Marcaje de leucocitos con 99mTc-exametazima y policitemia vera: un hallazgo casual / Leucocyte labelling with 99mTc-exametazime and polycythemia vera: a casual finding

La clasificación oficial de la Organización Mundial de la Salud de los tumores de tejidos hematopoyéticos y linfoides de 2016 introduce el concepto de policitemia vera (PV) enmascarada y revela que esta entidad ha sido infradiagnosticada en el pasado. Se presenta el caso de un varón de 49 años, fumador, intervenido de fractura de tobillo hace más de 15 años, remitido para valorar un posible proceso infeccioso asociado. Al no producirse separación previa de los hematíes por sedimentación durante el procedimiento de marcaje de leucocitos con 99mTc-exametazima se revisaron las causas de disminución de la velocidad de sedimentación globular. Entre ellas destacan la poliglobulia y el hábito tabáquico, ambas presentes en el paciente. Se recomendó realización de estudio hematológico que concluyó con el diagnóstico de PV. Las indicaciones del especialista en Radiofarmacia permitieron diagnosticar un caso no identificado hasta entonces, pese a que el paciente presentaba síntomas desde hacía años

The official World Health Organization classification of hematopoietic and lymphoid tissue tumors introduces in 2016 the concept of masked polycythemia vera (PV) and reveals that this entity has been underdiagnosed in the past. We present the case of a 49-year-old man, smoker, operated on for ankle fracture more than 15 years ago, referred to evaluate a possible associated infectious process. As there was no previous separation of the red blood cells by sedimentation during the leukocyte labelling procedure with 99mTc-exametazima, the causes of decreased erythrocyte sedimentation rate were reviewed. These include polyglobulia and smoking, both present in the patient. A haematological study was advised, which concluded with the diagnosis of PV. The indications of the specialist in Radiopharmacy allowed diagnosing a case not identified until then, although the patient had had symptoms for years

Ultrasound-guided injection of the cranial tibial artery for stem cell administration in horses.

BACKGROUND: A technique for intra-arterial injection of mesenchymal stem cells (MSC) has been established for front limbs with the use of the median artery. This approach has been proposed for treatment of soft tissue injuries of the equine distal limb. A technique has not been validated yet for hindlimb injection. OBJECTIVES: To assess the feasibility of injection of the cranial tibial artery in horses, and to evaluate the distribution and persistence of MSC after injection. STUDY DESIGN: In vivo experiment. METHODS: In a first phase, the cranial tibial arteries of both hindlimbs of three research horses were catheterised with ultrasound guidance under general anaesthesia and injected with iodinated contrast. In the second phase, iodinated contrast was injected in three standing sedated horses with ultrasound guidance. In the final phase, 99m Technetium-HMPAO labelled allogenic bone marrow derived equine MSC were injected under standing sedation with the same technique in three other horses. Scintigraphy was used to assess MSC distribution and persistence for 24 h. Ultrasound was performed 24 h after injection to assess vessel impairment. RESULTS: Arterial injection was achieved in all 18 limbs without any significant complications. Mild partial periarterial injection was observed in four limbs. Scintigraphic images demonstrated diffuse MSC distribution from the tarsal area to the foot. Persistence decreased over time but signal was still present at 24 h. MAIN LIMITATIONS: Limited retention of the radiolabel in the MSC. CONCLUSIONS: Ultrasound-guided injection of the cranial tibial artery can be performed both under general anaesthesia and standing sedation in horses. This technique could be used for MSC treatment of equine proximal suspensory desmopathy or other injuries in the distal hindlimb.

Protection by Inhaled Hydrogen Therapy in a Rat Model of Acute Lung Injury can be Tracked in vivo Using Molecular Imaging.

