Rem2-Targeted shRNAs Reduce Frequency of Miniature Excitatory Postsynaptic Currents without Altering Voltage-Gated Ca2+ Currents

Ca2+ influx through voltage-gated Ca2+ channels (VGCCs) plays important roles in neuronal cell development and function. Rem2 is a member of the RGK (Rad, Rem, Rem2, Gem/Kir) subfamily of small GTPases that confers potent inhibition upon VGCCs. The physiologic roles of RGK proteins, particularly in the brain, are poorly understood. Rem2 was implicated in synaptogenesis through an RNAi screen and proposed to regulate Ca2+ homeostasis in neurons. To test this hypothesis and uncover physiological roles for Rem2 in the brain, we investigated the molecular mechanisms by which Rem2 knockdown affected synaptogenesis and Ca2+ homeostasis in cultured rat hippocampal neurons. Expression of a cocktail of shRNAs targeting rat Rem2 (rRem2) reduced the frequency of miniature excitatory postsynaptic currents (mEPSCs) measured 10 d after transfection (14 d in vitro), but did not affect mEPSC amplitude. VGCC current amplitude after rRem2-targeted knockdown was not different from that in control cells, however, at either 4 or 10 d post transfection. Co-expression of a human Rem2 that was insensitive to the shRNAs targeting rRem2 was unable to prevent the reduction in mEPSC frequency after rRem2-targeted knockdown. Over-expression of rRem2 resulted in 50% reduction in VGCC current, but neither the mEPSC frequency nor amplitude was affected. Taken together, the observed effects upon synaptogenesis after shRNA treatment are more likely due to mechanisms other than modulation of VGCCs and Ca2+ homeostasis, and may be independent of Rem2. In addition, our results reveal a surprising lack of contribution of VGCCs to synaptogenesis during early development in cultured hippocampal neurons

Lipid and cholesterol oxidation in frozen stored pork, salame Milano and mortadella

An investigation has been carried out on oxidative processes of fresh and stored frozen pork and of two pork products (salame Milano and mortadella). Wholesale pork cuts (shoulder, ham trimmings, belly and backfat) were checked fresh and after frozen storage (1, 3 and 6 months). The streaky bacon was studied fresh and after scalding at 80\ub0C. Salame Milano and mortadella were produced with fresh and frozen raw materials and a few commercial samples were also part of the study. No real differences have been observed between commercial and experimentally produced salame and mortadella. Peroxide and TBARS values were low in all cases. Among the cholesterol oxides determined (7beta-hydroxycholesterol, 5,6a-epoxycholesterol, 7-ketocholesterol and 25-hydroxycholesterol) those dangerous for human health were either absent or their concentration was below toxic levels. The results are discussed on the light of literature reports and Italian pork products technologies

Oxidative stability and dietary treatment with vitamin E, oleic acid and copper of fresh and cooked pork chops

Research has been conducted on the effects of feed supplementation with vitamin E, oleic acid (sunflower oil) and copper, on some quality characteristics of pork chops. The study was based on the measurement of pH 45\u2019 and 24 hr post mortem, proximate composition, colour both objective and sensory, colour stability after packing in oxygen permeable film and protective atmosphere, copper and iron contents, vitamin E content in fresh and cooked chops, peroxide and TBARS value in raw and cooked meat, total cholesterol, cholesterol oxides and aldehydes in cooked meat, sensory evaluation of the eating quality of cooked chops and iodine number of subcutaneous fat. Vitamin E content was significantly higher with oil and vitamin supplementation. No variation has been observed in copper and iron contents. A higher stability of the colour of meat packed under modified atmosphere has been observed with increasing vitamin E content, but the same did not apply with meat packed in oxygen permeable film. No differences have been observed on fatty acid oxidative stability of fresh m. longissimus dorsi and cooked chops (peroxide and TBARS values), on cholesterol oxidation and aldehydes production of cooked chops with increasing vitamin E content. Iodine number in supplemented animals reached levels normally considere

Conservazione prolungata della materia prima e processi ossidativi nel salame e nella mortadella

Sono stati studiati gli effetti che una conservazione prolungata delle materie prime può avere sui fenomeni ossidativi dei lipidi nel salame Milano e nella mortadella. A tal fine sono state acquislate le materie prime (spalle, rifilature di prosciutto, pancettoni e lardo) che sono state confezionate in fogli di polietilene e cartoni, congelate a -20°C e corservate per un periodo di 6 mesi. Con tali materie prime sono stati prodotti salami e mortadelle a 0, l, 3 e 6 mesi di conservazione. La stabilità ossidativa dei lipidi è stata valutata sulle materie prime e sui prodotti alle stesse scadenze tramite la detetminazione del numero dei perossidi, del TBA e degli ossidi del colesterolo. Ne è emerso un quadro sostanzialmente positivo con valori del numero dei perossidi e del TBA piuttosto contenuti, con la assenza del 7-cheto e del 25-OH colesterolo, la presenza sporadica a livelli modesti del 7-OH-colesterolo e del 5,6a-epossi-colesteroleo con il 20-OHcolesterolo costantemente presente nelle materie prime e abbastanza frequente anche nei prodotti