Prenatal Risk Calculation (PRC) 3.0: An Extended DoE-Based First-Trimester Screening Algorithm Allowing For Early Blood Sampling.

AIM: Both previous versions of the German PRC algorithm developed by our group for routine first-trimester screening relied on the assumption that maternal blood sampling and fetal ultrasonography are performed at the same visit of a pregnant women. In this paper we present an extension of our method allowing also for constellations where this synchronization is abandoned through preponing blood sampling to dates before 11 weeks of gestation. METHODS: In contrast to the directly measured concentrations of the serum parameters PAPP-A and free ß-hCG, the logarithmically transformed values could be shown to admit the construction of reference bands covering the whole range from 16 to 84 mm CRL [corresponding to 63 to 98 days of gestation]. Prior to determining reference limits from which the DoEs for each individual patient had to be calculated, the log concentrations of all PAPP-A and free ß-hCG values were transformed once more using the calibration approach established in 1 for the elimination of the influence of maternal weight. RESULTS: Although that part of the database which was available for estimating the reference bands for blood sampling times prior to 11 weeks of gestation was comparatively sparse (898 out of 186 215 pregnancies with euploid outcome), the key statistical characteristics of the extended risk-calculation procedure turned out to be very satisfactory. Using the same cutoff value of 1:150 for the posterior risks of trisomy 21 and 13/18, the overall FPR (false positive rate) for diagnosing a T21 was found to be 3.42%. The corresponding DTR (detection rate) was obtained to be 86.8% and thus exceeded the DTR attained by PRC 2.0 for trisomy 21. For trisomies 13 and 18, the proportions of patients with calculated posterior risks exceeding the cutoff value of 1:150 were obtained to be 1.60% (=FPR) and 86.4% (=DTR). CONCLUSION: Transforming the measured concentrations of PAPP-A and free ß-hCG to the logarithmic scale allows one to extend the DoE-based algorithm developed by the FMF Germany for diagnosing trisomies 21 and 13/18 in such a way that it can be applied to constellations where blood sampling is done before 11 weeks of gestation.

Normal Doppler Reference Values of the Pericallosal Artery.

PURPOSE: To provide the normal reference values of the Doppler flow of the pericallosal artery in relation to gestational age from 18 to 41 weeks of gestation. MATERIALS AND METHODS: The pericallosal artery (PCA) was studied in 466 normal pregnancies. The pulsed Doppler evaluation of the pericallosal artery was done in A3 and A4 segments, and records from PI, RI and Vmax were studied. RESULTS: The resistance index of the pericallosal artery in A3/A4 segments exhibits a plateau from 18 to 31 weeks of gestation. After 31 weeks, a marked decrease becomes apparent. The pulsatility index of the pericallosal artery in A3/A4 segments shows a plateau until 36 weeks of gestation. During the final weeks of gestation, there is a decrease in the pulsatility index. Vmax exhibits a plateau for the maximal flow velocity in A3/A4 segments of the pericallosal artery from 18 to 28 weeks of gestation. After 28 weeks of gestation, there is a slight increase in Vmax. CONCLUSION: Normal reference values of the pericallosal artery might have an impact on clinical judgment during adaptive hemodynamic changes and regarding the progression of the fetal deterioration occurring in the presence of fetal hypoxia.

A comparison of region-of-interest measures for extracting whole brain data using survival analysis in alcoholism as an example.

