John Mertens

Prefrontal 5-HT2a receptor binding index, hopelessness and personality characteristics in attempted suicide

BACKGROUND Depression, hopelessness, impaired problem solving capacities and deficient serotonergic functions have been identified as major causes of suicidal behaviour. In general, the relation between biological markers of attempted suicide and psychological functions has been investigated using indirect peripheral markers of, e.g. the serotonergic system. Recently, functional neuroimaging techniques with radioligands allow direct in vivo assessment of the neurobiological status of the central nervous system. METHODS We studied the binding index of serotonin-(2a) (5-HT(2a)) receptors in the frontal cortex of attempted suicide patients (n=9) and normal controls (n=13) using [123I]5-I-R91150, a highly selective 5-HT(2a) receptor ligand. Moreover, we measured personality characteristics (using Cloningers Temperament and Character Inventory) and levels of hopelessness (using Becks Hopelessness Scale), and studied the association between 5-HT(2a) receptor binding index, hopelessness and these personality dimensions. RESULTS When compared to normal controls, attempted suicide patients had a significantly lower binding potential of frontal 5-HT(2a) receptors, a higher level of hopelessness, a higher score on the temperament dimension harm avoidance and lower scores on the character dimensions self-directedness and cooperativeness. A significant correlation was found between harm avoidance, hopelessness and binding index in the population of patients that attempted suicide. LIMITATIONS The limited number of patients and potential ingestion of psychotropic drugs may influence the results of the study. CONCLUSIONS Lower central serotonergic function, hopelessness and harm avoidance are interrelated phenomena, which may increase the probability of the occurrence of attempted suicide.

Estimates of regional cerebral blood flow and 5-HT2A receptor density in impulsive, aggressive dogs with 99mTc-ECD and 123I-5-I-R91150

Impulsive aggression in dogs has an important impact on human public health. Better insight into the pathophysiology of this phenomenon could lead to more adequate diagnosis and treatment. Indirect in vivo research on peripheral body fluids and post-mortem studies in impulsive animals and humans indicate a deficient serotonergic system in general and disturbances in the serotonin-2A (5-HT2A) receptor in particular. In this study, brain perfusion and the 5-HT2A receptors were examined in impulsive, aggressive dogs, in comparison with a group of normally behaving animals. In order to decide which dogs to include in this study, owners were asked to describe the general behaviour of the dogs, the circumstances in which aggression occurred and their conduct during aggressive acts. Finally, 19 dogs were retained for this study, showing, according to different behavioural specialists, disinhibited dominance aggression. Functional imaging studies were performed on all these dogs. Single-photon emission tomography (SPET) was used to measure regional brain perfusion using technetium-99m labelled ethyl cysteinate dimer (ECD). The 5-HT2A receptor binding properties were investigated using the selective radioligand iodine-123 labelled 5-I-R91150. A significant increase in uptake of the 5-HT2A radioligand was noted in all cortical areas. No significant alterations were found in regional cortical perfusion, indicating that the increased binding index was not a consequence of increased tracer delivery. This study supports a role for the serotonergic system in canine impulsive aggression.

New fast preparation of 123I labelled radiopharmaceuticals

A new fast kit preparation of 123I labelled radio-pharmaceuticals such as IMP, HIPDM, MIBG and Hippuran is proposed. A radiochemical yield >99% is obtained at 100°C within 10–30 min. The new labelling procedure is based on the nucleophilic exchange in presence of Cu(I) and an excess of reducing agents. The four kit prepared 123I-radiopharmaceuticals have been used with success in clinical studies involving 400 patients. The proposed method is also compared with earlier described methods which, yielding labelled side products and 123I2, do not fulfill the requirements for kit labelling.

Initial evaluation of123I-5-I-R91150, a selective 5-HT2A ligand for single-photon emission tomography, in healthy human subjects

