Human CYP1A1GFP expression in transgenic mice serves as a biomarker for environmental toxicant exposure.

The human CYP1A1 gene is regulated by the aryl hydrocarbon receptor (AhR), and induction of CYP1A1 is known to play an important role in xenobiotic metabolism. To examine the regulation of human CYP1A1 in vivo, we created a transgenic mouse strain (Tg-CYP1A1(GFP)) expressing a chimeric gene consisting of the entire human CYP1A1 gene (15 kb) fused with a GFP reporter gene. The treatment of Tg-CYP1A1(GFP) mice with a single intraperitoneal dose of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) or benzo[a]pyrene (B[a]P) led to the induction of CYP1A1(GFP) in both the liver and the lung as determined by fluorescence and Western blot analysis. The localization of induced fluorescence in liver also demonstrated the usefulness of cultured hepatocytes in examining the actions of AhR agonists toward induction of CYP1A1(GFP). Other routes of B[a]P administration, such as by oral exposure at 100 mg/kg for 3 days, led to reduced induction of CYP1A1(GFP) in liver and lung. In liver, expression of CYP1A1(GFP) was a sensitive marker for oral exposure, while mouse CYP1A1 was not induced at these doses. While first pass metabolism of B[a]P in the gastrointestinal tract reduces the potential of the AhR to induce CYP1A1(GFP) in the liver, adequate concentrations reach the hepatic circulation as demonstrated by induction of human UGT1A proteins in transgenic mice that express the human UGT1 locus. The capability to identify fluorescently labeled CYP1A1 in vivo provides a sensitive measurement of gene response and links exposure to potential environmental toxicants and activation of the AhR.     

Predicting age-appropriate pharmacokinetics of six volatile organic compounds in the rat utilizing physiologically based pharmacokinetic modeling.

The capability of physiologically based pharmacokinetic models to incorporate age-appropriate physiological and chemical-specific parameters was utilized to predict changes in internal dosimetry for six volatile organic compounds (VOCs) across different ages of rats. Typical 6-h animal inhalation exposures to 50 and 500 ppm perchloroethylene, trichloroethylene, benzene, chloroform, methylene chloride, or methyl ethyl ketone (MEK) were simulated for postnatal day 10 (PND10), 2-month-old (adult), and 2-year-old (aged) rats. With the exception of MEK, predicted venous blood concentrations of VOCs in the aged rat were equal or up to 1.5-fold higher when compared to the adult rat at both exposure levels, whereas levels were predicted to be up to 3.8-fold higher in the case of PND10 at 50 ppm. Predicted blood levels of MEK were similar in the adult and aged rat, but were more than 5-fold and 30-fold greater for PND10 rats at 500 and 50 ppm, respectively, reflecting high water solubility along with lower metabolic capability and faster ventilation rate per unit body weight (BW) of PND10 animals. Steady-state blood levels of VOCs, simulated by modeling constant exposure, were predicted to be achieved in the order PND10 > adult > aged, largely due to increasing fat volume. The dose metric, total amount metabolized per unit liver volume was generally much lower in PND10 than in adult rats. The blood:air partition coefficient, fat volume, and fat blood flow were identified as critical determinants for the predicted differences in venous blood concentrations between the adult and aged. The lower metabolic capability, largely due to a smaller liver size, and faster ventilation rate per unit BW of PND10 animals contribute the most to the differences between PND10 and adult rats. This study highlights the pharmacokinetic differences and the relevant parameters that may contribute to differential susceptibility to the toxic effects of VOCs across life stages of the rat.     

A Rationale for Using Synthetic Designs in Medical Education Research

The extent to which the results of a study can be attributed to the intervention under investigation(i.e., internal validity) is an important consideration in interpreting study findings. There are many threats to the internal validity of designs frequently used in medical education research. Synthetic designs, which involve the integration of two or more weak designs, or the addition of design elements, may afford investigators greater control over confounding variables in medical education research. A rationale for using synthetic designs is presented and two examples of their use in medical education settings are examined. The concluding proposition is that synthetic designs allow investigators flexibility in planning research that is feasible in medical education settings. In addition, they may permit stronger causal inferences between interventions and results than traditional research designs. This revised version was published online in June 2006 with corrections to the Cover Date.     

