DNA relaxation dynamics as a probe for the intracellular environment.

Investigations into the biophysical properties of single molecules traditionally involve well defined in vitro systems where parameters such as solvent viscosity and applied forces are known a priori. These systems provide means to develop models describing the polymers response to a variety of conditions, including the entropically driven relaxation of a stretched biopolymer upon release of the tension inducing force. While these techniques have proven instrumental for recent advancements in the fields of polymer physics and biophysics, how applicable they are to life inside the cell remains poorly understood. Here we report an investigation of in vivo stretched polymer relaxation dynamics using chromatin relaxation following the breakage of a dicentric chromosome subjected to microtubule-based spindle forces. Additionally, we have developed an in vitro system used to verify the conformations observed during the in vivo relaxation, including the predicted but previously unidentified taut conformation. These observations motivate our use of existing polymer models to determine both the in vivo viscosity as seen by the relaxing chromatin and the tension force applied by the microtubule-based spindle in vivo. As a result, the technique described herein may be used as a biophysical strategy to probe the intranuclear environment.

Dynamic instability of individual microtubules analyzed by video light microscopy: rate constants and transition frequencies.

We have developed video microscopy methods to visualize the assembly and disassembly of individual microtubules at 33-ms intervals. Porcine brain tubulin, free of microtubule-associated proteins, was assembled onto axoneme fragments at 37 degrees C, and the dynamic behavior of the plus and minus ends of microtubules was analyzed for tubulin concentrations between 7 and 15.5 microM. Elongation and rapid shortening were distinctly different phases. At each end, the elongation phase was characterized by a second order association and a substantial first order dissociation reaction. Association rate constants were 8.9 and 4.3 microM-1 s-1 for the plus and minus ends, respectively; and the corresponding dissociation rate constants were 44 and 23 s-1. For both ends, the rate of tubulin dissociation equaled the rate of tubulin association at 5 microM. The rate of rapid shortening was similar at the two ends (plus = 733 s-1; minus = 915 s-1), and did not vary with tubulin concentration. Transitions between phases were abrupt and stochastic. As the tubulin concentration was increased, catastrophe frequency decreased at both ends, and rescue frequency increased dramatically at the minus end. This resulted in fewer rapid shortening phases at higher tubulin concentrations for both ends and shorter rapid shortening phases at the minus end. At each concentration, the frequency of catastrophe was slightly greater at the plus end, and the frequency of rescue was greater at the minus end. Our data demonstrate that microtubules assembled from pure tubulin undergo dynamic instability over a twofold range of tubulin concentrations, and that the dynamic instability of the plus and minus ends of microtubules can be significantly different. Our analysis indicates that this difference could produce treadmilling, and establishes general limits on the effectiveness of length redistribution as a measure of dynamic instability. Our results are consistent with the existence of a GTP cap during elongation, but are not consistent with existing GTP cap models.

Induction of apoptosis by diethylstilbestrol in hormone-insensitive prostate cancer cells.

