The influence of emotional priming on the neural substrates of memory: a prospective fMRI study using portrait art stimuli.

Events coupled with an emotional context seem to be better retained than non-emotional events. The aim of our study was to investigate whether an emotional context could influence the neural substrates of memory associations with novel portrait art stimuli. In the current prospective fMRI study, we have investigated for one specific visual art form (modern artistic portraits with a high degree of abstraction) whether memory is influenced by priming with emotional facial pictures. In total forty healthy female volunteers in the same age range were recruited for the study. Twenty of these women participated in a prospective brain imaging memory paradigm and were asked to memorize a series of similar looking, but different portraits. After randomization, for twelve participants (Group 1), a third of the portraits was emotionally primed with approach-related pictures (smiling baby faces), a third with withdrawal-related pictures (baby faces with severe dermatological conditions), and another third with neutral images. Group 2 consisted of eight participants and they were not primed. Then, during an fMRI session 2h later, these portraits were viewed in random order intermixed with a set of new (previously unseen) ones, and the participants had to decide for each portrait whether or not they had already been seen. In a separate experiment, a different sample of twenty healthy females (Group 3) rated their mood after being exposed to the same art stimuli, without priming. The portraits did not evoke significant mood changes by themselves, supporting their initial neutral emotional character (Group 3). The correct decision on whether the portraits were Familiar of Unfamiliar led to similar neuronal activations in brain areas implicated in visual and attention processing for both groups (Groups 1 and 2). In contrast, whereas primed participants showed significant higher neuronal activities in the left midline superior frontal cortex (Brodmann area (BA) 6), unprimed volunteers displayed higher right medial frontal cortical (BA 10) activities. Furthermore, specifically in Group 1, correct retrieval of negatively primed portraits evoked increased neuronal activity in the left medial orbitofrontal cortex (BA 11) and in the right (posterior) insula, suggesting enhanced stress-related responses to the memory of withdrawal-related primed modern artistic portraits in this group. Our prospective memory data in healthy females indicate that, to reach a correct retrieval decision, different midline anterior neuronal networks are recruited for portraits that were emotionally primed than for the unprimed ones. Importantly, our results also suggest that the negative emotional context leads to the formation of associations that are reactivated during memory retrieval processes of the initially neutral art portraits. When correctly recognized, the portraits evoke neuronal activities consistent with the withdrawal-related character of the emotional visual stimuli with which they have been associated. Although our results show that abstract portrait art can be associated with emotional primes this doesn't mean that this effect is specific for art images.

Stress sensitive healthy females show less left amygdala activation in response to withdrawal-related visual stimuli under passive viewing conditions.

The amygdalae are key players in the processing of a variety of emotional stimuli. Especially aversive visual stimuli have been reported to attract attention and activate the amygdalae. However, as it has been argued that passively viewing withdrawal-related images could attenuate instead of activate amygdalae neuronal responses, its role under passive viewing conditions remains unclear. Furthermore, because individual sensitivity to stress reactions has been shown to modulate amygdalae processing, the aim of the current event-related fMRI study was to investigate whether individual differences in stress proneness could influence amygdala responses while passively viewing withdrawal and approach-related visual images. We presented 14 healthy female subjects with a random sequence of images of happy 'healthy' baby faces (approach-related) and baby faces disfigured by severe dermatological conditions (withdrawal-related). No instructions were given other than to watch the images attentively. We integrated individual perceived stress (PSS) scores in our analysis. The processing of withdrawal-related pictures resulted in less left amygdala activity in females scoring higher on perceived stress. Our findings suggest that stress-sensitive healthy females are less able to fully attend to withdrawal-related visual material and in essence avoid exposure to such images in an effort to reduce strong psychophysiological responses. Although the relatively small number of participants limits drawing firm conclusions, we suggest that in passive viewing emotional brain imaging paradigms, individual information on stress proneness should be included in the interpretation of amygdala neuronal processes.

Improved quantification in multiple-pinhole SPECT by anatomy-based reconstruction using microCT information.

