On scrapie interference and artificial prions

The mechanisms responsible for neuronal death in transmissible spongiform encephalopathies (TSEs) are still not completely understood, and at least two major hypotheses have been formulated, based on the peculiar aspects of prion protein biology. In fact, the neuronal spreading of the prion conformational change may lead either to gain toxic properties, or to loose the normal function of this protein. In order to investigate the relative contribution of these two opposite mechanisms, two theoretical approaches may be proposed: RNA interference (RNAi) and artificial prion engineering. In fact, RNAi techniques offer now an extremely exciting new tool for investigating the effects of gene silencing both in prion, and other neurological disorders. On the other hand, the gain-of-toxic-function hypothesis might be definitely evaluated by creating an artificial prion choosing a protein target whose loss of function could be bypassed in the experimental set. In this paper the two aforementioned strategies are outlined, briefly discussing the consequent implications for TSE therapy.     

Outcome of patients with cardiac diseases admitted to coronary care units: a report from Lazio, Italy

BACKGROUND: Coronary care units (CCUs) currently treat a variety of diseases, but little is known about the effectiveness of CCUs on heart conditions other than acute myocardial infarction. OBJECTIVES: The objectives of this study were to evaluate the association between direct admission to CCUs and the risk of inhospital death in patients with heart disease, to investigate factors affecting direct admission to a CCU, and to assess the effect of CCU admission on the use of invasive procedures in patients with arrhythmias. RESEARCH DESIGN: We conducted a retrospective analysis of discharge-abstract data from Lazio, Italy, hospitals. We used logistic regression, propensity score, and instrumental variable analysis to compare inhospital risk of death between patients admitted to CCUs and to ordinary wards in 13 different groups of heart disease. We used linear regression to study the association between the rate of CCU admission and the relative risk of death. RESULTS: The study included 181,049 heart disease admissions, of which 8620 were admitted to CCUs (4.8%). Risk of death was significantly lower in patients admitted directly to CCUs for "acute myocardial infarction" (odds ratio [OR], 0.57), "acute ischemic heart disease" (OR, 0.55), and "other arrhythmias" (OR, 0.56). Mortality ORs were inversely related to the rate of CCU admission. CCU patients with arrhythmias received more invasive procedures (OR, 2.70) than non-CCU patients. CONCLUSION: Direct admission to a CCU is associated with a decrease in mortality for patients with "acute myocardial infarction," "acute heart ischemia," and "other arrhythmias." Patients most likely to benefit from CCU care are preferentially admitted to CCUs. CCUs make larger use of invasive procedures than ordinary wards.     

Alcohol consumption and acute myocardial infarction: a benefit of alcohol consumed with meals?

BACKGROUND: The apparent favorable effect of alcohol on the risk of acute myocardial infarction (MI) may be related to its hypoinsulinemic effect when consumed with meals. We studied how the timing of alcohol consumption in relation to meals might affect the risk of MI in a population with relatively high regular alcohol consumption. METHODS: We conducted a case-control study between 1995 and 1999 in Milan, Italy. Cases were 507 subjects with a first episode of nonfatal acute MI, and controls were 478 patients admitted to hospitals for other acute diseases. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated by multiple logistic regression models. RESULTS: Compared with nondrinkers, an inverse trend in risk was observed when alcohol was consumed during meals only (for > or =3 drinks per day: OR = 0.50; 95% CI = 0.30-0.82). In contrast, no consistent trend in risk was found for subjects drinking outside of meals (for > or =3 drinks per day: 0.98; 0.49-1.96). The pattern of risk was similar when we considered people who drank only wine. CONCLUSIONS: Alcohol drinking during meals was inversely related with risk of acute MI, whereas alcohol drinking outside meals only was unrelated to risk.     

Dietary conjugated linoleic acid positively affects immunologic variables in lactating sows and piglets

We studied the effects of conjugated linoleic acid (CLA) on metabolic and immunologic variables in lactating sows and piglets. Gestating sows (n = 16) were assigned to 1 of 2 weight- and parity-matched groups supplemented with 0% (C) or 0.5% (T) of a CLA preparation containing 50% CLA isomers. Supplementation started in late pregnancy and continued throughout lactation. At weaning, 80 piglets, half from each group of sows, were assigned to 0% CLA (C) or 0.5% CLA (T). Thus, there were four groups of 20 piglets: C-C, C-T, T-T, and T-C. Body weight and the number of piglets per litter at birth and weaning, and the chemical composition of colostrum did not differ among the groups. CLA affected the fatty acid composition of colostrum fat; palmitoleic and gamma-linolenic acid were significantly lower compared with controls, whereas eicosenoic and eicosatrienoic acids were significantly higher. Feeding CLA increased (P < 0.05) colostrum IgG in sows. Sows fed CLA had higher (P < 0.05) serum leptin, IgG, and lysozyme. Nursing piglets from CLA-fed sows had significantly higher (P < 0.01) serum lysozyme and IgG. Consumption of CLA did not affect postweaning growth. Postweaning piglets fed CLA (T-T, C-T) had a higher IgG titer at 25 d (P < 0.05) and 35 d (P < 0.01) after weaning. Serum lysozyme was also higher at 25 d (P < 0.05) in CLA-fed piglets (T-T, C-T). At 35 d, serum alpha-1 acylglicoprotein was lower (P < 0.05) in piglets fed CLA. Dietary CLA had a positive effect on immunologic variables in lactating sows and piglets.     