BACKGROUND: Diethylstillbestrol (DES) and diethylstilbestrol diphosphate (DESdP) are effective agents for the treatment of advanced prostate cancers. Tumor-inhibiting effects of DES and DESdP are presumed secondary to suppression of androgen production in vivo. Little is known, however, about the direct cellular mechanisms of the tumor inhibition. Estrogens have been reported not only to stimulate growth but also to disrupt microtubule formation in prostate cancer cells. PURPOSE: The study was designed to examine and compare mechanisms of in vitro growth inhibition of DES and DESdP in human androgen-insensitive prostate cancer cells (DU145, 1-LN, and PC-3) and human androgen-sensitive prostate cancer cells (LNCaP) and to examine estrogen receptor modulation of such effects. METHODS: The cytotoxic effects of DES and DESdP were examined in vitro by use of a standard microculture tetrazolium assay to quantitate numbers of viable cells. Immunofluorescence microscopy, DNA fragmentation analysis, and fluorescence flow cytometry were used to investigate microtubules, the induction of apoptosis, and changes in cell cycle distribution. The degree of estrogen receptor positivity of untreated and treated cells was determined by immunohistochemistry and quantitative image analysis. RESULTS: LD50 levels (the dose at which 50% of cells are no longer viable) in the concentration range of 19-25 microM were observed for both DES and DESdP in all cell lines examined. DESdP-induced growth inhibition was found to be dependent on heat-labile phosphatases present in fetal calf serum. DES-induced cytotoxicity was not affected by the presence of 17 beta-estradiol, and it was not dependent on the presence of estrogen receptor. Estrogen receptor-positive cells and estrogen receptor-negative cells were equally responsive to DES. PC-3 cells stained with fluorescent anti-tubulin, phalloidin (actin stain), and 4',6-diamidino-2-phenylindole (DNA stain) showed no inhibition of microtubules or actin filaments but revealed the presence of apoptotic bodies in the nuclei. Fluorescence flow cytometry of nuclear DNA content of propidium iodide-stained nuclei from androgen-insensitive prostate cancer cells treated with 15 or 30 microM DES or DESdP revealed an increase in relative numbers of hypodiploid (apoptotic) nuclei, a depletion of G1- and S-phase cells, and an accumulation of cells in G2/M phase. Conversely, androgen-sensitive cells contained a lower percentage of hypodiploid nuclei but no accumulation of cells in G2/M phase. CONCLUSIONS: Direct cytotoxic effects of DES in prostate cancer cells are estrogen receptor independent and do not involve disruption of microtubule architecture but do involve the promotion of cell cycle arrest and apoptosis. These are the first data confirming direct cytotoxic effects of DES and DESdP in prostate cancer cells via an apoptotic mechanism. IMPLICATIONS. These results suggest that DES and DESdP have potential value as agents against androgen-insensitive prostate neoplasms through induction of an apoptotic cascade.

The trough-to-peak ratio as an instrument to evaluate antihypertensive drugs. The APTH Investigators. Ambulatory Blood Pressure and Treatment of Hypertension Trial.

The U.S. Food and Drug Administration designed the trough-to-peak ratio as an instrument for the evaluation of long-acting antihypertensive drugs, but the ratios are usually reported without accounting for interindividual variability. This study investigated how the trough-to-peak ratio would be affected by interindividual and intraindividual variability and by smoothing of the diurnal blood pressure profiles. The ambulatory blood pressure was recorded on placebo in 143 hypertensive patients (diastolic pressure on conventional measurement > 95 mm Hg). After 2 months, the recordings were repeated on 10 mg (n = 66) or 20 mg (n = 77) lisinopril given once daily between 7 and 11 PM. The baseline-adjusted trough-to-peak ratios were determined from diurnal blood pressure profiles with 1-hour precision. Lisinopril reduced (+/- SD) the 24-hour pressure by 16 +/- 17 mm Hg for systolic and 10 +/- 10 mm Hg for diastolic (P < .001). According to the usual approach, disregarding interindividual variability, the trough-to-peak ratio was 0.72 for systolic pressure and 0.67 for diastolic pressure. In the 143 patients the ratios were not normally distributed. They were the same on both lisinopril doses. When interindividual variability was accounted for, the median trough-to-peak ratio was 0.34 (P5 to P95 interval, -0.46 to 0.87) for systolic pressure and 0.26 (-0.44 to 0.84) for diastolic pressure. In 66 patients examined twice on 10 mg lisinopril at a median interval of 32 days, the trough-to-peak ratios were characterized by large intraindividual variability.(ABSTRACT TRUNCATED AT 250 WORDS)

Determining the trough-to-peak ratio in parallel-group trials. Systolic Hypertension in Europe (SYST-EUR) Trial Investigators.