PURPOSE: The aim of this study was to evaluate the potential of anatomy-based reconstruction, using microCT information, to improve quantitative accuracy in multiple-pinhole SPECT. METHODS: Multiple-pinhole SPECT and microCT images were acquired with the Micro Deluxe Phantom using both hot and cold rod inserts. The phantoms were filled with 3.7 MBq/ml of (99m)Tc. To improve microCT contrast, the phantoms were also filled with contrast agent. Emission images were reconstructed using a one-step-late (OSL) modification of the ordered subsets expectation maximization (OSEM) algorithm for incorporation of microCT information, to encourage smoothing within but not across boundaries. To allow quantification, the OSL OSEM algorithm takes into account imperfect camera motion, collimator response, angular variation of the sensitivity, intrinsic camera resolution, attenuation and scatter. For comparison, the emission images were also reconstructed by OSEM using post-reconstruction filtering and by OSL OSEM using a quadratic prior and an edge-preserving prior. In each rod of the phantoms the recovery coefficient (RC), defined as measured divided by the true activity concentration, was expressed as a function of the noise. Different noise levels were obtained by varying the amount of spatial filtering during or after reconstruction and by the use of binominal deviates. RESULTS: Compared to conventional OSEM using post-reconstruction filtering and compared to OSL OSEM using a quadratic prior, our study demonstrated that the use of anatomical information during reconstruction significantly improved the quantitative accuracy in both cold and hot rods with a diameter larger than or equal to 2.4 mm. When compared to the edge-preserving prior, the anatomical prior performs significantly better for hot rods with a diameter ≥ 2.4 mm. For the 4.0-mm hot rods for example, the RC averaged over the different noise levels was 0.67 ± 0.02 when multiple-pinhole SPECT images were reconstructed using anatomical information, compared to 0.54 ± 0.08, 0.60 ± 0.04 and 0.64 ± 0.02 when OSEM in combination with a post-reconstruction filter, OSL OSEM using a quadratic prior and OSL OSEM using a median root prior was used, respectively. For the 4.0-mm cold rods, the RC averaged over the different noise levels was 0.61 ± 0.03 when the multiple-pinhole SPECT images were reconstructed using anatomical information, compared to 0.54 ± 0.07, 0.53 ± 0.08 and 0.60 ± 0.03 when OSEM in combination with a post-reconstruction filter, OSL OSEM using a quadratic prior and OSL OSEM using a median root prior was used, respectively. CONCLUSION: Anatomy-based reconstruction using microCT information has the potential to improve quantitative accuracy in multiple-pinhole SPECT.

The effect of one left-sided dorsolateral prefrontal cortical HF-rTMS session on emotional brain processes in women.

Although repetitive Transcranial Magnetic Stimulation (rTMS) is frequently used to examine emotional changes in healthy volunteers, it remains largely unknown how rTMS is able to influence emotion.We carried out a sham-controlled single-blind crossover study using fMRI, we examined in 20 right-handed healthy female volunteers whether a single high frequency (HF)-rTMS session applied to the left dorsolateral prefrontal cortex (DLPFC) could influence emotional processing while focussing on blocks of positively and negatively valenced baby faces. A single HF-rTMS session selectively influenced the processing of positively and negatively valenced baby faces. In essence, our results indicate that the effects of one left-sided HF-rTMS sessions results in improved processing of positive emotions and reduced negative emotional processing in never depressed female subjects.

Time-course of contrast enhancement in spleen and liver with Exia 160, Fenestra LC, and VC.

OBJECTIVE: The purpose of this study was to compare the time-course of contrast-enhancement in spleen and liver using Exia 160 in comparison with Fenestra LC and VC in healthy mice. PROCEDURES: Healthy C57bl/6 mice were used in this study. Fenestra LC and VC was administered intravenously at a dose of 0.1 ml/20 g or 0.2 ml/20 g. Exia 160 at a dose of 0.05 ml/20 g or 0.1 ml/20 g. Each animal underwent a micro-CT scan before contrast injection (baseline) and immediately after contrast injection. Additional scans were performed at 1, 2, 3, 4, 24, and 48 h after contrast administration. The mice who received Exia 160 were also scanned after 15, 30, and 45 min. RESULTS: The peak enhancement of Exia 160 occurred after 15 min for the spleen and after 30 min for the liver. CONCLUSIONS: Exia 160 allows rapid spleen and liver enhancement. The high iodine content results in small injection volumes.

Improved quantification in single-pinhole and multiple-pinhole SPECT using micro-CT information.