The mapping of 5-HT2 receptors in the brain using functional imaging techniques has been limited by a relative lack of selective radioligands. Iodine-123 labelled 4-amino-N-[1-[3-(4-fluorophenoxy)propyl]-4-methyl-4-piperidinyl]-5-iodo-2-methoxybenzamide (123I-5-I-R91150 or123I-R93274) is a new ligand for single-photon emission tomography (SPET), with high affinity and selectivity for 5-HT2A receptors. This study reports on preliminary123I-5-I-R91150 SPET, wholebody and blood distribution findings in five healthy human volunteers. Maximal brain uptake was approximately 2% of total body counts at 180 min post injection (p.i.). Dynamic SPET sequences were acquired with the brain-dedicated, single-slice multi-detector system SEM-810 over 200 min p.i. Early peak uptake (at 5 min p.i.) was seen in the cerebellum, a region free from 5HT2A receptors. In contrast, radioligand binding in the frontal cortex increased steadily over time, up to a peak at approximately 100–120 min p.i. Frontal cortex-cerebellum activity ratios reached values of 1.4, and remained stable from approximately 100 min p.i. onwards. Multi-slice SPET sequences showed a pattern of regional variation of binding compatible with the autoradiographic data on the distribution of 5-HT2A receptors in (cerebral cortex>striatum>cerebellum). These findings suggest that123I-5-I-R91150 may be used for the imaging of 5-HT2A receptors in the living human brain with SPET.

The impact of HF-rTMS treatment on serotonin2A receptors in unipolar melancholic depression

BACKGROUND Currently, the underlying neurobiological mechanism as to how repetitive transcranial magnetic stimulation (rTMS) can alter depressive states remains unclear. Animal data suggest that its influence could occur at the neurotransmitter level, such as modulation of the serotonin system. METHODS Twenty-one antidepressant-free medication-resistant unipolar depressed patients, and 21 age- and gender-matched healthy subjects were studied. We examined the neurobiologic impact of 10 high-frequent (HF)-rTMS sessions applied to the left dorsolateral prefrontal cortex (DLPFC) on postsynaptic 5-HT(2A) receptor binding indices (BI) measured with ¹²³I-5-I-R91150 single photon emission computed tomography (SPECT) only in patients. RESULTS Compared with the control group, patients displayed significantly less bilateral dorsolateral prefrontal cortical and significantly higher left hippocampal baseline 5-HT(2A) receptor BI. Successful HF-rTMS treatment correlated positively with 5-HT(2A) receptor BI in the DLPFC bilaterally and correlated negatively with right hippocampal 5-HT(2A) receptor uptake values. CONCLUSIONS Our results indicate that HF-rTMS treatment affect the serotonergic system. Our data also suggest that this kind of treatment affects 5-HT(2A) receptor BI in the DLPFC and in the hippocampus in different ways.

Preclinical evaluation of [123I]R93274 as a SPECT radiotracer for imaging 5-HT2A receptors

Studies in rodents have suggested that the radioiodinated 5-HT2A receptor antagonist [123I]R93274 (123-iodine-N-[(3-p-fluorophenyl-1-propyl)-4-methyl-4-piperidinyl]-4-ami no- 5-iodo-2-methoxybenzamide) (Kd = 0.1 nM) might be a promising single photon emission computerized tomography (SPECT) radiotracer to image 5-HT2A receptors in the living human brain. In this study, we characterized the brain uptake of [123I]R93274 in baboons. Highest brain uptake was observed in cortical areas, while lower uptake was observed in the striatum and the cerebellum. Injection of pharmacological doses of the 5-HT2A receptor antagonist ketanserin resulted in reduction of cortical and striatal radioactivities to the level observed in the cerebellum. Injection of the selective dopamine D2 receptor antagonist raclopride did not affect [123I]R93274 brain uptake. Quantification of 5-HT2A receptors was achieved by measuring the binding potential of 5-HT2A receptors for [123I]R93274 (the binding potential is the product of the density and affinity of available receptors). Regional binding potential values were derived with a three-compartmental kinetic analysis of the time-activity curves in the brain and plasma. Binding potential values of 93 +/- 34 ml/g, 71 +/- 35 ml/g and 31 +/- 11 ml/g were measured in the occipital, temporal and striatal regions, respectively. Similar values were derived using a noncompartmental graphical analysis. These values were in accordance with the in vitro regional distribution of 5-HT2A receptors in primate brain. In conclusion, [123I]R93274 allows visualization and quantification of 5-HT2A receptors in the baboon brain with SPECT.

