Novel procedures for preparing 99mTc(III) complexes with tetradentate/monodentate coordination of varying lipophilicity and adaptation to 188Re analogues.

Improved methods are presented for the preparation of 99mTc and 188Re mixed-ligand complexes with tetradentate and monodentate ligands of the general formula [MIII(Lm)(Ln)] (M = Tc, Re; Lm = NS3 or NS3COOH; Ln = isocyanide or phosphine). To avoid the undesired formation of reduced-hydrolyzed species of both metals, the preparation of complexes is performed in a two-step procedure. At first the Tc(III)- or Re(III)-EDTA complex is formed which reacts in a second step with the tripodal ligand 2,2',2' '-nitrilotris(ethanethiol) (NS3) or its carboxyl derivative NS3COOH (a) and the monodentate phosphine ligands (triphenylphosphine L1, dimethylphenylphosphine L2) or isocyanides (tert-butyl isonitrile L3, methoxyisobutyl isonitrile L4, 4-isocyanomethylbenzoic acid-L-arginine L5, 4-isocyanomethylbenzoic acid-L-arginyl-L-arginine L6, 4-isocyanomethylbenzoic acid-neurotensin(8-13) L7) to the so-called '4+1' complex. Copper(I) isocyanide complexes are used for preparing the '4+1' complexes. That facilitates storage stability and allows kit formulations, and, moreover, enables the formation of 188Re complexes in acidic solution. Only micromolar amounts of the monodentate ligand are needed, and that results in high specific activity labeling of interesting molecules. The lipophilicity of complexes can be controlled by introducing a carboxyl group into the tetradentate ligand and/or derivatization of the monodentate ligands. Furthermore, the carboxyl group enables the conjugation of biomolecules. As an example, the neurotensin derivative CN-NT(8-13) was prepared and labeled with 99mTc according to the '4+1' approach, and its behavior in vivo was studied.

The influence of different external cooling methods on thermoregulatory responses before and after intense intermittent exercise in the heat.

The purpose of this study was to determine the effect of different cooling methods on thermoregulation before and after intermittent anaerobic exercise in the heat (38 degrees C). On separate days, 10 men completed 4 conditions consisting of 2 sets of six 30-second sprints (with 30 seconds of rest) at 125% of maximal aerobic power with each set of sprints followed by a cooling procedure. The 4 conditions were the following: passive cooling at room temperature (22 degrees C; PRC), fan cooling (4.0 m x s(-1), 22 degrees C; FAC), fan cooling with water spraying (50 ml x min(-1); FWC), and a noncooling passive recovery in the heat chamber (38 degrees C; PCC). Each set of 6 sprints was followed by a 12-minute cooling period; after the second 12-minute period, cooling continued until esophageal temperature (Tes) was reduced by 1.0 degrees C. Tes and mean skin temperatures (Tsk) were taken before and during exercise and during all cooling phases. Cooling rates (mean +/- SEM) after the second set of sprints (based on Tes) were greater (p < 0.05) in PRC (0.043 +/- 0.007) than in the other conditions (FWC = 0.027 degrees +/- 0.005 degrees, FAC = 0.03 degrees +/- 0.004 degrees, and PCC = 0.021 degrees +/- 0.003 degrees C per minute). Overall decreases in heat content, however, were greater in the FWC (-332.2 +/- 27.8) and FAC (-129.9 +/- 14.7 kJ) conditions compared with the PRC condition (29.0 +/- 14.9 kJ). The time required to lower Tes by 1.0 degrees C with PRC (22.8 +/- 1.8) was less than with FAC (30.4 +/- 2.7 minutes). Finally, the rate of increase in Tes during the second set of sprints was less in the FAC and FWC conditions (0.15 degrees +/- 0.01 degrees and 0.11 degrees +/- 0.01 degrees C per minute) compared with the PCC and PRC conditions (0.19 degrees +/- 0.01 degrees and 0.18 degrees +/- 0.01 degrees C per minute), suggesting differences in pre-exercise cooling. Based on cooling rates and the time required to lower Tes by 1.0 degrees C, PRC was the most effective method of cooling. The conclusion is different, however, when taking into account changes in heat content since the FAC and FWC conditions were more effective in dissipating heat and in preventing heat gain during the second set of sprints

Elevated DNA double strand breaks and apoptosis in the CNS of scid mutant mice.

