Genetic fusions of subunit c in the F0 sector of H+-transporting ATP synthase. Functional dimers and trimers and determination of stoichiometry by cross-linking analysis

The multicopy c subunit of the H+-transporting ATP synthase of Escherichia coli folds through the transmembrane F0 sector as a hairpin of two hydrophobic alpha-helices with the proton-translocating aspartyl-61 side chain centered in the second transmembrane helix. The number of subunits c in the F0 complex, which is thought to determine the H+-pumping/ATP stoichiometry, was previously not determined with exactness but thought to range from 9-12. The studies described here indicate that the exact number is 12. Based upon the precedent of the subunit c in vacuolar-type ATPases, which are composed of four transmembrane helices and seem to have evolved by gene duplication of an F0-type progenitor gene, we constructed genetically fused dimers and trimers of E. coli subunit c. Both the dimeric and trimeric forms proved to be functional. These results indicate that the total number of subunit c in F0 should be a multiple of 2 and 3. Based upon a previous study in which the oligomeric organization of c subunits in F0 was determined by cross-linking of Cys-substituted subunits (Jones, P. C. , Jiang, W., and Fillingame, R. H. (1998) J. Biol. Chem. 273, 17178-17185), we introduced Cys into the first and last transmembrane helices of subunit c monomers, dimers, and trimers and attempted to generate cross-linked products by oxidation with Cu(II)-(1,10-phenanthroline)2. Double Cys substitutions at two sets of positions gave rise to extensive cross-linked multimers. Multimers of the monomer that extended up to the position of c12 were correlated and calibrated with distinct cross-linked species of the appropriate doubly Cys-substituted dimers (i.e. c2, c4, . c12) and doubly Cys-substituted trimers (i.e. c3, c6, c9, c12). The results show that there are 12 copies of subunit c per F0 in E. coli, the exact number having both mechanistic and structural significance.     

Methods of measuring susceptibility of anaerobic bacteria to trovafloxacin, including quality control parameters

Three methods approved by the National Committee for Clinical Laboratory Standards for testing the susceptibility of anaerobic bacteria were used to evaluate the fluoroquinolone, trovafloxacin. The methods gave essentially comparable results with 126 anaerobes and with three quality control strains. A collaborative study defined the quality control range for trovafloxacin MICs. Trovafloxacin had good in vitro activity against the more common anaerobes (MIC 90 < = or 2.0 micrograms/ml).     

Acute arthritis of cats associated with feline calicivirus infection

Twelve specific pathogen-free cats were infected either by intra-articular inoculation or by contact exposure to one of two strains of feline calicivirus (FCV), either F65, a field strain originating from an outbreak of lameness in a group of cats, or a vaccine strain. Following either route of exposure, both strains induced signs typical of FCV infection including oral and nasal ulceration, conjunctivitis and ocular discharge. These signs were of equal severity for both virus strains, but overall, following either route of infection, F65 induced more severe disease than the vaccine strain, with marked pyrexia, lethargy and lameness. Vaccine virus only induced a relatively mild lameness following intra-articular inoculation. Gross pathological and histopathological lesions were seen in some of the joints, but again changes were more severe in the F65-exposed cats. Virus was isolated from both normal and affected joints from both groups of F65-exposed cats, and from a joint from each cat inoculated intra-articularly with vaccine virus. Mild transient lameness was also seen in one of two control cats inoculated intra-articularly, but no pathological changes were seen or virus isolated from joints. A cDNA probe used in RNA dot blot hybridisation experiments was found to be specific and more sensitive than virus isolation in detecting FCV in selected tissues. This may be useful in future studies on the pathogenesis of FCV disease and in studies on viral persistence in FCV carriers.     

Prevention of marital distress: Results of a German prospective longitudinal study.

This article reports on the development and long-term evaluation of a marital distress prevention program for German couples, the Ein Partnerschaftliches Lernprogramm (EPL, A Couple's Learning Program). The EPL is a 6-session program designed to teach couples effective communication and problem-solving skills. In the current article, the EPL is evaluated in a prospective, quasi-experimental, controlled trial. The results of the 3-year follow-up are reported, contrasting 55 EPL couples with a control group of 17 couples. Significant differences emerged with regard to the couples' dissolution rates, relationship satisfaction, and positive and negative communication behavior favoring the EPL couples. These results demonstrate the utility of the EPL program in assisting happy couples who are preparing for marriage. The implications of the findings for prevention research and for the dissemination of prevention programs are discussed. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Synthesis and characterization of cyclic pseudopeptide libraries containing thiomethylene and thiomethylene-sulfoxide amide bond surrogates

Resistance-modifying agents (RMAs) such as Verapamil have been proved to be effective in reversing multi-drug resistance (MDR) in many in vitro assays. In this study we have investigated the efficacy of Dex-Verapamil, the R-isomer of Verapamil, as a chemosensitizer in a murine leukemia cell line (P388) and in its resistant counterpart (P388/Dx) expressing a typical MDR phenotype. We have examined in vivo the effect of the co-administration of Dex-Verapamil and Doxorubicin in mice transplanted with P388 or P388/Dx cells. Mice treated with the combination of Doxorubicin plus RMA had a significant increase in survival rate as compared to controls; however, the effect was modest. On the contrary, in vitro Dex-Verapamil can enhance Doxorubicin cytotoxicity in P388/Dx cells with a much greater effect depending on the treatment scheme used, by increasing the intracellular content of drug. Taken together our data indicate that Dex-Verapamil can indeed increase the sensitivity to Doxorubicin in resistant cells, but the limited efficacy shown in vivo demonstrates that this phenomenon is strongly dependent on the treatment scheme used and on the maintenance of constantly elevated serum levels.     