PURPOSE: The purpose of this study was to demonstrate the feasibility of accurate quantification in pinhole SPECT using micro-CT information. METHODS: Pinhole SPECT scans were performed using a clinical dual-head gamma camera. Each pinhole SPECT scan was followed by a micro-CT acquisition. Functional and anatomical images were coregistered using six point sources visible with both modalities. Pinhole SPECT images were reconstructed iteratively. Attenuation correction was based on micro-CT information. Scatter correction was based on dual and triple-energy window methods. Phantom and animal experiments were performed. A phantom containing nine vials was filled with different concentrations of (99m)Tc. Three vials were also filled with CT contrast agent to increase attenuation. Activity concentrations measured on the pinhole SPECT images were compared with activity concentrations measured by the dose calibrator. In addition, 11 mice were injected with (99m)Tc-labelled Nanobodies. After acquiring functional and anatomical images, the animals were killed and the liver activity was measured using a gamma-counter. Activity concentrations measured on the reconstructed images were compared with activity concentrations measured with the gamma counter. RESULTS: The phantom experiments demonstrated an average error of -27.3 +/- 15.9% between the activity concentrations measured on the uncorrected pinhole SPECT images and in the dose calibrator. This error decreased significantly to -0.1 +/- 7.3% when corrections were applied for nonuniform attenuation and scatter. The animal experiment revealed an average error of -18.4 +/- 11.9% between the activity concentrations measured on the uncorrected pinhole SPECT images and measured with the gamma counter. This error decreased to -7.9 +/- 10.4% when attenuation and scatter correction was applied. CONCLUSION: Attenuation correction obtained from micro-CT data in combination with scatter correction allows accurate quantification in pinhole SPECT.

HF-rTMS treatment in medication-resistant melancholic depression: results from 18FDG-PET brain imaging.

INTRODUCTION: High frequency repetitive transcranial magnetic stimulation (HF-rTMS) of the left dorsolateral prefrontal cortex (DLPFC) might be a promising strategy to treat depression, but not all patients show a positive outcome. OBJECTIVE: In this open study, we evaluate whether a favorable HF-rTMS treatment outcome could be predicted by baseline prefrontal brain glucose metabolism (CMRglc), measured by 18fluorodeoxyglucose positron emission tomography (18FDG-PET). METHODS: A sample of 21 antidepressant-free, treatment-resistant depression (TRD) patients of the melancholic subtype received 10 sessions of HF-rTMS delivered on the left DLPFC. Patients underwent a static 18FDG-PET before and after HF-rTMS treatment. RESULTS: Forty-three percent of the patients showed a reduction of at least 50% on their Hamilton Rating Scale for Depression scores. Higher baseline metabolic activities in the DLPFC and the anterior cingulate cortex (ACC) were associated with better clinical outcome. Successful HF-rTMS treatment was related to metabolic changes in subdivisions of the ACC (Brodmann areas 24 and 32). CONCLUSION: This biological impact of HF-rTMS on regional brain CMRglc explains to some extent how HF-rTMS may improve moods in TRD patients. Larger sham-controlled HF-rTMS treatment studies are needed to confirm these results.

SPECT imaging with 99mTc-labeled EGFR-specific nanobody for in vivo monitoring of EGFR expression.

PURPOSE: Overexpression of the epidermal growth factor receptor (EGFR) occurs with high incidence in various carcinomas. The oncogenic expression of the receptor has been exploited for immunoglobulin-based diagnostics and therapeutics. We describe the use of a llama single-domain antibody fragment, termed Nanobody, for the in vivo radioimmunodetection of EGFR overexpressing tumors using single photon emission computed tomography (SPECT) in mice. METHODS: Fluorescence-activated cell sorting (FACS) analysis was performed to evaluate the specificity and selectivity of 8B6 Nanobody to bind EGFR on EGFR overexpressing cells. The Nanobody was then labeled with (99m)Tc via its C-terminal histidine tail. Uptake in normal organs and tissues was assessed by ex vivo analysis. In vivo tumor targeting of (99m)Tc-8B6 Nanobody was evaluated via pinhole SPECT in mice bearing xenografts of tumor cells with either high (A431) or moderate (DU145) overexpression of EGFR. RESULTS: FACS analysis indicated that the 8B6 Nanobody only recognizes cells overexpressing EGFR. In vivo blood clearance of (99m)Tc-8B6 Nanobody is relatively fast (half-life, 1.5 h) and mainly via the kidneys. At 3 h postinjection, total kidney accumulation is high (46.6+/-0.9%IA) compared to total liver uptake (18.9+/-0.6%IA). Pinhole SPECT imaging of mice bearing A431 xenografts showed higher average tumor uptake (5.2+/-0.5%IA/cm(3)) of (99m)Tc-8B6 Nanobody compared to DU145 xenografts (1.8+/-0.3%IA/cm(3), p<0.001). CONCLUSION: The EGFR-binding Nanobody investigated in this study shows high specificity and selectivity towards EGFR overexpressing cells. Pinhole SPECT analysis with (99m)Tc-8B6 Nanobody enabled in vivo discrimination between tumors with high and moderate EGFR overexpression. The favorable biodistribution further corroborates the suitability of Nanobodies for in vivo tumor imaging.

