Examining the effectiveness of a community-based self-help program to increase women's readiness for smoking cessation.

This study investigated the effectiveness of two components of a smoking cessation intervention: a reading manual and a series of televised programs. Female smokers in the Chicago metropolitan area with a high school education or less were interviewed at 4 different times: baseline, immediate postintervention, and 6 and 12 months. We examined the effects of several baseline measures (race, age, number of cigarettes smoked, and stage of readiness to change) and exposure to the intervention components on subsequent stage of change. Race, baseline smoking rate, baseline stage, and exposure to both intervention components had direct effects on stage at immediate postintervention, with both intervention components increasing readiness to quit. Furthermore, exposure to the manual interacted with baseline stage, such that the manual benefited women at earlier stages more than women at later stages. Effects of both components were sustained at 6 months, and the effects of the manual were sustained at 12 months.

The XcpR protein of Pseudomonas aeruginosa dimerizes via its N-terminus.

Extracellular protein secretion by the main terminal branch of the general secretory pathway in Pseudomonas aeruginosa requires a secretion machinery comprising the products of at least 12 genes. One of the components of this machinery, the XcpR protein, belongs to a large family of related proteins distinguished by the presence of a highly conserved nucleotide binding domain (Walker box A). The XcpR protein is essential for the process of extracellular secretion and amino acid substitutions within the Walker A sequence result in inactive XcpR. The same mutations exert a dominant negative effect on protein secretion when expressed in wild-type bacteria. Transdominance of XcpR mutants suggests that this protein is involved in interactions with other components of the secretion machinery or that it functions as a multimer. In this study, the amino-terminal portion of the cl repressor protein of phage lambda was used as a reporter of dimerization in Escherichia coli following fusion to full-length as well as a truncated form of XcpR. The cl-XcpR hybrid proteins were able to dimerize, as demonstrated by the immunity of bacteria expressing them to killing by lambda phage. The full-length XcpR as well as several deletion mutants of XcpR were able to disrupt the dimerization of the chimeric cl-XcpR protein. The disruption of cl-XcpR dimers using the deletion mutants of XcpR, combined with the analysis of their dominant negative effects on protein secretion, was used to map the minimal dimerization domain of XcpR, which is located within an 85 amino acid region in its N-terminal domain. Taken together, the data presented in this paper suggest that the XcpR protein dimerizes via its N-terminus and that this dimerization is essential for extracellular protein secretion.

Lower urinary changes over time in suprasacral spinal cord injury.

This paper reviews 179 patients (23 females and 156 males) with suprasacral spinal cord injury (SCI) who underwent videourodynamic evaluation to compare maximum detrusor pressure, compliance, and trabeculation with methods of bladder management, years post-injury, and age. The patients were divided into four groups based on mode of bladder management: clean intermittent catheterization (CIC), indwelling catheter (IND), external collector (EC), and voiding (V). Maximum pressure decreased significantly with increasing age for those using EC (P < 0.01) and CIC (P < 0.05). Maximum pressure also decreased significantly with years post-injury for patients on EC (P < 0.01) and was highest the first decade after injury and progressively decreased through the fifth decade. Post-hoc tests indicated more severe trabeculation in patients in the EC group than in either the CIC or IND groups. Age and trabeculation did not correlate in those on EC. We conclude that patients with long-standing suprasacral SCI using EC are more likely to have lower detrusor pressures than are those with less chronic SCI. This finding may reflect the effects of age as well as reduced survival in those using EC with chronically elevated detrusor pressure.

Mutations in the consensus ATP-binding sites of XcpR and PilB eliminate extracellular protein secretion and pilus biogenesis in Pseudomonas aeruginosa.

The process of extracellular secretion in Pseudomonas aeruginosa requires specialized machinery which is widely distributed among bacteria that actively secrete proteins to the extracellular medium. One of the components of this machinery is the product of the xcpR gene, which is homologous to pilB, a gene encoding a protein essential for the biogenesis of type IV pili. Both XcpR and PilB are characterized by the presence of a conserved ATP-binding motif (Walker sequence). The codons of highly conserved glycine residues within the Walker sequences of xcpR and pilB were altered to encode a serine, and the effects of these substitutions were examined. Bacteria expressing mutant XcpR or PilB were unable to secrete exotoxin A or assemble pili, respectively. In addition, high-level expression of mutant XcpR in wild-type P. aeruginosa led to a pleiotropic extracellular secretion defect, resulting in the periplasmic accumulation of enzymes that are normally secreted from the cell. These studies show that the putative ATP-binding sites of XcpR and PilB are essential for their functions in protein secretion and assembly of pili, respectively. Moreover, the observed dominant negative phenotype of mutant XcpR suggests that this protein functions as a multimer or, alternatively, interacts with another essential component of the extracellular protein secretion machinery.

Circular forms of developmentally excised DNA in Euplotes crassus have a heteroduplex junction.

Extensive DNA elimination via a DNA breakage and rejoining process occurs during macronuclear development in the hypotrich ciliate Euplotes crassus. The excision process involves the removal of short, unique segments of DNA (internal eliminated sequences; IESs) and at least two highly repetitive families of transposon-like elements (Tec elements). Previous studies have demonstrated that circular forms of both IESs and Tec elements are generated following their developmental excision and that some flanking DNA sequences are retained at the circle junctions. In this study we have further analyzed the circle junctions of IESs. Analysis of polymerase chain reaction (PCR) products derived from IES circle junctions indicates that at least two sequence arrangements can be present. The circle junctions contain both of the direct repeats that define the ends of the IES separated by either 2 bp flanking the right end of the IES and 8 bp from the left-flanking region, or 8 bp from the right and 2 bp from the left. Using a method that we have termed "strand-biased PCR," we obtained evidence that the junctions of free circular IESs have a 6-base heteroduplex at their center, such that one strand of the DNA is derived from the left-flanking region of the IES and the other from the right. Models of IES excision are presented that incorporate these results and those of previous studies on the excision process.

Sequence of a Euplotes crassus macronuclear DNA molecule encoding a protein with homology to a rat form-I phosphoinositide-specific phospholipase C.

A 604-base pair macronuclear DNA molecule from the hypotrichous ciliate Euplotes crassus was cloned and its DNA sequence determined. The DNA sequence contains an open reading frame capable of encoding a protein 141 amino acids in length. The putative protein contains significant sequence similarity to other eukaryotic proteins, including the rat form-I phosphoinositide-specific phospholipase-C.

High fidelity developmental excision of Tec1 transposons and internal eliminated sequences in Euplotes crassus.

Following the sexual phase of its life cycle, the hypotrichous ciliate Euplotes crassus transforms a copy of its chromosomal micronucleus into a transcriptionally active macronucleus containing short, linear, gene-sized DNA molecules. Tens of thousands of DNA breakage and joining, or splicing, events occur during macronuclear development. The DNA removed by such events includes transposon-like elements, referred to as Tec1 elements, as well as segments of unique sequence DNA, termed internal eliminated sequences (IESs). Both types of elements are bounded by short direct repeats. In the current study, a polymerase chain reaction (PCR) and DNA sequencing strategy has been used to examine the fidelity of excision of two Tec1 elements and three IESs. In all cases, the vast majority of excision events were found to be precise, with one copy of the terminal direct repeats retained at the empty site in the macronuclear DNA molecule. These results, in combination with previous studies that have characterized the excised DNA elements, indicate that the two products of excision (the free element and the macronuclear DNA molecule) share DNA sequences. This suggests that excision events are initiated by staggered cuts in the chromosomal DNA.