BACKGROUND: Aggregation of functional magnetic resonance imaging (fMRI) data in regions-of-interest (ROIs) is required for complex statistical analyses not implemented in standard fMRI software. Different data-aggregation measures assess various aspects of neural activation, including spatial extent and intensity. NEW METHOD: In this study, conducted within the framework of the PREDICT study, we compared different aggregation measures for voxel-wise fMRI activations to be used as prognostic factors for relapse in 49 abstinent alcohol-dependent individuals in an outpatient setting using a cue-reactivity task. We compared the importance of the data-aggregation measures as prognostic factors for treatment outcomes by calculating the proportion of explained variation. RESULTS AND COMPARISON WITH EXISTING METHOD(S): Relapse risk was associated with cue-induced brain activation during abstinence in the ventral striatum (VS) and in the orbitofrontal cortex (OFC). While various ROI measures proved appropriate for using fMRI cue-reactivity to predict relapse, on the descriptive level the most "important" prognostic factor was a measure defined as the sum of t-values exceeding an individually defined threshold. Data collected in the VS was superior to that from other regions. CONCLUSIONS: In conclusion, it seems that fMRI cue-reactivity, especially in the VS, can be used as prognostic factor for relapse in abstinent alcohol-dependent patients. Our findings suggest that data-aggregation measures that take both spatial extent and intensity of cue-induced brain activation into account make better biomarkers for predicting relapse than measures that consider an activation's spatial extent or intensity alone.

Advanced dynamic Doppler flow of the pulmonary artery in a normal population: reference values from 18 to 41 weeks of gestation calculated by automatic Doppler waveform analysis.

OBJECTIVE: The aim of the present cross-sectional prospective study was to establish Doppler reference ranges for blood flow velocity waveforms (V max, V mean and V min) and resistance indices (PI, RI) of the fetal pulmonary arteries using the new pulsed-wave color advanced dynamic flow (ADF) Doppler technique. METHOD: Data were collected in 206 low-risk pregnancies at 18-41 weeks of gestation. The measurements were obtained in the proximal pulmonary artery near the first bifurcation in the absence of fetal body or breathing movements. RESULTS: The pulsatility index (PI) in the pulmonary artery showed mean increases of 2.43-3.59 between gestational weeks 18 and 42. A similar pattern was observed for the resistance index (RI) with increases of 0.79-0.90. Increases in systolic (V max), mean (V mean) and end diastolic (V min) blood flow velocities of 36.0-63.3, 10.8-19.9 and 5.71-7.53 cm/s, respectively, were noted during the observation interval. CONCLUSIONS: The ranges for blood flow velocities and impedance indices in the fetal pulmonary artery calculated by the authors may serve as reference values to help distinguish a normal patient population from patients carrying fetuses at high risk for neonatal lung disease in antenatal examinations.

[The importance of pathology in the German prostate cancer study PREFERE]. / Die Bedeutung der Pathologie in der deutschen Prostatakrebsstudie PREFERE.

Prostate cancer is the most common carcinoma of elderly males and holds the third place in the ranking of cancer-specific mortality. However, total mortality rate of 3 % is low and half of the patients die from other diseases, which is for the most part due to significantly improved diagnostic methods and the increasing use of prostate-specific antigen (PSA) screening. This has led to a stage migration towards early tumor stages that are prognostically heterogeneous and require differentiated treatment. The German and European guidelines recommend four therapy options (i.e. radical prostatectomy, percutaneous irradiation, permanent seed implantation and active surveillance) for localized prostate cancer and from contemporary study data it is unclear which therapy is most beneficial. This will be the subject of the PREFERE trial, a prospective randomized multicentre trial which plans to recruit 7,600 patients and to observe them over a period of up to 17 years. The histopathological diagnosis of the primary biopsy plays a crucial role in the inclusion criteria, as this article outlines in detail.

Biometry of the fetal corpus callosum by three-dimensional ultrasound.

