Basis for the cardiac-related rhythm in muscle sympathetic nerve activity of humans

We tested the hypothesis that the cardiac-related rhythm in muscle sympathetic nerve activity (MSNA) of humans reflects entrainment of a central oscillator by pulse-synchronous baroreceptor nerve activity. Partial autospectral analysis was used to mathematically remove the portion of cardiac-related power in MSNA autospectra that was attributable to its linear relationship to the ECG. In 54 of 98 cases, > or =15% of cardiac-related power remained after partialization with the ECG; peak residual cardiac-related power was often at a frequency different than heart rate. When assessed on a cardiac-related burst-by-burst basis, there was a progressive and cyclic change in the ECG-MSNA interval (delay from R wave to peak of cardiac-related burst) on the time scale of respiration in four subjects. In these subjects, as well as in some in which the interval appeared to change randomly, there was an inverse relationship between the ECG-MSNA interval and cardiac-related burst amplitude. However, in 45% of the cases, these parameters were not related. These results support the view that the cardiac-related rhythm in MSNA reflects forcing of a nonlinear oscillator rather than periodic inhibition of unstructured, random activity.     

Kinetics of the initial steps of rabbit psoas myofibrillar ATPases studied by tryptophan and pyrene fluorescence stopped-flow and rapid flow-quench. Evidence that cross-bridge detachment is slower than ATP binding

The kinetics of the tryptophan fluorescence enhancement that occurs when myofibrils (rabbit psoas) are mixed with Mg-ATP were studied by stopped-flow in different solvents (water, 40% ethylene glycol, 20% methanol) at 4 degrees C. Under relaxing conditions (low Ca(2+)) in water (mu = 0.16 M, pH 7.4) and at high ATP concentrations, the transient was biphasic, giving a k(fast)(max) of 230 s(-)(1) and a k(slow)(max) of 15 s(-)(1). The kinetics of the two phases were compared with those obtained by chemical sampling using [gamma-(32)P]ATP and quenching in acid (P(i) burst experiments: these give unambiguously the ATP cleavage kinetics), or cold Mg-ATP (cold ATP chase: ATP binding kinetics). k(slow) is due to ATP cleavage, as with S1. Interestingly, k(fast) is slower than the ATP binding kinetics. Instead, this constant appears to report ATP-induced cross-bridge detachment from actin because (1) it was identical to the fluorescence transient obtained on addition of ATP to pyrene-labeled myofibrils; (2) when the initial filament overlap in the myofibrils was decreased, the amplitude of the fast phase decreased; (3) there was no fluorescent enhancement upon the addition of ADP to myofibrils. This is different from the situation with S1 or actoS1 where there was also a fast fluorescent ATP-induced transient but whose kinetics were identical to those of the tight ATP binding. To increase the time resolution and to confirm our results, we also carried out transient kinetics in ethylene glycol and methanol. We interpret our results by a scheme in which a rapid equilibrium between attached (AM.ATP) and detached (M.ATP) states is modulated by the fraction of myosin heads in rigor (AM) during the time of experiment.     

Medullary lateral tegmental field: an important source of basal sympathetic nerve discharge in the cat

We used blockade of excitatory amino acid (EAA) neurotransmission in the medullary lateral tegmental field (LTF) and rostral ventrolateral medulla (RVLM) to assess the roles of these regions in the control of inferior cardiac sympathetic nerve discharge (SND) and mean arterial pressure (MAP) in urethan-anesthetized, baroreceptor-denervated cats. Bilateral microinjection of a non-N-methyl-D-aspartate (NMDA)-receptor antagonist [1,2,3, 4-tetrahydro-6-nitro-2,3-dioxobenzo-[f]quinoxaline-7-sulfonamide (NBQX)] into the LTF significantly decreased SND to 46 +/- 4% of control (as demonstrated with power-density spectral analysis) and MAP by 16 +/- 6 mmHg. In contrast, bilateral microinjection of an NMDA-receptor antagonist [D(-)-2-amino-5-phosphonopentanoic acid (D-AP5)] into the LTF did not decrease SND or MAP. These results demonstrate that the LTF is an important synaptic relay in the pathway responsible for basal SND in the cat. Bilateral microinjection of NBQX or D-AP5 into the RVLM significantly decreased power in SND to 48 +/- 5 or 61 +/- 5% of control, respectively, and reduced MAP by 15 +/- 2 or 8 +/- 4 mmHg, respectively. These data indicate that EAA-mediated synaptic drive to RVLM-spinal sympathoexcitatory neurons accounts for a significant component of their basal activity.     

