p.Asn176Lys and p.Met137Thr dimorphisms of the PRNP gene significantly decrease the susceptibility to classical scrapie in ARQ/ARQ sheep.

In this study, we investigated the susceptibility to scrapie of Sarda breed sheep carrying the genotype ARQ/ARQ with additional polymorphisms at the PRNP gene. To do this, we examined 256 scrapie-affected sheep and 320 flock-mate negative controls from 24 flocks. Logistic regression analysis demonstrated that sheep carrying the ARQ/ARQ genotype with additional dimorphisms had lower risk of becoming scrapie affected when compared with those with ARQ/ARQ(wildtype) genotype. ARQ/ARQ genotypes that were detected with heterozygous or homozygous p.Asn176Lys and p.Met137Thr dimorphisms were associated with the lowest susceptibility to the disease. A significant lower risk was also associated with the p.Arg154His dimorphism, while p.Leu141Phe had a protective effect that was not statistically significant.

PrP(Sc) deposition in nervous tissues without lymphoid tissue involvement is frequently found in ARQ/ARQ Sarda breed sheep preclinically affected with natural scrapie.

The pathogenesis of natural scrapie in Sarda breed sheep was investigated in 1050 asymptomatic and 49 sick sheep from scrapie-affected flocks. Central and peripheral nervous system, along with lymphoreticular system (LRS) tissues, were subjected to immunohistochemistry (IHC) and Western-blotting (WB) for detection of pathological isoform of the prion protein (PrP(Sc)). A total of 69 of the 1050 clinically healthy sheep were found to be infected with scrapie, with PrP(Sc) being detected in both the central nervous system (CNS) and enteric nervous system (ENS) plexuses of 60 of the sheep, while IHC and WB yielded evidence of (PrP(Sc)) deposition only in lymphoid tissues of the remaining 9 clinically healthy sheep. PrP(Sc) was also detected in the CNS, as well as in ENS plexuses from all of the 49 clinically affected sheep. Nevertheless, 18 of the 69 clinically healthy animals (26%, 17 ARQ/ARQ and 1 ARQ/AHQ sheep), along with 3 ARQ/ARQ sheep (6%) of the clinically affected group, showed no IHC or WB evidence of PrP(Sc) in lymphoid tissues, but PrP(Sc) could be still detected in their CNS and ENS plexuses. The study demonstrates dual CNS and ENS PrP(Sc) deposition in Sarda sheep with scrapie, in spite of an apparent lack of lymphoid tissue involvement in a number of cases.

M-phase promoting factor (MPF) and mitogen activated protein kinases (MAPK) activities of domestic cat oocytes matured in vitro and in vivo.

This work was undertaken in order to examine M-phase promoting factor (MPF) and mitogen-activated protein kinases (MAPK) activities during meiotic progression of cat oocytes cultured in two different media for two different incubation times and preovulatory cat oocytes that reached MII in vivo. Oocytes recovered from ovaries of ovariectomized cats were cultured either in TCM 199 or SOF for 24 h and 40 h. In vivo matured oocytes were recovered by follicular aspiration from ovaries of domestic cats ovariectomized 24 h to 26 h after hormonal treatment. Results showed that the kinetic of MPF and MAPK activity was similar during meiotic progression of cat oocytes matured in TCM 199 and SOF. After 24 h of incubation, MII oocytes had significantly (p < 0.001) higher MPF and MAPK levels than MII oocytes cultured for 40 h in both culture media. MPF and MAPK activity was significantly (p < 0.01) lower in the oocytes matured in vitro than in those matured in vivo. This study provides evidence that the two different maturation media did not determine differences in MPF and MAPK fluctuations and levels during meiotic progression of cat oocytes and that the time of maturation influenced the level of the two kinases. Moreover, it shows that MPF and MPK activity is higher in in vivo matured oocytes than in in vitro matured oocytes, suggesting a possible incomplete cytoplasmic maturation after culture.

