[Cryoconservation of ovarian tissue: indications and outcome of the patients]. / Cryoconservation du tissu ovarien: indications et devenir des patientes.

OBJECTIVES: Ovarian Tissue Cryopreservation (OTC) is a very promising approach of fertility preservation for women and young patients who have to follow gonadotoxic treatments (chemotherapy, radiotherapy…). The aim of this study was to analyse the indications and the outcomes of the patients who had OTC in our center during the last 17 years. PATIENTS AND METHODS: The study is retrospective. Forty-six patients, who underwent OTC in the Laboratory of Reproductive Biology of the University Hospital of Clermont-Ferrand, between January 1997 and December 2009, were included. RESULTS: The average age on the day of ovarian tissue harvesting was 19.5 years. Fifty-two percent of the patients were minor. In order of decreasing frequency, the diseases for which OTC has been proposed were hematologic, ovarian tumors, sarcoma ou PNET and breast neoplasia. In 93.5 %, the harvesting of ovarian cortex was performed by laparoscopy. After OTC, 82.6 % of the patients were treated by chemotherapy. A bone marrow transplant was performed for 48 % of the study patients. At the time of data collection, 57 % of the patients who had evaluation of their ovarian function presented premature ovarian failure. Eight patients had one or more pregnancies after treatment. It was a natural pregnancy for five of them. The three others were obtained by medically assisted procreation (in vitro fertilization and oocyte donation). DISCUSSION AND CONCLUSION: We report a long-term follow-up of patients treated in our center for OTC. The originality of our study is to evaluate all aspects of OTC from the decision to propose the patients an OTC to their outcomes several years after the ovarian tissue harvesting. It is therefore a multidisciplinary approach both oncology, gynecological and pediatric whereas OTC is often considered restrictively in the literature. Finally, it seems to be essential to establish a specific medical care for these patients. This monitoring will allow an adequate assessment of pubertal development and ovarian function, management of estrogen deficiency and secondary infertility, supporting patients in their desire for motherhood.

Association between BoLA-DRB3 and somatic cell count in Holstein cattle from Argentina.

Different studies have proved that the resistance/susceptibility to mastitis is genetically determined. The major histocompatibility complex in cows is known as bovine lymphocyte antigen (BoLA). Genes from the BoLA have been associated with the occurrence of infectious diseases such as mastitis and leukosis, especially the BoLA-DRB gene. The object of the present study was to detect associations between BoLA-DRB3 alleles and somatic cell count (SCC), as an indicator of resistance/susceptibility to mastitis in Holstein cattle (N = 123) from La Pampa, Argentina. Fisher's exact test and Woolf-Haldane odds ratio were applied to study the association between SCC and BoLA-DRB3 allele frequencies. Significant association was noted between BoLA-DRB3.2*23 and *27 alleles (p < 0.05) and protective or susceptibility effects, respectively. In addition, alleles BoLA-DRB3.2*20 and *25 exhibit suggestive association with high SCC (p < 0.1). These results were partially in agreement with data reported from Japanese Holstein cattle, but differed from those published by other authors. A possible explanation for the contrasting results could be that the mastitis is a multifactor disease caused by different pathogens. Moreover, most of the studies were carried out using PCR-RFLP method, which has less resolution than PCR-SBT because PCR-RFLP defined alleles included more than one sequenced alleles.

Action of prolactin, IFN-gamma, TNF-alpha and LPS on heme oxygenase-1 expression and VEGF release in human monocytes/macrophages.

The pituitary hormone prolactin (PRL) has recently been regarded as a local regulator of macrophage responses. Our goal in this study was to investigate the regulatory interaction between PRL, interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha) and lipopolysaccharide (LPS) in the heme oxygenase-1 (HO-1) expression and the vascular endothelial growth factor (VEGF) production in human monocytes/macrophages (HMMs). In vitro treatment of HMMs with PRL, IFN-gamma, TNF-alpha and LPS was found to increase both HO-1 expression and protein synthesis in a time-dependent manner. HMMs treated with PRL, IFN-gamma, TNF-alpha and LPS also showed an enhanced release of VEGF. Moreover, co-stimulation of PRL with LPS caused activation of HMMs functions, enhancement of HO-1 expression and induction of VEGF release, whereas addition of PRL inhibited up-regulation of HO-1 or VEGF induced by IFN-gamma or TNF-alpha. Our results demonstrate that PRL, IFN-gamma, TNF-alpha and LPS modulate the expression of angiogenic factors providing additional information about the regulatory mechanism, which controls the angiogenic function of macrophages.