Quantification of organic eluates from polymerized resin-based dental restorative materials by use of GC/MS

Residual monomers, additives and degradation products from resin-based dental restorative materials eluted into the oral cavity may influence the biocompatibility of these materials. Emphasis has been placed on studies addressing cytotoxic, genotoxic and estrogenic potential of these substances. A prerequisite for analyzing the potential of exposure to eluted compounds from dental materials is reliable quantification methods, both real time and accelerated measurements. The purpose of the present study was to quantify nine eluates; 2-hydroxyethyl methacrylate (HEMA), hydroquinone monomethyl ether (MEHQ), camphorquinone (CQ), butylated hydroxytoluene (BHT), ethyl 4-(dimethylamino)benzoate (DMABEE), triethylene glycoldimethacrylate (TEGDMA), trimethylolpropane trimethacrylate (TMPTMA), oxybenzone (HMBP) and drometrizole (TIN P) leaching from specimens of four commonly used resin-based dental materials in ethanol and an aqueous solution. All analyses were performed by use of GC/MS, each component was quantified separately and the results presented in microg mm(-2). This study has shown that elution from various materials differs significantly, not only in the types of eluates, but also regarding amounts of total and of single components. A high amount of HMBP, a UV stabilizer with potential estrogenic activity, was detected from one material in both solutions.     

Interactions of a few azole derivatives with a transport protein: role of heteroatoms

The interaction of a few azole derivatives, 2‐(4′‐N,N‐dimethylaminophenyl)benzimidazole, 2‐(4′‐N,N‐dimethylaminophenyl)benzoxazole, 2‐(4′‐N,N‐dimethylaminophenyl)oxazolo[4,5‐b]pyridine with bovine serum albumin (BSA) were examined by absorption and fluorescence spectroscopy. The results were compared with the previously studied imidazopyridine derivative 2‐(4′‐N,N‐dimethylaminophenyl)imidazo[4,5‐b]pyridine. Displacement studies were carried out with site selective probes to locate the binding site of these ligands. The spectral shifts and the binding constant vary depending on the nature of the ligand. The fluorescence intensity of both oxazole derivatives 2‐(4′‐N,N‐dimethylaminophenyl)benzoxazole and 2‐(4′‐N,N‐dimethylaminophenyl) oxazolo[4,5‐b]pyridine increases substantially in the presence of BSA, whereas the intensity of 2‐(4′‐N,N‐dimethylaminophenyl)benzimidazole decreases. However, hypsochromic shift is observed in presence of BSA. The results obtained from the docking studies are also in good agreement with the experimental results. The location and orientation of binding depend upon the nature of the ligand. The studies revealed that apart from hydrophobic interaction, hydrogen bonding also plays a vital role in the molecular binding. Oxazoles have higher binding affinity than imidazoles and substitution of extra nitrogen further increases the binding affinity. Copyright © 2015 John Wiley & Sons, Ltd.     