Isolation of chromosomal regions controlling intersex development in a marsupial.

A marsupial (Sminthopsis douglasi) with bilateral intersexuality had a hemiscrotum on the right side and a hemi-pouch with nipples on the left. A normal female karyotype (2n = 14, XX) was present in cells from the right (male) side, while cells from the left (female) side initially had a female karyotype plus two dot-like chromosomes (2n = 14, XX + 2B). It is proposed that the dots represented a region deleted from the X chromosome that contains the "pouch-mammary/scrotum" (PMS) switch gene whose dosage determines development of a pouch and teats (two doses) or a scrotum (one dose). Mis-segregation early in embryonic development produced a lineage with one normal X and one deleted X (male side), and a lineage with a normal and deleted X, plus two copies of the deleted region (female side). The origin of the supernumerary elements was therefore investigated in the expectation that they may contain the long-sought pouch-mammary/scrotum switch gene. Several elements were microdissected, and amplified DNA was used for in situ hybridization, producing signals in five different chromosome regions including the X. This could represent a region of the X that contains, as well as PMS, repetitive DNA that is present also at other chromosomal sites.

Clinical relevance blood pressure variability.

Blood pressure fluctuates continuously over time, either spontaneously or in response to a variety of external stimulations. The occurrence of these continuous and often marked blood pressure variations is not only of pathophysiologic interest, but it may also have a clinical relevance. Indeed, it has been shown that the occurrence of pronounced blood pressure changes at the time of the physician's visit may introduce errors in the diagnosis of hypertension and in the assessment of the efficacy of antihypertensive treatment. Moreover, several studies have reported that the end-organ damage of hypertension is significantly and independently related to the degree of blood pressure variability during the day and night. This was shown by reports that assessed blood pressure variability by a variety of different methods, i.e. by computing the 24 h or daytime blood pressure standard deviation, the degree of morning blood pressure rise or that of night-time blood pressure fall, the frequency of blood pressure peaks over the 24 h, and the blood pressure increases under stressful conditions or during physical exercise. Results from a recent follow-up study have provided evidence that the degree of blood pressure variability may also have prognostic relevance in hypertensive patients. Thus, optimal antihypertensive treatment might also need to reduce the degree of blood pressure fluctuations together with the 24 h average blood pressure levels. Until recently, however, available antihypertensive drugs have been ineffective in buffering blood pressure variability or have even been responsible for an increase in the degree of blood pressure fluctuations. Further studies are needed to assess whether recently developed antihypertensive agents, and in particular those able to induce a smooth reduction in blood pressure over the 24 h or to modulate the sympathetic influences exerted on the cardiovascular system, may represent better tools to reduce the magnitude of an enhanced blood pressure variability in hypertensive patients over the 24 h. Recent progress in technology has offered us more powerful tools to address this issue. They include devices for continuous noninvasive ambulatory blood pressure monitoring (Portapres, TNO), and techniques for a more comprehensive analysis of all components which contribute to overall blood pressure variability (broad-band spectral analysis).

Difference between clinic and daytime blood pressure is not a measure of the white coat effect.