The Role of Stromal Cell Heparan Sulphate in Regulating Haemopoiesis

Intimate contact between haemopoietic progenitor cells and elements of the bone marrow stroma is required for progenitor cell proliferation and differentiation. It is believed that the stroma provides particular niches for the development of haemopoietic cells of different lineages. Cytokines, stromal cell surface molecules and molecules of the stromal extracellular matrix all contribute to defining these mi-croenvironmental niches. Data obtained using an in vitro model of haemopoiesis support the view that progenitor cell adhesion to stroma is mediated by multiple receptor-ligand interactions. The possibility of a tethering step, mediated by the engagement of stromal cell heparan sulphate with its ligands on the progenitor cells, preceding stable cell adhesion is discussed. The role of stromal cell heparan sulphate is likely to include cytokine presentation to progenitors as well as the tethering of progenitors to stroma. It is proposed that intracellular signals induced by progenitor cell adhesion to stroma act in association with cytokine induced signals to regulate progenitor cell proliferation and differentiation.     

Validating the performance of the mammary sentinel lymph node team

BACKGROUND AND OBJECTIVES: The mammary sentinel lymph node (SLN) procedure has the potential to improve the accuracy and lower the morbidity of axillary staging in breast cancer patients, but results are closely linked to experience and can vary widely between institutions. Standardized performance measures need to be established in order to optimize the transition to SLN biopsy only. METHODS: Performance data were prospectively collected for the first 156 mammary SLN procedures performed by three surgeons in our institution. RESULTS: Seventy-five cases were required to achieve an SLN visualization rate of > 80% on preoperative lymphoscintigraphy. The SLN visualization rate was 90% for the last 52 cases. Two surgeons required 25 cases before consistently achieving a > or = 90% SLN identification rate in the operating room and one required 15 cases. The metastasis detection rate increased from 22% for the first 52 cases to 31% for the last 52 cases. The false negative rate for the procedure was 5%. CONCLUSIONS: The following performance criteria and benchmarks are suggested for validating the performance of the SLN team: (1) SLN visualization rate on preoperative lymphoscintigraphy > or = 80%, (2) SLN identification rate in the operating room > or = 90%, (3) False negative rate for the procedure 5%. Thirty procedures per surgeon were sufficient to achieve these benchmarks in our group.     

Inter-arm differences in blood pressure: when are they clinically significant?

OBJECTIVE : To determine whether there is significant disparity in blood pressure between the two arms. DESIGN : Prospective, observational study. SETTING : One general hospital in Birmingham, England. PARTICIPANTS : Four hundred participants [age 56.3 +/- 19.7 years (mean +/- SD), 50% male] were recruited from staff and patients. Simultaneous bilateral blood pressure measurements were obtained using Omron HEM-705CP automated oscillatory devices; with two measurements taken in each arm. MAIN OUTCOME MEASURES : Mean inter-arm blood pressure differences and frequency of clinically important disparities. RESULTS : Mean +/- SD inter-arm differences in systolic and diastolic blood pressure were 1.81 +/- 8.6 mmHg and -0.23 +/- 8.3 mmHg, respectively. The analogous figures for mean +/- SD absolute differences were 6.32 +/- 6.12 mmHg and 5.06 +/- 6.57 mmHg, respectively. Significant differences were present between the mean right and left arm systolic blood pressure [t(399) = 4.20, P < 0.0001], and the mean absolute difference for both systolic [t(399) = 20.65; P < 0.0001] and diastolic [t(399) = 15.39; P < 0.0001] blood pressure. The variation in mean inter-arm blood pressure was unrelated to age, sex, ethnicity, arm circumference, handedness, being hypertensive, diabetic, or previous history of cardiovascular disease. Clinically significant inter-arm differences in systolic blood pressure of > 10 and > 20 mmHg were found in 20 and 3.5%, respectively; diastolic differences of > 10 and > 20 mmHg were present in 11 and 3.5%, respectively. Age was the only significant predictor of clinically significant variations in inter-arm blood pressures and mean absolute blood pressure differences. CONCLUSIONS : Significant differences in mean inter-arm systolic blood pressure, and mean absolute inter-arm systolic and diastolic blood pressure are present. This emphasizes the importance of measuring blood pressure in both arms initially to prevent this misdiagnosis of hypertension, due to normal differences in blood pressure between the arms.     

Predictors of relapse after treatment for methamphetamine use

The purpose of this study is to describe treatment utilization and relapse and examine possible predictors of time to relapse after treatment for methamphetamine (MA) use. This analysis is based on natural history interview data from 98 subjects treated for MA use in publicly-funded programs in Los Angeles County in 1995-97 and interviewed two to three years following their treatment admission. Results showed that half of the subjects had resumed MA use: 36% within six months of the end of treatment, and 15% more within seven to 19 months. Survival analysis methods showed significant predictors of (shorter) time to relapse were shorter length of treatment, older age of first substance use, and involvement in selling MA; ethnicity (being Hispanic) and more previous time in treatment had weaker effects.