Effect of sex hormone status on chloroform nephrotoxicity and renal mixed function oxidases in mice

In mice, only makes are susceptible to chloroform (CHCl₃) nephrotoxicity and the susceptibility appears to be related to renal mixed function oxidase activity. There were sex-related differences of renal cytochrome P-450 and b₅ concentrations and of ethoxycoumarin O-deethylase activity in mouse kidneys; in all cases activity was higher in males. Castration of male mice eliminated susceptibility to ChCl₃ nephrotoxicity and reduced renal mixed function oxidases to concentrations observed in female mice. Treatment of male and female mice with testosterone increased the susceptibility to ChCl₃ nephrotoxicity and increased renal mixed function oxidases to similar activities in both sexes. Previous data have suggested that CHCl₃ is metabolized in situ by the kidney, possibly by a mechanism similar to that occurring in the liver. The data from this investigation are consistent with the concept that CHCl₃ is metabolized by a cytochrome P-450-dependent mechanism in the kidney.     

Immuno-stimulating complexes prepared by ethanol injection

This study describes the formulation of immuno-stimulating complexes (ISCOMs) utilising the ethanol injection technique. Cholesterol and phosphatidylcholine were dissolved in ethanol and the resulting solution was rapidly injected into a stirred, aqueous solution of the triterpene-saponin mixture Quil-A. The reversed experiment was also carried out by adding the aqueous Quil-A solution to a solution of cholesterol/phosphatidylcholine dissolved in ethanol. This was done by either rapid injection or dropwise addition of the aqueous Quil-A solution. The colloidal dispersions obtained by ethanol injection and reversed addition were compared with formulations obtained by the dialysis and lipid-film hydration methods. In a further experiment, the preparation of ISCOMs from liposomes as precursor structures was investigated. Transmission electron microscopy was used to analyse the resulting colloidal dispersions. Samples were also compared with respect to homogeneity of obtained particle species. The ethanol injection technique led to formation of ISCOMs in high numbers within 2 h post formulation. The reversed rapid injection resulted in a similar colloidal dispersion, differing from the former mainly due to the presence of some helical micellar structures. The reversed, dropwise addition led to the formation of helices as the predominant colloidal structure. Of the three previously established methods, only dialysis led to the formation of ISCOMs within 48 h. The lipid-film hydration method and the approach using liposomes as precursor structures did not produce ISCOMs under the conditions and within the time periods investigated. However, it is known that dispersions prepared by the hydration method equilibrate towards ISCOMs after longer storage. Ethanol injection and reversed rapid injection are simple, cost-effective and quick methods to produce ISCOMs.     

Malaria in the Nuba Mountains of Sudan: baseline genotypic resistance and efficacy of the artesunate plus sulfadoxine-pyrimethamine and artesunate plus amodiaquine combinations

Both northern and southern Sudan are deploying artemisinin-based combinations against uncomplicated Plasmodium falciparum malaria (artesunate+sulfadoxine-pyrimethamine [AS+SP] in the north, artesunate+amodiaquine [AS+AQ] in the south). In 2003, we tested the efficacy of 3 day AS+SP and AS+AQ regimens in vivo in the isolated, seasonally endemic Nuba Mountains region (the first study of AS combinations in southern Sudan). We also analysed pre-treatment blood samples for mutations at the P. falciparum chloroquine transporter (Pfcrt) gene (associated with CQ resistance), and at the dihydrofolate reductase (Dhfr) gene (associated with pyrimethamine resistance). Among 161 randomized children under 5 years, PCR-corrected cure rates after 28 days were 91.2% (52/57, 95% CI 80.7-97.1) for AS+SP and 92.7% (51/55, 95% CI 82.4-98.0) for AS+AQ, with equally rapid parasite and fever clearance. The Pfcrt K76T mutation occurred in 90.0% (144/160) of infections, suggesting CQ would work poorly in this region. Overall, 82.5% (132/160) carried mutations at Dhfr (N51I, C59R or S108N, but not I164L), but triple mutants (more predictive of in vivo SP failure) were rare (3.1%). CQ use should be rapidly discontinued in this region. SP resistance may propagate rapidly, and AS+AQ is likely to be a better long-term option, provided AQ use is limited to the combination.     