Inhaled hydrogen gas (H2) provides protection in rat models of human acute lung injury (ALI). We previously reported that biomarker imaging can detect oxidative stress and endothelial cell death in vivo in a rat model of ALI. Our objective was to evaluate the ability of Tc-hexamethylpropyleneamineoxime (HMPAO) and Tc-duramycin to track the effectiveness of H2 therapy in vivo in the hyperoxia rat model of ALI. Rats were exposed to room air (normoxia), 98% O2 + 2% N2 (hyperoxia) or 98% O2 + 2% H2 (hyperoxia+H2) for up to 60 h. In vivo scintigraphy images were acquired following injection of Tc-HMPAO or Tc-duramycin. For hyperoxia rats, Tc-HMPAO and Tc-duramycin lung uptake increased in a time-dependent manner, reaching a maximum increase of 270% and 150% at 60 h, respectively. These increases were reduced to 120% and 70%, respectively, in hyperoxia+H2 rats. Hyperoxia exposure increased glutathione content in lung homogenate (36%) more than hyperoxia+H2 (21%), consistent with increases measured in Tc-HMPAO lung uptake. In 60-h hyperoxia rats, pleural effusion, which was undetectable in normoxia rats, averaged 9.3 gram/rat, and lung tissue 3-nitrotyrosine expression increased by 790%. Increases were reduced by 69% and 59%, respectively, in 60-h hyperoxia+H2 rats. This study detects and tracks the anti-oxidant and anti-apoptotic properties of H2 therapy in vivo after as early as 24 h of hyperoxia exposure. The results suggest the potential utility of these SPECT biomarkers for in vivo assessment of key cellular pathways in the pathogenesis of ALI and for monitoring responses to therapies.

99mTc-labeled White Blood Cell Scan as a Guide to Open Biopsy in the Management of Hip and Knee Prosthesis Infection: Preliminary Results.

OBJECTIVE: The aim of the present prospective study was to evaluate the usefulness of labeled leukocyte scan as a guide to open biopsy for the management of hip and knee prosthesis infection in patients without loosening of orthopedic device. METHODS: Twenty-six patients with suspected hip (24) and knee (2) prosthesis infection underwent routine analysis of blood, plain radiography and 99mTc-HMPAO labelled leukocyte scan (WBCS). On these basis, patients were subdivided in the following groups: bone infection without loosening (n°=14), septic loosening (n°=8), superficial infection (n°=2), no infection (n°=2). Patients with septic loosening underwent empirical antibiotic therapy in order to avoid two-stage reimplantation. When the medical treatment was effective patients were submitted to one-stage operation. Patients without loosening of prosthesis but positive WBCS results underwent open biopsy: bone samples and periprosthetic tissues were taken from the regions showing pathological leukocyte uptake at the scan. Samples were submitted to microbiological examination and antibiotic treatments were undertaken in cases of bacterial growth. A 24-months clinical and instrumental follow-up was carried out in all patients. RESULTS: WBCS showed 22 patients affected by bone infection, 2 by superficial infection and 2 not infected. Height out of the 22 patients affected by deep infection had a septic loosening. In these cases, the medical treatment was inadequate in 6 patients and effective in 2. Fourteen patients with bone infection without loosening were submitted to open biopsy: in 9 cases a complete remission of the disease was found. Two patients, without infection, underwent single-stage surgery for mechanical problems. Superficial infection was assessed and successfully treated in 2 patients. CONCLUSION: The obtained results indicate that a multidisciplinary approach to infection of orthopedic prostheses, characterized by the combined use of open biopsy, WBC, and microbiological examination, produced positive outcome in 9 out of 14 patients.

[Determination of the efficiency of single-photon emission computed tomography with 99mTc-HMPAO-labelled leukocytes in the diagnosis of myocarditis: Comparison of scintigraphic and histological data].

OBJECTIVE: to determine the efficiency of single-photon emission computed tomography (SPECT) with 99mTc-HMPAO- labelled leukocytes in diagnosing myocarditis, by comparing scintigraphic and histological data. MATERIAL AND METHODS: The investigation enrolled 35 patients with suspected myocarditis, who were planned to undergo coronaroventriculography or intervention with endomyocardial biopsy. Prior to endomyocardial biopsy, all the patients underwent myocardial scintigraphy using 99mTC-exametazime-labelled leukocytes. The results of myocardial scintigraphic and histological examinations were compared. RESULTS: Abnormal myocardial 99mTc-HMPAO-labelled leukocyte accumulation was detected in 7 (20%) examinees. Myocarditis was histologically verified in 9 (25.7%) persons. Our findings showed that the sensitivity of 99mTc-HMPAO-labelled leukocyte SPECT in diagnosing myocardial inflammatory changes was 62%; its specificity and diagnostic accuracy were 92% and 85%, respectively. Conclusion. 99mTc- HMPAO-labelled leukocyte scintigraphy is today one of a few procedures for the primary noninvasive diagnosis of myocardial inflammation. However, in view of its sufficiently low sensitivity and laboriousness and the sigh cost of consumables, the technique is irrationally used in routine clinical practice.