Genetic approaches have provided evidence that DNA end-joining problems serve an essential role in neuronal survival during development of mammalian embryos. In the present study, we tested whether the DNA repair enzyme, DNA dependent protein kinase, plays an important role in the survival of cerebral cortical neurons in mice. DNA-PK is comprised of a DNA-binding subunit called Ku and a catalytic subunit called DNA-PKcs. In mice with the scid mutation, DNA-PKcs is truncated near the kinase domain, which causes loss of kinase activity. We compared the spatial and temporal aspects of neuronal cell death in scid versus isogenic wild-type embryos and found a significant increase in dying cells in scid mice, as assessed by nuclear changes, DNA fragmentation and caspase-3 activity. Additional biochemical and immunocytochemical studies indicated that of several DNA repair enzymes investigated, only PARP was increased in scid mice, possibly in response to elevated DNA strand breaks.

Clinical relevance of the primary findings of the MTA: success rates based on severity of ADHD and ODD symptoms at the end of treatment.

OBJECTIVES: To develop a categorical outcome measure related to clinical decisions and to perform secondary analyses to supplement the primary analyses of the NIMH Collaborative Multisite Multimodal Treatment Study of Children With Attention-Deficit/Hyperactivity Disorder (MTA). METHOD: End-of-treatment status was summarized by averaging the parent and teacher ratings of attention-deficit/hyperactivity disorder and oppositional defiant disorder symptoms on the Swanson, Nolan, and Pelham, version IV (SNAP-IV) scale, and low symptom-severity ("Just a Little") on this continuous measure was set as a clinical cutoff to form a categorical outcome measure reflecting successful treatment. Three orthogonal comparisons of the treatment groups (combined treatment [Comb], medication management [MedMgt], behavioral treatment [Beh], and community comparison [CC]) evaluated hypotheses about the MTA medication algorithm ("Comb + MedMgt versus Beh + CC"), multimodality superiority ("Comb versus MedMgt"), and psychosocial substitution ("Beh versus CC"). RESULTS: The summary of SNAP-IV ratings across sources and domains increased the precision of measurement by 30%. The secondary analyses of group differences in success rates (Comb = 68%; MedMgt = 56%; Beh = 34%; CC = 25%) confirmed the large effect of the MTA medication algorithm and a smaller effect of multimodality superiority, which was now statistically significant (p < .05). The psychosocial substitution effect remained negligible and nonsignificant. CONCLUSION: These secondary analyses confirm the primary findings and clarify clinical decisions about the choice between multimodal and unimodal treatment with medication.

Psychosocial treatment strategies in the MTA study: rationale, methods, and critical issues in design and implementation.

The Collaborative Multimodal Treatment Study of Children with Attention Deficit Hyperactivity Disorder (ADHD), the MTA, is the first multisite, cooperative agreement treatment study of children, and the largest psychiatric/psychological treatment trial ever conducted by the National Institute of Mental Health. It examines the effectiveness of Medication vs. Psychosocial treatment vs. their combination for treatment of ADHD and compares these experimental arms to each other and to routine community care. In a parallel group design, 579 (male and female) ADHD children, aged 7-9 years, 11 months, were randomly assigned to one of the four experimental arms, and then received 14 months of prescribed treatment (or community care) with periodic reassessments. After delineating the theoretical and empirical rationales for Psychosocial treatment of ADHD, we describe the MTA's Psychosocial Treatment strategy applied to all children in two of the four experimental arms (Psychosocial treatment alone; Combined treatment). Psychosocial treatment consisted of three major components: a Parent Training component, a two-part School Intervention component, and a child treatment component anchored in an intensive Summer Treatment Program. Components were selected based on evidence of treatment efficacy and because they address comprehensive symptom targets, settings, comorbidities, and functional domains. We delineate key conceptual and logistical issues faced by clinical researchers in design and implementation of Psychosocial research with examples of how these issues were addressed in the MTA study.

Diagnosis of cellular states of microbial organisms using proteomics.

Two-dimensional (2-D) polyacrylamide gel electrophoresis has much to contribute to experimental analysis of the proteomes of microbial organisms, since this method separates most cellular proteins and allows synthesis rates to be determined quantitatively. Databases generated using 2-D gels can grow to be very large from even just a few experiments, since each sample provides the data for a field (or column) in the database for several hundreds to even thousands of records (or rows), each of which represents a single polypeptide species. The value of such databases for generating an encyclopedia of how each of the cell's proteins behave in different conditions (protein phenotypes) has been recognized for some time. The potential exists, however, to glean even more valuable information from such databases. Because the measurements of each protein are made in the context of all other proteins, a comprehensive glimpse of the cell's physiological state is theoretically achievable with each 2-D gel. By examining enough conditions (and 2-D gels), expression patterns of subsets of proteins (proteomic signatures) can be found that correlate with the cell's state. This type of information can provide a unique contribution to proteomic analysis, and should be a major focus of such analyses.