The purpose of the present study was to evaluate whether the difference between blood pressure measured in the clinic or physician's office and the average daytime blood pressure accurately reflects the blood pressure response of the patient to the physician ("white coat effect" or "white coat hypertension"). We studied 28 hypertensive outpatients (mean age, 41.8+/-11.2 years; age range, 21 to 64 years) of 35 consecutive patients attending our hypertension clinic, in whom (1) continuous noninvasive finger blood pressure was recorded before and during the visit, (2) blood pressure was measured according to the Riva-Rocci-Korotkoff method (mercury sphygmomanometer) with the patient in the supine position, and (3) daytime ambulatory blood pressure was monitored with a SpaceLabs 90207 device. The peak blood pressure increase recorded directly during the visit was compared with the difference between clinic and daytime average ambulatory blood pressures. Compared with previsit values, peak increases in finger systolic and diastolic blood pressures during the visit to the clinic were 38.2+/-3.1 and 20.7+/-1.6 mm Hg, respectively (mean+/-SEM, P<.01 for both). Daytime average systolic and diastolic blood pressures were 135.5+/-2.5 and 89.2+/-1.9 mm Hg, with both lower than the corresponding clinic blood pressure values (146.6+/-3.6 and 94.9+/-2.2 mm Hg, P<.01). These differences, however, were <30% of the peak finger blood pressure increases during the physician's visit, to which these increases showed no relation. Although the visit to the physician's office was associated with tachycardia (9.0+/-1.6 bpm, P<.01), there was no difference between clinic and daytime average heart rates. These data indicate that the clinic-daytime average blood pressure difference does not reflect the alerting reaction and the pressure response elicited by the physician's visit and thus is not a reliable measure of the white coat effect.

Hemodilution reduces clinic and ambulatory blood pressure in polycythemic patients.

Limited information is available for humans on whether blood viscosity affects total peripheral resistance and, hence, blood pressure. Our study was aimed at assessing the effects of acute changes in blood viscosity on both clinic and 24-hour ambulatory blood pressure (BP) values. In 22 normotensive and hypertensive patients with polycythemia, clinic and 24-hour ambulatory BPs were measured before and 7 to 10 days after isovolumic hemodilution; this was performed through the withdrawal of 400 to 700 mL of blood, with concomitant infusion of an equivalent volume of saline-albumin solution. Hematocrit, plasma renin activity, plasma endothelin-1, right atrial diameter (echocardiography), and blood viscosity were measured under both conditions. Plasma renin activity and right atrial diameter were used as indirect markers of blood volume changes. Plasma endothelin-1 was used to obtain information on a vasomotor substance possibly stimulated by our intervention, which could counteract vasomotor effects. Isovolumic hemodilution reduced hematocrit from 0.53+/-0.05 to 0.49+/-0.05 (P<.01). Plasma renin activity, plasma endothelin-1 and right atrial diameter were unchanged. Clinic blood pressure was reduced by hemodilution (systolic, 144.3+/-5.4 to 136.0+/-3.9 mm Hg[mean+/-SEM]; diastolic, 87.0+/-2.8 to 82.1+/-2.6 mm Hg, P<.05 for both) and a reduction was observed also for 24-hour average ABP (systolic, 133.6+/-2.9 to 129.5+/-2.7 mm Hg; diastolic, 80.0+/-2.0 to 77.3+/-1.7 mm Hg, P<.05 for both). The reduction was consistent in hypertensive patients (n = 12), whereas in normotensive patients (n = 10) it was small and not significant. Both clinic and 24-hour average heart rates were unaffected by the hemodilution. Thus, in polycythemia, reduction in blood viscosity without changing blood volume causes a significant fall in both clinic and 24-hour ambulatory BPs; this is particularly true when, as can often happen, blood pressure is elevated. This emphasizes the importance this variable may have in the determination of blood pressure and the potential therapeutic value of its correction when altered.

Ambulatory blood pressure in hypertension with particular reference to the kidney.

Ambulatory blood pressure is more closely related to the end organ damage of hypertension than clinic blood pressure is. This is the case not only for left ventricular hypertrophy, but also for an index of renal involvement such as microalbuminuria. The closer correlation of ambulatory blood pressure with end organ damage characterises not only the 24 hour average value but also, to a similar extent, day-time and night-time average blood pressure, while the clinical importance of the difference between day and night blood pressure is still a matter of controversy. A more promising index derived from ambulatory blood pressure recordings seems to be blood pressure variability, which in preliminary studies has been shown to display an independent correlation with the end organ damage of hypertension.

Blood pressures other than the one at the clinic.