We explored how in parallel-group trials interindividual variability, correction for placebo effects, and smoothing of blood pressure profiles can be handled in measuring the trough-to-peak ratio in 244 individuals with isolated systolic hypertension (> or = 60 years) enrolled in the placebo-controlled Systolic Hypertension in europe Trial. Net treatment effects were computed by subtracting the mean changes from baseline during placebo (n = 133) from those during active treatment (n = 111). At entry, systolic/diastolic pressures averaged 176/86 mm Hg in the clinic and 149/80 mm Hg on 24-hour ambulatory monitoring. With corrections applied for baseline and placebo, nitrendipine (10 to 40 mg/d), with the possible addition of enalapril (5 to 20 mg/d) and/or hydrochlorothiazide (12.5 to 25 mg/d), reduced (P < .001) these blood pressure values by 16.6/7.3 and 9.8/4.7 mm Hg, respectively. The net trough-to-peak ratios were first determined from blood pressure profiles (12 hours) with 1-hour precision, synchronized by the morning and evening doses of the double-blind medication. According to the usual approach, disregarding interindividual variability, the systolic/diastolic net trough-to-peak ratios were 0.46/0.40 in the morning and 0.77/0.99 in the evening. In individual subjects, the baseline-adjusted trough-to-peak ratios were nonnormally distributed. We therefore used a nonparametric technique to calculate the net trough-to-peak ratios from the results in individual subjects. In the morning, these ratios averaged 0.25 systolic (95% confidence interval, 0.09 to 0.41) and 0.15 diastolic (95% confidence interval, 0.00 to 0.31) and in the evening, 0.19 and 0.36 (95% confidence intervals, 0.00 to 0.38 and 0.14 to 0.56), respectively. When the blood pressure profiles were smoothed by substituting the 1-hour averages by moving or fixed 2-hour averages or by Fourier modeling, the trough-to-peak ratios remained unchanged after the morning dose (0.20/0.13, 0.20/0.14, and 0.16/0.21, respectively) but tended to increase in the evening (0.32/0.38, 0.28/0.40, and 0.48/0.49). In conclusion, the parallel-group analysis proposed makes it possible for one to correct the trough-to-peak ratio for baseline as well as placebo, to account for interindividual variability, and to calculate a confidence interval for the net trough-to-peak ratio. Accounting for interindividual variability reduces the trough-to-peak ratio. Smoothing affects the individualized net trough-to-peak ratios in an unpredictable way and should therefore be avoided.

Ambulatory monitoring uncorrected for placebo overestimates long-term antihypertensive action. Systolic Hypertension in Europe (SYST-EUR) Trial Investigators.

This study compares blood pressure (BP) changes during active antihypertensive treatment and placebo as assessed by conventional and ambulatory BP measurement. Older patients (> or = 60 years, n=337) with isolated systolic hypertension by conventional sphygmomanometry at the clinic were randomized to placebo or active treatment consisting of nitrendipine (10 to 40 mg/d), with the possible addition of enalapril (5 to 20 mg/d) and/or hydrochlorothiazide (12.5 to 25 mg/d). At baseline, clinic systolic/diastolic BP averaged 175/86 mm Hg and 24-hour and daytime ambulatory BPs averaged 148/80 and 154/85 mm Hg, respectively. After 13 months (median) of active treatment, clinic BP had dropped by 22.7/7.0 mm Hg and 24-hour and daytime BPs by 10.5/4.5 and 9.7/4.3 mm Hg, respectively (P<.001 for all). However, clinic (9.8/1.6 mm Hg), 24-hour (2.1/1.1 mm Hg), and daytime (2.9/1.0 mm Hg) BPs decreased also during placebo (P<.05, except for daytime diastolic BP); these decreases represented 43%/23%, 20%/24%, and 30%/23% of the corresponding BP fall during active treatment. After subtraction of placebo effects, the net BP reductions during active treatment averaged only 12.9/5.4, 8.3/3.4, and 6.8/3.2 mm Hg for clinic, 24-hour, and daytime BPs, respectively. The effect of active treatment was also subject to diurnal variation (P<.05). Changes during placebo in hourly systolic and diastolic BP means amounted to (median) 21% (range, -1% to 42%) and 25% (-3% to 72%), respectively, of the corresponding changes during active treatment. In conclusion, expressed in millimeters of mercury, the effect of antihypertensive treatment on BP is larger with conventional than with ambulatory measurement. Regardless of whether BP is measured by conventional sphygmomanometry or ambulatory monitoring, a substantial proportion of the long-term BP changes observed during active treatment may be attributed to placebo effects. Thus, ambulatory monitoring uncorrected for placebo or control observations, like conventional sphygmomanometry, overestimates BP responses in clinical trials of long duration.