National Institute of Mental Health Collaborative Multimodal Treatment Study of Children with ADHD (the MTA). Design challenges and choices.

The Collaborative Multimodal Treatment Study of Children with Attention Deficit Hyperactivity Disorder (ADHD), the MTA, is the first child multisite cooperative agreement treatment study of children conducted by the National Institute of Mental Health, Rockville, Md. It examines the long-term effectiveness of medication vs behavioral treatment vs both for treatment of ADHD and compares state-of-the-art treatment with routine community care. In a parallel-groups design, 576 children (age, 7-9 years) with ADHD (96 at each site) are thoroughly assessed and randomized to 4 conditions: (1) medication alone, (2) psychosocial treatment alone, (3) the combination of both, (4) or community comparison. The first 3 groups are treated for 14 months and all are reassessed periodically for 24 months. Designers met the following challenges: framing clinically relevant primary questions; defining the target population; choice, intensity, and integration and combination of treatments for fair comparisons; combining scientific controls and standardization with clinical flexibility; and implementing a controlled clinical trial in a nonclinical setting (school) controlled by others. Innovative solutions included extensive decision algorithms and manualized adaptations of treatments to specific needs.

What we know about using research findings: implications for improving special education practice.

This article discusses the alternative roles and responsibilities that researchers might assume to bridge the gap between special education research and classroom practice. Literature is reviewed that addresses why some research-based practices fail, and the factors likely to lead to sustained use of research-based practices. The roles of researchers as collaborators, facilitators and coaches, interpreters of the culture and dynamics of the school, and critics are described in an attempt to promote researchers as leaders in efforts to improve practices in the field of special education.

Quantification of reactive oxygen species generated by alveolar macrophages using lucigenin-enhanced chemiluminescence--methodical aspects.

Alveolar macrophages lavaged from bovine lungs using Ca2(+)- and Mg2(+)-deprived saline containing EGTA for calcium chelation were cultivated in RPMI-1640 medium. The generation of reactive oxygen species (ROS) was assessed by determination of lucigenin-enhanced chemiluminescence (LUC-CL) using a LB 9505 C Biolumat. Monitoring of LUC-CL depends on various methodical parameters: besides medium constituents, the method of cell harvesting and the time-schedule of the protocol appear to be essential parameters that influence ROS-generation. In addition, the cell number (cell density) as well as the ratio of cell number to particle mass influence the amount of ROS-generation. Following exposure of cells to micronized quartz, we observed a dose-related increase in the generation of ROS.

[Dose intensified carboplatin monotherapy in advanced ovarian cancer]. / Dosisintensivierte Carboplatinmonotherapie beim fortgeschrittenen Ovarialkarzinom.

Platinum derivatives are known as the most effective cytostatic agents in ovarian carcinoma. The steep dose-response curves described by Hryniuk and Levin [5] make them the ideal substances for dose escalation. We treated patients with advanced ovarian carcinoma with dose intensified carboplatinmonotherapy six times at AUC 5 (= +/- 400 mg/m2 in patients with normal renal function). Dose intensification was not done by increasing single doses, but by shortening the intervals between therapies (23, 21, 17, 14 days). In order to prevent leucopenia G-CSF was administered at a dose of 5 micrograms/kg s.c. Besides mild emesis no extramedullary toxicity especially no alopezia was documented. Up to an interval of 17 days therapy was safe; no thrombocytopenia < 49,000 x 10(6)/L and no leucopenia < 1000 x 10(6) was observed. With the intention to arrive at a 14-day interval and to attain further dose escalation we will treat future patients with a slightly reduced dose of carboplatin (AUC 4) in combination with escalating doses of cis-platinum, which is known for its low myelotoxicity.

[Ultrastructure of the liver and endocrine system in aging rats: effect of piracetam]. / Ultrastruktur von Leber und Inkretsystem alter Ratten: Einfluss von Piracetam.

Lipogenic pigmentation is described in thyroid follicular, parathyroid, and pineal cells, as well as in adrenocortical macrophages. Piracetam activated pituitary prolactin cells and increased paracrystals in thyroid follicular cells; smooth endoplasmic reticulum was increased and glycogen content decreased in numerous hepatocytes randomly distributed over the lobule.

Erythromycin therapy for subclinical intrauterine infections in threatened preterm delivery--a preliminary report.

