Delta opioid receptor on equine sperm cells: subcellular localization and involvement in sperm motility analyzed by computer assisted sperm analyzer (CASA)

<p>Abstract</p> <p>Background</p> <p>Opioid receptors and endogenous opioid peptides act not only in the control of nociceptive pathways, indeed several reports demonstrate the effects of opiates on sperm cell motility and morphology suggesting the importance of these receptors in the modulation of reproduction in mammals. In this study we investigated the expression of delta opioid receptors on equine spermatozoa by western blot/indirect immunofluorescence and its relationship with sperm cell physiology.</p> <p>Methods</p> <p>We analyzed viability, motility, capacitation, acrosome reaction and mitochondrial activity in the presence of naltrindole and DPDPE by means of a computer assisted sperm analyzer and a fluorescent confocal microscope. The evaluation of viability, capacitation and acrosome reaction was carried out by the double CTC/Hoechst staining, whereas mitochondrial activity was assessed by means of MitoTracker Orange dye.</p> <p>Results</p> <p>We showed that in equine sperm cells, delta opioid receptor is expressed as a doublet of 65 and 50 kDa molecular mass and is localized in the mid piece of tail; we also demonstrated that naltrindole, a delta opioid receptor antagonist, could be utilized in modulating several physiological parameters of the equine spermatozoon in a dose-dependent way. We also found that low concentrations of the antagonist increase sperm motility whereas high concentrations show the opposite effect. Moreover low concentrations hamper capacitation, acrosome reaction and viability even if the percentage of cells with active mitochondria seems to be increased; the opposite effect is exerted at high concentrations. We have also observed that the delta opioid receptor agonist DPDPE is scarcely involved in affecting the same parameters at the employed concentrations.</p> <p>Conclusions</p> <p>The results described in this paper add new important details in the comprehension of the mammalian sperm physiology and suggest new insights for improving reproduction and for optimizing equine breeding.</p

Galactosylated Prodrugs: A Strategy to Improve the Profile of Nonsteroidal Anti-Inflammatory Drugs

Carbohydrates are one of the most abundant and important classes of biomolecules. The variety in their structures makes them valuable carriers that can improve the pharmaceutical phase, pharmacokinetics and pharmacodynamics of well-known drugs. D-galactose is a simple, naturally occurring monosaccharide sugar that has been extensively studied for use as a carrier and has proven to be valuable in this role. With the aim of validating the galactose-prodrug approach, we have investigated the galactosylated prodrugs ibuprofen, ketoprofen, flurbiprofen and indomethacin, which we have named IbuGAL, OkyGAL, FluGAL and IndoGAL, respectively. Their physicochemical profiles in terms of lipophilicity, solubility and chemical stability have been evaluated at different physiological pH values, as have human serum stability and serum protein binding. Ex vivo intestinal permeation experiments were performed to provide preliminary insights into the oral bioavailability of the galactosylated prodrugs. Finally, their anti-inflammatory, analgesic and ulcerogenic activities were investigated in vivo in mice after oral treatment. The present results, taken together with those of previous studies, undoubtedly validate the galactosylated prodrug strategy as a problem-solving technique that can overcome the disadvantages of NSAIDs

Influence of mycotoxin zearalenone and its derivatives (alpha and beta zearalenol) on apoptosis and proliferation of cultured granulosa cells from equine ovaries