Immunologic Network and Response to Intramyocardial CD34+ Stem Cell Therapy in Patients With Dilated Cardiomyopathy.

BACKGROUND: Although stem cell therapy (SCT) is emerging as a potential treatment for patients with dilated cardiomyopathy (DCM), clinical response remains variable. Our objective was to determine whether baseline differences in circulating immunologic and nonimmunologic biomarkers may help to identify patients more likely to respond to intramyocardial injection of CD34(+)-based SCT. METHODS AND RESULTS: We enrolled from January 3, 2011 to March 5, 2012 37 patients with longstanding DCM (left ventricular ejection fraction [LVEF] <40%, New York Heart Association functional class III) who underwent peripheral CD34(+) stem cell mobilization with granulocyte colony-stimulating factor (G-CSF) and collection by means of apheresis. CD34(+) cells were labeled with (99m)Tc-hexamethylpropyleneamine oxime to allow assessment of stem cell retention at 18 hours. Response to SCT was predefined as an increase in LVEF of ≥5% at 3 months. The majority (84%) of patients were male with an overall mean LVEF of 27 ± 7% and a median N-terminal pro-B-type natriuretic peptide (NT-proBNP) level of 2,774 pg/mL. Nineteen patients (51%) were responders to SCT. There was no significant difference between responders and nonresponders regarding to age, sex, baseline LVEF, NT-proBNP levels, or 6-minute walking distance. With the use of a partial least squares (PLS) predictive model, we identified 9 baseline factors that were associated with both stem cell response and stem cell retention (mechanistic validation). Among the baseline factors positively associated with both clinical response and stem cell retention were G-CSF, SDF-1, LIF, MCP-1, and MCP-3. Among baseline factors negatively associated with both clinical response and retention were IL-12p70, FASL, ICAM-1, and GGT. A decrease in G-CSF at 3-month follow-up was also observed in responders compared with nonresponders (P = .02). CONCLUSIONS: If further validated, baseline immunologic and nonimmunologic biomarkers may help to identify patients with DCM who are more likely to respond to CD34(+)-based SCT.

[Possibilities of a software-based hybrid single photon emission computed tomography/magnetic resonance imaging in the diagnosis of complicated diabetic foot syndrome].

OBJECTIVE: To give the results of a software-based hybrid single photon emission computed tomography/magnetic resonance imaging (SPECT/MRI) in detecting osteomyelitis (OM) in patients with diabetic foot syndrome (DFS). MATERIAL AND METHODS: Seventy-six patients (35 men and 41 women) (mean age, 59.4 +/- 7.1 years) with type 1 and 2 diabetes mellitus and suspected OM were examined. The investigation enrolled patients with neuropathic (n = 25), ischemic (n = 13), and mixed (n = 38) DFS. All the patients underwent (99m)Tc-HMPAO/ (99m)Tc-technefit labeled leukocyte scintigraphy; magnetic resonance imaging was performed in 30 patients. The results were combined using RView 9.06 software (Colin Studholme). RESULTS: Labeled leukocyte SPECT to Diagnose OM yielded 255 true positive (TP), 38 true negative (TN), 12 false negative (FP), and 1 false negative (FN) results. The accuracy of the technique was 82.9%. The FP results were due to the low resolution of the technique and to the small sizes of the object under study. One FN result was detected in a patient with ischemic DFS because of reduced blood flow. MRI to identify OM in patients with DFS provided 20 TP, 16 TN, 4 FP, and 2 FN results. Its diagnostic accuracy was 85.7%. The relative low specificity of MRI was associated with the presence of FP results due to the complexity of differential diagnosis of bone marrow edema and inflammatory infiltration. Assessing 42 hybrid SPECT/MR-images revealed 21 TP, 17 TN, 3 FP, and I FN results. The diagnostic accuracy was equal to 95.9%. CONCLUSION: Thus, comparing MRI (90.9% sensitivity and 80.0% specificity), labeled leukocyte scintigraphy (96.2% sensitivity and 76.0% specificity), and hybrid SPECT/MRI (95.5% sensitivity and 85.0% specificity) showed the high diagnostic efficiency of the latter.