Application of the case-crossover design to reduce unmeasured confounding in studies of condom effectiveness

This analysis examined how unmeasured confounding affects estimates of the effectiveness of condoms in preventing sexually transmitted infections. Data were analyzed from a prospective cohort study of 1,122 female sexually transmitted disease clinic patients in Alabama (1992-1995), wherein participants were evaluated for sexually transmitted infections at six 1-month intervals. Associations between condom use and incident gonorrhea and chlamydia infection were compared between case-crossover and cohort analyses. In a case-crossover analysis of 228 follow-up visits ending in gonorrhea/chlamydia ("case intervals") and 743 self-matched follow-up visits not ending in gonorrhea/chlamydia ("noncase intervals") (183 women), consistent condom use without breakage or slippage was associated with significantly reduced risk of infection relative to nonuse (adjusted risk odds ratio = 0.49, 95% confidence interval: 0.26, 0.92). Conversely, a cohort analysis of 245 case intervals and 3,896 noncase intervals (919 women) revealed no significant reduction in infection risk from consistent use of condoms (adjusted risk odds ratio = 0.79, 95% confidence interval: 0.53, 1.17). Dose-response relations between the number of unprotected sex acts and infection were stronger in the case-crossover analysis (p for trend = 0.009) than in the cohort analysis (p for trend = 0.18). These findings suggest that epidemiologic studies confounded by unmeasured differences between condom users and nonusers underestimate condom effectiveness against these infections. The case-crossover method provides an additional technique for reducing unmeasured confounding in studies of condom effectiveness.     

The Terri Schiavo saga: the making of a tragedy and lessons learned

The recent case of Terri Schiavo has been an important medical, legal, and ethical controversy. However, much of the public discussion of the tragedy has been based on inaccurate information regarding the facts of the case and the actual legal and ethical issues involved. This article reviews the pertinent aspects of the case and the ethical and legal questions raised and highlights the lessons we should learn from this unique story.     

Monitoring recovery after injury: procedures for deriving the optimal test window

Researchers studying the impact of treatments designed to facilitate recovery after neural injury face competing demands. On the one hand, because treatment effects often emerge slowly over days, and because researchers seek evidence of stable long-term effects, it is common practice to observe experimental subjects for many weeks after treatment. On the other hand, the cost of performing studies and the need to evaluate a multitude of alternative treatment procedures requires optimal efficiency, pushing researchers towards shorter test procedures. With these issues in mind, researchers have appeared to derive a test window based on previously published methodologies and inspection of their recovery curves, with testing terminated after the recovery curve reaches asymptote (approaches a slope of 0). An alternative procedure is introduced here that evaluates the stability of the data set over time. Using correlational techniques, researchers can determine whether (1) testing should be continued for additional days; or (2) equivalent statistical power can be achieved in fewer days. This provides a rational decision rule to help researchers balance competing demands. Applying these techniques to a procedure that evaluates the impact of acute treatments on recovery from spinal cord injury, it is shown that equal statistical power can be achieved in half the time, greatly increasing the efficiency with which alternative treatments can be evaluated.     

The behavioral deficit observed following noncontingent shock in spinalized rats is prevented by the protein synthesis inhibitor cycloheximide

Spinalized rats that receive shock when 1 hind limb is extended (contingent shock) exhibit an increase in flexion duration, a simple form of instrumental learning. Rats that receive shock independent of leg position (noncontingent shock) do not exhibit an increase in flexion duration and fail to learn when tested with contingent shock 24 hr later. It appears that noncontingent shock induces an intraspinal modification that inhibits the capacity to learn. The authors propose that the mechanisms that underlie this effect depend on de novo protein synthesis. To evaluate this hypothesis, the authors gave spinalized rats the protein synthesis inhibitor Cycloheximide (CXM) or saline intrathecally prior to, or immediately after, noncontingent shock exposure. Twenty-four hours later, rats were tested with contingent shock. Rats that received the vehicle and noncontingent shock failed to learn. CXM-treated shocked rats learned normally, suggesting that the learning deficit depends on protein synthesis within the spinal cord.