BACKGROUND: The mycotoxin zearalenone (ZEA) and its derivatives, alpha and beta-zearalenol (alpha and beta-ZOL), synthesized by genera Fusarium, often occur as contaminants in cereal grains and animal feeds. The importance of ZEA on reproductive disorders is well known in domestic animals species, particularly in swine and cattle. In the horse, limited data are available to date on the influence of dietary exposure to ZEA on reproductive health and on its in vitro effects on reproductive cells. The aim of this study was to evaluate the effects of ZEA and its derivatives, alpha and beta-ZOL, on granulosa cells (GCs) from the ovaries of cycling mares. METHODS: The cell proliferation was evaluated by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test after 3 days exposure at different concentrations of ZEA and its derivatives (from 1 × 10-7 to 0.1 microM). The apoptosis induction was evaluated after 1 day exposure, by DNA analysis using flow cytometry. RESULTS: An increase in cell proliferation with respect to the control was observed in the presence of ZEA at 1 × 10-3 and 1 × 10-4 microM and apoptosis was induced by all mycotoxins at different concentrations. CONCLUSION: The simultaneous presence of apoptosis and proliferation in GC cultures treated with zearalenones could indicate that these mycotoxins could be effective in inducing follicular atresia. These effects of zearalenones may result from both direct interaction with oestrogen-receptors as well as interaction with the enzymes 3alpha (beta)-hydroxysteroid dehydrogenase (HSD), involved in the synthesis and metabolism of endogenous steroid hormones. These cellular disturbances, described for the first time in equine GCs cultured in vitro, could be hypothesized as referred to reproductive failures of unknown ethiology in the mare

Intelligenza territoriale per l' area Stadio di Fuorigrotta, Napoli( Italia)

Alla domanda rivolta in presentazione del Convegno , se "Il territorio, che sempre paga i costi delle strategie politiche sociali ed economiche, potrebbe contribuire a superare alcune contraddizioni e a delineare la centralità di un benessere che sia di ciascuno e di tutti? La comunicazione che si intende presentare, offre una proposta progettuale che evidenzi l'importanza di avanzare verso l' incremento dell'intelligenza territoriale. Specificatamente: l'obiettivo principale è quello di individuare un sistema di governance che attraverso la sostenibilità del territorio nelle dimensioni ambientale, sociale ed economica possa affermare il criterio di migliorare la qualità della vita in un'area territoriale specifica e circoscritta. Poichè E' ormai sotto gli occhi di tutti, quanto drammatico sia stato il consumo del suolo napoletano che rende difficilissimo e a volte impossibile interventi di aggiustamento e quanto danno, detto territorio ipercementificato e con super pressione demografica abbia subito per le scellerate scelte delle strategie politico- economiche; oggi proprio l' intelligenza territoriale può offrire un assennato governo del territorio a vantaggio di tutti gli Stakeholder. L' obiettivo operativo della comunicazione sarà implementato in un modello di ricerca ad una realtà più precisa ,il quartiere di Fuorigrotta in Napoli. Metologicamente saranno adottati indicatori(metodi quantitativi) e interviste (metodi qualitativi) non tralasciando la descrizione dell'esistente

Intelligenza territoriale per l' area Stadio di Fuorigrotta, Napoli( Italia)

Alla domanda rivolta in presentazione del Convegno , se "Il territorio, che sempre paga i costi delle strategie politiche sociali ed economiche, potrebbe contribuire a superare alcune contraddizioni e a delineare la centralità di un benessere che sia di ciascuno e di tutti? La comunicazione che si intende presentare, offre una proposta progettuale che evidenzi l'importanza di avanzare verso l' incremento dell'intelligenza territoriale. Specificatamente: l'obiettivo principale è quello di individuare un sistema di governance che attraverso la sostenibilità del territorio nelle dimensioni ambientale, sociale ed economica possa affermare il criterio di migliorare la qualità della vita in un'area territoriale specifica e circoscritta. Poichè E' ormai sotto gli occhi di tutti, quanto drammatico sia stato il consumo del suolo napoletano che rende difficilissimo e a volte impossibile interventi di aggiustamento e quanto danno, detto territorio ipercementificato e con super pressione demografica abbia subito per le scellerate scelte delle strategie politico- economiche; oggi proprio l' intelligenza territoriale può offrire un assennato governo del territorio a vantaggio di tutti gli Stakeholder. L' obiettivo operativo della comunicazione sarà implementato in un modello di ricerca ad una realtà più precisa ,il quartiere di Fuorigrotta in Napoli. Metologicamente saranno adottati indicatori(metodi quantitativi) e interviste (metodi qualitativi) non tralasciando la descrizione dell'esistente