Scintigraphic tracking of mesenchymal stem cells after intravenous regional limb perfusion and subcutaneous administration in the standing horse.

OBJECTIVE: To assess distribution, uptake, and persistence of radiolabeled mesenchymal stem cells (MSC) using scintigraphy after intravenous regional limb perfusion (RLP) and subcutaneous injections in standing, sedated horses. STUDY DESIGN: Experimental study. ANIMALS: Horses (n = 12). METHODS: Six horses had RLP performed through the cephalic vein on 1 limb and subcutaneous injection in the metacarpal area in the opposite limb. The other 6 horses had RLP performed through the lateral palmar digital vein and subcutaneous injection in the coronary band. A pneumatic tourniquet was used for the RLP. MSC were labeled with technetium-HMPAO. Scintigraphic images were obtained at the time of injection, 1, 6, and 24 hours later. Results of RLP were compared with results from previous studies where similar injections were performed in anesthetized horses. RESULTS: Both RLP techniques led to greater variability, lower uptake, lower persistence, and poorer distribution when compared to results previously reported for horses under general anesthesia. The subcutaneous injections in the metacarpal area and coronary band resulted in MSC loss to the general circulation but no evidence of local migration. CONCLUSION: Due to partial or complete failure of the tourniquet, RLP performed in the standing horse as described is less efficient than performed under general anesthesia. Further work is needed to optimize the use of tourniquets to perform RLP for MSC administration in standing patients. The subcutaneous injections did not result in local migration in these normal horses.

In vitro assessment of cytotoxicity and labeling efficiency of 99mTc-HMPAO with stromal vascular fraction of adipose tissue.

INTRODUCTION: Noninvasive radionuclide imaging of cells using technetium99m-hexamethylpropyleneamine oxime ((99m)Tc-HMPAO) is a potential diagnostic tool for several applications. Herein we aimed to evaluate the labeling efficiency and cellular toxicity of (99m)Tc-HMPAO with Stromal Vascular Fraction (SVF) of adipose tissue to develop a process tool for theranostic purposes, in particular imaging cardiac stem cell therapy. METHODS: Ten million cells of SVF were labeled with (99m)Tc-HMPAO complex and excess radiolabel was cleared off through washing in PBS. The labeling efficiency of (99m)Tc-HMPAO was detected in labeled cells and their subsequent supernatant wash using isotope dose calibrator and gamma camera. The cytotoxicity was assessed for the comparative reactive oxygen species (ROS) by H2DCFDDA, apoptotic events by annexin-V and TUNEL assay and mitochondrial potential by JC-1. RESULTS: An encouraging labeling efficiency of 33% was observed with (99m)Tc-HMPAO complex. The radionuclide labeling of SVF demonstrated significant safety profile as evaluated by apoptotic assays. CONCLUSION: (99m)Tc-HMPAO labeling efficiency of 33% of total SV fraction would produce sufficient radioactive signals that would enable for in vivo tracking of cells by SPECT-CT. The radionuclide did not demonstrate any significant impact on the structural or functional organization of the labeled cells. Our study indicates that SVF can be safely labeled with (99m)Tc-HMPAO without adverse cytotoxic events and for its potential role in imaging cardiac stem cell therapy.

Scintigraphic comparison of intra-arterial injection and distal intravenous regional limb perfusion for administration of mesenchymal stem cells to the equine foot.