Short report: ambulatory blood pressure in normotensive compared with hypertensive subjects. The Ad-Hoc Working Group.

OBJECTIVE: To delineate more precisely an operational threshold for making clinical decisions based on ambulatory blood pressure (ABP) measurement by studying the ABP in subjects who were diagnosed as either normotensive or hypertensive by conventional blood pressure (CBP) measurement. SUBJECTS: Twenty-four research groups recruited 7069 subjects. Of these, 4577 were normotensive (CBP < or = 140/90 mmHg), 719 were borderline hypertensive (systolic CBP 141-159 mmHg or diastolic CBP 91-94 mmHg) and 1773 were definitely hypertensive. Of the subjects in the last of these categories, 1324 had systolic hypertension (systolic CBP > or = 160 mmHg) and 1310 had diastolic hypertension (diastolic CBP > or = 95 mmHg). Hypertension had been diagnosed from the mean of two to nine (median two) CBP measurements obtained at one to three (median two) visits. RESULTS: The 95th centiles of the 24-h ABP distributions in the normotensive subjects were (systolic and diastolic, respectively) 133 and 82 mmHg. Of the subjects with systolic hypertension, 24% had 24-h systolic ABP < 133 mmHg. Similarly, 30% of those with diastolic hypertension had 24-h diastolic ABP < 82 mmHg. The probability that hypertensive subjects had 24-h ABP below these thresholds tended to increase with age and was two- to fourfold greater if the CBP of the subject had been measured at only one visit and if fewer than three CBP measurements had been averaged for establishing the diagnosis of hypertension. By contrast, for each 10-mmHg increment in systolic CBP, this probability decreased by 54% for 24-h systolic ABP and by 26% for 24-h diastolic ABP, and for each 5-mmHg increment in diastolic CBP it decreased by 6 and 9%, respectively. CONCLUSIONS: The ABP distributions of the normotensive subjects included in the present international database were not materially different from those in previous reports in the literature. One-fifth to more than one-third of hypertensive subjects had an ABP which was below the 95th centile of the ABP of normotensive subjects, but this proportion decreased if the hypertensive subjects had shown a higher CBP upon repeated measurement. The prognostic implications of elevated CBP in the presence of normal ABP remain to be determined.

A method of quantifying retinal microvascular alterations associated with blood pressure and age.

OBJECTIVE: To find an objective, sensitive method for quantifying microvascular alterations associated with level of blood pressure and age. DESIGN: A prospective cross-sectional study. SUBJECTS AND METHODS: Seventy-four previously untreated hypertensive patients, referred to a hospital outpatients department, and 26 normotensive volunteers participated. Twenty-four-hour ambulatory blood pressure monitoring and bilateral fundal photography were performed. The fundal photographs were projected on a screen such that the optic disc filled a circle of radius 5 cm. Microvessels crossing the border of a concentric circle of radius 20 cm were identified as arteriolar or venular, counted and their luminal diameters measured. MAIN OUTCOME MEASURES: Arteriolar and venular numbers, mean diameters and vascularities (arteriolar and venular vascularities defined as the sum of arteriolar and venular diameters, respectively). RESULTS: The technique was reproducible. As blood pressure increased, arteriolar vascularity declined and venular vascularity increased. These associations resulted in a strong inverse correlation between blood pressure level and the ratio arteriolar vascularity: venular vascularity (r = 0.48, P < 0.001). Arteriolar number declined with increasing diastolic blood pressure (r = 0.22, P < 0.05). Mean arteriolar diameter appeared to have a U-shaped relationship with diastolic blood pressure levels (r = 0.27, P < 0.05). Venular dilation was associated with increasing blood pressure levels (r = 0.22, P < 0.05). Mean arteriolar and venular diameters declined significantly with age (r = 0.33 and 0.26, respectively; P < 0.01) and there was no association between arteriolar vascularity:venular vascularity ratio and age. CONCLUSIONS: The method detected disparate retinal microvascular alterations with age and blood pressure. The arteriolar vascularity:venular vascularity ratio shows promise as a non-invasive, prognostic and therapeutic guide in hypertension.

