Ferroxidase kinetics of human liver apoferritin, recombinant H-chain apoferritin, and site-directed mutants

A detailed study of the kinetics of iron(II) oxidation by molecular oxygen in natural and recombinant human apoferritins has been carried out using electrode oximetry to better understand the ferroxidase activity of the protein shell. A comparative study of recombinant L-chain ferritin (rLF), recombinant H-chain ferritin (rHF), and variants has shown that (1) rLF lacks a ferroxidase activity, confirming the results of previous studies; (2) the ferroxidase site of rHF involves Glu-62 and His-65, presumably as Fe2+ ligands, since mutation of these residues abolishes most of the oxidase activity, in agreement with previous studies; and (3) mutation of both the putative ferroxidase and nucleation site ligands in rHF renders the protein totally incapable of catalyzing the oxidation of Fe2+ whereas mutation of nucleation site ligands alone (Glu-61, Glu-64, and Glu-67) decreases the activity only slightly. Analysis of the kinetics of rHF and natural human liver ferritin (HLF) (4% H-chain, 96% L-chain) gave the following apparent parameters at pH 7: Km,O2 = 6 +/- 2 microM, Km,Fe = 80 +/- 10 microM, and kcat = 201 +/- 14 min-1 for rHF and Km,O2 = 60 +/- 12 microM, Km,Fe = 50 +/- 10 microM, and kcat = 31.2 +/- 0.6 min-1 for HLF. Furthermore, Zn2+ was shown to be a noncompetitive inhibitor of Fe2+ oxidation in rHF but a mixed inhibitor in HLF. These different forms of Zn2+ inhibition in the two proteins and the higher activity of HLF than expected, based on its H-chain composition as well as differences in their enzyme kinetic parameters, suggest that H- and L-chains cooperate in modulating the ferroxidase activity of the apoferritin even though the L-subunit lacks a ferroxidase site itself.     

Cotrel-Dubousset instrumentation. Results in 52 patients

Fifty-two posterior spinal fusions were performed for pediatric idiopathic, congenital, and neuromuscular scoliotic curves. Cotrel-Dubousset instrumentation was used in all patients. Nine had prior anterior spinal releases and fusions. The patterns were mixed, with a predominance of right thoracic curvatures. The average preoperative curve measured 60.6 degrees, with correction to 29. Seven patients required revision surgery, and 17 wore orthoses after operation. There were 17 complications in this group, including hook pullout, prominent hardware, infection, pseudarthrosis, and two cases of broken Cotrel-Dubousset instrumentation rods. Fatigue failure of this instrumentation, secondary to pseudarthrosis, has not been reported previously, and these two cases are presented in detail. The operative morbidity and difficulty were increased in the larger idiopathic curves and in neuromuscular and congenital scoliosis. Cotrel-Dubousset instrumentation is an overall excellent tool for the multiplanar correction of scoliosis and is amenable to revision surgery.     

Neurologic evaluation of the equine head and neurogenic dysphagia

Neurologic evaluation should be performed in horses with diseases of the head. Although neurologic examination should focus on assessing behavior, mental status, and cranial nerve evaluation, evaluation of neurologic function of other body regions should be performed. Neurologic evaluation of the head can be performed expediently by practitioners to provide useful diagnostic and prognostic information. The numerous causes of dysphagia can be classified as obstructive, painful, or neurogenic. Common causes of neurogenic dysphagia are summarized, and methods for initial diagnosis and management are described. Maintaining adequate nutrition and preventing aspiration pneumonia are principal concerns in managing horses with neurogenic dysphagia.     

Increase of aminotransferases

Elevated aminotransferases activities are frequent in medical practice. In acute elevations, the mains causes are generally easily found (viral, drug-induced, toxic, ischemic). In moderate or prolonged elevations, the most frequent causes are steatosis (alcoholic, diabetes, obesity) and chronic hepatitis (viral B, D, C, drug-induced and auto-immune diseases.     

Willing and able to fake emotions: A closer examination of the link between emotional dissonance and employee well-being.

