Alcohol relapse repetition, gender, and predictive validity

Earlier exploratory work on a scoring algorithm for the Reasons for Drinking Questionnaire [Zywiak, W. H., Westerberg, V. S., Connors, G. J., & Maisto, S. A. (2003). Exploratory findings from the Reasons for Drinking Questionnaire. Journal of Substance Abuse Treatment, 25, 287–292.] presented a number of interesting findings, but was limited by a fairly low consistency in type of relapse between the first relapse and the second relapse (63%). This scoring algorithm objectively classifies alcohol relapses into one of three types (negative affect, social pressure, or craving/cued). While examining gender differences in the type of first relapse, evidence indicating that relapses were more consistent for men (81%) than for women (44%) was uncovered. For initial posttreatment relapses, women were more likely to have negative affect relapses, and men were more likely to have social pressure relapses. For men, negative affect relapses were predicted by the Beck Depression Inventory score. For women, negative affect relapses were predicted by the Alcohol Dependence Scale score, and craving/cued relapses were predicted by situational craving.     

LC/MS/MS method for the quantitation of trans-2-hexenal-derived exocyclic 1,N(2)-propanodeoxyguanosine in DNA

trans-2-Hexenal is an alpha,beta-unsaturated aldehyde to which humans are exposed daily in small amounts. Hexenal has demonstrated mutagenicity and genotoxicity in vitro and reacts with deoxyguanosine to form diastereomeric hexenal-derived exocyclic 1,N(2)-propanodeoxyguanosine (Hex-PdG) adducts. A highly sensitive and specific method for the measurement of Hex-PdG in DNA has not previously been available. An LC/MS/MS assay for the quantitation of Hex-PdG, using [(13)C4(15)N2]Hex-PdG as an internal standard, was developed, to assess binding of hexenal to DNA. Samples were purified prior to analysis by centrifuge filtration and solid phase extraction and analyzed by LC/MS/MS in the selected reaction monitoring (SRM) mode (SRM m/z 366.2 --> 250.2 for Hex-PdG; SRM m/z 372.2 --> 256.2 for [(13)C4(15)N2]Hex-PdG). Recovery of standards was 89% or greater, and quantitation was unaffected by the addition of increasing concentrations of calf thymus DNA (ctDNA). The limit of quantitation, determined in samples of 200 microg of ctDNA spiked with analyte standard, was 0.015 fmol/microg DNA, which corresponds to approximately 5 Hex-PdG/10(9) unmodified nucleotides. Hex-PdG was detected in ctDNA treated with 0.021 microM, 0.21 microM, or 2.1 mM hexenal but not in untreated DNA. Furthermore, Hex-PdG was not detected in DNA exposed to reactive oxygen species-mediated deoxyribose attack and lipid peroxidation, which resulted in a significant increase in the malondialdehyde-derived pyrimido[1,2-a]purin-10(3H)one. Hex-PdG was not detected in DNA of untreated rat liver, but Hex-PdG in hexenal-treated calf thymus DNA was quantifiable when spiked into the rat liver DNA at 0.035 or 0.35 fmol/microg DNA. These data indicate that Hex-PdG is formed following hexenal treatment and that this method is suitable for in vitro or in vivo assessment of Hex-PdG formation.     