Ambulatory pressure decreases on long-term placebo treatment in older patients with isolated systolic hypertension. Syst-Eur Investigators.

OBJECTIVE: This long-term study investigated the widely accepted hypothesis that ambulatory pressure does not decrease in patients given placebo. METHODS: One hundred and twelve older (> or = 60 years) outpatients with isolated systolic hypertension were recruited. Treatment consisted of a placebo during a 3-month baseline period and long-term follow-up. RESULTS: At baseline, on placebo treatment, clinic systolic/diastolic (SBP/DBP) blood pressure (+/- SD) averaged 176 +/- 12/86 +/- 7 mmHg and 24-h SBP/DBP 151 +/- 15/81 +/- 10 mmHg. These pressures were unaltered in 51 patients in whom the baseline measurements were repeated after a further month on placebo. After the 112 patients had received placebo for 1 year (median), clinic SBP/DBP fell by 6.6 +/- 15.9 (P < 0.001)/1.4 +/- 7.4 (P = 0.06)mmHg and 24-h SBP by 2.4 +/- 10.7 mmHg (P < 0.05), whereas 24-h DBP did not change significantly. The 24-h SBP decreased more with higher baseline level and longer follow-up (5-21 months). CONCLUSIONS: These findings in older patients with isolated systolic hypertension suggest that in long-term studies the ambulatory pressure may slightly but significantly decrease on a placebo. Like those using conventional sphygmomanometry, long-term studies using non-invasive ambulatory monitoring require a placebo-controlled design.

Mechanism of action of the marine natural product stypoldione: evidence for reaction with sulfhydryl groups.

Stypoldione, a marine natural product that possesses an o-quinone functional group, has been shown to inhibit a variety of biological processes including cell division. We found that stypoldione binds covalently to sulfhydryl groups of thiol-containing compounds via addition of sulfur to the C-4' position of the quinone ring. We examined the ability of stypoldione to add to sulfhydryl groups of a number of thiol-containing substances, including glutathione, thiophenol, beta-mercaptoethanol, and the protein tubulin. We suggest that the biological actions of stypoldione may be caused by the addition of this compound to thiol groups of biological molecules.

Localization of myocilin to the golgi apparatus in Schlemm's canal cells.

PURPOSE: Biochemical and genetic evidence suggests that overexpression of or mutations in myocilin within the cells of the aqueous humor outflow pathway play a significant role in the development of steroid-induced and several other open-angle glaucomas. As a baseline to understanding the normal and pathologic function of myocilin, we determined the subcellular localization of myocilin in steroid-treated human Schlemm's canal endothelial (SC) cells. METHODS: SC cells were grown to confluence, treated with dexamethasone for 10 days, and then stained using antibodies against myocilin, tubulin, or beta-COP (a specific golgi protein) or vital stains for endoplasmic reticulum (ER) and golgi. Brefeldin A (BFA) and nocodazol (NZ) were used to disrupt the golgi or microtubules. RESULTS: The authors found that myocilin staining was (a) always centered around the centrosome, (b) very similar to the pattern seen with NBD-ceramide, (c) was disrupted in characteristic ways by BFA and NZ and (d) showed extensive colocalization with beta-COP. CONCLUSIONS: Results indicate that myocilin is localized to the golgi in SC cells. Such localization is consistent with myocilin being processed for secretion but is also consistent with sequence analysis and other data that suggest that myocilin or myocilin mutations might be targeted to the cytoplasmic face of the golgi, and under some circumstances play a role in or interfere with golgi or vesicle function. How such interference could eventually lead to open angle glaucoma is discussed.

