Transcripts of antibacterial peptides in chicken erythrocytes infected with Marek's disease virus.

BACKGROUND: Chicken erythrocytes are involved in immunity through binding of toll-like receptors (TLRs) with their ligands to activate downstream signaling and lead to cytokine production in erythrocytes. Some avian ß-defensins (AvBDs) are constitutively expressed in tissues and some others can be induced by various bacteria and viruses. However, the expression of AvBDs in erythrocytes has not yet been studied extensively. RESULTS: The transcripts of eight AvBDs (AvBD1 to AvBD7, and AvBD9) and liver-expressed antimicrobial peptide-2 (LEAP-2) were found in normal chicken erythrocytes. The expression levels of AvBD2, 4 and 7 were significantly increased (P < 0.01), whereas the levels of AvBD1, 6 and 9 were significantly decreased (P < 0.01) after Marek's disease virus (MDV) infection. The mRNA expression level of LEAP-2 was not significantly changed after MDV infection. Highest viral nucleic acid (VNA) of MDV in the feather tips among the tested time points was found at 14 days post-infection (d.p.i.). In addition, 35 MD5-related gene segments were detected in the erythrocytes at 14 d.p.i. by transcriptome sequencing. CONCLUSIONS: These results suggest that the AvBDs in chicken erythrocytes may participate in MDV-induced host immune responses.

Oxygen-initiated chain mechanism for hydride transfer between NADH and NAD+ models. Reaction of 1-benzyl-3-cyanoquinolinium ion with N-methyl-9,10-dihydroacridine in acetonitrile.

A reinvestigation of the formal hydride transfer reaction of 1-benzyl-3-cyanoquinolinium ion (BQCN(+)) with N-methyl-9,10-dihydroacridine (MAH) in acetonitrile (AN) confirmed that the reaction takes place in more than one step and revealed a new mechanism that had not previously been considered. These facts are unequivocally established on the basis of conventional pseudo-first-order kinetics. It was observed that even residual oxygen under glovebox conditions initiates a chain process leading to the same products and under some conditions is accompanied by a large increase in the apparent rate constant for product formation with time. The efficiency of the latter process, when reactions are carried out in AN with rigorous attempts to remove air, is low but appears to be much more pronounced when MAH is the reactant in large excess. On the other hand, the intentional presence of air in AN ([air] = half-saturated) leads to a much greater proportion of the chain pathway, which is still favored by high concentrations of MAH. The latter observation suggests that a reaction intermediate reacts with oxygen to initiate the chain process in which MAH participates. Kinetic studies at short times show that there is no kinetic isotope effect on the initial step in the reaction, which is the same for the two competing processes. Our observation of the chain pathway of an NADH model compound under aerobic conditions is likely to be of importance in similar biological processes where air is always present.

A thermodynamic and kinetic study of hydride transfer of a caffeine derivative.

One representative type of heterocyclic compound that can release a hydride ion is 7,8-dihydro-9-methylcaffeine (CAFH). The one-electron oxidation potential of CAFH [-0.294 (V vs Fc(+/0))] and the one-electron reduction potential of CAF(+) [-2.120 (V vs Fc(+/0))] were obtained using two different methods, CV and OSWV. Applying titration calorimetry data in thermodynamic cycles, the enthalpies of CAFH releasing a hydride ion [57.6 kcal/mol] and releasing a hydrogen atom [80.3 kcal/mol] and of its radical cation CAFH(•+) releasing a proton [33.0 kcal/mol] and releasing a hydrogen atom [38.4 kcal/mol] have been determined. Several conclusions can be drawn from the thermodynamic results: (1) CAFH is a very good single-electron donor whose single-electron oxidation potential is much less positive than that of NAD(P)H model compound BNAH [E(ox) = 0.219 V vs Fc(+/0)]. (2) The single-electron reduction potential of CAF(+) is much more negative than that of BNA(+) [E(red) = -1.419 V], which means that CAF(+) is not a good electron acceptor. Furthermore, CAFH is a very good hydride donor compared to BNAH. The results of non-steady-state kinetic studies, for the reaction of CAFH and AcrH(+)ClO(4)(-), show that the ratio of t(0.50)/t(0.05) is larger than 13.5 and the ratio of k(init)/k(pfo) is larger than 1. The pseudo-first-order rate constants obtained at different reaction stages decrease with the time, and the kinetic isotope was observed to be small at a short reaction time and slowly increases to 3.72 with the progress of the reaction. These kinetic results clearly display that the hydride transfer of CAFH to AcrH(+) in acetonitrile is not a one-step mechanism, while the thermodynamic results show that CAFH is a very good electron donor. The combination of the kinetic results with the thermodynamics analysis shows that the hydride transfer of the caffeine derivative CAFH takes place by a two-step reversible mechanism and there is an intermediate in the reaction.

The reversible consecutive mechanism for the reaction of trinitroanisole with methoxide ion.

Although the competitive mechanism for Meisenheimer complex formation during the reaction of 2,4,6-trinitroanisole with methoxide ion in methanol is generally accepted, no kinetic evidence has been presented to rule out a reversible consecutive mechanism. Simulation of the competitive mechanism revealed that a fractional order in [MeO(-)] is predicted by the latter. Conventional pseudo-first-order analysis of the kinetics resulted in cleanly first-order in [MeO(-)], which rules out the competitive mechanism. The kinetic data are consistent with the reversible consecutive mechanism, which is proposed for this important reaction. An intermediate is required for this mechanism, and we propose that a dianion complex (III) is formed reversibly from the initial 1,3-σ complex (I). The trimethoxy complex (III), the (1)H NMR spectrum of which was observed earlier by Servis (Servis, K. L. J. Am. Chem. Soc. 1965, 87, 5495; 1967, 89, 1508), then eliminates methoxide ion reversibly to form the 1,1-σ complex product (II).