While assessing the cardiovascular risk of hypertensive patients, different types of blood pressure measurements can be regarded as suitable surrogate endpoints. In this context the possible role of clinic, stress, exercise, basal, home and ambulatory blood pressures is briefly discussed. The clinical value of night-time blood pressure, of the clinic-daytime blood pressure difference and of blood pressure variability is also addressed.

Clinical value of blood pressure variability.

Blood pressure (BP) is known to continuously fluctuate over time and both the amplitude and frequency of these changes have been reported to be clinically relevant. Indeed, it has been shown that the occurrence of pronounced BP changes at the time of the physician's visit may introduce errors in the diagnosis of hypertension and in the assessment of the efficacy of antihypertensive treatment. Moreover, several studies have provided evidence that the end organ damage of hypertension is not only more closely related to 24h average BP values than to casual BP readings, it is also significantly and independently related to the degree of BP variability during the day and night. Results from a recent follow-up study support the possibility that the degree of BP variability may also have prognostic relevance in hypertensive patients. Recent progress in technology has offered us non-invasive techniques for the assessment of beat-to-beat finger blood pressure variability and for the detailed quantification of all its frequency components from the fastest to the slowest ones. Their role as additional surrogate endpoints in essential hypertension needs to be addressed in future studies.

[The difference between clinical ambulatory measured blood pressure and daily monitored pressure does not reflect the "white coat effect"]. / La differenza tra la pressione misurata in ambulatorio e la pressione diurna monitorata non riflette l' "effetto camice bianco".

The difference between clinic and average daytime ambulatory blood pressure is frequently used to identify patients with "white coat" hypertension (i.e. with a pronounced pressor response to the clinical evaluation) although there is no evidence that this difference is indeed due to a white coat effect. In 28 mild hypertensive outpatients, the blood pressure was continuously recorded by a noninvasive finger device before and during the doctor's visit. The peak blood pressure increase, recorded during the visit was compared with the difference between clinic and daytime average ambulatory blood pressure. Peak increases in systolic and diastolic finger blood pressure during the doctor's visit were 38.2 +/- 3.1 mmHg and 20.7 +/- 1.6 mmHg, respectively compared to pre visit values (means +/- standard error, both p < 0.01). Daytime average systolic and diastolic blood pressure were 135.5 +/- 2.5 mmHg and 89.2 +/- 1.9 mmHg, both being lower than the corresponding clinic blood pressure values (146.6 +/- 3.6 mmHg and 94.9 +/- 2.2 mmHg, p < 0.01). Their differences, however, were < 30% of the peak finger blood pressure increase during the physician's visit. While the physician's visit was associated with tachycardia (+9.0 +/- 1.6 b/min, p < 0.01) there was no difference between clinic and daytime average heart rate. The alerting reaction and the pressor response induced by the physician's visit is not reflected by the difference between clinic and daytime average blood pressure. Such a difference is not therefore a reliable measure of the white coat effect.

Reproducibility of blood pressure measurements.

Clinic blood pressure readings are less reproducible than 24 h and daytime average ambulatory blood pressures. This may depend on (1) the continuous and spontaneous variability which characterizes daily life blood pressure, (2) the well-known 'white-coat' effect and (3) other methodological problems affecting traditional sphygmomanometric readings, such as the observer's bias and digit preference. The higher reproducibility of average 24 h and daytime blood pressures is also, at least in part, a function of the greater number of blood pressure readings available. On the other hand, the various components of the 24 h blood pressure profile appear, in general, to be less reproducible than the 24 h average value. This is the case for hourly values, for the day-night blood pressure difference and for a number of statistical parameters employed for modelling the 24 h blood pressure curve. An alternative and simple way to obtain reproducible blood pressure values, without making use of ambulatory blood pressure monitoring techniques, may be offered by the average of 20-25 blood pressure readings obtained automatically or semi-automatically under standardized conditions in outpatient clinics. The implications of these findings for clinical trials aimed at evaluating the features of the effects of antihypertensive drugs on 24 h blood pressure are discussed.

Blood pressure variability, cardiovascular risk and antihypertensive treatment.