Effect of atherogenic diet on chicken plasma lipids and lipoproteins

Adult male White Leghorn chickens were used in an experimental model system to study atherogenesis, and the effects of an atherogenic diet on plasma lipoprotein composition including carotenoids were determined. This model also included treatment with diazepam, a drug known to reduce formation of atherogenic plaques. After 6 wk consumption of a high-cholesterol, high-triglyceride diet, chickens had mean total plasma cholesterol, triglyceride, and carotenoid concentrations that were significantly increased over those from chicks that consumed the standard diet. Diazepam treatment had no significant effect on whole plasma concentrations of these lipids. Total body weight gain was unaffected by diet, but liver weight expressed as percentage of body weight was significantly increased in chickens that consumed the atherogenic diet. High density lipoprotein (HDL) and low density lipoprotein (LDL) fractions were isolated from plasma samples by ultracentrifugation. The atherogenic diet increased the carotenoid, cholesterol, and protein content of the LDL fractions but not the HDL fractions.     

p53 expression in neurofibroma and malignant peripheral nerve sheath tumor. An immunohistochemical study of sporadic and NF1-associated tumors

OBJECTIVES--(1) to evaluate regional cerebral blood flow (rCBF) with single photon emission computed tomography and 99mTc-hexamethylpropyleneamine oxime in patients with the idiopathic adult hydrocephalus syndrome (IAHS); (2) to examine regional cerebral blood flow (rCBF), gait, and psychometric functions before and after CSF removal (CSF tap test); (3) to assess abnormalities in subcortical white matter by MRI. METHODS--Thirty one patients fulfilling the criteria for IAHS (according to history and clinical and neuroradiological examination) were studied. Quantified gait measurements, psychometric testing, and rCBF before and after removal of CSF were obtained. Pressure of CSF and CSF outflow conductance were investigated with a constant pressure infusion method. Brain MRI was used to quantify the severity of white matter lesions and periventricular hyperintensities. In IAHS a widespread rCBF hypoperfusion pattern was depicted, with a caudal frontal and temporal grey matter and subcortical white matter reduction of rCBF as the dominant feature. Removal of CSF was not accompanied by a concomitant increase in rCBF. Significant white matter lesions were detected only in a minority of patients by MRI. An altered CSF hydrodynamic state with a higher CSF pressure and lower conductance was confirmed. IAHS is characterised by an abnormal CSF hydrodynamic state, associated with a widespread rCBF reduction with preference for subcortical white matter and frontal-temporal cortical regions. Furthermore in most patients MRI did not show white matter changes suggestive of a coexistent subcortical arteriosclerotic encephalopathy. At least in the idiopathic group of patients with AHS, measurements of rCBF before and after temporary relief of the CSF hydrodynamic disturbance will not provide additional information that would be helpful in the preoperative evaluation but is suggestive of a preserved autoregulation of rCBF.     

Publication and citation of alcohol and drug articles in journals of the American Psychological Association.

Examined publishing practices of the American Psychological Association (APA) in the alcohol and drug area. Although the total number of articles published by APA increased only 7.6% between 1984 and 1990, the number of alcohol and drug articles more than doubled. Despite the disproportionate growth of alcohol and drug articles between 1984 and 1990, only 3.8% of 1,408 APA articles published in 1990 addressed alcohol and drug use issues. An earlier investigation by M. O. Howard and D. A. Howard (1992) reported that 541 articles published in 1984 in 12 alcohol and drug specialty journals received an average of 3.48 citations by the end of 1988. Alcohol and drug articles published by APA in 1984 received more than 4 times as many citations during this period. Also, alcohol and drug articles published by APA received significantly more citations between 1984 and 1988 than those published in specialty journals. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Thermodynamic equations of state for hydrazine and monomethylhydrazine

Thermodynamic equations of state are evaluated for the aerospace fuels hydrazine and monomethylhydrazine using Peng-Robinson (PR) and Soave-Redlich-Kwong (SRK) formulations. The PR formulation is shown to be the best fit for hydrazine, and the SRK formulation to be the best fit for monomethylhydrazine, based on available critical property data and evaluations of thermodynamic consistency. The adequacy of the differing property data for these fuels in the literature is discussed, and the methodology used to validate the formulations is outlined. The importance of using appropriate real fluid equations of state in thermodynamic safety and hazards analysis of fuel systems is demonstrated by considering an adiabatic compression of gaseous fuels previously postulated in accident scenarios of aerospace propulsion systems. Calculation of isentropic compression temperatures for pure components using ideal gas constant heat capacity, ideal gas with variable heat capacity, and real fluid equations of state are compared to illustrate the need for real fluid equations of state. In addition, three separate approaches are used for estimating isentropic compression temperatures for mixtures involving these fuels, again illustrating the importance of treating these mixtures as real fluids for design and safety analysis.     

Infant affect and affect regulation during the still-face paradigm with mothers and fathers: the role of infant characteristics and parental sensitivity

This laboratory study examined mothers' and fathers' sensitivity during face-to-face interactions with their infants as well as infants' affective and regulatory responses during mother-infant versus father-infant still face (SF). The degree to which infant gender and temperament as well as parental sensitivity predicted SF responses was also examined. Participants included 94 healthy, primarily White, middle-class 4-month-olds and their parents. Results indicated that mothers and fathers were equally sensitive toward their infants. Infants' affect and regulatory behaviors were also significantly stable across mother- and father-infant SF situations, although several differences in mean levels of regulation emerged. Finally, the extent to which exogenous and endogenous variables predicted infant SF responses differed as a function of which affect or regulatory variable was being examined and with which parent the infant was experiencing SF.