In vivo binding of 2,3,6,2′,3′,6′-hexachlorobiphenyl and 2,4,5,2′,4′,5′-hexachlorobiphenyl to mouse liver macromolecules

The role of metabolic activation in the binding of polychlorinated biphenyls (PCBs) to cellular macromolecules was investigated in vivo by comparing the relative binding of 2,4,5,2′,4′,5′-[U-¹⁴C]hexachlorobiphenyl (2,4,5), a slowly metabolized PCB, with that of 2,3,6,2′,3′,6′-[U-¹⁴C]hexachlorobiphenyl (2,3,6), a rapidly metabolized PCB, and the appropriate controls. Each hexachlorobiphenyl was administered to mice, orally for 5 days (7.28 mg/kg/day). Following the dosing schedule, animals were killed at 1, 5 and 8 days. The concentration of each PCB was determined in liver, muscle and kidney and in purified macromolecules isolated from those tissues. The concentration of 2,4,5 was consistently higher than the concentration of 2,3,6 in all tissues studied. However, the amount of 2,3,6 bound to the purified macromolecules was consistently at least one order of magnitude greater than that of 2,4,5. The greatest binding was observed in RNA followed by protein and DNA, respectively. The purity of the macromolecules and the presence of PCB-derived radioactivity at the monomer level were confirmed. This is the first report of ¹⁴C-labeled PCB being bound to purified RNA, DNA, and proteins isolated from the tissues of animals treated in vivo. The binding is thought to be covalent and to be the result of metabolic activation.     

The capybara (Hydrochoerus hydrochaeris) as a reservoir host for Trypanosoma evansi

Discovery of two ill horses and three dogs naturally infected with Trypanosoma evansi near an experimental station in the Eastern Plains of Colombia led to a search for reservoir hosts of the parasite. Infection was detected in 8/33 healthy capybaras (Hydrochoerus hydrochaeris), none of the remaining 14 horses, and none of 32 Zebu cattle (Bos indicus), 18 paca (Cuniculus paca) and 20 spiny rats (Proechimys sp.). Contrary to common opinion, the results indicated a carrier state in the capybara. Diagnosis was based on morphology, behaviour in albino rats, and pathogenicity and host range in domestic animals.     

States of myosin subfragment-1 studied by catalyzed ascorbate reduction of bound spin label.

The second-order rate constant, k, whereby ascorbate reduces spin label, N-(1-oxyl-2, 2,6,6-tetramethyl-4-piperidyl) iodoacetamide, bound to the fast-reacting (SH₁) thiol groups of heavy meromyosin (HMM) has been compared with the k whereby ascorbate reduces free spin label in the same solvent. It is clear that the k of protein-bound spin label is primarily determined by conditions “on-board” subfragment-1 (S-1), rather than by properties of the solvent. First, in saturating [STP] the k of HMM-bound spin label was much greater than the k of free spin label, and both k's were independent of [KCl], from 0.05 to 1 m. Second, in the absence of ATP, or even in the presence of ADP, the k of HMM-bound spin label was less than the k of free spin label at l m KCl, and much more in a 0.05 m KCl. The organized structure of S-1 is required for observing the change of k with ATP, because the change of k disappeared on denaturing HMM with either guanidine hydrochloride or urea.Measuring k can be a “probe” to specify HMM states. However, the parameter, k, is conceptually dissimilar to measuring peak heights on an EPR spectrum. Experimentally we have observed that when [KCl] is increased, while [MgATP] = 0, spectral peak height is constant, but k varies remarkably. At no [KCl] did excess F-actin affect k. Quantitative examination of metal contamination (e.g., Cu, Fe) in HMM showed that changes in the k of HMM-bound spin label cannot arise from changes in proximity to contaminating metal redox catalysts bound to HMM.An intramolecular participant in the reaction of ascorbate with bound nitroxyl half inhibits the Ca²⁺-ATPase of spin labeled HMM, so signal annihilation and ATPase activity are closely correlated in time. The rate of signal annihilation is unaffected by prior reaction of the “SH₂” thiols with N-ethylmaleimide.     

Defining the "fast-reacting" thiols of myosin by reaction with 1, 5 IAEDANS.

