Antiacquaporin 4 antibodies detection by different techniques in neuromyelitis optica patients.

BACKGROUND: Antibodies against aquaporin-4 (AQP4), a water channel particularly expressed on perivascular astrocytic podocytes, are proposed as a marker for the diagnosis of neuromyelitis optica (NMO). However, a consensus on seroprevalence and optimal detection method has not yet been reached. OBJECTIVES: To investigate the performance of different assays to detect anti-AQP4 antibodies. METHODS: We set up five different assays. Two of them were capable to detect perivascular IgG reactivity on brain tissue by immunofluorescence (NMO-IgG). Other three assays have been set to detect anti-AQP4 antibodies: immunofluorescence and flow cytometry on AQP4-expressing cells, and a radioimmunoprecipitation assay. We assessed sensitivity and specificity of these assays by interrogating sera of 33 NMO patients, 13 patients at high risk to develop NMO (hrNMO), 6 patients affected by acute partial transverse myelitis (APTM), 20 patients with multiple sclerosis (MS), and 67 age- and sex-matched healthy controls. RESULTS: We found that the presence of serum NMO-IgG and anti-AQP4 reactivity is almost exclusively restricted to patients with NMO and hrNMO. Seroprevalence and sensitivity ranged from 30 to 47%, depending on the assay. Specificity ranged from 95 to 100%. Comparing results obtained in the five assays, we noticed lack of concordance in some samples. CONCLUSIONS: Detection of NMO-IgG or anti-AQP4 antibodies may represent a valuable tool to assist neurologists in the differential diagnosis between patients with NMO, hrNMO, APTM, or MS. The current lack of a gold standard to detect anti-AQP4 antibodies implies the necessity to standardize the detection of these antibodies.

Quantitative ultrasound calcaneous measurements: normative data for the Italian population. the ESOPO study.

Quantitative ultrasound (QUS) is a reliable technique to evaluate skeletal status, to identify osteoporotic subjects, and to estimate the risk of fractures. The purpose of this study was to generate QUS normative data for Italian females and males aged 60-79 yr participating in the Epidemiologic Study on the Prevalence of Osteoporosis (ESOPO) study, using the Achilles Plus apparatus. ESOPO is a cross-sectional study conducted in 2000, aiming at assessing risk of osteoporosis in a random sample of 11,011 women and 4981 men, representative of the Italian population. All participants were administered a questionnaire on the most relevant risk factors for osteoporosis and fractures; 3 QUS parameters were also measured: broadband ultrasound attenuation (BUA); speed of sound (SOS); and Stiffness Index (SI). We studied the age-dependent changes in QUS values, and their correlation with body size. For both men and women, weight was the variable with the highest correlation with BUA and SI; for SOS, age among women and body mass index (BMI) among men presented the highest correlation coefficients. Average decreases of 3.0% in BUA, 0.8% in SOS, and 9.1% in SI from 60 to 79 yr were detected for females, whereas no significant changes with age in males were observed. Our data show lower QUS values for women, and a decline at a greater rate than in men.

Association of quantitative heel ultrasound with history of osteoporotic fractures in elderly men: the ESOPO study.

In order to evaluate the usefulness of calcaneal quantitative ultrasound (QUS) in the assessment of male osteoporosis, a cross-sectional, population-based study was performed. A cohort of 4,832 men, randomly selected, community-dwelling, aged 60-80 years and representative of the general older male Italian population was recruited. QUS measurements were assessed in 83 centers distributed all over Italy and equipped with an Achilles device (GE-Lunar, Madison, Wisconsin, USA). All participants were administered a questionnaire covering lifestyle variables and medical history. Low-energy fractures that had occurred since age 50 were recorded. Overall, 43 subjects reported a previous hip fracture and 455 subjects reported other non-spinal fractures. Univariate analysis showed that fractured subjects were older, with a lower level of outdoor physical activity and a more frequent history of prolonged bedridden periods in comparison with unfractured subjects. Men reporting non-spinal fractures showed a higher prevalence of smoking, while no difference was found among groups in anthropometric measures and calcium intake. QUS measurements showed that all QUS parameters were significantly lower in both fracture groups (p<0.001). Multiple logistic regression analysis demonstrated that each SD reduction in QUS measures was associated with an approximate doubling of the risk for hip fracture, independent of age and other clinical variables (broadband ultrasound attenuation [BUA]: odds ratio [OR]=2.24; 95% confidence interval [CI] 1.61-3.08; stiffness index: OR=2.19; CI 1.56-3.11; speed of sound [SOS]: OR=1.71; CI 1.18-3.24) and with an increase of the risk of other non-spinal fractures (BUA: 1.38; CI 1.22-1.59; stiffness index: OR=1.27; CI 1.17-1.38; SOS: OR=1.14; CI 0.96-1.40). It can be concluded that calcaneal QUS measurement is associated with the risk for hip fracture and any non-spinal fractures among a community-dwelling cohort of elderly men. The strength of the association between QUS measurement and fracture is similar to that observed in elderly women.