Intra-individual comparison of the human biodistribution and dosimetry of the D and L isomers of 2-[123I]iodo-phenylalanine.

PURPOSE: Several authors have shown in animal studies that D-enantiomeric amino acid analogues can have better tumour imaging properties compared to their L-analogues. In our group, the D and L isomers of 2-[I]iodo-phenylalanine were identified as tumour-specific tracers in rat and mouse tumour models, with an advantage for the D-isomer. Here we compare, intra-individually, the normal biodistribution and dosimetry of both tracers in healthy human subjects. METHODS: Five male volunteers received both the L- and D-enantiomers, ranging from 84 to 114 MBq, with a 1 week interval between the tracers, allowing intra-individual comparison. Whole-body scans were performed and blood and urine samples were collected and analysed up to 24 h. Dosimetry was calculated using OLINDA 1.0 software. RESULTS: The biodistributions of the tracers are comparable as both show a moderate uptake in heart and the liver, a marked uptake in muscle tissue and clearance via the renal system. However, due to faster clearance, from 2.5 h, the uptake of the D-enantiomer was significantly lower compared to the L-isomer in all organs. The radiation dose estimations showed an effective dose of, respectively, 0.0120+/-0.0020 mSv x Bq(-1) and 0.0106+/-0.0038 mSv x Bq(-1) for 2-123I-L-Phe and 2-123I-D-Phe (P=0.18). In both cases the organ receiving the highest dose was the bladder wall. CONCLUSION: Both 2-123I-L-Phe and 2-123I-D-Phe show comparable moderate uptake in all organs. 2-123I-D-Phe is the more promising tracer, as it shows a faster clearance resulting in a lower dose and a lower background, favouring tumour imaging with respect to the tumour/background ratio.

Comparison of the uptake of [123/125I]-2-iodo-D-tyrosine and [123/125I]-2-iodo-L-tyrosine in R1M rhabdomyosarcoma cells in vitro and in R1M tumor-bearing Wag/Rij rats in vivo.

INTRODUCTION: Recently, promising results concerning uptake in vivo in tumors of D-amino acids have been published. Therefore, we decided to evaluate the tumor uptake of the D-analogue of [(123)I]-2-iodo-L-tyrosine, a tracer recently introduced by our group into clinical trials. The uptake of 2-amino-3-(4-hydroxy-2-[(123/125)I]iodophenyl)-D-propanoic acid (2-iodo-D-tyrosine) was studied in vitro in LAT1-expressing R1M rat rhabdomyosarcoma cells and in vivo in R1M tumor-bearing Wag/Rij rats. METHODS: The uptake of [(125)I]-2-iodo-L-tyrosine and [(125)I]-2-iodo-D-tyrosine into R1M cells was determined in appropriate buffers, allowing the study of the involved transport systems. In vivo, the biodistribution in R1M-bearing rats of [(123)I]-2-iodo-L-tyrosine and [(123)I]-2-iodo-D-tyrosine was performed by both dynamic and static planar imaging with a gamma camera. RESULTS: In in vitro conditions, the uptake of both [(125)I]-2-iodo-L-tyrosine and [(125)I]-2-iodo-D-tyrosine in the HEPES buffer was 25% higher in the presence of Na(+) ions. In the absence of Na(+) ions, [(125)I]-2-iodo-D-tyrosine was taken up reversibly in the R1M cells, with an apparent accumulation, probably for the larger part by the LAT1 system. Dynamic planar imaging showed that the uptake in the tumors of [(123)I]-2-iodo-D-tyrosine was somewhat lower than that of [(123)I]-2-iodo-L-tyrosine. At 30 min postinjection, the mean differential uptake ratio values of the L- and D-enantiomers are 2.5+/-0.7 and 1.7+/-0.6, respectively. Although the uptake of the D-isomer is lower, probably due to a faster clearance from the blood, the tumor-background ratio is the same as that of the l-analogue. CONCLUSION: A large part (75%) of [(125)I]-2-iodo-D-tyrosine in vitro and [(123)I]-2-iodo-D-tyrosine in vivo is reversibly highly taken up in R1M tumor cells by Na(+)-independent LAT transport systems, more likely by the LAT1. The clearance from the blood of [(123)I]-2-iodo-D-tyrosine in the rats is faster than that of the L-analogue, resulting in a slightly lower tumor uptake but with the same tumor-background ratio.

