Biodistribution of bone marrow mononuclear cells in chronic chagasic cardiomyopathy after intracoronary injection.

BACKGROUND: Animal and human clinical studies have indicated that bone marrow (BM) mononuclear cell (MNC) therapy for Chagasic Cardiomyopathy (ChC) is feasible, safe and potentially efficacious. Nevertheless, little is known about the retention of these cells after intracoronary (IC) infusion. METHODS: Our study investigated the homing of technetium-99m ((99m)Tc) labeled BM MNCs and compared it to thallium-201 ((201)Tl) myocardial perfusion images using the standard 17-segment model. Six patients with congestive heart failure of chagasic etiology were included. RESULTS: Scintigraphic images revealed an uptake of 5.4%±1.7, 4.3%±1.5 and 2.3%±0.6 of the total infused radioactivity in the heart after 1, 3 and 24h, respectively. The remaining activity was distributed mainly to the liver and spleen. Of 102 segments analyzed, homing took place in 36%. Segments with perfusion had greater homing (58.6%) than those with decreased or no perfusion (6.8%), p<0.0001. There was no correlation between the number of injected cells and the number of segments with homing for each patient (r=-0.172, p=0.774). CONCLUSIONS: These results indicate that (99m)Tc-BM MNCs delivered by IC injection homed to the chagasic myocardium. However, cell biodistribution was heterogeneous and limited, being strongly associated with the myocardial perfusion pattern at rest. These initial data suggest that the IC route may present limitations in chagasic patients and that alternative routes of cell administration may be necessary.

Assessment of intra-arterial injected autologous bone marrow mononuclear cell distribution by radioactive labeling in acute ischemic stroke.

OBJECTIVE: To evaluate the feasibility of monitoring the autologous mononuclear bone marrow (ABMMN) cells implanted into the brain after acute ischemic stroke by the technique of labeling with Tc-99m-HMPAO. CASE REPORT: A 37-year-old man presented with aphasia, right-side hypoesthesia, and right homonymous hemianopsia after an acute ischemic stroke of the left middle cerebral artery. He was included in an autologous bone marrow mononuclear cell-based therapy research protocol about the safety of intra-arterial autologous bone marrow mononuclear cell transplantation for acute ischemic stroke. Nine days after the stroke he received 3.0 x 10(7) ABMMN cells delivered into the left cerebral middle artery via a balloon catheter. Approximately 1% of these cells were labeled with 150 MBq (4 mCi) Tc-99m by incubation with hexamethylpropylene amine oxime (HMPAO). RESULTS: Brain perfusion images with Tc-99m ECD demonstrated hypoperfusion in the left temporal and parietal regions. The perfusion brain images were compared with tomographic views of the brain obtained 8 hours after ABMMN-labeled cell delivery, revealing intense accumulation of the ABMMN-labeled cells in the ipsilateral hemisphere. A whole-body scan was done and showed left brain, liver, and spleen uptake. CONCLUSIONS: Our results showed that Tc-99m HMPAO can be used to label ABMMN cells for in vivo cell visualization, and that brain SPECT imaging with labeled ABMMN cells is a feasible noninvasive method for studying the fate of transplanted cells in vivo. Additionally, our findings demonstrate the localization of these intra-arterially injected cells.

Use of 99mTc-mononuclear leukocyte scintigraphy in nosocomial fever.

PURPOSE: To determine the overall diagnostic accuracy of mononuclear leukocyte- 99mTc scintigraphy in the routine detection of infectious lesions and fever of unknown origin (FUO) in inpatients. MATERIAL AND METHODS: The use of mononuclear leukocyte 99mTc scintigraphy is presented in 87 patients who fulfilled the Durack and Street diagnostic criteria of nosocomial FUO; 66 patients were suspected of having infectious lesions (myocarditis, endocarditis, infected catheters, diabetic foot, and osteomyelitis) and 21 patients presented with unknown causes of FUO. Scans were carried out 1, 3, and 24 h after injection of labeled leukocytes. RESULTS: In three cases (3/27) where scintigraphs were negative, biopsies were positive. There were two (2/87) false-positive scintigrams. We found a 95.8% sensitivity and 92.3% specificity. PPV was 93.8%, PPN 94.7%, and accuracy 94.2%. CONCLUSION: Mononuclear leukocyte 99mTc scintigraphy showed high sensitivity, specificity, positive and negative predictive values in patients with nosocomial FUO. These results suggest an important role for nuclear medicine in the management of patients with infection/inflammation.