Long-term effects of neonatal surgery on adulthood pain behavior

The long-term consequences of neonatal noxious stimulation on adulthood pain behavior were investigated in male and female mice. On the day of birth, mouse pups were exposed to a laparotomy under cold anesthesia followed by an analgesic dose of morphine (10 mg/kg) post-operatively, or a saline control. An additional group of subjects was exposed to the non-noxious aspects of the surgical procedure (cold exposure, separation from the dam, injection) comprising a 'sham' surgery control group, whereas another group of control subjects was administered an injection of saline or morphine, but was otherwise undisturbed. Behavioral observations of the pups immediately following the procedure indicated that the laparotomy produced increased distress vocalizations in the ultrasonic range (40 kHz) compared to both groups of control subjects. During 90 min observations periods following the surgery and 1-week later, maternal care did not vary among treatment conditions. In adulthood, offspring were tested for nociceptive sensitivity on the hot-plate (HP; 53 degrees C), tail-withdrawal (TW; 50 degrees C) and acetic acid abdominal constriction test (AC). On both the TW and the AC tests, neonatal surgery decreased pain behavior relative to both groups of control subjects, an effect that was reversed by post-operative morphine treatment. On the HP test, both groups of subjects exposed to the stressful aspects of neonatal surgery (laparotomy or sham surgery) exhibited decreased pain behavior in adulthood. These findings suggest that early exposure to noxious and/or stressful stimuli may induce long-lasting changes in pain behavior, perhaps mediated by alterations in the stress-axis and antinociceptive circuitry.     

Pregnancy in women with congenital heart disease: the importance of evaluation and counselling

Issues surrounding the growing population of pregnant women with congenital heart disease are becoming increasingly important as more women with surgically corrected heart conditions reach childbearing age.     

Single-sample diagnosis of recent rubella by fractionation of antibody on Sephadex G-200 column

To facilitate the diagnosis of recent rubella infection, rubella haemagglutination inhibiting antibody has been determined in four fractions obtained by Sephadex G-200 gel filtration of samples of serum. All the 21 samples collected at the convalescent stage of the disease had varying proportions of haemagglutination inhibiting antibody in fraction 1, representing the major portion of IgM antibody whereas all but three out of 22 sera from persons with no history of recent rubella had negative titres in this fraction. The haemagglutination inhibiting titres in the three positive sera in the second group was very low as compared to the other fractions. Fractionation of sera on a Sephadex G-200 column coupled with the rubella haemagglutination inhibition test can, therefore, be used to diagnose recent rubella infection.     

Molecular fingerprinting of Legionella species by repetitive element PCR.

Repetitive element PCR (rep-PCR) uses outward-facing primers to amplify multiple segments of DNA located between conserved repeated sequences interspersed along the bacterial chromosome. Polymorphisms of rep-PCR amplification products can serve as strain-specific molecular fingerprints. Primers directed at the repetitive extragenic palindromic element were used to characterize isolates of Legionella pneumophila and other Legionella species. Substantial variation was seen among the rep-PCR fingerprints of different Legionella species and serogroups. More limited, but distinct, polymorphisms of the rep-PCR fingerprint were evident among epidemiologically unrelated isolates of L. pneumophila serogroup 1. Previously characterized Legionella isolates from nosocomial outbreaks were correctly clustered by this method. These results suggest the presence of repetitive extragenic palindromic-like elements within the genomes of members of the family Legionellaceae that can be used to discriminate between strains within a serogroup of L. pneumophila and between different Legionella species. rep-PCR appears to be a useful technique for the molecular fingerprinting of Legionella species.     

Anogenital distance measured at weaning is correlated with measures of blood chemistry and behaviors in 450-day-old female mice

In female mice, anogenital distance (AGD), measured at weaning, provides an estimate of uterine exposure to testosterone from flanking male mouse littermates. A variant of the anogenital distance index (AGDI) that uses the residual value of AGD after accounting for the effect of weight by regression (AGDWTRES) was measured at weaning in F(2) female mice from a C57BL/6J x DBA2/J cross. AGDWTRES was used to examine the relationship between intrauterine environment and blood chemistry variables and activity-related behaviors when the females were 450 days old. Longer AGDWTRES values correlated with lower levels of calcium, cholesterol, phosphorus, iron, and protein, which is opposite to the expected direction, based on underlying sex differences for blood chemistry. A positive correlation was found between AGDWTRES and two activity-related measures (the number of rears in a test of exploration, and the number of sectors of a rod that are entered by the mouse). These findings suggest that in utero proximity to males, as indexed by AGDWTRES, may have effects on fundamental aspects of blood chemistry and behavior that extend well into mouse middle age, and could play an important role in health.