REASONS FOR PERFORMING STUDY: Intra-arterial (i.a.) and intravenous (i.v.) regional limb perfusions (RLP) through the median artery and cephalic vein, respectively, have been previously investigated for administration of mesenchymal stem cells (MSCs) to the equine distal limb. Limitations due to thrombosis of the arteries after i.a. RLP and poor distribution of MSCs to the foot with i.v. RLP were observed. These techniques need to be modified for clinical use. OBJECTIVES: Evaluate the distribution, uptake and persistence of radiolabelled MSCs after i.a. injection through the median artery without a tourniquet and after i.v. RLP through the lateral palmar digital vein. STUDY DESIGN: In vivo experimental study. METHODS: (99m) Tc-HMPAO-labelled MSCs were injected through the median artery of one limb and the lateral palmar digital vein of the other limb of 6 horses under general anaesthesia. No tourniquet was used for the i.a. injection. A pneumatic tourniquet was placed on the metacarpus for i.v. injection. Scintigraphic images were obtained up to 24 h after injection. RESULTS: Intra-arterial injection resulted in MSCs retention within the limb despite the absence of a tourniquet and no thrombosis was observed. Both i.a. injection and i.v. RLP led to distribution of MSCs to the foot. The i.a. injection resulted in a more homogeneous distribution. The MSC uptake was higher with i.v. RLP at the initial timepoints, but no significant difference was present at 24 h. CONCLUSIONS: Both i.a. injection through the median artery without a tourniquet and i.v. RLP performed through the lateral palmar digital vein under general anaesthesia are safe and reliable methods for administration of MSCs to the equine foot. The i.a. technique is preferred owing to the better distribution, but is technically more challenging. The feasibility of performing these techniques on standing horses remains to be investigated.

Distribution and persistence of technetium-99 hexamethyl propylene amine oxime-labelled bone marrow-derived mesenchymal stem cells in experimentally induced tendon lesions after intratendinous injection and regional perfusion of the equine distal limb.

REASONS FOR PERFORMING STUDY: Intralesional (i.l.) injection is currently the most commonly used technique for stem cell therapy in equine tendon injury. A comparison of different techniques of injection of mesenchymal stem cells for the treatment of tendon lesions is required. OBJECTIVES: We hypothesised that vascular perfusion of the equine distal limb with mesenchymal stem cells (MSCs) would result in preferential distribution of MSCs to acute tendon injuries. STUDY DESIGN: In vivo experimental study. METHODS: Lesions were surgically induced in forelimb superficial digital flexor tendons of 8 horses. Three or 10 days after lesion induction, technetium-99 hexamethyl propylene amine oxime-labelled MSCs were injected via i.v. or intra-arterial (i.a.) regional limb perfusion (RLP) at the level of the distal antebrachium and compared to i.l. injection. Mesenchymal stem cell persistence and distribution within the forelimb and tendon lesions was assessed with scintigraphy for 24 h. RESULTS: Lesion uptake was higher with i.l. injection than with RLP, but MSC persistence decreased similarly over time in all 3 techniques. Intra-arterial RLP resulted in a better distribution of MSCs and a higher uptake at the lesion site than i.v. RLP. Limbs perfused i.a. on Day 10 showed greater accumulation of MSCs in the lesion than limbs perfused on Day 3. Arterial thrombosis occurred in 50% of the i.v. RLP limbs and in 100% of the i.a. RLP limbs, which led to clinical complications in one horse. CONCLUSIONS AND POTENTIAL RELEVANCE: Compared with i.l. injection, RLP results in lower uptake but similar persistence of MSCs at the site of tendon lesions. A time dependent accumulation of MSCs was identified with i.a. RLP. The i.a. RLP appears more advantageous than the i.v. RLP in terms of distribution and uptake. However, the described i.a. technique produced arterial thrombosis and thus cannot currently be recommended for clinical use.

Noninvasive MRI measurement of CBF: evaluating an arterial spin labelling sequence with 99mTc-HMPAO CBF autoradiography in a rat stroke model.