Rapid formation and slow collapse of a carbocation-anion pair to a neutral molecule.

The 4,4',4' '-trimethoxytrityl cation (TMT(+)) was observed to react with acetate ion in acetic acid reversibly to give the corresponding ester (TMT-OAc). The rate of the reaction was found to be independent of [NaOAc] over a 25000-fold range. Similar results were observed in the presence of Bu(4)N(+) in acetic acid as well as in HOAc/AN (1/1). It was concluded that {TMT(+) (HOAc/AcO(-))} is an ion pair that forms essentially completely from free TMT(+) and HOAc/AcO(-) during the time of mixing under stopped-flow conditions. The process which was studied kinetically is the intramolecular collapse of the ion pair to TMT-OAc which takes place in two steps involving a kinetically significant intermediate. The remarkably close resemblance of this reaction to the Winstein scheme for solvolysis reactions is noted. In analogy to the Winstein scheme, it was proposed that the intermediate could be an intimate ion pair formed upon extrusion of solvent from the solvent separated ion pair. The product-forming step could then correspond to the intimate ion pair reacting further to form a covalent bond between the two moieties within the complex. The values of the thermodynamic and the activation parameters as well as the apparent rate constants for the reaction in the presence of either sodium or tetrabutylammonium ions suggest that these counterions play insignificant roles in the reactions. However, the equilibrium constant for the intramolecular step (K(4)) was observed to be two times greater in the presence of Bu(4)N(+) than in the presence of Na(+). The rate of the reaction in HOAc was observed to be about four times as great as that in HOAc/AN (1/1).

Determination of N-NO bond dissociation energies of N-methyl-N-nitrosobenzenesulfonamides in acetonitrile and application in the mechanism analyses on NO transfer.

The heterolytic and homolytic N-NO bond dissociation energies of seven substituted N-methyl-N-nitrosobenzenesulfonamides (abbreviated as G-MNBS, G = p-OCH(3), p-CH(3), p-H, p-Cl, p-Br, 2,5-2Cl, m-NO(2)) in acetonitrile solution were evaluated for the first time by using titration calorimetry and relative thermodynamic cycles according to Hess' law. The results show that the energetic scales of the heterolytic and homolytic N-NO bond dissociation energies of G-MNBS in acetonitrile solution cover the ranges from 44.3 to 49.5 and from 33.0 to 34.9 kcal/mol for the neutral G-MNBS, respectively, which indicates that N-methyl-N-nitrosobenzenesulfonamides are much easier to release a NO radical (NO(*)) than to release a NO cation (NO(+)). The estimation of the heterolytic and homolytic (N-NO)(-)(*) bond dissociation energies of the seven G-MNBS radical anions in acetonitrile solution gives the energetic ranges of -15.8 to -12.9 and -3.1 to 1.8 kcal/mol for the (N-NO)(-)(*) bond homolysis and heterolysis, respectively, which means that G-MNBS radical anions are very unstable at room temperature and able to spontaneously or easily release a NO radical or NO anion (NO(-)), but releasing a NO radical is easier than releasing NO anion. These determined N-NO bond dissociation energies of G-MNBS and their radical anions have been successfully used in the mechanism analyses of NO transfer from G-MNBS to 3,6-dibromocarbazole and the reactions of NO with the substituted N-methyl-benzenesulfonamide nitranions (G-MBSN(-)) in acetonitrile solution.

Dissociation energies and charge distribution of the Co-NO bond for nitrosyl-alpha,beta,gamma,delta-tetraphenylporphinatocobalt(II) and nitrosyl-alpha,beta,gamma,delta-tetraphenylporphinatocobalt(III) in benzonitrile solution.

The first two series of Co-NO bond dissociation enthalpies in benzonitrile solution were determined for 12 cobalt(II) nitrosyl porphyrins and for 12 cobalt(III) nitrosyl porphyrins by titration calorimetry with suitable thermodynamic cycles. The results display that the energy scales of the heterolytic Co(III)-NO bond dissociation, the homolytic Co(III)-NO bond dissociation, and the homolytic Co(II)-NO bond dissociation are 14.7-23.2, 15.1-17.5, and 20.8-24.6 kcal/mol in benzonitrile solution, respectively, which not only indicates that the thermodynamic stability of cobalt(II) nitrosyl porphyrins is larger than that of the corresponding cobalt(III) nitrosyl porphyrins for homolysis in benzonitrile solution but also suggests that both cobalt(III) nitrosyl porphyrins and cobalt(II) nitrosyl porphyrins are excellent NO donors, and in addition, cobalt(III) nitrosyl porphyrins are also excellent NO(+) contributors. Hammett-type linear free energy analyses suggest that the nitrosyl group carries negative charges of 0.49 +/- 0.06 and 0.27 +/- 0.04 in T(G)PPCo(II)NO and in T(G)PPCo(III)NO, respectively, which indicates that nitric oxide is an electron-withdrawing group both in T(G)PPCo(II)NO and in T(G)PPCo(III)NO, behaving in a manner similar to Lewis acids rather than to Lewis bases. The energetic and structural information disclosed in the present work is believed to furnish hints to the understanding of cobalt nitrosyl porphyrins' biological functions in vivo.