Temporal relationship between emitted and endogenous floral scent volatiles in summer‐ and winter‐blooming Jasminum species

Jasminum spp. is cultivated for their fragrant flowers used in essential oil production and cosmetic uses. An attempt was made to study the temporal variations in floral scent volatiles composition including emitted, free endogenous and glycosyl‐linked volatile compounds from two summer‐blooming species namely, Jasminum auriculatum and Jasminum grandiflorum as well as from two winter‐blooming species namely, Jasminum multiflorum and Jasminum malabaricum. The overall emitted volatile organic compounds (VOCs) were found to be highest when the matrix Porapak Q 80/100 was used with dichloromethane (DCM) as elution solvent. The floral volatile emission from bud to senescence exhibited nocturnal maxima pattern for both the summer‐blooming species. Both the winter‐blooming species emitted its highest concentration at noon. The free endogenous concentrations of all VOCs were low when corresponding emitted concentrations were high. Enzymatic treatment of petal extract revealed that several aromatic volatiles including aromatic alcohols and monoterpenols are synthesized and stored in the flowers as water‐soluble glycosides; these compounds were shown to accumulate in higher amounts in flowers at late bud stage. These findings indicate the utilization of the precursors, i.e. the volatile‐conjugates, through hydrolysis followed by their release as free‐volatiles at flower opening stage. The outcome as a whole suggests a linkage among the temporal pattern of emitted volatiles, free‐endogenous volatiles and glycoside‐bound volatile compounds in all above studied Jasminum spp. and provided an overview of their floral volatilome.     

Protective efficacy of maternal antibodies induced by Salmonella toxoid (vaccine)

An attempt was made to evaluate the protective efficacy of maternal antibodies in chicks against salmonellosis. Layer chicks ageing 21 days were individually vaccinated with 100 microg of Salmonella enterica subspecies enterica serovar Weltevreden (BM 1643) toxoid adjuvanted with vitamin E subcutaneously. After 90 days of the primary vaccination the birds were given booster dose of the vaccine. The saline extract of the yolk of eggs laid by the vaccinated birds yielded agglutination and ELISA titres ranging from 43.2 +/- 5.33 to 75.2 +/- 6.26 and 4.987 x 10(3) +/- 0.54 to 5.89x103 +/- 0.56, respectively. Sera of chicks hatched from eggs laid by the vaccinated layers were also subjected to agglutination and ELISA. Agglutination and ELISA titres on the 5th day--post hatching (dph) were 21.6 +/- 1.75 and 4.025 x 10(2) +/- 0.59, while on the 10th dph titers were 13.6 +/- 1.65 and 1.21 x 10(2) +/- 0.60, respectively. It was also observed that only one out of 6 chicks died when challenged with 2 x 10(9) CFU of S. serovar Gallirarum at the age of 7 days showing 83.33% protection. Thus it can be concluded that passive immunity confided by Salmonella enterica subspecies enterica serovar Weltevreden (BM 1643) toxoid can protect chicks against salmonellosis during their early days of life.     

Role of transmembrane domain and cytoplasmic tail amino acid sequences of influenza a virus neuraminidase in raft association and virus budding

Influenza virus neuraminidase (NA), a type II transmembrane glycoprotein, possesses receptor-destroying activity and thereby facilitates virus release from the cell surface. Among the influenza A viruses, both the cytoplasmic tail (CT) and transmembrane domain (TMD) amino acid sequences of NA are highly conserved, yet their function(s) in virus biology remains unknown. To investigate the role of amino acid sequences of the CT and TMD on the virus life cycle, we systematically mutagenized the entire CT and TMD of NA by converting two to five contiguous amino acids to alanine. In addition, we also made two chimeric NA by replacing the CT proximal one-third amino acids of the NA TMD [NA(1T2N)NA] and the entire NA TMD (NATRNA) with that of human transferrin receptor (TR) (a type II transmembrane glycoprotein). We rescued transfectant mutant viruses by reverse genetics and examined their phenotypes. Our results show that all mutated and chimeric NAs could be rescued into transfectant viruses. Different mutants showed pleiotropic effects on virus growth and replication. Some mutants (NA2A5, NA3A7, and NA4A10) had little effect on virus growth while others (NA3A2, NA5A27, and NA5A31) produced about 50- to 100-fold-less infectious virus and still some others (NA5A14, NA4A19, and NA4A23) exhibited an intermediate phenotype. In general, mutations towards the ectodomain-proximal sequences of TMD progressively caused reduction in NA enzyme activity, affected lipid raft association, and attenuated virus growth. Electron microscopic analysis showed that these mutant viruses remained aggregated and bound to infected cell surfaces and could be released from the infected cells by bacterial NA treatment. Moreover, viruses containing mutations in the extreme N terminus of the CT (NA3A2) as well as chimeric NA containing the TMD replaced partially [NA(1T2N)NA] or fully (NATRNA) with TR TMD caused reduction in virus growth and exhibited the morphological phenotype of elongated particles. These results show that although the sequences of NA CT and TMD per se are not absolutely essential for the virus life cycle, specific amino acid sequences play a critical role in providing structural stability, enzyme activity, and lipid raft association of NA. In addition, aberrant morphogenesis including elongated particle formation of some mutant viruses indicates the involvement of NA in virus morphogenesis and budding.     