OBJECTIVES: To construct reference ranges of quantitative characteristics of the fetal corpus callosum. METHODS: Women referred to a tertiary center for sonographic examination were recruited to undergo a detailed fetal scan from 17 to 41 weeks of gestation. Three-dimensional (3D) sonographic volumes of normal fetal brains were acquired and analyzed offline. We obtained three different measurements of the corpus callosal length, as well as the height (/thickness) of its segments, namely the rostrum, genu, body and splenium. RESULTS: Initially we recruited 604 pregnant women, of whom 138 were excluded because of various disorders/abnormalities, multiple pregnancy or gestational age < 18 weeks. Thus, included in the analysis were 466 sonographic volumes of normal fetal brains from singleton pregnancies, acquired by transabdominal ( n = 170) or transvaginal (n = 296) ultrasound. The corpus callosum was visualized as a hypoechoic structure. Reference ranges were established for the following parameters: curved corpus callosal length, inner-inner corpus callosal length, outer-outer corpus callosal length, rostrum height, genu height, body height and splenium height. We observed non-linear growth and an approximately four-fold increase in all corpus callosal lengths, a three-fold increase in rostrum height, a four-fold increase in genu height, a two-fold increase in body height and a three-fold increase in splenium height between 18 and 41 weeks. The growth patterns of rostrum and body height appeared to be similar: there was rapid development until 24 and 22 weeks of gestation, respectively, and growth slowed beyond this period. The growth patterns of genu and splenium were also similar, being characterized by progressive growth throughout gestation. CONCLUSIONS: Using 3D ultrasound, we have constructed reference charts for measurements of the corpus callosum. Knowledge of the normal growth pattern may be useful for evaluation of abnormal development of the corpus callosum, and so help in the accurate diagnosis of pathologies such as hypogenesis and dysgenesis.

[PREFERE - the German prostatic cancer study: questions and claims surrounding study initiation in January 2013]. / PREFERE - die Deutsche Prostatakrebsstudie : Fragen und Behauptungen rund um den Studienbeginn im Januar 2013.

The PREFERE study is a multicenter randomized study of patients with low or early intermediate risk for prostatic cancer. The four treatment options, radical prostatectomy, percutaneous irradiation therapy, permanent seed implantation and active surveillance recommended by the German S3 guidelines and international guidelines will be tested and compared with respect to effectiveness and potential side effects. Over a period of 4 years a total of 7,600 patients are to be recruited and assigned to 1 of these 4 therapy forms according to personal preference (by possible exclusion of 1 or 2 therapy options) in a 2-4 arm study design by randomization.

A combination of galantamine and memantine modifies cognitive function in subjects with amnestic MCI.

OBJECTIVES: Mild cognitive impairment (MCI) is etiologically heterogeneous, and a substantial proportion of MCI subjects will develop different dementia disorders. One subtype of this syndrome, amnestic MCI, occurs preferentially but not exclusively in prodromal AD and is characterized by defined deficits of episodic memory. DESIGN, SETTING AND PARTICIPANTS: For a 2-year, double-blinded, placebo-controlled study MCI patients, presenting with an amnestic syndrome but not necessarily based on presumed prodromal AD were randomized. INTERVENTION: Patients received (a) a combination of 16 mg galantamine plus 20 mg memantine, or (b) 16 mg galantamine alone or (c) placebo. MEASUREMENTS: The primary objective was to explore the differential impact of these interventions on the progression to dementia and on cognitive changes as measured by the ADAScog. RESULTS: After recruitment of 232 subjects, the trial was halted before reaching the planned sample size, because safety concerns arose in other studies with galantamine in MCI. This resulted in a variable treatment duration of 2-52 weeks. The statistical analysis plan was amended for studying cognitive effects of discontinuing the study medication, which was done separately for galantamine and memantine, and under double-blind conditions. There was one death, no unexpected severe adverse events, and no differences of severe adverse events between the treatment arms. The cognitive changes on the ADAScog were not different among the groups. Only for the subgroup of amnestic MCI with presumed AD etiology, a significant improvement of ADAScog score over placebo before the discontinuation of medication was observed, while amnestic MCI presumably due to other etiologies showed no cognitive changes with broad variation. Cognitive improvement was numerically larger in the combination treatment group than under galantamine alone. Patients who received placebo declined as expected. Discontinuation of galantamine, either as part of the combination regimen or as mono treatment, resulted in a transient decline of the ADAScog score in amnestic MCI of presumed AD etiology, while discontinuation of Memantine did not change the cognitive status. CONCLUSION: In an interrupted trial with amnestic MCI subjects the combination of galantamine plus memantine were generally well tolerated. In the subgroup of MCI subjects with presumed AD etiology, a cognitive benefit of a short-term combination treatment of galantamine plus memantine was observed, and cognitive decline occurred after discontinuation of galantamine.