The specificity of the fluorescent reagent N-iodoacetyl-N-(5-sulfo-1-naphthyl)ethylenediamine (1,5 IAEDANS) for a specific thiol group of myosin has been characterized by a comparison with iodoacetamide (IAA) and by observing maximal enhancement of the Ca²⁺-ATPase activity and inhibition of the K⁺-EDTA-ATPase activity of myosin. The stoichiometry of the [³H]1,5 IAEDANS bound to myosin indicates the presence of two fast-reacting thiols which correspond to the “SH₁” groups responsible for the catalytic properties of myosin. Moreover, it has been unequivocally demonstrated by gel electrophoresis that the fast-reacting thiol is located on the myosin heavy chain. A single radioactivity-labeled thiol peptide obtained from tryptic digests of myosin labeled with [³H]1,5 IAEDANS or iodo[1-¹⁴C]acetamide indicates strongly that the identical thiol was labeled by both reagents.     

A synaptic model for the kindling effect

In this work we present a model of the kindling effect based on the Hopf bifurcation for a system of ordinary differential equations. The model shows how quantitative changes in the physiological parameters at the microscopic, synaptic scale, produce the afterdischarge which is a macroscopic effect at the neuronal network scale. The presynaptic mechanisms are based on the vesicular hypothesis, or more generally on the quantal theory of synaptic transmission. The postsynaptic processes rely on Granit's law. This model gives a consistent framework which organizes and explains several experimental observations.     

Asynchrony of labyrinthine receptors as a physical factor in motion sickness

Evidence is presented to show that despite their effectiveness in producing motion sickness the actual frequencies and accelerations involved in ship motion are rather low. It is suggested that the disparity is due to the fact that ship motion is such as to cause an asynchrony between signals from the labyrinth and those from other motion receptors.     

ZAKβ antagonizes and ameliorates the cardiac hypertrophic and apoptotic effects induced by ZAKα

ZAK (sterile alpha motif and leucine zipper containing kinase AZK), a serine/threonine kinase with multiple biochemical functions, has been associated with various cell processes, including cell proliferation, cell differentiation, and cardiac hypertrophy. In our previous reports, we found that the activation of ZAKα signaling was critical for cardiac hypertrophy. In this study, we show that the expression of ZAKα activated apoptosis through both a FAS‐dependent pathway and a mitochondria‐dependent pathway by subsequently inducing caspase‐3. ZAKβ, an isoform of ZAKα, is dramatically expressed during cardiac hypertrophy and apoptosis. The interaction between ZAKα and ZAKβ was demonstrated here using immunoprecipitation. The results show that ZAKβ has the ability to diminish the expression level of ZAKα. These findings reveal an inherent regulatory role of ZAKβ to antagonize ZAKα and to subsequently downregulate the cardiac hypertrophy and apoptosis induced by ZAKα.     

Global phylogeny of Hadrosauridae (Dinosauria: Ornithopoda) using parsimony and Bayesian methods

The late Cretaceous hadrosaurids were the most specialized and diverse clade of ornithopod dinosaurs. Parsimony and Bayesian methods were implemented to elucidate the phylogenetic relationships of all hadrosaurid species. Traditional and geometric morphometrics were applied to discover patterns of variation containing phylogenetic information. In total, 286 phylogenetically informative characters (196 cranial and 90 postcranial) were defined and documented: the most extensive character data set ever constructed for hadrosaurid dinosaurs. Of these, 136 characters were used for the first time in phylogenetic analysis of these ornithopods, and 93 were modified from those of other authors. Parsimony and the Bayesian analysis (using the Mk model without the gamma parameter) confirmed the split of hadrosaurids into Saurolophinae and Lambeosaurinae. Saurolophines included a major clade composed of the Prosaurolophus–Saurolophus and the Kritosaurus–Gryposaurus–Secernosaurus subclades. Edmontosaurus and Shantungosaurus were recovered outside the major clade of saurolophines. The Brachylophosaurus clade was recovered as the most basal clade of saurolophines in the parsimony analysis, whereas following the Bayesian analysis it was recovered as the sister clade to the Kritosaurus–Gryposaurus–Secernosaurus clade. These two analyses resulted in a Lambeosaurinae composed of a succession of Eurasian sister taxa to two major clades: the Parasaurolophus clade and the Hypacrosaurs altispinus–Corythosaurus clade. In contrast, the Bayesian analysis using the Mk model with the gamma parameter included, resulted in an unbalanced hadrosauroid tree, with a paraphyletic Saurolophinae, and with the Prosaurolophus clade, Edmontosaurus, and Shantungosaurus as successively closer sister taxa to Lambeosaurinae. Based on the strict reduced consensus tree derived from the parsimony analysis, Hadrosauridae was redefined as the clade stemming from the most recent common ancestor of Hadrosaurus foulkii and Parasaurolophus walkeri. © 2010 The Linnean Society of London, Zoological Journal of the Linnean Society, 2010, 159 , 435–502.