Hypertension in the elderly: an overview.

Hypertension in the elderly is associated with a high incidence of cardiovascular disease. There is no convincing data which prove that treatment of mild to moderate hypertension in this age group improves prognosis, but a review of available data suggests that treatment is beneficial. It is suggested that a thiazide diuretic or a beta-adrenoceptor blocking drug should be used as first-line therapy. These drugs may be combined if an inadequate response is seen and other drugs may be used if the blood pressure is not controlled.

Arterial hypotension: prevalence of low blood pressure in the general population using ambulatory blood pressure monitoring.

BACKGROUND: Chronic constitutional hypotension has been described in a proportion of the population, and has a symptom complex ascribed to it. The true prevalence of low blood pressure in the normal population has not been defined. AIM OF STUDY: This study was undertaken to determine the prevalence of low blood pressure states, as measured using ambulatory blood pressure monitoring, in a general population cohort, and to determine the association between low blood pressure and clinical and demographic variables. PATIENT POPULATION: The population enrolled were a cohort of mainly urban dwelling Irish subjects, either employees or spouses of employees of a major national bank. METHODS: Subjects had an ambulatory blood pressure monitor fitted between 09.00 and 12.00 and wore the monitor for 24 hours. The subjects also filled out a detailed lifestyle questionnaire, and kept an activity diary. Blood was drawn for serum electrolyte estimation. RESULTS: A total of 254 subjects were included, 49% of whom demonstrated hypotensive events. Hypotensive means and individual hypotensive values were more frequently found in women, and occurred in a group of individuals with a distinct body habitus, specifically thin subjects, with a lower creatinine suggesting a smaller muscle mass. Hypotensive events in these subjects were associated with a low risk cardiovascular profile, in that subjects who displayed these events had a lower blood pressure, a lower weight and were less likely to have a positive family history of hypertension or vascular disease. CONCLUSION: Hypotension is common in the general population and is associated with a distinct body habitus. It carries a generally benign cardiovascular risk factor profile.

A consistent reference frame for ambulatory blood pressure monitoring is found in different populations.

This study investigated the consistency of a reference frame for ambulatory pressure monitoring, which using various approaches was determined in two different populations. The two reference groups were 718 subjects randomly selected from the population and 895 bank employees. The reference values derived in these two groups were subsequently tested in 591 untreated hypertensive patients. The ambulatory pressures equivalent to a conventional pressure of 140 mmHg systolic and 90 mmHg diastolic were calculated by regression analysis in all subjects. In addition, in subjects who were normotensive by conventional sphygmomanometry, the mean +2 and +3 standard deviations and the 90th, 95th and 99th percentiles of the ambulatory measurements were determined. The distributions of the ambulatory measurements were similar in the two reference groups and the aforementioned parameters therefore agreed within 4 mmHg in the two populations. There was considerable overlap in the ambulatory pressures between the two reference groups and the hypertensive patients. Classification of the patients according to the means +3 standard deviations and the regression limits gave the same results because in both reference groups these boundaries approximated to each other within 1 mmHg. For the 24 h pressures in the population sample these boundaries were 140 mmHg systolic and 88 mmHg diastolic. Of the patients with systolic hypertension (> or = 160 mmHg on conventional measurement), 39% had a 24 h systolic pressure of < 140 mmHg and of those with diastolic hypertension (> or = 95 mmHg), 44% had a 24 h diastolic pressure of < 88 mmHg; if the corresponding boundaries derived in the bank employees (143/90 mmHg) were applied, these proportions were 47% and 44%, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Is elevation of clinic blood pressure in patients with white coat hypertension who have normal ambulatory blood pressure associated with target organ changes?