Blood flow velocity waveforms of the umbilical artery in a normal population: reference values from 18 weeks to 42 weeks of gestation.

PURPOSE: The aim of the present study was to calculate new Doppler reference ranges for systolic, intensity-weighted mean, and end-diastolic blood flow velocities (Vmax, Vmean, Vmin) and fetal heart rate and impedance indices (PI, RI) of the umbilical artery by automatic waveform analysis. MATERIALS AND METHODS: A cross-sectional prospective study of 1926 low-risk pregnancies was performed at 18 - 42 weeks of gestation. The measurements were carried out in the region of a free floating loop of cord. Reference ranges for the individual measuring parameters were constructed based on a growth function from a four-parameter class of monotonic continuous functions according to the smallest square principle. The intraobserver reliability was also assessed. RESULTS: A significant increase in umbilical blood flow velocity was observed at 18 - 42 weeks of gestation (Vmax = 33.7 cm/sec to 65 cm/sec (p < 0.001), Vmean = 18.2 cm/sec to 38.7 cm/sec (p < 0.001) and Vmin = 8.2 to 31 cm/sec (p < 0.001). Reference curves for the pulsatility and the resistance index were associated with significant decreases with increasing gestational age (PI: 18 weeks: 1.3; 28 weeks: 1.1; 42 weeks: 0.8 and RI: 18 weeks: 0.7; 28 weeks: 0.6; 42 weeks: 0.5). The fetal heart rate decreased from 147 - 134 bpm. The intraobserver reliability was low, but of no clinical relevance. CONCLUSION: The reference ranges for blood flow velocities and resistance indices in the umbilical artery calculated by the authors serve as the basis for Doppler ultrasound antenatal examinations in a normal patient population and enable the early diagnosis of fetal risk.

Reference values for blood flow velocity in the uterine artery in normal pregnancies from 18 weeks to 42 weeks of gestation calculated by automatic Doppler waveform analysis.

PURPOSE: The goal of the present study was to establish new Doppler reference ranges for maternal heart rate, intensity-weighted mean blood flow velocities (Vmean) and impedance indices (PI, RI) for the uterine artery by automated waveform analysis. MATERIALS AND METHODS: A cross-sectional prospective study of 921 low-risk pregnancies was performed at 18 - 42 weeks of gestation. Uterine blood flow velocities were derived with pulsed-wave color Doppler. Measurements were carried out 1 to 2 cm above the crossing of the uterine and external iliac arteries. Reference ranges for the individual measuring parameters were constructed based on a growth function from a four-parameter class of monotonic continuous functions according to the smallest square principle. RESULTS: A significant increase in intensity-weighted mean uterine blood flow velocities was observed at 18 - 42 weeks of gestation (Vmean = 43 cm/s to 50 cm/s (p < 0.001)). Reference curves for the pulsatility and resistance indices (PI, RI) significantly decreased with progressing gestation (PI: 18 weeks: 0.89; 42 weeks: 0.65 and RI: 18 weeks: 0.45; 42 weeks: 0.35). No significant PI and RI differences were observed when different placental locations were compared. The maternal heart rate decreased from 88 bpm to 77 bpm. CONCLUSION: Normal ranges for blood flow velocities and impedance indices in the uterine artery were established by Doppler ultrasound antenatal examinations of a large population of low-risk pregnancies. The data are proposed as reference curves to allow the early diagnosis of maternal and fetal risks.

Fetal lung volume measurements using 3D ultrasonography.