Factors affecting bone response around loaded titanium dental implants: A literature review.

This paper was designed as a review of the knowledge concerning bone adaptation around dental implants. Current literature concerning the biomechanics of bone and titanium dental implants as a main focus and pertinent to key aspects of the review was taken into consideration. Implant stiffness, design and surface were considered as factors affecting bone response to loaded dental implants. The emerging data allows the conclusion that implant design influences force transmission to periimplant bone, but seems to have poor power in preventing time-related marginal bone crest resorption. Mechanical stimulation such as prosthetic stress-dependent strains, affects bone tissue adaptation. Therefore, it is crucial to have high biomechanical control on implants to preserve implant ankilosis and to promote periimplant bone deposition.

Prolactin increases HO-1 expression and induces VEGF production in human macrophages.

The pituitary hormone prolactin (PRL) is a multifunctional polypeptide which exerts a role on cell proliferation and may also contribute to cell differentiation. PRL is also produced by immune cells and is regarded as a key component of the neuroendocrine-immune loop and as a local regulator of macrophage response. The involvement of PRL in regulating monocyte/macrophage functions is suggested by the presence of PRL receptors in these cells. It has been shown that PRL possess both angiogenic and antiangiogenic effects. Recently, we revealed that augmentation of HO-1 activity enhances PRL-mediated angiogenesis in human endothelial cells. Since macrophages are key participants in angiogenesis our objective was to investigate the effect of PRL also in human macrophages. In vitro treatment of macrophages with PRL was found to increase both heme oxygenase-1 (HO-1) expression and protein synthesis in a time and dose dependent manner as quantified respectively by reverse-transcriptase real-time polymerase chain reaction and Western blot analysis. PRL-treated macrophages also showed an enhanced release of vascular endothelial growth factor (VEGF) as demonstrated by ELISA assay. Furthermore, to determine whether PRL-induced HO-1 activity was required for VEGF production by macrophages, the effect of PRL on the induction of VEGF was studied in the presence of an inducer stannic chloride (SnCl(2)) and of an inhibitor stannic mesoporphyrin (SnMP) of HO activity. Our observations suggest that PRL may regulate monocyte activation and influences not only immune function but also angiogenesis.

In vitro and in vivo follow-up of titanium transmucosal implants with a zirconia collar.

The advantages of transmucosal healing implants with a bioactive zirconia collar as a support for partially fixed prosthodontic restorations are optimal peri-implant marginal tissue sealing, reduction in plaque accumulation and satisfactory aesthetic results. The zirconia used in this study evidenced not only optimal clinical performances, but also good biocompatibility. The results from this study demonstrated that zirconia coating enhances fibroblasts and osteoblast-like cell adhesion, spreading and proliferation, favoring microscopic tissue/cell in-growth and clinical implant fixation improvement. From clinical analysis, it emerged that the treatment group obtained better scores in every peri-implant parameter. This evidence attests faster stabilization of soft and hard tissues around both the transmucosal zirconia collar and at the crestal level of the implant. A reduced plaque accumulation around the implant with zirconia collar could provide a better peri-implant microbiological en-vironment by allowing the soft tissues expression of optimal sealing and good bone adaptation to loading. From these clinical and radiographic comparative analyzes, it emerged that in the treatment group the mean values were always similarly low. A rapid stabilization of both hard and soft peri-implant tissues was documented in the 1st yr. In the treatment group, there was the formation of stable tissue sealing the zirconia collar, which could preserve mucosal and bone levels. In conclusion, 2-yr clin-ical results demonstrated that implants supporting fixed restorations using transmucosal healing implants with a zirconia collar appeared a valid method, reporting 100% implant survival rates. Moreover, in vivo results obtained using strict parame-ters to assess the peri-implant status affirmed that a zirconia collar offers excellent biological acceptance. Our preliminary in vitro results statistically evidenced increased fibroblast and osteoblast adhesion and proliferation to zirconia compared to tita-nium, and an index of enhanced material integration with bone and soft tissue cells. (Journal of Applied Biomaterials & Biomechanics 2004; 2: 143-50).

