Observation of the iron-sulfur cluster in Escherichia coli biotin synthase by nanoflow electrospray mass spectrometry

Biotin synthase from Escherichia coli was analyzed by nanoflow electrospray ionization mass spectrometry. From solution conditions in which the protein is in its native state, a distribution of monomeric, dimeric, and tetrameric species was observed. The distribution of these species was sensitive to changes in ionic strength: in the positive ion spectrum, biotin synthase at low ionic strength (pH 7.0-8.5) yielded less than 10% dimer. The masses of the monomeric species were consistent with the presence of a [2Fe-2S] cluster with a mass difference of 175.3 Da from the apomonomer with one disulfide bond. Despite the molecular mass of the noncovalent dimer (77 kDa), it was possible to observe a dimeric species containing one iron-sulfur cluster in both positive and negative ion spectra. Additionally, observation of a series of charge states assigned to the apodimer indicated that binding of the iron-sulfur cluster was not required to maintain the dimer. Binding of Cu2+ to biotin synthase was also observed; in the presence of excess chelating agent, free metals were removed and the iron-sulfur cluster remained intact. Evidence for the coordination of the iron-sulfur cluster in biotin synthase was obtained in a tandem mass spectrometry experiment. A single charge state containing the cluster at m/z 2416.9 was isolated, and collision-induced dissociation resulted in sequential loss of sulfur and retention of Fe3+.     

Development of an intravascular impedance catheter for detection of fatty lesions in arteries

Tissue angiotensin II (AngII) is increased in the infarcted rat heart, where it may have autocrine or paracrine properties that influence cellular protein synthesis and growth and therefore tissue repair. It was our hypothesis that treatment with an AT1 receptor antagonist would attenuate fibrous tissue formation after myocardial infarction (MI). To investigate a role for local AngII in the regulation of connective tissue formation during early and late wound healing that follows MI, this study was undertaken. Animals were randomized into two groups in which rats were or were not treated with the AT1 receptor antagonist losartan (10 mg x kg(-1) daily gavage). At 1 and 4 weeks after experimental MI was induced by coronary artery ligation, rat hearts were examined. Infarct size, infarct area, and collagen volume fraction at the site of infarction and in noninfarcted myocardium were determined by picrosirius red staining with videodensitometry. Quantitative in vitro autoradiography was used to detect AngII receptor binding density ((125)I-(Sar1,Ile8)AngII). Compared with an untreated MI control group, in losartan-treated rats we found (1) infarct size was comparable in both groups at weeks 1 and 4, (2) infarct area was comparable between groups at week 1 but was significantly reduced (p < 0.05) at week 4 in losartan-treated rats, (3) a detectable reduction in collagen volume fraction at the site of MI was not found at week 1 but was reduced (p < 0.05) at remote sites at week 4, (4) AngII receptor binding density was reduced (p < 0.05) by 50% at the site of MI at both weeks 1 and 4 in keeping with delivery of losartan to this site of injury. Thus AT1 receptor antagonism appears to influence late phase wound healing at and remote to the site of MI and suggests an association between AngII and the fibrogenic response that appears in the injured rat heart. Although still speculative, an attenuation in fibrosis after MI may account for less ventricular dysfunction and geometric remodeling of right and left ventricles and ventricular arrhythmias that have been observed in such rats treated with angiotensin converting enzyme inhibitor or AT1 receptor antagonist.     

A Social Relations Model analysis of parent and child play.

Introduces the Social Relations Model (Kenny & LaVoie, 1984) as a new approach for understanding reciprocity during parent–child and sibling play. 32 12-month-old infants and their 3- to 4-year-old siblings were videotaped during separate dyadic sessions in a laboratory playroom with their mothers, their fathers, and each other. Each partner's play was coded using a detailed classification system consisting of six discrete play behaviors. Multivariate analyses indicated that mother–child dyads engaged in more instructive play than father–child dyads, although both parents demonstrated age-appropriate adjustments in their play with their two children. Infants and preschoolers engaged in more elaborate types of play with their parents than with their less socially competent siblings. Social Relations Model analyses revealed that the unique adjustments that family members made to one another outweighed family members' general tendencies to behave in particular ways and the partners' tendencies to elicit particular behaviors from family members. Furthermore, these analyses suggested that parents may be primarily responsible for much of the structuring occurring in children's early social interactions. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Effects of Gac aril microwave processing conditions on oil extraction efficiency,and β-carotene and lycopene contents

The effects of Gac oil extraction conditions including microwave power, microwave time, steaming time and hydraulic pressure on extraction efficiency (EE), and β-carotene and lycopene contents were studied. It was found that the EE, and β-carotene and lycopene contents could be enhanced by suitable extraction conditions. Microwave drying was found to be better than air drying for pretreatment. Moisture content after drying and steaming between 8% and 11% (wt/wt) were best for pressing. Results showed that the most suitable conditions for Gac oil extraction from 900 g samples were microwave power of 630 W, microwave time of 65 min, steaming time of 20 min and hydraulic pressure of 170 kg/cm². Under these conditions, the highest EE of 93% was achieved while Gac oil contained the highest content of β-carotene and lycopene at 140 and 414 mg/100 mL, respectively.