Hepatitis E virus (HEV) in sewage from treatment plants of Messina University Hospital and of Messina City Council.

Samples of sewage from treatment plants at the "G. Martino" University Hospital of Messina (AOU) and that of Messina City Council were analysed to detect the hepatits E virus. Samples were taken on sewage entering and exiting the treatment plants on a monthly basis over a one-year period from both the hospital plant (24 samples) and the municipal plant (22 samples). All sewage samples were pretreated by ultrafiltraton and concentration and finally processed by the PCR method to amplify gene material. A total of three samples tested positive: two (8.33%) entering the AOU treatment plant and one (4.5%) entering the municipal plant while no cases of HEV were detected in samples of treated sewage. These findings confirm the presence of the virus in the city of Messina and showed that the two treatment plants to be working efficiently when tested.

Comparison between a conventional subunit vaccine and the MF59-adjuvanted subunit influenza vaccine in the elderly: an evaluation of the safety, tolerability and immunogenicity.

The objective of this study was to evaluate and compare the safety, tolerability and immunogenicity for two seasonal influenza subunit vaccines, one with MF59 adjuvant (Fluad) and one without an adjuvant (Agrippal). A total of 195 subjects aged > or = 65 years were enrolled to receive one dose of vaccine intramuscularly, 96 were vaccinated with Fluad, 99 received Agrippal. Blood samples were taken from all subjects in order to assess their antibody titre by the haemagglutination inhibition assay (HI), before (Time 0) and after (Time 1: 28 +/- 7 days) vaccination, against the A/H3N2 (A/Moscow/10/99), A/H1N1 (A/New Caledonia/20/99) and B/Shandong/7/97 antigens contained in the influenza vaccine in the 2002/2003 influenza season for the northern hemisphere. A good humoral antibody response was detected for both vaccines, meeting all the criteria of EMEA. The number of subjects in whom > or = 4-fold increase in antibody titre was recorded, in comparison with the pre-vaccination value, proved to be lower in the group vaccinated with AgrippaPl than in those vaccinated with the adjuvated preparation. Fluad" exhibited better immunogenicity than Agrippal. This difference was probably linked to the potentiated immune stimulation exerted by the adjuvant molecules. These results take on a particular importance if we consider that the immune system is weaker in the elderly; the administration of an adjuvated vaccine in such subjects is clearly preferable in that it provides greater and more prolonged protection. Both vaccines were generally well tolerated; no severe adverse events occurred in any of the subjects vaccinated, confirming the excellent safety profile of Fluad and Agrippal.

Eliminating congenital rubella: a seroepidemiological study on women of childbearing age and MMR vaccine coverage in newborns.

INTRODUCTION: Rubella can have particularly serious effects on the product of conception if contracted during pregnancy. Thus, the main aim of rubella vaccination programmes is to prevent infection during pregnancy. MATERIALS AND METHODS: A seroepidemiological study was conducted from July 2006 to December 2007 on 1000 women of childbearing age, 15 to 45 years old, using specific rubivirus antibody assays, IgG and IgM. A questionnaire administered at the same time allowed us to survey how much women knew about this disease. In addition, MMR vaccine coverage rates were analysed for cohorts born in the local health districts of Messina for the period 1993-2006. RESULTS: An analysis of the replies given to the questionnaire showed an estimated 42.8% of the women to have immunity from rubella, while the serological study showed an immunity coverage rate of80.6%. Vaccination coverage in the local health districts regarding the first dose of MMR was 81% (cohorts 1993-2005), while the rate was only 24% for the second dose (cohorts 1993-2002). CONCLUSIONS: Both immunity coverage in women of childbearing age and that for newborns (for the cohort considered)fall below the 95% target set by the National Elimination Plan for Measles and Congenital Rubella (PNEM). It is therefore necessary to provide women with adequate information about the risks of rubella during pregnancy and about the benefits of vaccination, as well as to recoup subjects at risk or those whose immune status is unknown. Public health authorities also need to make continued efforts to increase the number of MMR vaccinations throughout the region.

Effect of estrogen agonists and antagonists on induction of progesterone receptor in a rat hypothalamic cell line.