PURPOSE: The aim of the study was to establish fetal lung, thoracic and heart volume nomograms using three-dimensional (3D) ultrasonographic measurements. MATERIALS AND METHODS: For this purpose a total of 300 fetuses were examined between 18 and 34 weeks of gestation using Voluson 530 ME and Voluson 730 PRO (General Electic, USA) ultrasound devices with 5 MHz three-dimensional annular volume transducers. To determine fetal lung volume, each lung was measured separately using a transversal sectional plane in the multiplanar mode. Measurements were performed by area tracing around the fetal lung in cross-sectional planes in different slices. The distance between two represented slices was calculated by computer. Calculated volume data was plotted against gestational age in order to obtain nomograms of fetal lung, heart and thorax volumes. RESULTS: Our nomograms revealed increasing lung, heart and thoracic volume growth between 22 and 34 weeks of gestation. Furthermore, we could demonstrate that there is a statistically significant difference between the growth of the right and left lung volume (right > left). In a group of 12 fetuses with skeletal dysplasias or hydrothorax pulmonary hypoplasia was suspected by ultrasound. Comparing two-dimensional (2D) sonographic measurements of oblique lung diameter and 3D lung volumetry, it was found that these two methods were complementary for the recognition of pulmonary hypoplasia before the 24 weeks of gestation. CONCLUSION: Using 3D ultrasound it is easy to perform fetal lung volumetry especially before 30 weeks of gestation. The encouraging results suggest that this method could be useful for the early detection of pulmonary hypoplasia even before 24 weeks of gestation.

Individualized correction for maternal weight in calculating the risk of chromosomal abnormalities with first-trimester screening data.

AIM: In the algorithm developed by the Fetal Medicine Foundation (FMF) Germany designed to evaluate the findings of routine first-trimester screening, the false-positive rate (FPR) was determined for the entire study group without stratification by maternal weight. Based on the data received from the continuous audit we were able to identify an increase in the FPR for the weight-related subgroups of patients, particularly for patients with extremely high body weights. The aim of this study was to demonstrate that the variability of the FPR can be reduced through adjusting the concentrations of free ß-HCG and PAPP-A measured in the maternal serum by means of a nonlinear regression function modeling the dependence of these values on maternal weight. MATERIAL AND METHODS: The database used to establish a version of the algorithm enabling control of the FPR over the whole range of maternal weight consisted of n = 123 546 pregnancies resulting in the birth of a child without chromosomal anomalies. The group with positive outcomes covered n = 500 cases of trisomy 21 and n = 159 trisomies 13 or 18. The dependency of the serum parameters free ß-HCG and PAPP-A on maternal weight was analyzed in the sample of negative outcomes by means of nonlinear regression. The fitted regression curve was of exponential form with negative slope. Using this model, all individual measurements were corrected through multiplication with a factor obtained as the ratio of the concentration level predicted by the model to belong to the average maternal body weight of 68.2 kg, over the ordinate of that point on the regression curve which belongs to the weight actually measured. Subsequently, the totality of all values of free ß-HCG and PAPP-A corrected for deviation from average weight were used as input data for carrying out the construction of diagnostic discrimination rules described in our recent paper for a database to which no corrections for over- or under-weight had been applied. This entailed in particular the construction of new reference bands for the corrected biochemical values as the basis for calculating the degree of extremeness (DOE) measures to replace the more traditional MOMs. In the final and most crucial step, stratified FPRs were computed and compared over a set of intervals partitioning the whole range of maternal weight into 18 classes. RESULTS: For the posterior risks of both trisomy 21 and 13 / 18 computed from the weight-corrected database, the use of a cutoff value of 1:150 turned out to be an appropriate choice. For T 21, the overall FPR obtained through comparing the individual risks with this cutoff was found to be 3.51 %. The corresponding proportion of ascertained cases of trisomy 21 detected by means of the new algorithm was 86.2 %. For the trisomy 13 / 18 group, the analogous results were a FPR of 2.07 % and a detection rate (DTR) of 83.0 %, respectively. A comparison between the FPRs obtained for the 18 intervals into which the range of maternal weight had been partitioned, showed the deviation of the strata-specific from the overall FPR to be fairly small: for T 21, the FPR ranged from 2.72 to 4.86 %, and the maximum was found in the group of 87.5 - 95.0 kg. For women with a weight of more than 120 kg, the FPR was only slightly above the FPR for the total sample (3.69 as compared to 3.51 %). Similar results were obtained for the discrimination rule constructed for diagnosing T 13 / 18: here, the minimum FPR (1.17 %) was found for patients weighing more than 120 kg, whereas the maximum (2.66 %) occurred in the interval 75.0 - 77.5 kg. CONCLUSION: In this study we demonstrated that the new algorithm developed by the FMF Germany to estimate risks for fetal trisomies 21 and 13 / 18 combines very good misclassification rates with a far-reaching stability of the false-positive rate against even extreme deviations from the average maternal weight.