Evidence suggests that subclinical intrauterine infections which can be indicated by elevated maternal CRP-values may cause preterm labor and that Ureaplasma urealyticum might play a role in the pathogenesis of preterm delivery. Since these organisms are sensitive to erythromycin, 43 patients with threatened preterm labor were at the time of tocolysis treated with erythromycin, or placebo in a randomized study. Treatment with erythromycin resulted in a greater mean delay of delivery for 9 days than among the placebo-treated women. This effect is statistically significant in patients with intact membranes and cervical dilatation: in contrast to the placebo group these women achieved an increased delay of 23 days before delivering. Moreover, successful erythromycin treatment was significantly associated with the isolation of ureaplasmas from vaginal swabs. Our results emphasize that in cases with cervical dilatation, vaginal microorganisms, especially ureaplasmas, are more liely to ascend and might be involved in the initiation of preterm labor. Therefore, additional treatment with erythromycin was beneficial only in these women.

[Premature labor in gestosis]. / Frühgeburtlichkeit bei Gestose.

796 pregnancies complicated by preeclampsia and 1,299 pregnancies without toxemia of the years 1981 to 1985 have been compared with regard to prematurity. Prematurity rates were 12.4 respectively 13.8 per cent, hypotrophy rates were 20 resp. 13 per cent, acidosis morbidity was 75 resp. 20 per cent and Apgar values below 8 were 63 resp. 24 per cent. Morbidity rate of respiratory distress syndrome was 8 resp. 12 per cent, of sepsis 2 resp. 7 per cent, intrauterine death rate 5 resp. 2 per cent, but survival rate overall was 93 resp. 90 per cent. Prematurity was influenced by severity of preeclampsia, time of onset and prenatal care. Prolongation of pregnancy by tocolysis is possible principally, but influenced in its effect by maternal and fetal symptoms and the necessity of termination of pregnancy by these factors.

[Clinical value of determining C-reactive protein in the maternal serum in pregnancies with threatened premature labor]. / Zur klinischen Wertigkeit der Bestimmung des C-reaktiven Proteins im mütterlichen Serum bei Schwangerschaften mit drohender Frühgeburt.

In this article, because of the importance of subclinical infections for triggering premature labor, we have investigated the maternal serum levels of C-reactive protein (CRP) in 88 pregnancies with comparable gestational ages at the onset of premature birth symptoms. We found certain associations between possible prolongation of gestation by tocolysis and absence or presence of pathologic values of this acute-phase-protein. Positive CRP-values are associated with significantly lowered prolongation of pregnancy by tocolysis and subsequently lowered gestational age at birth. Though there are no correlations to maternal temperature and white blood cell counts, antenatal CTG-results, amniotic fluid properties and parameters of the neonate, we found probable relations to the detection of Ureaplasmas in the vaginal swabs. Once more we point out to possible associations between mycoplasmas and premature birth.

Mating preference inSchistosoma mansoni.

We investigated the suitability of an in vitro culture system for measurement of mating behavior ofSchistosoma mansoni. The criteria used to evaluate this system were the level of phosphorylated nucleotides, egg production, and mating status of parasites. The level of ATP, ADP, AMP, and G6-P was measured at different time intervals during cultivation of worm pairs and remained essentially the same as that of control worms for up to 6 days. Egg production was observed in this system during 19 days of cultivation. Peak egg production occurred on day 4 with 72% of the total eggs being laid during the first week of cultivation. The variability in the number of eggs produced by different pairs ofS. mansoni necessitated the selection and matching of tubes with the same number of eggs after 48 hr. This permitted the detection of small changes in egg production by decreasing intertube variation. Mating recognition between male and femaleS. mansoni was evaluated by culturing separated adult worms with their original partner or with a different partner. During the first 24 hr, mating occurred among a greater percentage of worm pairs comprised of their original partner than among worm pairs comprised of different partners (P < 0.001). After 48 and 72 hr of cultivation, these differences were not statistically significant. Similar results were obtained with a culture of mixed males and females. Two drugs were studied for their effects on the mating ofS. mansoni in vitro. Aminoglutethimide (AG) at a concentration of 1 × 10(-4) had no effect on the frequency of mating whereas diethyldithiocarbamate (DDC) completely inhibited mating at a concentration of 3 × 10(-6) M and reduced the level of ATP in these worms.

Extraction of intersexual chemoattractants fromSchistosoma mansoni.

FemaleSchistosoma mansoni and their excretory-secretory (ES) products were extracted with a series of solvents to provide fractions of varying polarity. These fractions were assayed for chemoattractivity to males in vitro. One major component of these mixtures was found to be nonattractive and was identified as cholesterol.n-Pentane- and ether-soluble fractions derived from ES products exhibited chemoattractive activity comparable to that possessed by whole-worm extracts, but appeared to be simpler mixtures.