Arterial spin labelling (ASL) is increasingly available for noninvasive cerebral blood flow (CBF) measurement in stroke research. Here, a pseudo-continuous ASL technique (pCASL) was evaluated against (99m)Tc-D, L-hexamethylpropyleneamine oxime ((99m)Tc-HMPAO) autoradiography in a rat stroke model. The (99m)Tc-HMPAO was injected (intravenously, 225 MBq) during pCASL acquisition. The pCASL and (99m)Tc-HMPAO autoradiography CBF measures, relative to the contralateral hemisphere, were in good agreement across the spectrum of flow values in normal and ischemic tissues. The pCASL-derived quantitative regional CBF values (contralateral: 157 to 177 mL/100 g per minute; ipsilateral: 9 to 104 mL/100 g per minute) were consistent with the literature values. The data show the potential utility of pCASL for CBF assessment in a rat stroke model.

Scintigraphic evaluation of intra-arterial and intravenous regional limb perfusion of allogeneic bone marrow-derived mesenchymal stem cells in the normal equine distal limb using (99m) Tc-HMPAO.

REASONS FOR PERFORMING STUDY: Mesenchymal stem cells (MSCs) are commonly injected intralesionally for treatment of soft tissue injuries in the horse. Alternative routes of administration would be beneficial for treatment of lesions that cannot be accessed directly or to limit needle-induced iatrogenic damage to the surrounding tissue. OBJECTIVES: The purpose of our study was to evaluate MSC distribution after intra-arterial (IA) and intravenous (IV) regional limb perfusions (RLP) using scintigraphy. We hypothesised that MSCs would persist in the distal limb after tourniquet removal and that both techniques would lead to diffuse MSC distribution. METHODS: Six horses were used in the study. MSCs were labelled with hexamethyl propylene amine oxime (HMPAO) and technetium-99m. RLP was performed through the median artery of one forelimb and the cephalic vein of the opposite limb under general anaesthesia. The tourniquet was left in place for 45 min. Scintigraphic images were obtained at 0, 45, 75 min, 6 h and 24 h post injection. RESULTS: Distribution of labelled MSCs through the entire distal limb was achieved with all 6 IA RLP, but 3 out of 6 IV RLP showed poor or absent uptake distal to the metacarpus. Mesenchymal stem cell persistence was 39% (30-60%) and 28% (14-50%) (median [minimum-maximum]) at 6 h for IA and IV RLP, respectively. Severe arterial thrombosis occurred in one horse after IA RLP. CONCLUSIONS: Both IA and IV RLP of the distal limb result in MSC persistence in perfused tissues. The IA perfusion resulted in more reliable cell distribution to the pastern and foot area. POTENTIAL RELEVANCE: Regional limb perfusion of MSCs might be used in cases where intralesional injection is not possible or in order to avoid iatrogenic needle damage. Further work is needed to assess the safety of IA RLP before its clinical use.

Applicability of a high-resolution small semiconductor gamma camera to small animal imaging.

OBJECTIVE: Recently, small semiconductor gamma cameras (SSGCs) with high resolution and sensitivity, which are much more convenient to use as compared with SPECT and PET, have been developed for mapping the sentinel lymph node. The high resolution and sensitivity of the SSGCs may make them useful for small animal imaging. Therefore, we assessed the applicability of the SSGC to small animal imaging using a rat model of focal cerebral ischaemia. METHODS: The right middle cerebral artery (MCA) of anaesthetized rats was occluded intraluminally with a nylon monofilament. Twenty-four hours after the occlusion, 99mTc-HMPAO (3.7 MBq) was injected and a static acquisition (5 min) was performed using the SSGC. Regions of interest (ROIs) were set on each hemisphere of the horizontal brain images. After the acquisition, the brains were removed and the radioactivity in each hemisphere was measured using an NaI scintillation counter. RESULTS: Reduced CBF in the right MCA territory was clearly visualized with the SSGC in vivo. The radioactivity in the ROIs determined by the SSGC was significantly correlated with that determined by the ex vivo counting method (P<0.001, R2=0.74). Furthermore, in both of the in-vivo imaging and ex-vivo counting methods, the right to left count ratio (R/L ratio) was significantly lower in the MCA-occluded rats than that in normal rats (MCA-occluded rats: 0.77+/-0.08, normal rats: 1.01+/-0.07, P<0.005). CONCLUSIONS: The SSGC clearly visualized and quantitatively detected the reduced CBF in MCA-occluded rats. Furthermore, these high resolution and sensitivity of SSGC can avoid the disadvantage of small animal imaging with PET and SPECT, such as a large mass injected tracer and the exposure of investigators to radiation. Thus, the high resolution and sensitivity of the SSGC make it useful for small animal imaging.