A new approach to calculating the risk of chromosomal abnormalities with first-trimester screening data.

PURPOSE: First-trimester screening at 11 - 14 weeks has been proven to be very useful in the early detection of chromosomal defects. The aim of this project was to develop a CE-certified new risk calculation program (PRC = Prenatal Risk Calculation) using a nationwide database. MATERIALS AND METHODS: The database underlying the new risk calculation procedure was established in Germany from 2003 through 2006. Overall, the database includes measurements from 70,030 pregnant women having given birth to healthy children. Following consideration of all pregnancies associated with a chromosomally abnormal outcome, the sample size was 451. The algorithm used for calculating the risk of a chromosomally abnormal outcome comprises the following variables: maternal age, crown-rump length (CRL) (restricted to a range from 45 - 84 mm or, equivalently, 11 + 1 - 14 + 0 weeks of gestation), nuchal translucency (NT), as well as the maternal serum parameters PAPP-A (pregnancy associated plasma protein A) and free beta-hCG (free human chorionic gonadotropin). In a preliminary cross-validation study, we applied both the new algorithm and the FMF UK program to an independent sample containing n = 40,568 pregnancies with negative outcome, n = 187 cases of trisomy 21, n = 34 trisomies 18 and n = 13 trisomies 13. RESULTS: Using the primary sample of 70,030 pregnancies with a negative outcome, reference bands were constructed for the sonographic parameter fetal nuchal translucency and the biochemical parameters PAPP-A and free beta-HCG. Instead of MoM values we used "degree of extremeness" (DoE) values. This statistical parameter has been proven to give more precise results than the MoM measure because it assesses the deviation of the actual measurement value from the centre of the reference band expressed as a multiple of the width of the respective band section. The result of the risk calculation is visualized by means of a traffic light graph which allows the patient to comprehend her individual risk at first glance. The red color indicates a high risk, green a low risk, and yellow represents a moderate risk. In our preliminary cross-validation study the detection rate obtained for the German algorithm was 86.6 % for trisomy 21, 94.1 % for trisomy 18 and 92.4 for trisomy 13. The corresponding detection rates obtained with the same data by the FMF UK program were 86.1 %, 82.3 % and 69.2 % throughout. The false-positive rate was 5.0 % throughout. CONCLUSION: The new risk calculation procedure of the FMF Germany (PRC) has been made available as a CE-certified computer program. In screening for trisomy 21 it yields results comparable to those of the program used by the FMF UK. Regarding the diagnosis of trisomy 13 and 18, even higher detection rates are currently achieved with the German algorithm. Program, data base and license key are available free of charge to registered members of the FMF Germany.

A new approach to sample size calculation for reference interval studies.

A new criterion is proposed for determining the sample size required for a study performed for the purpose of establishing reference intervals. The basic idea behind the criterion is to compare the empirical coverage (i.e. the probability content) of the reference region obtained from the sample with its target value (e.g. 95 per cent) and to set suitable limits delta1, delta2 to the difference between both quantities which must not be exceeded with sufficiently large probability beta (e.g. beta=90 per cent). For the most frequently used parametric and distribution-free methods of estimating univariate reference limits, implicit formulae are derived relating the sample size to the design parameters delta1, delta2 and beta. For symmetric specification of (delta1, delta2), explicit approximation formulae for the computation of n are given. Exact values obtained by means of suitable numerical techniques are presented in a set of tables covering specifications of delta1, delta2 and beta which can be recommended for real applications. The tables can be used both for one- and two-sided reference intervals.

