The extent of drug availability is often measured by the area under the concentration-time curve. In animal studies, experimental constraints can limit the number of observations available on each animal. Estimation of area under the curve and its standard error are straightforward when each animal is measured at each time point. Bailer and Nedelman et al., have described techniques for estimating the area under the curve and its standard error when each animal is measured once. Yeh has described a technique for the hybrid case where animals are measured more than once, but not at all time points. We describe a method for estimating area under the curve and its standard error which is applicable to all three types of designs. We give formulas for testing treatment differences, including dose trends and dose proportionality, in area under the curve for designs containing an arbitrary number of treatments. A jackknife estimator is also described.

Three frequently used and cited formulas used to rate correct the QT interval (Bazett's, Fridericia's, and Van de Water's) were compared and ranked using a large population-based cohort of beagle dogs (99 males and 99 females). In addition, analysis of covariance was used to derive a flexible method to rate correct the QT.interval for heart rate. The method is flexible in that it utilizes pretest or control data to determine the degree of correction. In addition, it can also be used to evaluate whether treatment alters the association between heart rate and QT. Specifically, pretest QT (unadjusted) and heart rate data were used to estimate coefficients in the linear regression log(QT) = alpha + beta log(HR). The estimated slope (beta) from the pretest data was used to heart rate correct the QT interval in the formula log(QT)ca = log(QT) - beta *[log(HR - log(HRm)]. The term "log(HRm)" is included to standardize QTca to a reference value, either a fixed value or an average heart rate for the data set being analyzed. These formulas were retrospectively compared under a typical toxicity study paradigm with a class III antiarrhythmic agent (L-768,673) that selectively prolongs the QT interval by blocking the slow activating component of the delayed rectifying potassium channel (lks). Based on their ability to dissociate the effects of heart rate on the QT interval, the formulas received the following ranking: Covariate Adjustment (preferred) = Van De Water's > Fridericia's > Bazett's (not recommended). Analysis of covariance based on pretest or control data is preferred for moderate to large studies where there are adequate data for estimation of the slope parameter beta, the investigator does not have sufficient control over HR, or treatment alters the association between HR and the QT interval. Conversely, for smaller studies a fixed rate adjustment formula from the literature (such as Van de Water's or Fridericia's equations) may be preferable since the bias from using a fixed formula is likely to be smaller than the variance resulting from estimating beta from a small sample.

The effects of alkaline pH and elevated sodium concentrations in culture medium on rat bladder explants for 1, 2, and 3 weeks were investigated by continuous BrdU labeling and histopathology. Increasing the sodium chloride concentration of normal medium by 50 or 100 mM caused slight urothelial hyperplasia with statistically significant increases in labeling in week 2 (50 mM) and at all time points with 100 mM NaCl. Cytotoxicity was seen in the high salt group. Increasing the pH from 7.2 to 7.8 and 8.2 also caused a slight hyperplastic response with significant increases in labeling and cytotoxicity at pH 8.2. However, bladder explants treated at pH 7.8 or 8.2 with excess sodium concentrations of 50 to 100 mM had a more marked hyperplastic response with evidence of cytotoxicity as well. There were significant increases in the labeling index (6.4- to 15.0-fold relative to control) after 1 week, with the maximum response at 100 mM sodium/pH 8.2. These results suggest that alkaline pH and elevated sodium concentration have a direct mitogenic effect on rat urothelial cells with some cytotoxicity-induced regenerative cell proliferation as well. These in vitro results in an organ culture system are in agreement with in vivo studies that have shown an important role for elevated urinary cation concentrations and pH in the stimulation of DNA synthesis, induction of hyperplasia, and tumor promotion in rat bladder epithelium.

The objective of this study was to determine the effects of 2 different 5-alpha reductase inhibitors (finasteride and MK-0434) on the glandular and stromal compartments of hyperplastic canine prostates. In this study, dogs received 1 of the 2 compounds orally, at a dose of 1 mg/kg/day for 16 weeks; control dogs received a placebo. The morphological changes in the glandular and stromal compartments in the prostate were quantitated by a point-counting method on Masson's trichrome-stained sections. Treatment with 5-alpha reductase inhibitors resulted in significant (P < or = 0.05) decreases in mean prostatic volumes, microscopic evidence of prostatic atrophy, and significant (P < or = 0.05) decreases in the absolute volumes of the prostatic glandular and stromal compartments compared to controls. In finasteride-treated dogs, the mean percent change from baseline was: epithelium, -52; lumens, -58; fibrovascular stroma, -41; and smooth muscle, -29. In MK-0434-treated dogs, the mean percent change from baseline was: epithelium, -77; lumens, -58; fibrovascular stroma, -38; and smooth muscle, -42. The effect on the glandular compartment in dogs treated with MK-0434 was slightly greater than in dogs treated with finasteride; however, the effect on the stroma was similar. These results clearly demonstrate that inhibition of 5-alpha reductase enzyme activity affects growth and maintenance of both glandular and stromal compartments of dog hyperplastic prostates. It is likely that the decrease in size of the prostate in finasteride-treated (Proscar) men is due to shrinkage of both glandular and stromal compartments.