Early events of secretory granule formation in the rat parotid acinar cell under the influence of isoproterenol. An ultrastructural and lectin cytochemical study.

The events involved in the maturation process of acinar secretory granules of rat parotid gland were investigated ultrastructurally and cytochemically by using a battery of four lectins [Triticum vulgaris agglutinin (WGA), Ulex europaeus agglutinin I (UEA-I), Glycine max agglutinin (SBA), Arachys hypogaea agglutinin (PNA)]. In order to facilitate the study, parotid glands were chronically stimulated with isoproterenol to induce secretion. Specimens were embedded in the Lowicryl K4M resin. The trans-Golgi network (TGN) derived secretory granules, which we refer to as immature secretory granules, were found to be intermediate structures in the biogenesis process of the secretory granules in the rat parotid acinar cell. These early structures do not seem to be the immediate precursor of the mature secretory granules: in fact, a subsequent interaction process between these early immature granule forms and TGN elements seems to occur, leading, finally, to the mature granules. These findings could explain the origin of the polymorphic subpopulations of the secretory granules in the normal acinar cells of the rat parotid gland. The lectin staining patterns were characteristic of each lectin. Immature and mature secretory granules were labelled with WGA, SBA, PNA, and lightly with UEA-I. Cis and intermediate cisternae of the Golgi apparatus were labelled with WGA, and trans cisternae with WGA and SBA.

Lectin binding sites in parotid acinar secretory granules of normal and isoproterenol treated rat.

Lectin staining patterns in secretory granules of rat parotid gland acinar cell of untreated and isoproterenol-injected animals were examined by electron microscopy. We used four lectin-gold complexes: Ulex europaeus agglutinin I (UEA-I), Helix pomatia agglutinin (HPA), wheat germ agglutinin (WGA), Glycine max agglutinin (SBA). Specimens were low temperature embedded in the hydrophilic Lowicryl K4M resin. The normal acinar cells produced glycoconjugates which were positive for all of the lectins used and with a characteristic topographic distribution in relation to the morphological type of granule. The cells of isoproterenol-treated rat showed marked ultrastructural changes in the size and structure of granules; significant changes in lectin binding sites in the granules were also observed.

Antibodies to beta2-glycoprotein I: a potential marker for clinical features of antiphospholipid antibody syndrome in patients with systemic lupus erythematosus.

OBJECTIVE: To clarify risk factors for the development of clinical features of antiphospholipid syndrome (APS) in patients with anticardiolipin antibodies (aCL) in systemic lupus erythematosus (SLE). METHODS: We studied 65 SLE patients, all with positive IgG and/or IgM aCL. Patients were divided into 2 groups; I: 29 SLE patients with features of APS (SLE/APS) and II: 36 aCL positive SLE patients without any feature of APS (SLE/aCL). Serum samples were collected from our serum bank. Anti-beta2-glycoprotein I (anti-beta2-GPI) were tested by ELISA using irradiated plates in the absence of cardiolipin. Anti-dsDNA antibodies were tested by standard Farr assay. RESULTS: There were no major differences between SLE clinical manifestations in both groups. However, the frequency of IgG anti-beta2-GPI was markedly increased in SLE/APS (18/29, 62%) than in SLE/aCL (4/36, 11%) (chi-squared 18.6, p=0.0001). The levels of anti-dsDNA antibodies in the same samples were slightly lower in SLE/APS. CONCLUSION: Our data suggest that increased levels of IgG anti-beta2-GPI may be a specific feature of SLE/APS patients rather than reflecting a polyclonal B cell activation.

Lipoid proteinosis: a case report.