Endothelin-1: expression and role in human corpus luteum.

PROBLEM: Several recent data suggest an involvement of endothelin (ET)-1, a powerful vasoconstrictor peptide, in reproductive function. This study was designed to investigate the presence and role of ET-1 in human corpus luteum. METHOD OF STUDY: Purified luteal cells were incubated for different times with ET-1 or ET-3 alone or associated with human chorionic gonadotropin. In another set of experiments cells were treated with ET-1 and BQ485, an ET-A receptor antagonist, or with phorbol 12-myristate-13 acetate (PMA), an activator of protein kinase C. RESULTS: ET-1 reduced both basal and human chorionic gonadotropin-induced progesterone production at all examined times, similarly PMA inhibited basal progesterone synthesis. BQ485 prevented the inhibitory effect of ET-1, while no effect was observed with ET-3. Finally, ET-1 mRNA was detected in the luteal cells. CONCLUSION: ET-1 is expressed by human luteal cells and reduces basal and human chorionic gonadotropin-induced progesterone synthesis through the ET-A receptors and the protein kinase C pathway. Conversely, ET-3 does not affect luteal steroidogenesis.

Endothelin-1 inhibits basal and human chorionic gonadotrophin-stimulated progesterone production.

Endothelin-1 (ET-1) is a peptide classically produced by endothelial cells and known for its powerful vasoconstrictor activity. However, recent data suggest an involvement of ET-1 also in reproductive function. This study was designed to examine the possible presence and role of ET-1 in human luteal cells. Purified luteal cells were incubated for different times with ET-1 (10(-9)-10(-6) M) or ET-3 (10(-9)-10(-6)) alone or associated with human chorionic gonadotrophin (HCG) (100 ng/ml). Both basal and HCG-induced progesterone production were significantly reduced by ET-1 at all examined times whereas preincubation of luteal cells with BQ485 (10(-9)-10(-6) M), an ET-A receptor antagonist, prevented the inhibitory effect of ET-1. Conversely, no effect on progesterone synthesis was observed when ET-3 was added to the cultures. Luteal cells were then incubated for 24 h with phorbol 12-myristate-13 acetate (PMA) (100 ng/ml), an activator of protein kinase C. Inhibition of progesterone synthesis by PMA was similar to that induced by ET-1 alone. This study demonstrates that ET-1 negatively affects, at physiological concentrations, basal and HCG-induced progesterone synthesis. These effects seem to be exerted through the ET-A receptors and the protein kinase C pathway. Conversely, ET-3 was not able to influence human luteal steroidogenesis.

Control of human luteal steroidogenesis: role of growth hormone-releasing hormone, vasoactive intestinal peptide, and pituitary adenylate cyclase-activating peptide.

OBJECTIVE: To examine the possible effect of growth hormone-releasing hormone (GHRH), vasoactive intestinal peptide, and pituitary adenylate cyclase-activating peptide on basal and hCG-stimulated P production by human luteal cells. DESIGN: Cultures of human luteal cells from the early and midluteal phase. SETTING: All corpora lutea were obtained from the Obstetrics and Gynecology Department of the Università Cattolica, a public care center. PATIENT(S): Ten nonpregnant women between 35 and 47 years of age underwent surgery for various nonendocrine disorders, such as leiomyomatosis. INTERVENTION(S): Corpora lutea were obtained at the time of hysterectomy. MAIN OUTCOME MEASURE(S): Luteal cells were incubated with GHRH, vasoactive intestinal peptide, and pituitary adenylate cyclase-activating peptide with or without hCG at different concentrations. RESULT(S): Pituitary adenylate cyclase-activating peptide stimulated P production in a dose- and time-dependent manner, whereas GHRH and vasoactive intestinal peptide did not affect luteal steroidogenesis. None of the three peptides were found to synergize with hCG. CONCLUSION(S): Pituitary adenylate cyclase-activating peptide can influence human luteal steroidogenesis.