Sigma receptor scintigraphy with N-[2-(1'-piperidinyl)ethyl]-3-(123)I-iodo-4-methoxybenzamide of patients with suspected primary breast cancer: first clinical results.

UNLABELLED: The aim of this study was to investigate the potential of a new iodobenzamide, N-[2-(1'-piperidinyl)ethyl]-3-(123)I-iodo-4-methoxybenzamide (P-(123)I-MBA), to visualize primary breast tumor in humans in vivo. Tumor accumulation of benzamides is based on a preferential binding to sigma receptors that are overexpressed on breast cancer cells. METHODS: P-(123)I-MBA (148-185 MBq) was administered to 12 patients with a mammographically suspicious breast mass. Two hours after administration, whole-body and spot images of the healthy and the diseased breast were obtained. RESULTS: A focal increased tracer accumulation was observed in 8 of 10 patients with histologically confirmed breast cancer (mean tumor-to-background ratio, 2.04). No uptake was seen in a case of lymphatic adenitis. CONCLUSION: This preliminary patient study shows that P-(123)I-MBA accumulates in most breast tumors in vivo. Future work should focus on the relationship between P-(123)I-MBA uptake and the proliferative activity of cells to anticipate use of this technique as a tool to noninvasively assess the degree of tumor proliferation.

Comparative biodistribution of iodinated amino acids in rats: selection of the optimal analog for oncologic imaging outside the brain.

UNLABELLED: 3-(123)I-Iodo-alpha-methyltyrosine ((123)I-3-IMT) is used for the detection of residual and recurrent brain tumors. The application of (123)I-3-IMT for the study of extracerebral malignancies is limited by its marked and rapid renal uptake. In this study, we compared the tumor uptake, biodistribution, and specificity of 5 structurally related iodinated amino acids with those of (123)I-3-IMT. The aim was to select the optimal analog for oncologic imaging outside the brain. METHODS: We studied 3-(123)I-iodotyrosine ((123)I-3-IT), 2-(123)I-iodotyrosine ((123)I-2-IT), (123)I-iodo-azatyrosine ((123)I-IAzaT), 2-(123)I-iodophenylalanine ((123)I-2-IPhe), and 4-(123)I-iodophenylalanine ((123)I-4-IPhe). Tumor uptake and renal uptake in sarcoma-bearing rats were measured by use of in vivo dynamic imaging. The differential uptake ratio (average counts per pixel of the region of interest divided by the average counts per pixel inside the total body) and rates of tracer accumulation (K(1) values) were calculated. Results were compared with the values obtained for (123)I-3-IMT in the same rat. Tracers that demonstrated high tumor uptake were labeled with (125)I and coinjected with (18)F-FDG in rats with turpentine-induced acute inflammation. After 30 min, the rats were sacrificed and dissected. Amino acid tracer uptake in organs and tissues was measured, and the increase in uptake in the inflamed muscle was expressed relative to the increase in (18)F-FDG uptake. RESULTS: Tumor uptake and K(1) values for (123)I-2-IT and (123)I-2-IPhe were comparable to those for (123)I-3-IMT. (123)I-4-IPhe showed high tumor uptake but a reduced K(1) value because of high blood-pool activity. (123)I-3-IT and (123)I-IAzaT did not accumulate markedly in tumor tissue. Renal accumulation of (123)I-2-IT, (123)I-2-IPhe, and (123)I-4-IPhe was at least 6 times lower than that of (123)I-3-IMT. (18)F-FDG uptake was markedly increased in areas of acute inflammation (215%). The increases for (125)I-3-IMT and (125)I-4-IPhe were 35.5% and 22.2%, respectively, of the increase for (18)F-FDG. Almost no increase was found for (125)I-2-IT (3.3%) and (125)I-2-IPhe (2.8%). CONCLUSION: (123)I-2-IT and (123)I-2-IPhe are promising tracers for oncologic imaging outside the brain. (123)I-2-IT has the advantage of an established kit for radiosynthesis.