99mTc-HMPAO labelling inhibits cell motility and cell proliferation and induces apoptosis of NC-NC cells.

(99m)Tc-hexamethyl-propylenamine-oxime ((99m)Tc-HMPAO)-labelled leukocytes have been used in standard diagnostic procedures for the detection of infection and inflammation. Although some investigators have already pointed out that labelling of leukocytes with (99m)Tc-HMPAO has detrimental effects on the cells, still very little is known regarding the effects of ionizing radiation on lymphocyte function. The effects of (99m)Tc-HMPAO-labelling on lymphocyte adhesion, proliferation, mitotic index, migration and apoptosis were evaluated. The lymphoblastoid cell line NC-NC was used as the lymphocyte population. (99m)Tc-HMPAO-labelling decreased cell adhesion, proliferation, mitotic index and motility, whereas it induced apoptosis and cell-cycle arrest. The rate of decrease in cell proliferation was up to 70% (P<0.001) by day 4 after labelling. (99m)Tc-HMPAO-labelling led a 35% decrease (P<0.001) in adhesion ability of the cells on fibronectin at 16h. Using the Boyden chamber motility assay, it was shown that both spontaneous and monocyte chemotactic protein (MCP-1)-induced lymphocyte motility were strongly reduced by (99m)Tc-HMPAO-labelling. The decrease in motility was approximately five-fold (P<0.05). In addition, a 12-fold increase (P<0.05) was observed in apoptosis of the (99m)Tc-HMPAO-treated cells compared with control cells. Besides, it was shown that cell-cycle arrest was induced starting from the 3rd day after treatment with (99m)Tc-HMPAO. Our observations indicate that (99m)Tc-HMPAO-labelling has damaging effects on lymphocyte function including cell adhesion, proliferation, mitotic index, motility and cell cycle under in vitro conditions.

Effect of linearization correction on statistical parametric mapping (SPM): a 99mTc-HMPAO brain perfusion SPECT study in mild Alzheimer's disease.

OBJECTIVE: Statistical parametric mapping (SPM) was employed to investigate the regional decline in cerebral blood flow (rCBF) as measured by 99mTc-hexamethyl propylene amine oxime (HMPAO) single photon emission computed tomography (SPECT) in mild Alzheimer's disease (AD). However, the role of the post reconstruction image processing on the interpretation of SPM, which detects rCBF pattern, has not been precisely studied. We performed 99mTc-HMPAO SPECT in mild AD patients and analyzed the effect of linearization correction for washout of the tracer on the detectability of abnormal perfusion. METHODS: Eleven mild AD (NINCDS-ADRDA, male/female, 5/6; mean+/-SD age, 70.6+/-6.2 years; mean+/-SD mini-mental state examination score, 23.9+/-3.41; clinical dementia rating score, 1) and eleven normal control subjects (male/female, 4/7; mean+/-SD age, 66.8+/-8.4 years) were enrolled in this study. 99mTc-HMPAO SPECT was performed with a four-head rotating gamma camera. We employed linearization uncorrected (LU) and linearization corrected (LC) images for the patients and controls. The pattern of hypoperfusion in mild AD on LU and LC images was detected by SPM99 applying the same image standardization and analytical parameters. A statistical inter image-group analysis (LU vs. LC) was also performed. RESULTS: Clear differences were observed between the interpretation of SPM with LU and LC images. Significant hypoperfusion in mild AD was found on the LU images in the left posterior cingulate gyrus, right precuneus, left hippocampus, left uncus, and left superior temporal gyrus (cluster level, corrected p < 0.005). With the LC images, significant hypoperfusion in AD was found only in the bilateral posterior cingulate gyrus and left precuneus (cluster level, corrected p < 0.005). A pattern of greater rCBF distribution at the high flow cortices and low flow cortices was observed on LC and LU images, respectively, in the case of both controls and mild AD patients. CONCLUSION: Hippocampal hypoperfusion could be detected by means of SPM in the LU images but not in the LC images. The results of SPM may vary in 99mTc-HMPAO SPECT with or without linearization correction, which should be carefully evaluated when interpreting the pattern of rCBF changes in mild Alzheimer's disease.