Fractal properties of human muscle sympathetic nerve activity

Muscle sympathetic nerve activity (MSNA) in resting humans is characterized by cardiac-related bursts of variable amplitude that occur sporadically or in clusters. The present study was designed to characterize the fluctuations in the number of MSNA bursts, interburst interval, and burst amplitude recorded from the peroneal nerve of 15 awake, healthy human subjects. For this purpose, we used the Allan and Fano factor analysis and dispersional analysis to test whether the fluctuations were time-scale invariant (i.e., fractal) or random in occurrence. Specifically, we measured the slopes of the power laws in the Allan factor, Fano factor, and dispersional analysis curves. In addition, the Hurst exponent was calculated from the slope of the power law in the Allan factor curve. Whether the original time series contained fractal fluctuations was decided on the basis of a comparison of the values of these parameters with those for surrogate data blocks. The results can be summarized as follows. Fluctuations in the number of MSNA bursts and interburst interval were fractal in each of the subjects, and fluctuations in burst amplitude were fractal in four of the subjects. We also found that fluctuations in the number of heartbeats and heart period (R-R interval) were fractal in each of the subjects. These results demonstrate for the first time that apparently random fluctuations in human MSNA are, in fact, dictated by a time-scale-invariant process that imparts "long-term memory" to the sequence of cardiac-related bursts. Whether sympathetic outflow to the heart also is fractal and contributes to the fractal component of heart rate variability remains an open question.     

Estrogen-induced contraction of coronary arteries is mediated by superoxide generated in vascular smooth muscle

Although previous studies demonstrated beneficial effects of estrogen on cardiovascular function, the Women's Health Initiative has reported an increased incidence of coronary heart disease and stroke in postmenopausal women taking hormone replacement therapy. The objective of the present study was to identify a molecular mechanism whereby estrogen, a vasodilatory hormone, could possibly increase the risk of cardiovascular disease. Isometric contractile force recordings were performed on endothelium-denuded porcine coronary arteries, whereas molecular and fluorescence studies identified estrogen signaling molecules in coronary smooth muscle. Estrogen (1-1,000 nM) relaxed arteries in an endothelium-independent fashion; however, when arteries were pretreated with agents to uncouple nitric oxide (NO) production from NO synthase (NOS), estrogen contracted coronary arteries with an EC(50) of 7.3 +/- 4 nM. Estrogen-induced contraction was attenuated by reducing superoxide (O(2)(-)). Estrogen-stimulated O(2)(-) production was detected in NOS-uncoupled coronary myocytes. Interestingly, only the type 1 neuronal NOS isoform (nNOS) was detected in myocytes, making this protein a likely target mediating both estrogen-induced relaxation and contraction of endothelium-denuded coronary arteries. Estrogen-induced contraction was completely inhibited by 1 muM nifedipine or 10 muM indomethacin, indicating involvement of dihydropyridine-sensitive calcium channels and contractile prostaglandins. We propose that a single molecular mechanism can mediate the dual and opposite effect of estrogen on coronary arteries: by stimulating type 1 nNOS in coronary arteries, estrogen produces either vasodilation via NO or vasoconstriction via O(2)(-).