Increased tumor uptake of 3-(123)I-Iodo-L-alpha-methyltyrosine after preloading with amino acids: an in vivo animal imaging study.

UNLABELLED: 3-(123)I-Iodo-L-alpha-methyltyrosine (3-IMT) is an amino acid analog used for tumor imaging. Specific accumulation is mediated mainly by the system L amino acid transport system. System L activity is known to increase when cells are loaded with amino acids. The aim of our study was to measure the effects of amino acid preload on (123)I-3-IMT tumor uptake and image contrast in a rat tumor model using in vivo dynamic imaging. METHODS: Rhabdomyosarcoma (R1M) tumor-bearing rats underwent 2 dynamic (123)I-3-IMT studies on separate days: 1 baseline study and 1 after intraperitoneal injection (0.25 mmol/kg) of a single amino acid (arginine, proline, glutamate, asparagine, tryptophan, or phenylalanine) administered 30 min before intravenous injection of 18.5 MBq (123)I-3-IMT. A (99m)Tc-labeled human serum albumin study was performed on each rat for the calculation of the blood-pool activity inside the tumor. Time-activity curves were generated for tumor, contralateral background region, kidney, heart, and total body. Tumor uptake was corrected for blood-pool and background activity. Image contrast was calculated as the ratio between tumor and background activity. The rate (K(1)) of tracer entering the tumor was obtained using Patlak analysis. A displacement study was performed on a separate group of rats, in which a high dose of phenylalanine was administered 40 min after (123)I-3-IMT injection. RESULTS: (123)I-3-IMT accumulation in tumor reached a plateau 10 min after injection. Tumor uptake on the baseline scans correlated well with tumor size (r = 0.92). After preloading, tumor uptake and contrast increased in all conditions: arginine, +26% and +26%; proline, +15% and +13%; glutamate, +14% and +9%; asparagine, +19% and +15%; tryptophan, +36% and 11%; phenylalanine, +22% and + 13%. K(1) values also increased. Administration of an afterload with phenylalanine induced a significant displacement of (123)I-3-IMT tumor accumulation. CONCLUSION: Prior amino acid administration increases (123)I-3-IMT tumor accumulation and image contrast. This effect can be explained by the increased antiporter activity of the amino acid transport system L in preloaded conditions. Our results indicate that the fasted state might not be the optimal metabolic condition to study tumor accumulation of L-transported tracers such as (123)I-3-IMT. Amino acid administration before (123)I-3-IMT injection could improve tumor uptake and image contrast.

SPECT and PET amino acid tracer influx via system L (h4F2hc-hLAT1) and its transstimulation.

UNLABELLED: System L amino acid transport is increased in various types of cancer. The tracer 123I-2-iodotyrosine (2IT), which is accumulated via system L, could thus serve to allow visualization of cancer in vivo. Here, we studied the transport of 125I-2IT by h4F2hc-hLAT1, the major transporter subserving system L in growing cells, using the Xenopus laevis oocyte expression system. We compared the apparent affinity of 125I-2IT with that of tyrosine, tested the influence of intracellular methionine availability on the influx rate of this substrate, and then compared the transport of 2IT with that of the other tracers-iodo-alpha-methyltyrosine (IMT), fluoroethyltyrosine (FET), and 2-fluorotyrosine (2FT)-by measuring their transstimulating effect on phenylalanine efflux. METHODS: Transport experiments were performed with Xenopus laevis oocytes expressing h4F2hc-hLAT1 (the functional transporter) and oocytes expressing only h4F2hc (negative control). The values obtained for the functional transporter were corrected for endogenous background transport by subtracting the values for the negative controls. RESULTS: The apparent affinity for 125I-2IT and 3H-tyrosine was 29.3 +/- 9.3 micromol/L and 21.2 +/- 4.2 micromol/L, respectively. The influx rate of 125I-2IT was, similarly to that of 3H-phenylalanine, transstimulated by a factor of > or =3 when the oocytes were preinjected with methionine or phenylalanine. The proportion of preinjected 3H-phenylalanine that effluxed within 90 s in the presence of an extracellular 2IT concentration of 0.1 mmol/L was 4.1% +/- 0.5%, compared with 3.3% +/- 0.4% for extracellular IMT, 1.3% +/- 0.3% for FET, 9.3% +/- 0.8% for 2FT, and 9.1% +/- 0.5% for phenylalanine. CONCLUSION: 2IT has a high affinity for h4F2hc-hLAT1, comparable to that of natural tyrosine, and its influx rate is transstimulated by intracellular amino acids. The 2IT influx rate is comparable to that of IMT but lower than that of phenylalanine. In contrast to FET, which is only poorly transported, 2FT displays a high influx rate equal to that of phenylalanine.