Estrogen is essential in the hypothalamus for the central regulation of reproduction. To understand the molecular mechanism(s) of estrogen action in the hypothalamus, immortalized rat embryonic hypothalamic cell lines were characterized for steroid receptors and subcloned. Scatchard analysis of the D12 subclone demonstrated one high affinity estrogen receptor-binding site (Kd = 31.3+/-1.9 pM) with a Bmax of 30.8+/-0.8 fmol/mg. Estrogen receptor-alpha protein was identified by Western blot and gel shift analyses. Treatment with estradiol (48 h) stimulated progesterone receptor (PR) messenger RNA expression and binding to [3H]R5020, a synthetic progestin. Because the agonist or antagonist activity of estrogen mimetics can be cell type dependent, the activities of various estrogen mimetics were determined in D12 cells. ICI 182,780 (IC50 = 0.63 nM), raloxifene (IC50 = 1 nM), enclomiphene (IC50 = 77 nM), and tamoxifen (IC50 = 174 nM) inhibited the induction of PR by estradiol, and none of these compounds significantly stimulated PR when given alone. In contrast, 17alpha-ethynyl estradiol (EC50 = 0.014 nM), zuclomiphene (EC50 = 100 nM), and genistein (EC50 = 17.5 nM) functioned as estrogen agonists in these cells. In addition, the estrogen-induced progesterone receptor activated a progesterone response element reporter construct in response to progestins. Thus, the D12 rat hypothalamic cell line provides a useful model for characterizing tissue-selective estrogenic compounds, identifying estrogen- and progesterone-regulated hypothalamic genes, and understanding the molecular mechanisms of steroid action in various physiological processes mediated by the hypothalamus.

Expression of estrogen receptor-beta protein in rodent ovary.

Estrogen is an essential hormone for the LH surge and ovulation. The primary source of estrogen is from ovarian granulosa cells and in rats, estrogen, in turn, increases granulosa cell number and enhances FSH-stimulated gene expression in these cells. Thus, rat granulosa cells both respond to and synthesize estrogen. To further elucidate the mechanisms mediating the actions of estrogen in granulosa cells, we have identified and characterized the estrogen receptor-beta (ER-beta) subtype in rodent granulosa cells. ER-beta protein was localized to the nuclei of rat granulosa cells in preantral and antral follicles by immunocytochemistry, coincident with the location of ER-beta messenger RNA (mRNA). Immunoprecipitation and Western blot analysis using ER-beta specific antisera demonstrated a protein of approximately 60 kDa in granulosa cells prepared from PMSG-primed immature mice and estrogen-treated immature rats. Extracts from granulosa cells specifically bound an estrogen response element and the complex was recognized by antisera to ER-beta. A synthetic steroid estrogen radioligand, [125I]-17alpha-iodovinyl-11beta-methoxyestradiol ([125I]-VME2), bound to cytosolic granulosa cell preparations with high affinity (estimated K(D) value of 401 +/- 83 pM, and Bmax value of 102 +/- 9 fmol/mg protein). ER-beta protein levels rapidly declined following hCG treatment consistent with the reported decrease in binding activity and ER-beta mRNA levels by high levels of gonadotropins. Overall, we have demonstrated that 1) ER-beta protein is the dominant estrogen receptor subtype present in rodent granulosa cells, 2) this receptor is functional, and 3) it is regulated by ovulatory doses of gonadotropins. Thus, ER-beta is likely to be a mediator of estrogen action in rodent granulosa cells during follicular development.

Expression of growth differentiation factor-9 messenger ribonucleic acid in ovarian and nonovarian rodent and human tissues.

Growth differentiation factor-9 (GDF-9) is a member of the transforming growth factor-beta family that is reported to be expressed exclusively in the ovary, specifically in the oocyte. Female mice deficient in GDF-9 are infertile, suggesting that GDF-9 receptor agonists and antagonists may specifically modulate fertility. We now report that GDF-9 messenger RNA (mRNA) is expressed in nonovarian tissues in mice, rats, and humans. GDF-9 mRNA was detected in mouse and rat ovary, testis, and hypothalamus by Northern blot and RT-PCR analyses. The localization of GDF-9 mRNA specifically in oocytes of the mouse ovary was confirmed by in situ hybridization histochemistry. In mouse testis, although localization in Sertoli cell cytoplasm could not be ruled out, mRNA expression was observed in large pachytene spermatocytes and round spermatids. The expression of GDF-9 mRNA in human tissues was assessed by Northern blot and RT-PCR analyses. GDF-9 mRNA was observed in ovary and testis and, surprisingly, in diverse nongonadal tissues, including pituitary, uterus, and bone marrow. Therefore, GDF-9 mRNA expression in rodents is not exclusive to the ovary, but includes the testis and hypothalamus. Furthermore, human GDF-9 mRNA is expressed not only in the gonads, but also in several extragonadal tissues. The function and relevance of nongonadal GDF-9 mRNA are not known, but may affect strategies for contraception and fertility that are based on GDF-9 activity.