndrome. METHODS: A review of the epidemiology, pathophysiology, risk factors, classification, clinical features, and treatment and prevention of OHSS. CONCLUSION: OHSS can be thought of as the loss of control over the hyperstimulation of the ovaries. Although the prevalence of the severe form of OHSS is small, it is important to remember that OHSS is usually an iatrogenic complication of a nonvital treatment that has the potential for a fatal outcome. Therefore, critical care physicians play an integral part in the care of these patients and therefore should be familiar with and recognize the various clinical manifestations and potential outcomes of this entity.

Critical Care↗