Dorsal prefrontal cortical serotonin 2A receptor binding indices are differentially related to individual scores on harm avoidance.

Although the serotonergic system has been implicated in healthy as well as in pathological emotional states, knowledge about its involvement in personality is limited. Earlier research on this topic suggests that post-synaptic 5-HT2A receptors could be involved in particular in frontal cortical areas. In drug-naïve healthy individuals, we examined the relationship between these 5-HT2A receptors and the temperament dimension harm avoidance (HA) using 123I-5-I-R91150 single photon emission computed tomography (SPECT). HA is a personality feature closely related to stress, anxiety and depression proneness, and it is thought to be mediated by the serotonergic system. We focused on the prefrontal cortices as these regions are frequently implicated in cognitive processes related to a variety of affective disorders. We found a positive relationship between dorsal prefrontal cortical (DPFC) 5-HT2A receptor binding indices (BI) and individual HA scores. Further, our results suggest that those individuals with a tendency to worry or to ruminate are particularly prone to display significantly higher 5-HT2A receptor BI in the left DPFC. Although we only examined psychologically healthy individuals, this relationship suggests a possible vulnerability for affective disorders.

Is treatment-resistance in unipolar melancholic depression characterized by decreased serotonin2A receptors in the dorsal prefrontal - anterior cingulate cortex?

OBJECTIVES: Quite a number of patients diagnosed with major depression are resistant to several well carried-out psychopharmacological interventions. It remains unclear as to how the serotonergic system is implicated in the phenomenon of treatment-resistance. METHODS: We examined the involvement of post-synaptic 5-HT(2A) receptors in the pathophysiology of treatment-resistance in unipolar melancholic major depression with (123)I-5-I-R91150 SPECT. 15 antidepressant-naïve (ADN) first-episode depressed patients, 15 antidepressant-free treatment-resistant depressed (TRD) patients and 15 never-depressed individuals, matched for age and gender were studied. RESULTS: Compared to ADN patients and healthy controls, TRD patients displayed significantly lower 5-HT(2A) receptor binding index (BI) in the dorsal regions of the prefrontal and the anterior cingulate cortex. No significant 5-HT(2A) receptor BI differences between ADN patients and controls were observed. CONCLUSIONS: At the cortical level, 5-HT(2A) receptor BI does not significantly differ in first-episode melancholic depressed patients compared to healthy controls. This observation might imply a limited short-term impact on the serotonergic system in first episode depression. Our results also suggest that when encountered with treatment-resistance, the 5-HT(2A) receptors in the DPFC-ACC axis are significantly down-regulated. However, whether this assumed underlying pathophysiological mechanism is due solely to abnormalities in the serotonergic system remains to be answered. This article is part of a Special Issue entitled 'Anxiety and Depression'.

Inhibition of firefly luciferase by general anesthetics: effect on in vitro and in vivo bioluminescence imaging.

UNLABELLED: Bioluminescence imaging is routinely performed in anesthetized mice. Often isoflurane anesthesia is used because of its ease of use and fast induction/recovery. However, general anesthetics have been described as important inhibitors of the luciferase enzyme reaction. AIM: To investigate frequently used mouse anesthetics for their direct effect on the luciferase reaction, both in vitro and in vivo. MATERIALS AND METHODS: isoflurane, sevoflurane, desflurane, ketamine, xylazine, medetomidine, pentobarbital and avertin were tested in vitro on luciferase-expressing intact cells, and for non-volatile anesthetics on intact cells and cell lysates. In vivo, isoflurane was compared to unanesthetized animals and different anesthetics. Differences in maximal photon emission and time-to-peak photon emission were analyzed. RESULTS: All volatile anesthetics showed a clear inhibitory effect on the luciferase activity of 50% at physiological concentrations. Avertin had a stronger inhibitory effect of 80%. For ketamine and xylazine, increased photon emission was observed in intact cells, but this was not present in cell lysate assays, and was most likely due to cell toxicity and increased cell membrane permeability. In vivo, the highest signal intensities were measured in unanesthetized mice and pentobarbital anesthetized mice, followed by avertin. Isoflurane and ketamine/medetomidine anesthetized mice showed the lowest photon emission (40% of unanesthetized), with significantly longer time-to-peak than unanesthetized, pentobarbital or avertin-anesthetized mice. We conclude that, although strong inhibitory effects of anesthetics are present in vitro, their effect on in vivo BLI quantification is mainly due to their hemodynamic effects on mice and only to a lesser extent due to the direct inhibitory effect.