TWENTY-FOUR HOUR BLOOD PRESSURE PARAMETERS: The use of ambulatory blood pressure monitoring techniques has shown clearly that 24-h average blood pressure is more closely related to the end-organ damage of hypertension than isolated office blood pressure readings. It has also provided evidence that the degree of blood pressure variability over a 24-h period may be independently related to the cardiovascular complications of hypertension. However, all the available data on this issue come from cross-sectional studies, and prospective evidence on the actual prognostic value of 24-h blood pressure parameters has only recently been provided for daytime blood pressure variability. There is still no prospective evidence concerning overall 24-h blood pressure variability. ANTIHYPERTENSIVE AGENTS AND BLOOD PRESSURE VARIABILITY: Available antihypertensive agents are unable to effectively buffer blood pressure variability. However, drugs with a long-lasting antihypertensive effect and an optimal trough: peak ratio may at least prevent further iatrogenic increases in the amplitude of blood pressure fluctuations. BEAT-TO-BEAT BLOOD PRESSURE MONITORING: The ability of antihypertensive agents to actually reduce 24-h blood pressure variability needs to be demonstrated in future studies, using beat-to-beat blood pressure monitoring which is now possible by means of non-invasive techniques.

Ambulatory blood pressure monitoring in the evaluation of antihypertensive drugs.

CLINIC VERSUS AMBULATORY BLOOD PRESSURE MEASUREMENT: Clinic blood pressure measurements do not give the best estimate of the efficacy of antihypertensive drugs because (1) they provide readings for one time-point only, (2) they are subject to a 'white-coat' effect and also show a significant placebo effect and (3) they are poorly reproducible. Ambulatory blood pressure monitoring overcomes these problems and offers the possibility of obtaining reliable, reproducible and detailed information on the time-course and magnitude of the effect of antihypertensive treatment on blood pressure over 24 h. MEAN 24-H VERSUS MEAN HOURLY AMBULATORY BLOOD PRESSURE VALUES: Compared with mean 24-h values, reproducibility is poorer for mean hourly ambulatory values, which show greater variability when tested under different conditions. Thus, caution must be exercised when the effects of antihypertensive drugs on hourly blood pressure profiles are analysed and the trough:peak ratio is calculated. FURTHER PROBLEMS WITH AMBULATORY MONITORING: There are further reasons for caution: (1) ambulatory blood pressure readings can be inaccurate, (2) there is only preliminary information on normal reference values and (3) there have been no longitudinal studies demonstrating the prognostic value of ambulatory blood pressure monitoring, which introduces uncertainty into attempts to evaluate the optimal effect of antihypertensive treatment by this method. CONCLUSIONS: These limitations do not detract from the use of ambulatory blood pressure monitoring in drug studies, but they do suggest that this measurement technique should not be used routinely for assessing antihypertensive treatment in clinical practice.

Blood pressure variability: clinical implications and effects of antihypertensive treatment.

HYPOTHESIS ON RELATIONSHIP BETWEEN BLOOD PRESSURE VARIABILITY AND END-ORGAN DAMAGE: Several studies have shown that the cardiovascular complications of hypertension are more closely related to ambulatory 24-h or daytime average blood pressure than to office readings. A few studies have also provided evidence that in hypertensive patients, not only average ambulatory blood pressure but also the degree of blood pressure variability is significantly and independently related to the end-organ damage associated with hypertension. LIMITATIONS OF PREVIOUS STUDIES: A common limitation of previous studies is that they were based on cross-sectional or retrospective observations, so that the correlative evidence they provide does not allow the relationship between blood pressure variability and end-organ damage to be interpreted causally. EVIDENCE FROM RECENT STUDIES: Recent evidence from follow-up observations has strongly supported the hypothesis that blood pressure variability is prognostically important in hypertensive patients. These findings suggest that optimal antihypertensive treatment should aim not only to reduce mean blood pressure levels, but also to reduce the degree of blood pressure fluctuation. EFFECTS OF ANTIHYPERTENSIVE DRUGS: Unfortunately, while most new antihypertensive drugs seem to be effective in reducing 24-h mean blood pressure levels, they are frequently unable to reduce 24-h blood pressure variability, which is often increased during treatment when expressed in normalized units. The development of drugs that guarantee a constant and uniform reduction in blood pressure over 24 h may, in principle, offer a further advantage by preventing the increase in 24-h blood pressure fluctuations that may follow the administration of short-acting antihypertensive agents. TROUGH: PEAK MEASUREMENTS OF BLOOD PRESSURE: The trough: peak ratio, proposed as an arithmetic indicator of the duration of the antihypertensive effect of a drug, may be a useful measure of the occurrence of a smooth reduction in blood pressure over 24 h. The possibility of obtaining an additional reduction in cardiovascular risk for hypertensive patients by minimizing the net trough: peak effect of antihypertensive drugs is thus an important issue for future studies.