BACKGROUND: The issue as to whether white coat hypertension is a pathologically significant entity, with associated target organ changes, or that the condition carries the same risk for target organ involvement as normotension, is undecided. Previous studies which have shown pathological correlates between white coat hypertension and target organ damage have not controlled for the most obvious confounder, mean 24 h blood pressure (BP). METHODS AND RESULTS: In this study we retrospectively identified 33 age and sex-matched pairs, one group with normal BP, the other with white coat hypertension. The white coat hypertensive group showed significantly greater left ventricular mass indexed for body surface area than normal controls (99.0 g/m2 vs 78.3 g/m2, P < 0.001). The population was then further matched for 24-h mean BP (20 pairs), and was again compared for cardiac muscle changes. The significantly increased left ventricular mass index in the white coat population remained after controlling for 24-h mean BP (101.1 g/m2 vs 81.0 g/m2, P < 0.021). CONCLUSION: White coat hypertension is indeed associated with a larger left ventricular muscle mass than normotensives and these changes are independent of the actual 24-h BP load, and may reflect increased BP lability, sympathetic nervous system derangement, or a genetic propensity in people with white coat hypertension to stress-related hypertensive reactions, as part of a pre-hypertensive state.

Reference values for the ambulatory blood pressure and the blood pressure measured at home: a population study.

In order to determine reference values for the ambulatory blood pressure, a population sample of 328 subjects, aged 20-81 years, who reported themselves to be in good health, was investigated. The ambulatory blood pressure was recorded over 24 h, taking measurements at 20 min intervals from 8 am to 10 pm, and at 45 min intervals from 10 pm to 8 am. Blood pressure was also measured by trained observers on each of two separate home visits (5 readings per visit). The ambulatory blood pressure in the 328 subjects averaged 118/71 mmHg over 24 h, 124/76 mmHg during the day (10 am-8 pm), and 108/62 mmHg at night (0 am-6 am). Blood pressure measured by an observer at the occasion of the second home visit was 4/5 mmHg lower (P less than 0.001) than the daytime ambulatory blood pressure. The 95th centiles for the daytime ambulatory pressures were 144/95 mmHg in 85 men below age 50; 154/90 mmHg in 74 men aged greater than or equal to 50 years; 132/85 mmHg in 96 women below age 50; and 151/91 mmHg in 73 women aged greater than or equal to 50 years. The 95th centiles for the nighttime pressures in these four sex-age groups were 124/79, 140/83, 121/70, and 132/72 mmHg, respectively.

Ambulatory blood pressure monitoring and postprandial hypotension in elderly patients with isolated systolic hypertension. Systolic Hypertension in Europe (SYST-EUR) Trial Investigators.

The present analysis was undertaken to evaluate postprandial (PP) changes in blood pressure (BP) assessed with ambulatory BP monitoring (ABPM) in elderly subjects with isolated systolic hypertension (ISH) on conventional measurement. A total of 530 patients (335 women and 195 men, aged 60-100 years, median 70 years) who performed an ABPM during the placebo run-in period of the Syst-Eur trial were included into the analysis. The PP changes in BP and heart rate (HR) were calculated by subtracting the mean systolic BP (SBP), diastolic BP (DBP) and HR in the 2 h preceding the main meal from the corresponding means covering the 2 h after the meal. The reproducibility of the postprandial fall in BP and heart rate (PPH) was assessed by contrasting the first and second ABPM in a subgroup of 147 patients who performed two ABPM's during the placebo run-in period. The mean SBP and DBP decreased and reached the nadir 2 h after the main meal while HR did not change. When PPH was assessed by comparing BP in the 2 h before and after the meal, both SBP and DBP decreased significantly (respectively -6.6 mm Hg, -5.4 mm Hg; P < 0.001). In 67.6% of all patients a decrease in SBP was observed and in 24.1% it exceeded 16 mm Hg. The corresponding values for DBP were 71.3% and 24.5% (DBP decreased more than 12 mm Hg). A greater fall in DBP was associated with a greater decrease in HR (r = 0.20, P < 0.001), while changes in SBP and HR were not interrelated. Regression analysis did not identify any significant covariate of PPH. Group means of PPH could be reproduced without significant changes in their values, but the within-subject reproducibility of the PP changes was low. There were no differences in PPH according to the place of residence of the patients. In conclusion, the descriptive analysis of the meal-induced changes in ABPM in elderly subjects with ISH showed that in every day circumstances most of them experience falls in both SBP and DBP within 2 h after the meal.