A significant correlation exists between average daily food consumption and 2-yr survival in control ad libitum (AL)-fed Sprague-Dawley (SD) rats. SD rats were fed Purina Rodent Chow 5002 or a modified chow, 5002-9, with lower protein, fat, metabolizable energy and increased fiber AL or by dietary restriction (DR) to 65% of the AL amount by measurement or time (6.5 hr). At 52 wk, food consumption and key pathology biomarkers correlated with 106-wk survival. The modified chow, 5002-9 fed AL, did not significantly improve survival. SD rats fed either diet AL consumed the greatest amount of feed and kcal/rat but consumed the same amount of feed per gram body weight as DR-fed rats. At 52 wk, AL rats fed either diet had the same brain weights as DR rats, but the AL-fed rats had greater body weight and body fat content and increased heart, lung, kidney, liver, adrenal, thyroid, and pituitary weights as well as an increased incidence and severity of degenerative and/or proliferative lesions in these organs. This study demonstrates that overfeeding best correlates with low 2-yr survival in SD rats and that simple DR by caloric restriction modifies key pathology biomarkers in the pituitary, mammary gland, kidney, and heart of SD rats at 52 wk that are predictive of 106-wk survival.

A 30-year experience with 83 patients, median age two years, with Children's Cancer Study Group (CCSG) Stages I, II, and III localized neuroblastoma was studied to determine factors that influence outcome. In addition, histology was reclassified in all patients based on the Shimada system, which is divided into five subtypes according to age and cytohistologic criteria. A multivariant survival analysis was carried out and patients were considered to have failed if they relapsed or died from any cause. Initial analysis determined that CCSG stage, Shimada histologic classification, and presence of disease in lymph nodes were statistically significant predictors of failure. Histology was the most important factor with Shimada subtypes 1, 2, and 4 having good outcome and 3 and 5 poor outcome. The latter three variables were combined to create four prognostic groups that had distinctly different rates of survival. Further analysis showed that after controlling for prognostic groups, extent of surgery was a statistically significant predictor. Patients who had more complete surgical resection had better disease-free survival.

The dorsal horn of the cat spinal cord contains substance P and somatostatin within nerve endings which arise from cells located in dorsal root ganglia and from cells within the neuraxis. Previous studies from this laboratory have demonstrated that dorsal rhizotomy depletes both peptides from the dorsal horn. However, the changes in the two peptides differ. Substance P is at first severely depleted by dorsal rhizotomy and then recovers in part, whereas somatostatin is diminished less but does not recover. In the present experiments the validity of these conclusions which were based on anatomical observations has been evaluated quantitatively with the use of radioimmunoassay. After a 74% reduction at 10-14 days postoperative, substance P immunoreactivity in the deafferented dorsal horn shows a small, statistically significant recovery by 30 days to 60% of normal values. In contrast, somatostatin is reduced by 46% at 10-14 days but does not return significantly. As previously suggested by immunocytochemistry, dorsal rhizotomy produces no significant decline of either peptide in the ventral horn. The differing response of the two peptides is consistent with the hypothesis that intrinsic spinal substance P-containing neurons increase their projections (or their production of substance P) in the deafferented dorsal horn, but that somatostatin-containing neurons do not. Because synaptic number returns to normal in at least the deafferented lamina II of the cat yet substance P recovers only partially, it is likely that axons which contain transmitters other than substance P or somatostatin also increase the numbers of their terminals in response to dorsal rhizotomy.

The response of total nonsulfated serum bile acids, cholylglycine, and chenodeoxycholyl species was examined every 20 min for 3 h in 6 subjects. Noncaloric feeding led to a progressive decline or no change in bile acids, while there was a progressive rise in response to a standard liquid meal. After reaching a peak at 60 min, total bile acids declined progressively but cholylglycine and chenodeoxycholyl species remained elevated. Continuous infusion of cholecystokinin led to significantly greater levels most probably due to more rapid enterohepatic recirculation. Oral administration of 250 mg of chenodeoxycholic acid with water resulted in a rise in total bile acids and chenodeoxycholyl species, but not cholylglycine, indicating the rise was due to the administered bile acid and not gallbladder contraction. Administration of a meal and chenodeoxycholic acid simultaneously caused no greater rise of total serum bile acids or cholylglycine than either stimulus alone. Peak response and area under the curve were compared for each patient. The increase for chenodeoxycholyl species was additive for the two stimuli, suggesting that free chenodeoxycholic acid, when administered with a meal, decreased the absorption of endogenous conjugated bile acids. This study is compatible with the thesis that serum bile acids accurately reflect enterohepatic cycling and that administration of chenodeoxycholic acid with a meal may decrease its efficacy because exogenous chenodeoxycholic acid may compete with endogenous bile acids for absorption.