Cerebral activation during withholding urine with full bladder in healthy men using 99mTc-HMPAO SPECT.

UNLABELLED: The aim of this study was to identify the brain areas that control urinary continence in healthy men by 99mTc-hexamethylpropyleneamine oxime (99mTc-HMPAO) SPECT. METHODS: SPECT scans were performed on 15 right-handed healthy male volunteers, 24-45 y old. Each subject was scanned twice without movement in a supine position, and 444 and 555 MBq 99mTc-HMPAO were separately injected intravenously during the following 2 conditions: resting state with an empty bladder and urine-withholding state with a full bladder. The final image during urine withholding was obtained by subtracting the first scan data from the second scan data. The images were analyzed by statistical image analysis software and displayed on Z-score images at a significance threshold of P < 0.05 with correction for multiple comparisons. RESULTS: In the urine-withholding state, as compared with resting, there was a significant increase in tracer activity in the bilateral inferior frontal gyri and the right superior and the middle temporal gyri. Among the regions, the right inferior frontal gyrus was distinctly prominent. When the threshold value was decreased to P < 0.005 without correction, there was a vast network of cortical and subcortical regions involved during urine withholding. CONCLUSION: Our results suggest that the right inferior frontal gyrus plays an important role in brain control of urinary continence. This study also suggests that brain control of continence can be confirmed by statistical image analysis software using SPECT.

The contribution of statistical parametric mapping in the assessment of precuneal and medial temporal lobe perfusion by 99mTc-HMPAO SPECT in mild Alzheimer's and Lewy body dementia.

AIM: To assess the role of 99mTc-hexamethylpropyleneamine oxime single-photon emission computed tomography (99mTc-HMPAO SPECT) imaging of the precuneus and medial temporal lobe in the individual patient with mild Alzheimer's disease and dementia with Lewy bodies (DLB) using statistical parametric mapping and visual image interpretation. METHODS: Thirty-four patients with mild late-onset Alzheimer's disease, 20 patients with early-onset Alzheimer's disease, 15 patients with DLB and 31 healthy controls were studied. All patients fulfilled appropriate clinical criteria; the DLB patients also had evidence of dopaminergic presynaptic terminal loss on 123I-N-omega-fluoropropyl-2beta-carbomethoxy-3beta-(4-iodophenyl)-tropane imaging. 99mTc-HMPAO SPECT brain scans were acquired on a multidetector gamma camera and images were assessed separately by visual interpretation and with SPM99. RESULTS: Statistical parametric maps were significantly more accurate than visual image interpretation in all disease categories. In patients with mild late-onset Alzheimer's disease, statistical parametric mapping demonstrated significant hypoperfusion to the precuneus in 59% and to the medial temporal lobe in 53%. Seventy-six per cent of these patients had a defect in either location. No controls had precuneal or medial temporal lobe hypoperfusion (specificity, 100%). Statistical parametric mapping also demonstrated 73% of patients with DLB to have precuneal abnormalities, but only 6% had medial temporal lobe involvement. CONCLUSION: These findings illustrate the capability of statistical parametric mapping to demonstrate reliable abnormalities in the majority, but not all, patients with either mild Alzheimer's disease or DLB. Precuneal hypoperfusion is not specific to Alzheimer's disease and is equally likely to be found in DLB. In this study, medial temporal hypoperfusion was significantly more common in Alzheimer's disease than in DLB. Statistical parametric maps appear to be considerably more reliable than simple visual interpretation of 99mTc-HMPAO images for these regions.