Methoctramine analogues inhibit responses to capsaicin and protons in rat dorsal root ganglion neurons

We have investigated the possibility that vanilloid receptors have a binding site for polyamines and determined the consequences of binding to such a site. Whole-cell and single-channel patch-clamp recordings were used to investigate the effect of the tetraamine, methoctramine, and 16 of its analogues on capsaicin and proton induced responses of foetal rat dorsal root ganglion neurons. All but two methoctramine analogues inhibited responses to 10 microM capsaicin with IC50 values in the range of 2-70 microM at a holding potential of -100 mV. Inhibition was generally non-competitive and voltage-dependent. Methoctramine at 10 microM reduced the single channel mean open time (>3-fold), but also increased the mean closed time (1.7-fold). Sustained responses to pH 5.4 were antagonized by methoctramine with similar potency to capsaicin responses. Similar data were obtained with adult rat dorsal root ganglion neurons. These data indicate that methoctramine analogues bind to vanilloid receptors to inhibit their function.     

The N-terminal of icatibant and bradykinin interact with the same Asp residues in the human B2 receptor

The pharmacology of peptide and non-peptide bradykinin B2 receptor ligands was evaluated in the inositol phosphate (IP) production assay in CHO cells expressing the human bradykinin B2 receptor. The effect of single and double alanine mutation of D266 and D284 residues at the human bradykinin B2 receptor was evaluated on the agonist profile of bradykinin (H-Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg-OH) and the synthetic agonist FR190997 (8-[2,6-dichloro-3-[N-methylcarbamoyl)cinnamidoacetyl]-N-methylamino]benzyloxy]-2-methyl-4-(2-pyridylmethoxy)quinoline). Bradykinin potency (EC50 0.5 nM at the wild-type receptor) was reduced by 16-fold at D266A and D284A mutants and by 2300-fold at the D266A/D284A double mutant. None of the mutants affected the potency or the efficacy of FR190997. Peptide antagonists, Icatibant (H-DArg-Arg-Pro-Hyp-Gly-Thi-Ser-Dtic-Oic-Arg-OH) and MEN11270 (H-DArg-Arg-Pro-Hyp-Gly-Thi-c(Dab-DTic-Oic-Arg)c(7gamma-10alpha)) (100 nM) similarly antagonized the concentration-response curve to bradykinin or FR190997 (pA2 values 8.5 and 8.4 versus bradykinin and 8.2 and 8.4 versus FR190997) at the wild-type receptor. Non-peptide antagonists FR173657 ((E)-3-(6-acetamido-3-pyridyl)-N-[N-[2,4-dichloro-3-[(2-methyl-8-quinolinyl) oxymethyl]phenyl]-N-methylaminocarbonyl methyl]acrylamide) and LF16-0687 (1-[[2,4-dichloro-3-[(2,4-dimethylquinolin-8-yl)oxy] methyl]-phenyl]sulfonyl]-N-[3-[[4-(aminoiminomethyl)-phenyl]carbonylamino]propyl]-(S)-pyrrolidine carboxamide) (100 nM) showed an equivalent potency values in blocking the IP production induced by bradykinin or FR190997 (pA2 values 8.7 and 8.8 versus bradykinin and 8.8 and 8.6 versus FR190997). Whilst the antagonist potency of FR173657 and LF16-0687 was not affected by D266A/D284A double mutation (IP production induced by the synthetic agonist), that of Icatibant and MEN11270 was reduced by 50- and 200-fold. The antagonist potency of [Ala1]-Icatibant and [Ala2]-Icatibant (pA2 values at wild-type 7.7 and 6.4) was significantly less reduced (20-fold and 13-fold, respectively) by the D266A/D284A double mutation. Our results highlight a crucial role for two aspartic residues, D266 and D284, located at the top of transmembrane segments 6 and 7, in the high-affinity interaction of peptide antagonists with the human bradykinin B2 receptor. An interaction of these receptor residues with the N-terminal basic residues of Icatibant is hypothesized.