In vitro characterization of the influx of 3-[125I]iodo-L-α-methyltyrosine and 2-[125I]iodo-L-tyrosine into U266 human myeloma cells: Evidence for System T transport

The aim of this study was to investigate the cellular uptake mechanisms responsible for the accumulation of 3-[(125)I]iodo-L-alpha-methyltyrosine ((125)I-3-IMT) and 2-[(125)I]iodo-L-tyrosine ((125)I-2-IT), two radiotracers for metabolic tumor imaging, using single-photon emission tomography, into U266 human myeloma cancer cells. Time course and concentration dependency of (125)I-3-IMT uptake was assessed. Kinetic parameters were calculated using an Eadie Hofstee plot. A set of competitive inhibitors of the main amino acid transport systems was used for the discrimination of the transporters responsible for the uptake of (125)I-3-IMT and (125)I-2-IT. Protein incorporation of both tracers was determined using acid precipitation. The measured maximum velocity for (125)I-3-IMT transport was 4.199 nmol per mg protein 20 s(-1), and the Michaelis constant was 107.9 microM. Addition of 2-aminobicyclo[2,2,1]heptane-2-carboxylic acid (BCH), a competitive inhibitor of System L, reduced the influx by 39.0+/-3.3% for (125)I-3-IMT and 66.3+/-0.9% for (125)I-2-IT. The BCH-insensitive influx was further reduced by Tryptophan (Trp) by 43.8+/-3.5% for (125)I-3-IMT and 15.3+/-1.3% for (125)I-2-IT. This suggests involvement of System T transport. We measured <2% of radioactivity in the acid precipitable fractions of both tracers with no increase in time. We conclude that the influx of (125)I-3-IMT and (125)I-2-IT into U266 human myeloma cells is mediated by both System L and System T amino acid transporters. The kinetic parameters suggest that elevated plasma levels of aromatic amino acids will reduce (123)I-3-IMT uptake in myeloma patients. Both tracers do not enter protein synthesis significantly.

The Use of an Ammonium Selective Electrode for the Automatic Determination of Ammonia in Boiler Feed-Waters

Abstract An automatic potentiometric method for the determination of ammonia in boiler feed-waters is presented. An ammonium selective membrane electrode of the neutral carrier type was used as sensor-electrode. The accuracy and reproducibility were very good. The method allows a sample rate of 30 samples an hour.

New high yield Cu(I) assisted 123I radioiodination of 15(p-I-phenyl)-9-methyl pentadecanoic acid, a potential myocardial tracer.

A new high yield 123I radioiodination (97%), based on the Cu(I) assisted isotopic exchange in an ethanol water mixture, of 15(p-I-phenyl)-9 methyl pentadecanoic acid, a potential myocardial tracer, is proposed. The method allows a true kit preparation of radioiodinated phenyl fatty acids for a substrate concentration of ±10-6 moles. High specific activities (>0.1 mCi/μg) can be obtained when coupling the labelling method to HPLC purification with an overall radiochemical yield of 75%.

I-123/125-labelled 2-iodo-L-phenylalanine and 2-iodo-D-phenylalanine : comparative uptake in various tumour types and biodistribution in mice

PurposeIn vitro in the R1M cell model and in vivo in the R1M tumour-bearing athymic model, both [123I]-2-iodo-L-phenylalanine and [123I]-2-iodo-D-phenylalanine have shown promising results as tumour diagnostic agents for SPECT. In order to compare these two amino acid analogues and to examine whether the observed characteristics could be generalised, both isomers were evaluated in various tumour models.MethodsTransport type characterisation in vitro in A549, A2058, C6, C32, Capan2, EF43fgf4, HT29 and R1M cells with [123I]-2-iodo-L-phenylalanine was performed using the method described by Shotwell et al. Subsequently, [123I]-2-iodo-L-phenylalanine and [123I]-2-iodo-D-phenylalanine tumour uptake and biodistribution were evaluated using dynamic planar imaging and/or dissection in A549, A2058, C6, C32, Capan2, EF43fgf4, HT29 and R1M inoculated athymic mice. Two-compartment blood modelling of the imaging results was performed.ResultsIn vitro testing demonstrated that [123I]-2-iodo-L-phenylalanine was transported in all tumour cell lines by LAT1. In all tumour models, the two amino acid analogues showed the same general biodistribution characteristics: high and specific tumour uptake and renal tracer clearance. Two-compartment modelling revealed that the D-isomer showed a faster blood clearance together with a faster distribution to the peripheral compartment in comparison with [123I]-2-iodo-L-phenylalanine.Conclusion[123I]-2-iodo-L-phenylalanine and its D-isomer are promising tumour diagnostic agents for dynamic planar imaging. They showed a high and similar uptake in all tested tumours. [123I]-2-iodo-D-phenylalanine showed better tracer characteristics concerning radiation dose to other organs.