Emotional dissonance resulting from an employee's emotional labor is usually considered to lead to negative employee outcomes, such as job dissatisfaction and emotional exhaustion. Drawing on Festinger's (1957) cognitive dissonance theory, we argue that the relationship between service employees' surface acting and job dissatisfaction and emotional exhaustion is moderated by 2 aspects of a service worker's self-concept: the importance of displaying authentic emotions (reflecting the self-concept's self-liking dimension) and the employee's self-efficacy when faking emotions (reflecting the self-competence dimension). A survey of 528 frontline employees from a wide variety of service jobs provides support for the moderating role of both self-concept dimensions, which moderate 3 out of 4 relationships. Theoretical and practical implications are discussed from the perspectives of cognitive dissonance and emotional labor theories. (PsycINFO Database Record (c) 2011 APA, all rights reserved)     

Effect of increased afterload on cardiac lipoprotein lipase and VLDL receptor expression

Fatty acids are a major source of fuel for energy production by myocytes. Lipoprotein lipase (LPL) and very low density lipoprotein (VLDL) receptor are abundantly expressed by the heart and skeletal muscles. LPL and possibly VLDL receptor represent the primary route of access to fatty acids contained in circulating triglyceride-rich lipoproteins. Physical exercise and thyroid hormone, which promote energy consumption, upregulate LPL expression in skeletal muscles. This study tested the hypothesis that increased cardiac workload might modulate myocardial LPL and/or VLDL receptor expressions. Accordingly, cardiac tissue LPL activity, LPL and VLDL receptor proteins and mRNA abundance were studied in Sprague-Dawley rats 4 weeks after induction of severe thoracic aorta constriction or sham operation. Elevation of afterload with thoracic aortic constriction led to a significant cardiomegaly and a marked upregulation of cardiac LPL activity, LPL mRNA and LPL protein abundance, but did not modify VLDL receptor mRNA or protein abundance. Thus, increased cardiac workload in this model results in upregulation of myocardial LPL expression which can enhance fatty acid availability to accommodate the heart's increased energy requirement.     

Activation of p38 mitogen-activated protein kinase by lipopolysaccharide in human neutrophils requires nitric oxide-dependent cGMP accumulation

This study examined the signal transduction pathway(s) leading to phosphorylation of p38 in human neutrophils stimulated with lipopolysaccharide and formyl peptides. Blockade of the nitric oxide (NO) pathway in neutrophils with the NO synthase inhibitor N-nitro-L-arginine methyl ester or by treatment with the NO scavenger 2-phenyl-tetramethylimidazoline-1-oxyl-3-oxide attenuated phosphorylation of the mitogen-activated protein kinase p38 in response to lipopolysaccharide but not fMet-Leu-Phe. Using the NO releasing agents S-nitroso-N-acetylpenicillamine and sodium nitroprusside it was determined that nitric oxide is sufficient to cause an increase in phosphorylation of p38. Increasing cellular cGMP with phosphodiesterase inhibitors, by stimulation of soluble guanylyl cyclase with YC-1 or with exogenous dibutyryl cGMP resulted in mitogen-activated protein kinase/extracellular signal-regulated kinase kinase 3,6 (MEK3,6) activation and phosphorylation of p38. This phenomenon was specific for MEK3,6, because these agents had no effect on the phosphorylation state of MEK1,2. A role for protein kinase G but not protein kinase A downstream of lipopolysaccharide but not formylmethionylleucylphenylalanine was shown using the specific inhibitors KT5823 and H89, respectively. These data indicate that activation of p38 by fMet-Leu-Phe and lipopolysaccharide involve different mechanisms, and that activation of protein kinase G by NO-dependent stimulation of guanylyl cyclase is necessary and sufficient for phosphorylation of p38 downstream of lipopolysaccharide.     

Typicality effects in attitudes toward social policies: A concept-mapping approach.

Social typicality effects occur when people apply their attitudes more consistently toward typical than toward atypical category members presumably because attitudes are directed toward the prototypic category member. Four studies tested whether individuals also apply social policy attitudes more consistently toward typical than toward atypical persons affected by the policy. Study 1 used attitude concept maps to assess how extensively individuals spontaneously reference the typical person affected by a social policy. Death penalty proponents did so more than did opponents; welfare proponents and opponents did so equally. In 3 other studies, death penalty proponents displayed greater typicality effects in sentencing "convicted murderers" than did opponents, welfare proponents and opponents displayed equal typicality effects in evaluating cases of suspected welfare fraud, and individuals whose attitude concept maps about welfare referred more to persons displayed greater typicality effects than did individuals whose maps referred more to principles. (PsycINFO Database Record (c) 2010 APA, all rights reserved)