Oxidative DNA damage in circulating mononuclear blood cells after ingestion of blackcurrant juice or anthocyanin-rich drink.

Berry extracts possess antioxidant activity in cell free systems, whereas cell culture and animal experimental systems have produced mixed outcomes. Our aim was to investigate the effects of blackcurrant juice and specifically blackcurrant anthocyanins on the steady state level of oxidative DNA damage in mononuclear blood cells (MNBC) of humans, determined as strand breaks (SB) as well as endonuclease III (Endo III) and formamidopyrimidine DNA glycosylase (Fpg) sensitive sites by the comet assay. Fifty-seven healthy humans completed a 3-week controlled parallel intervention study with three groups randomized to supplementation with blackcurrant juice, anthocyanin drink, or a control drink. The daily doses ranged from 475 to 1000ml/d according to body weight (mean anthocyanin intakes in blackcurrant juice and anthocyanin drink groups were 397 and 365g/d, respectively) and they were ingested during three daily meals while all volunteers were on the same strictly controlled low-flavonoid diet. Fasting venous blood samples were obtained at baseline and after 3-week of supplementation. The baseline level of oxidative DNA damage was low (e.g. less than 200 Fpg lesions per diploid cell). Fpg sensitive sites increased during the intervention within the blackcurrant juice group, whereas there were no differences between treatments in any of the DNA damage markers. In conclusion, this study shows that even large amounts of dietary antioxidants did not decrease the already low steady state levels of oxidative DNA damage in healthy adequately nourished humans.

Detection of early atherosclerosis with radiolabeled monocyte chemoattractant protein-1 in prediabeteic Zucker rats.

BACKGROUND: Migration of monocytes into the arterial wall is an early finding of atherosclerosis. Monocytes are attracted to sites of vascular endothelial cell injury, the initiating event in the development of atheromatous disease, by a chemokine known as monocyte chemoattractant protein-1 (MCP-1). Injured vascular endothelial and smooth muscle cells selectively secrete MCP-1. OBJECTIVE: This study was performed to determine if radiolabeled MCP-1 would co-localize at sites of monocyte/macrophage concentration in an experimental model of transplant-induced vasculopathy in diabetic animals. MATERIALS AND METHODS: Hearts from 3-month-old male Zucker rats, heterozygote (Lean) or homozygote (Fat) for the diabetes-associated gene fa, were transplanted into the abdomens of genetically matched recipients. Lean and Fat animals were then fed normal or high-fat diets for 90 days. RESULTS: At 90 days significant increases (P < 0.013) of MCP-1 graft uptake were seen at imaging and confirmed on scintillation gamma well counting studies in Lean (n = 5) and Fat (n = 12) animals, regardless of diet, 400 % and 40 %, above control values, respectively. MCP-1 uptake of native and grafted hearts correlated with increased numbers of perivascular macrophages (P < 0.02), as seen by immunostaining with an antibody specific for macrophages (ED 2). CONCLUSION: Radiolabeled MCP-1 can detect abnormally increased numbers of perivascular mononuclear cells in native and grafted hearts in prediabetic rats. MCP-1 may be useful in the screening of diabetic children for early atherosclerotic disease.

Effect of omeprazole, lansoprazole, and ranitidine on the DNA synthesis of mononuclear cells.

OBJECTIVE: To examine and compare the effects of omeprazole, lansoprazole, and ranitidine on the DNA synthesis of peripheral blood mononuclear cells. DESIGN: Ex vivo laboratory study. SETTING: Clinical research laboratory of an academic medical center. SUBJECTS: Healthy volunteers. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Venous blood was collected from normal subjects and peripheral blood mononuclear cells (PBMCs) were isolated using centrifugation techniques over a Ficoll-Hypaque density gradient. PBMCs were added to 12-well culture plates in four groups of media: a) control; b) control plus lansoprazole (25 microg/mL); c) control plus omeprazole (0.35 microg/mL); and d) control plus ranitidine (50 microg/mL). PBMCs were exposed to the drug for 96 hrs, with addition of phytohemagglutinin (2.5 microg/ mL) for the last 48 hrs, and 3H-thymidine (1 microCi) during the final 6 hrs. PBMCs were filtered onto glass-fiber filter paper and the radioactivity was determined by scintillation counting. Since radioactivity is measured only in those cells undergoing DNA synthesis or cell division, results are expressed as quantification of 3H-thymidine uptake. Median disintegrations per min (DPM)/number of PBMCs per well+/-SEM are reported: control 68.3+/-37.8; ranitidine 38.4 +/-94.2; lansoprazole 14.6+/-84.4; and omeprazole 15.1+/-48.9. There was a significant difference between lansoprazole vs. ranitidine (p< .01), and omeprazole vs. ranitidine (p< .05), and no significant difference between lansoprazole and omeprazole. CONCLUSIONS: This is the first study to compare the potential immunomodulating effects of these commonly used agents. Ranitidine caused increased DNA synthesis in PBMCs when compared with lansoprazole and omeprazole. This phenomenon may be an important, often disregarded, effect of histamine-2-receptor antagonists when used in postsurgical or trauma patients who have T-lymphocyte-mediated immune suppression.