An 8-year-old Italian boy, born to consanguineous parents, with clinical, histopathologic, and ultrastructural findings of lipoid proteinosis is reported. The main signs of the syndrome-papulonodular, hyperkeratotic and verrucous lesions distributed over the skin of the head and extremities, hoarseness, and dysphagia-were present in the child. The mother had papulonodular lesions on her hands and also complained of slowness in healing.

Effects of corticotropin-releasing hormone on ovarian estrogen production in vitro.

It is a common clinical observation that stress is accompanied by dysfunction of the hypothalamic-pituitary-ovarian axis, and there is mounting experimental evidence that CRH, the principal regulator of ACTH release and the central coordinator of the stress response, is able to suppress gonadal function by inhibiting hypothalamic GnRH release. Recently, it has been shown that immunoreactive CRH, CRH messenger RNA, and CRH receptors are also present in the ovary. This prompted us to examine the role of CRH on ovarian function. To accomplish this, we studied the effects of this neuropeptide on estrogen production and cAMP intracellular content from rat granulosa and human granulosa-luteal cells. We also evaluated the activity of the enzyme aromatase by measuring the production of tritiated water from homogenates of cultured rat granulosa cells. CRH inhibited FSH-stimulated estrogen production from rat granulosa cells in a dose-dependent fashion. The maximal effect was achieved at a concentration of 10(-8) M, which suppressed estrogen production by about 30%. Low concentrations of CRH (10(-10) M), incapable of modulating maximal estrogen production in response to FSH, provoked a right-ward shift of the estrogen dose-response curve to FSH. CRH (10(-8) M) suppressed the production of tritiated water (equivalent to estrogen production) from homogenates of rat granulosa cells incubated with a half-maximal concentration of FSH. Basal estrogen production by human granulosa-luteal cells was also inhibited by CRH at a concentration of 10(-10) M. The maximal effect was achieved with a concentration of 10(-8) M, which lowered estrogen production by 25%. The CRH receptor antagonist alpha-helical CRH-(9-41) antagonized the inhibitory effect of CRH on estrogen production from rat granulosa and human granulosa-luteal cells, whereas alone it had no effect. CRH did not have any effect on the intracellular cAMP content of rat granulosa and human granulosa-luteal cells. In conclusion, these results suggest that CRH is able to suppress estrogen production from rat and human granulosa cells in vitro. This effect seems to be linked to inhibition of aromatase activity in the rat and is independent of cAMP generation. We speculate that CRH may also interfere with hypothalamic-pituitary-gonadal axis function by acting directly at the ovarian level.

Effects of a tryptophan deficient diet on the morphology of skeletal muscle fibers of the rat. Preliminary observations at neuroendocrinological and submicroscopical levels.

It is proposed an animal model consisting of young male, L-tryptophan-deprived, namely 5-HT-free rats since their ontogenesis. This was obtained by feeding their mothers with a L-tryptophan-free (tf) diet since the day 1 of pregnancy. They were studied and compared with control rats of the same ages fed with a complete diet. Already at birth tf-litters were significantly underdeveloped as compared to the control newborn rats. Postnatal growth was in the tf-rats so poor that it worsened into a stricking dwarfism characterized by physical immaturity, muscular hypotrophy with alterations of motor activity and impairment of the hypothalamo-pituitary-axis. A radioimmunological study of growth hormone (GH) showed in tf-rats dramatic low plasma levels of the hormone, thus confirming the existence of serotonergic hypothalamo-pituitary pathways for GH in normal animals. By histological and ultrastructural examinations, hypotrophy and degenerative alterations of the muscle fibers could be observed. The possible causes for this finding are extensively considered and discussed.

Fragile X expression in normal and mentally retarded subjects: effect of treatment with an antifolic agent.

Expression of the fragile site fra(X) (q27.3) in peripheral lymphocytes was evaluated in mentally retarded patients and in normal control individuals before and after administration of the antifolic agent trimethoprim for 7 days. This treatment was effective in converting the status of some individuals from fra(X)-negative to fra(X)-positive. However, the induced level of fra(X) expression was very low and not significantly different in patients and in control subjects and did not increase in those individuals where it was already present before treatment. These data support the contention that fra(X)(q27.3) is a common fragile site and that treatment in vivo with an antifolic agent is not effective in enhancing its degree of expression in vitro. Therefore, such treatment seems to be of no diagnostic value in those cases where the fra(X) syndrome is suspected clinically, but where there is no or very low cytogenetic expression of the fra(X).