Localization, mechanism and reduction of renal retention of technetium-99m labeled epidermal growth factor receptor-specific nanobody in mice.

BACKGROUND: Nanobodies are single-domain antigen binding fragments derived from functional heavy-chain antibodies elicited in Camelidae. They are powerful probes for radioimmunoimaging, but their renal uptake is relatively high. In this study we have evaluated the role of megalin on the renal uptake of anti-EGFR (99m)Tc-7C12 nanobody and the potency of gelofusine and/or lysine to reduce renal uptake of (99m)Tc-7C12. METHODS: First we compared the renal uptake of (99m)Tc-7C12 in megalin-deficient and megalin-wild-type mice using pinhole SPECT/microCT and ex vivo analysis. The effect of gelofusine and lysine administration on renal accumulation of (99m)Tc-7C12 was analyzed in CD-1 mice divided into lysine preload at 30 min before tracer injection (LysPreload), LysPreload + gelofusine coadministration (LysPreload + GeloCoad), lysine coadministration (LysCoad), gelofusine coadministration (GeloCoad) and LysCoad + GeloCoad. The combined effect of gelofusine and lysine on tumor uptake was tested in mice xenografts. RESULTS: Renal uptake of (99m)Tc-7C12 was 44.22 ± 3.46% lower in megalin-deficient compared with megalin-wild-type mice. In CD-1 mice, lysine preload had no effect on the renal retention whereas coinjection of lysine or gelofusine with the tracer resulted in 25.12 ± 2.99 and 36.22 ± 3.07% reduction, respectively. The combined effect of gelofusine and lysine was the most effective, namely a reduction of renal retention of 45.24 ± 2.09%. Gelofusine and lysine coadministration improved tumor uptake. CONCLUSION: Megalin contributes to the renal accumulation of (99m)Tc-7C12. Gelofusine and lysine coinjection with the tracer reduces the renal uptake while tumor uptake is improved. Although this methodology allows for optimization of imaging protocol using nanobodies, further improvements are needed before using these molecules for radionuclide therapy.

The impact of HF-rTMS treatment on serotonin(2A) 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.

Plasma protein binding of luciferase substrates influences sensitivity and accuracy of bioluminescence imaging.

INTRODUCTION: Potential confounding factors in bioluminescence imaging (BLI)-quantification need to be identified to improve its accuracy and repeatability. PURPOSE: The aim of this study was to study the degree to which plasma protein binding of BLI substrates influences signal intensity and quantification, both in vitro and in vivo. PROCEDURES: Protein binding was assessed using ultrafiltration and spectrophotometry. Effects of increasing serum protein concentrations were studied in vitro, in intact cells expressing Firefly or Renilla luciferase, and in vivo, in a doxorubicin-induced hypoalbuminemia mouse model. RESULTS: Increasing concentrations of serum proteins showed an important negative effect on BLI signal intensity, both for D-luciferin and coelenterazine-dependent reactions. This was due to a decrease in the free fraction of the substrate in the presence of serum proteins of up to 88%. In vivo, hypoalbuminemia was associated with higher BLI signal intensity, although this was only significant in animals with a major decrease in albumin (<2 g/dL). Important kinetic differences, including earlier peak luminescence and a more rapid decline in luminescence, were observed for coelenterazine-dependent BLI under hypoalbuminemic conditions. Changes in protein concentrations can significantly influence BLI quantification and its presence decreases sensitivity in vitro. In vivo, effects on signal intensity and kinetics are only noted at severe hypoalbuminemia and can be neglected in most experimental designs. These findings again emphasize the need for careful interpretation of BLI quantification data.