Cell irradiation caused by diagnostic nuclear medicine procedures: dose heterogeneity and biological consequences.

Most radionuclides used for diagnostic imaging emit Auger electrons (technetium-99m, iodine-123, indium-111, gallium-67 and thallium-201). Their very short range in biological tissues may lead to dose heterogeneity at the cellular level with radiobiological consequences. This report describes the dosimetric models used to calculate the mean dose absorbed by the cell nucleus from Auger radionuclides. The techniques used to determine the biodistribution of radiopharmaceuticals at the subcellular level are also described and compared. Published examples of cellular dosimetry computations performed with radiotracers are reviewed in various clinical settings. Finally, the biological implications of the subcellular localization of Auger emitters are examined. While a number of efforts have been made to obtain dosimetric models and to estimate subcellular distribution of radioactivity, little is known of the cellular dosimetry of most radiopharmaceuticals used in diagnostic imaging. However, biological examples of selective radiotracer uptake have been shown, leading to extremely strong cell-cell dose heterogeneity. Furthermore, radiobiological experiments show that the biological effects of Auger emitters incorporated into DNA can be severe, with relative biological effectiveness greater than 1 compared with external X-rays. These findings clearly show that the assessment of biological risks associated with internal administration of diagnostic radiopharmaceuticals must focus not only on target organs as a whole, but also on the cellular level. This review proposes the most appropriate model for dosimetric computations (cellular or conventional) according to the subcellular distribution of radiotracers. The radionuclide of choice and the general strategy used to design new diagnostic radiopharmaceuticals are also discussed.

Elimination of adult T cell leukemia cells by ultrasound in the presence of porfimer sodium.

Sonodynamic effects using porfimer sodium (Photofrin; Pf) on leukemic and normal cells were evaluated. The purpose of this experiment was to compare cell survival among MT-2 cells, normal peripheral mononuclear cells (PMNCs) and adult T cell leukemia (ATL) patients' PMNCs after sonodynamic treatment. Cells were exposed to 450 kHz ultrasound at an intensity of 500 mW/cm2. The survival rate of MT-2 cells exposed to ultrasound alone for 80 s was 20.1 +/- 4.8%, whereas survival rates exposed to ultrasound in combination with 25, 50 and 100 microg/ml of Pf resulted in 11.5 +/- 2.9, 3.2 +/- 1.6 and 1.6 +/- 1.4%, respectively. There was a significant difference of cell survival between the group exposed to ultrasound alone and the Pf-combined groups (n = 6, p < 0.05). On the other hand, in the normal human PMNCs, no significant differences of cell survival rates were found between ultrasound-treated groups with and without Pf. We similarly examined the survival rate of PMNCs in the peripheral blood of five acute-type ATL patients (n = 5) after ultrasound (60 s, 300 mW/cm2) exposure with or without 100 microg/ml of Pf. Comparison of cell survival rate between ultrasound alone and ultrasound plus Pf showed significant differences (69.4 +/- 22.5 and 30.0 +/- 23.0%, respectively). There were no significant cytotoxicities in all Pf alone treated groups of the MT-2 cells, the normal PMNCs and the ATL patients' PMNCs (p < 0.05). It was suggested from this study that there was a specific selectivity of sonodynamic effects to MT-2 cell lines and ATL patients' PMNCs. It is anticipated that this new method of treatment, i.e. sonodynamic therapy, could be used for extracorporeal blood treatment of acute-type ATL patients.