The fragile X in Sicily: an epidemiological survey.

We have studied a group of 349 institutionalized propositi with mental retardation, and found 12 fra(X)-positive cases among 155 males (7.7%) and 8 fra(X)-positive cases among 194 females (4.1%). The males had characteristic manifestations of the Martin-Bell syndrome. Another 7 males, who were initially considered "borderline", having expression of fra(X) less than 4% and a non-characteristic phenotype, were eventually considered negative. Among 5,624 patients (2,764 males and 2,860 females) that were admitted to the Pediatric Department of the University of Catania during the period July 1986 - June 1987, 210 (120 males and 90 females) had mental retardation. Of these, 75 were analyzed for the presence of fra(X) (q27.3); 5 males (0.18% of all males) and 2 females (0.07% of all females) were fra(X)-positive. The males had the Martin Bell syndrome phenotype. The presence of fra(X) (q27) was confirmed in another 4 male propositi that were referred to our outpatient services with a clinical diagnosis of Martin-Bell syndrome.

A dermatoglyphic study of a group of Sicilian children with fragile-X syndrome.

In a dermatoglyphic study of 14 fra(X) boys (compared with a control group of 191 normal schoolboys), we observed the following statistically significant (p less than 0.01) differences: 1) lower frequency of ulnar loops on the fingertips, particularly on the 2nd and 3rd fingers, with a corresponding increase of whorls; 2) transverse course of main line A; 3) increased frequency of abnormal palmar creases. The log score index of Simpson et al [1984] identified 71.4% of our patients and that of Rodewald et al [1986] 64.2%. The different values of these indexes can probably be attributed to ethnic differences. We think that by combining the results of dermatoglyphic analysis from several centers a more discriminatory log score index can be obtained.

Prevalence of a novel epileptogenic EEG pattern in the Martin-Bell syndrome.

In this paper, we describe a study aiming at establishing the prevalence, specificity, and the sensitivity of a characteristic sleep EEG pattern in patients with Martin-Bell syndrome, in comparison with a sample of etiologically different mentally retarded patients. The estimation of the prevalence (11% among the total sample), the specificity, and the sensitivity, allows us to propose this pattern as an important "marker", useful in the diagnosis of the Martin-Bell syndrome.

IVIC syndrome: report of a second family.

The IVIC syndrome derives its name from the Instituto Venezolano de Investigaciones Cientìficas, where it was described by Arias et al. [Am J Med Genet 6:25-59, 1980]. We report on several individuals in a family with the IVIC syndrome, the second described in the literature. In this family there are 3 affected individuals in 2 generations. This observation shows that the IVIC syndrome is not a private syndrome, and confirms that it is due to an autosomal dominant mutation.

Facial midline defect in the fetal alcohol syndrome: embryogenetic considerations in two clinical cases.

We report on two unrelated patients with fetal alcohol syndrome with hypoplasia of the periocular region, resulting in a low and narrow forehead and hypotelorism. Other typical manifestations of the syndrome involving the facial midline are also present. These observations can be added to clinical and experimental evidence from other authors, supporting the concept that the facial anomalies of the fetal alcohol syndrome are the expression of a midline defect originating from the disruption of the ordered development of midline mesoderm cells during early embryogenesis.

Fragile-X syndrome: a particular epileptogenic EEG pattern.

A clinical and EEG study of 12 fragile-X syndrome subjects (six with epilepsy) is presented. All subjects had clinical-family history examinations, EEG evaluations, and karyotyping. Spikes were present in the sleep EEG of one nonepileptic and four epileptic subjects: these spikes were similar in location, occurrence, voltage, frequency, and morphology (and similar to those of the Rolandic spikes). These data, together with the clinical similarities (type of epilepsy, responses to drugs, ages of seizure onset, etc.), have resulted in the postulation of EEG characteristics of epileptic and nonepileptic fragile-X patients. However, further studies with fragile-X patients are needed to confirm this hypothesis.