Statistical confirmation of negative results of association studies in genetic epidemiology.

One of the most important reasons warranting the common reservation about publishing association studies with negative results is due to the fundamental fact that an insignificant result of a statistical testing procedure tailored for establishing an association, fails to provide conclusive evidence of the contrary. In this contribution, we show how the methodology of equivalence testing, as provided by modern biostatistics, can be exploited for removing this basic flaw. In order to keep the exposition as simple as possible, we restrict discussion to the setting of a study which aims at ruling out that some given single SNP has a relevant impact on the risk of developing the disease under consideration. As solutions to the problem of how to perform a valid confirmatory analysis of a trial of this type, two different approaches are presented: (i) exact Fisher type test for equivalence of an odds ratio to unity; (ii) distribution-free test for equivalence of cases and controls with respect to the full (i.e., trinomial) genotype distributions. The practical implementation of both testing procedures are described in detail and illustrated with examples taken from recent studies of the genetic epidemiology of psychiatric disorders.

Relationship between cortisol and serotonin metabolites and transporters in alcoholism [correction of alcolholism].

BACKGROUND: Stress hormone activation may induce clinical depression via an interference with central serotonergic neurotransmission. In alcoholics, a reduction in serotonin transporters was associated with clinical depression, and an activation of cortisol secretion is frequently found during detoxification. We assessed the interaction between stress hormone activation, serotonin transporters, monoamine metabolites in the cerebrospinal fluid (CSF), and mood states in male and female alcoholics and healthy control subjects. METHODS: The availability of serotonin transporters was measured with [I-123]beta-CIT and SPECT in the raphe area of the brainstem in 31 alcoholics after four weeks of abstinence and in 25 age-matched healthy volunteers. Concentrations of plasma cortisol were measured on the day of the SPECT scan. Within one week after the SPECT scan, we assessed monoamine metabolites and corticotropin-releasing factor (CRF) in the CSF. RESULTS: Clinical depression was associated with a reduction in serotonin transporter availability among male alcoholics. Among male alcoholics and healthy volunteers, CSF 5-HIAA and plasma cortisol concentrations were inversely correlated with the availability of raphe serotonin transporters and positively correlated with the severity of clinical depression. No significant correlations were observed between raphe serotonin transporters and HVA, MHPG and CRF concentrations in the CSF. CONCLUSION: Our findings support the hypothesis of an interaction between reduced serotonin transporters, stress hormone activation and clinical depression. They confirm the hypothesis that serotonergic neurotransmission dysfunction in alcoholism is limited to male alcoholics. The observed interactions between high cortisol concentrations and reduced serotonin transporter availability warrant further studies in major depression and other neuropsychiatric diseases with implied cortisol activation and serotonergic dysfunction.

Age-related reference regions for longitudinal measurements of growth characteristics.

Most studies on age-related reference centiles published up to now have adopted a strictly cross-sectional perspective. Clearly, the results of studies of that type do not provide a tool for the diagnostic assessment of whole series of measurements taken sequentially over time in the same individual. In this paper, the approach of Wellek & Merz (1995) to the construction of age-dependent reference ranges for cross-sectional measurements is generalized in such a way that data sets containing time series of arbitrary length varying between subjects can be accommodated. Since repeated measurements on the same subject are typically correlated, the regression function to be used as the central line for the reference band eventually obtained is determined by fitting a nonlinear mixed model describing the dependence of conditional means on age by growth functions of the same class we proposed in the case of cross-sectional data. Estimation of the parameters of this mixed model is done in a way closely related to the population-averaged GEE approach by Zeger et al. (1988). Given the regression line, the reference band is constructed by means of an iterative procedure guaranteeing that the proportion of observed profiles which nowhere leave the band, has some prespecified value (frequently set equal to 90% in practice). The approach is illustrated with two examples taken from child psychiatry and prenatal sonography.