The antihypertensive efficacy of verapamil in the elderly evaluated by ambulatory blood pressure measurement.

To assess the efficacy, tolerability and pharmacokinetics of verapamil in the elderly, ten patients with blood pressure greater than 160/90 mmHg were studied in a randomized double-blind placebo-controlled cross-over trial. Nine patients aged 75 (+/- 4.9) years completed the study. After titration, doses of verapamil varying from 40 to 120 mg (40 mg in four, 80 mg in one and 120 mg in four patients) twice daily for six weeks were taken. Mean (+/- SEM) clinic lying blood pressure was reduced on verapamil from 187 +/- 6.8/100 +/- 4.1 to 167 +/- 4.6/86 +/- 3.1 mmHg, [P less than 0.001). Mean ambulatory blood pressure was reduced from 174 +/- 1.4/95 +/- 1.0 to 169 +/- 1.3/90 +/- 0.8 mmHg, (P less than 0.01). Lying heart rate was significantly reduced but glomerular filtration rate, renal blood flow and mental function, were not altered by treatment. The mean plasma half-life of verapamil was 6.9 +/- 1.1 hours. Side effects were minimal. We conclude that verapamil is an effective blood pressure lowering agent in the elderly.

Remodeling of the femoral neck after in situ pinning for slipped capital femoral epiphysis.

During a period of thirty-six years, sixty-two patients with seventy slipped capital femoral epiphyses were treated by pinning in situ. Twelve of these patients, ten years and eight months to sixteen years and one month old, were treated for moderate to severe slipping by pinning in situ. After follow-ups ranging from two to seventeen years, all but two patients had satisfactory remodeling of the femoral head and neck and were asymptomatic. The two with incomplete or no remodeling had no symptoms. It was concluded that the effects of remodeling have been largely ignored and that pinning in situ when possible, followed if necessary by osteoplasty or osteotomy through the lesser trochanter, is a safe and effective treatment.

Ethacrynic acid disrupts steady state microtubules in vitro.

PURPOSE: Ethacrynic acid (ECA) has been shown to increase facility of aqueous outflow in whole eyes and perfused anterior segments, to open up spaces between cells in the trabecular meshwork and inner wall of Schlemm's canal, and to cause separation and retraction of trabecular meshwork and endothelial cells in culture. One mechanism by which ECA has been proposed to act in cells is via disruption of microtubules, leading to cell retraction. Although it is known that ECA can inhibit de novo assembly of microtubules from tubulin subunits in vitro, we wanted to determine, as a better correlate to the proposed effect of ECA in cells, whether ECA could disrupt microtubule polymers that had reached steady state. We also wanted to determine whether calcium ion could enhance this process. METHODS: We therefore assembled purified and crude porcine brain tubulin to steady state at 37 degrees C and then added ECA and/or calcium. Reaction kinetics were followed spectrophotometrically. RESULTS: We found that ECA effectively disrupted assembled microtubules in vitro. Although 0.8-1.0 mM ECA was required to produce a half-maximal effect in pure tubulin microtubules and 0.2-0.3 mM ECA was necessary with crude microtubule protein, significant disassembly also occurred in the 0.01-0.2 mM range. Calcium had a greater maximal effect than ECA, and was more potent on a molar basis, showing half maximal effect between 2 and 12 microM free calcium ion. Combination experiments showed that ECA did not act synergistically with calcium to increase microtubule disassembly. CONCLUSIONS: Our results are consistent with the proposed disruptive action of ECA on the assembled microtubules of outflow pathway cells, but do not support a rise in intracellular calcium as being an added factor.