Blood pressure variability and organ damage.

Several studies have now shown that hypertension-induced end-organ damage is more closely related to 24-h average blood pressure than to clinic measurements. Furthermore, the degree of variability of blood pressure during a 24-h period bears a relation to organ damage that is independent of average blood pressure value. The measurement of blood pressure variability is a complex task, however, because data from automatic ambulatory blood pressure monitoring should be interpreted with caution, especially if the interval between blood pressure measurements is more than 15 min, and different types of blood pressure variability (e.g., short-term and long-term) can make calculation of variability by standard deviation of 24-h blood pressure values difficult, which further complicates measurement procedures and interpretation. Evidence is growing to suggest that blood pressure variability in hypertension is clinically significant. We have recently shown that over a 7.5-year period, end-organ damage is independently related to the initial blood pressure variability. Although information on the effect of antihypertensive treatment on 24-h blood pressure variability is limited, the available data suggest more of an effect on 24-h average blood pressure levels than on 24-h blood pressure changes. Further studies should investigate treatment effects on different types of blood pressure variability and the impact of treatment on patient protection and prognosis.

Clinical value of ambulatory blood pressure monitoring.

This report reviews the evidence for and against clinical use of ambulatory blood pressure monitoring (ABPM) on a large scale. Such monitoring is supported by a number of data, among which is that the end-organ damage associated with hypertension correlates more with 24-h average blood pressure than with clinic blood pressure, the correlation becoming even closer with addition of blood pressure variability values. However, the evidence thus far collected is largely cross-sectional. Furthermore, ABPM devices have limited accuracy and the procedure has a high cost. Therefore, while prospective studies on the prognostic value of ambulatory blood pressure are awaited, use of this approach should be restricted to a limited number of clinical circumstances (e.g., identification of white-coat hypertension) and used only in specialized centers.

Radial artery compliance in essential hypertension: effects of antihypertensive therapy with lacidipine.

OBJECTIVE: To evaluate the effects of antihypertensive therapy with lacidipine on the increase in radial artery compliance observed in mild essential hypertensive patients. METHODS: The study was performed in eight mild to moderate essential hypertensive patients in whom clinic blood pressure, radial artery diameter and radial artery compliance were evaluated before and after 3 months' administration of lacidipine, at a single daily dose of 4 mg. Radial artery diameter and compliance were evaluated by means of a high precision echo-tracking device able to assess arterial compliance over the blood pressure oscillations that characterize the cardiac cycle. RESULTS: Lacidipine treatment caused a significant reduction in clinic systolic and diastolic blood pressure, while the heart rate was not modified by the drug. Radial artery diameter and compliance were both reduced by lacidipine over the entire systolodiastolic blood pressure range. CONCLUSIONS: Chronic administration of lacidipine seems to reverse the increase in compliance observed in essential hypertension at the radial artery level. We suggest that lacidipine treatment can reverse an increase in the smooth muscle component in the arterial wall induced by hypertension.