Insulin-like growth factor (IGF)-I and IGF-II stimulate progesterone production by human luteal cells: role of IGF-I as mediator of growth hormone action.

OBJECTIVE: To examine the possible direct effect of insulin-like growth factor (IGF)-I and IGF-II on basal and hCG-stimulated P production by cultured human luteal cells. The possible role of IGF-I as mediator of GH action on luteal steroidogenesis also was investigated. DESIGN: Cultures of human luteal cells from early and midluteal phase. SETTING: All corpora lutea were obtained from the Obstetrics and Gynecology Department of the Universita Cattolica, a public care center. PATIENTS: Eight nonpregnant women between 35 and 47 years of age underwent surgery for various nonendocrine disorders such as leiomyomatosis. INTERVENTIONS: Corpora lutea were obtained at the time of hysterectomy. MAIN OUTCOME MEASURES: Luteal cells were incubated with IGF-I or IGF-II with or without hCG at different concentrations. Growth hormone also was used alone and with an anti-IGF-I-antibody. RESULTS: We found that IGF-I and IGF-II were able to stimulate directly the P production at all used concentrations and that both of them significantly amplified the steroidogenic hCG effect. Finally, IGF-I was shown to mediate the positive GH action on P synthesis.

Molecular alterations of the AKT2 oncogene in ovarian and breast carcinomas.

The AKT2 gene is one of the human homologues of v-akt, the transduced oncogene of the AKT8 virus, which induces lymphomas in mice. In previous studies, AKT2, which codes for a serine-threonine protein kinase, was shown to be amplified and overexpressed in some human ovarian carcinoma cell lines and amplified in primary tumors of the ovary. To confirm and extend these findings, we conducted a large-scale, multicenter study of AKT2 alterations in ovarian and breast cancer. Southern-blot analysis demonstrated AKT2 amplification in 16 of 132 (12.1%) ovarian carcinomas and in 3 of 106 (2.8%) breast carcinomas. No AKT2 alteration was detected in 24 benign or borderline tumors. Northern-blot analysis revealed overexpression of AKT2 in 3 of 25 fresh ovarian carcinomas which were negative for AKT2 amplification. The difference in the incidence of AKT2 alterations in ovarian and breast cancer suggests a specific role for this gene in ovarian oncogenesis. No significant association was found between AKT2 amplification and amplification of the proto-oncogenes MYC and ERBB2, suggesting that amplification of AKT2 defines an independent subset of breast and ovarian cancers. Ovarian cancer patients with AKT2 alterations appear to have a poor prognosis. Amplification of AKT2 was especially frequent in undifferentiated tumors (4 of 8, p = 0.019), suggesting that AKT2 alterations may be associated with tumor aggressiveness.

Post-traumatic hypopituitarism due to a hypothalamic lesion.

Two patients were studied who suffered from severe head trauma with skull fracture. Hypopituitarism developed in both. Because of elevated serum prolactin levels and because of preserved response of some of the pituitary hormones to exogenous thyrotropin and gonadotropin-releasing hormones, the responsible lesion in both cases was most likely suprasellar. Findings of interest included slightly elevated thyroid-stimulating hormone (TSH) levels in the presence of hypothyroidism in one of the patients, and dissociation of the luteinizing hormone (LH) and follicle-stimulating hormone (FSH) responsiveness to both gonadotropin-releasing hormone and clomiphene citrate in the other patient. It is suggested that the diagnosis of post-traumatic hypopituitarism be considered in patients with head trauma and that periodically appropriate laboratory testing be performed to confirm the diagnosis.

Intraventricular craniopharyngioma.

An unusual case of an adult with a craniopharyngioma within the third ventricle is reported. The patient complained of headaches, had a history suggestive of diabetes insipidus, and presented with a severe dementia. A brain scan revealed the suprasellar midline lesion, and a pneumoencephalogram confirmed its location within the third ventricle. Therapy included partial surgical excision, followed by a ventriculoatrial shunt and radiation.