Search

The effects of fat on gastric emptying (GE), gut hormones, and energy intake are dependent on digestion to free fatty acids (FFAs). In animals, small intestinal oleic acid inhibits energy intake more potently than the triacylglyceride (TG) triolein, but there is limited information about the comparative effects of FFA and TG in human beings. We compared the effects of FFA and TG on GE, gut hormone secretion, appetite, and energy intake in healthy males.Nine men (age, 23 +/- 2 y; body mass index, 22 +/- 1 kg/m(2)) were studied on 3 occasions to evaluate the effects of (1) 40 g oleic acid (FFA, 1830 kJ), (2) 40 g macadamia oil (TG, 1856 kJ; both 600-mL oil-in-water emulsions stabilized with 4% milk protein and labeled with 15 MBq (123)I), or (3) 600 mL 4% milk protein (control, 352 kJ), administered intragastrically, on GE, plasma cholecystokinin (CCK) and peptide-YY (PYY) levels, appetite perceptions, and subsequent energy intake.GE of FFA was much slower than that of TG (P.05), with greater retention of FFA, than TG, in the proximal stomach (P.001). Hunger was less (P.05), and fullness was greater (P.05), after FFA when compared with control and TG. Increases in plasma CCK and PYY levels were greater after FFA than TG or control (P.05). Energy intake tended to be less after FFA compared with TG (control, 4754 +/- 610 kJ; TG, 5463 +/- 662 kJ; FFA, 4199 +/- 410 kJ).FFAs empty from the stomach more slowly, but stimulate CCK and PYY and suppress appetite more potently than TG in healthy human beings.

To confirm the pharmacokinetics and biodistribution of 99mTc aprotinin in normal volunteers and to determine the optimum time for scanning post-injection, prior to further investigations of 99mTc aprotinin as an imaging agent for amyloidosis.Five patients (three men and two women, average age 49 years, age range 38-66 years) without a history of amyloidosis or any of the associated diseases, were included in this prospective study. Blood and urine were collected and images were performed of the whole body and wrists.Normal biodistribution of 99mTc aprotinin includes early cardiac and lung activity in the blood pool phase with subsequent hepatic activity and renal excretion with variable splenic activity. There is variable bowel uptake on later images. The best quality images were obtained 90 min post-intravenous administration, and this is likely to be the optimum time for clinical imaging.

Early identification of tumor responses to treatment is crucial for devising more effective and safer cancer treatments. No widely applicable, noninvasive method currently exists for specifically detecting tumor cell death after cytotoxic treatment and thus for predicting treatment outcomes. Methods: We have further characterized the targeting of the murine monoclonal antibody DAB4 specifically to dead tumor cells in vitro, in vivo, and in clinical samples. We found that sustained DAB4 binding to treated cells was closely associated with markers of intrinsic apoptosis and DNA double-strand break formation. In a competition binding assay, DAB4 bound EL4 murine thymic lymphoma cells in preference to the normal counterpart of murine thymocytes. Defective in vivo clearance of apoptotic cells augmented in vivo accumulation of DAB4 in tumors particularly after chemotherapy but was unchanged in normal tissues. Tumor targeting of DAB4 was selective for syngeneic murine tumors and for human tumor xenografts of prostate cancer (PC-3) and pancreatic cancer (Panc-1) before and more so after chemotherapy. Furthermore, DAB4 was shown to bind to dead primary acute lymphoblastic leukemic blasts cultured with cytotoxic drugs and dead epithelial cancer cells isolated from peripheral blood of small cell lung carcinoma patients given chemotherapy. Conclusion: Collectively, these results further demonstrate the selectivity of DAB4 for chemotherapy-induced dead tumor cells. This postchemotherapy selectivity is related to a relative increase in the availability of DAB4-binding targets in tumor tissue rather than in normal tissues. The in vitro findings were translated in vivo to human xenograft models and to ex vivo analyses of clinical samples, providing further evidence of the potential of DAB4 as a marker of tumor cell death after DNA-damaging cytotoxic treatment that could be harnessed as a predictive marker of treatment responses.

Background: Antineoplastic therapy may impair the survival of malignant cells to produce cell death. Consequently, direct measurement of tumor cell death in vivo is a highly desirable component of therapy response monitoring. We have previously shown that APOMAB® representing the DAB4 clone of a La/SSB-specific murine monoclonal autoantibody is a malignant cell-death ligand, which accumulates preferentially in tumors in an antigen-specific and dose-dependent manner after DNA-damaging chemotherapy. Here, we aim to image tumor uptake of APOMAB® (DAB4) and to define its biological correlates. Methodology/Principal Findings: Brisk tumor cell apoptosis is induced in the syngeneic EL4 lymphoma model after treatment of tumor-bearing mice with DNA-damaging cyclophosphamide/etoposide chemotherapy. Tumor and normal organ accumulation of Indium 111 (111In)-labeled La-specific DAB4 mAb as whole IgG or IgG fragments was quantified by whole-body static imaging and organ assay in tumor-bearing mice. Immunohistochemical measurements of tumor caspase-3 activation and PARP-1 cleavage, which are indicators of early and late apoptosis, respectively, were correlated with tumor accumulation of DAB4. Increased tumor accumulation of DAB4 was associated directly with both the extent of chemotherapy-induced tumor cell death and DAB4 binding per dead tumor cell. Tumor DAB4 accumulation correlated with cumulative caspase-3 activation and PARP-1 cleavage as tumor biomarkers of apoptosis and was directly related to the extended median survival time of tumor-bearing mice. Conclusions/Significance: Radiolabeled La-specific monoclonal antibody, DAB4, detected dead tumor cells after chemotherapy, rather than chemosensitive normal tissues of gut and bone marrow. DAB4 identified late apoptotic tumor cells in vivo. Hence, radiolabeled DAB4 may usefully image responses to human carcinoma therapy because DAB4 would capture the protracted cell death of carcinoma. We believe that the ability of radiolabeled DAB4 to rapidly assess the apoptotic tumor response and, consequently, to potentially predict extended survival justifies its future clinical development as a radioimmunoscintigraphic agent. This article is part I of a two-part series providing proof-of-concept for the the diagnostic and therapeutic use of a La-specific monoclonal antibody, the DAB4 clone of which is represented by the registered trademark, APOMAB®.

The purpose of this study was to validate a rapid quality control method for the lymphoscintigraphic tracer 99mTc-antimony trisulfide colloid (99mTc-ATC).ATC was labeled with 99mTc according to the manufacturer's instructions as well as by alternative heating conditions designed to provide a range of percentages of radiochemical purity (RCP): the tracer was prepared in a dry block heater with heating cavities of different sizes, the temperature of the heating block was varied from 70 degrees C to 115 degrees C, or the duration of heating was varied from 15 to 35 min. Anion-exchange minicolumns were trialled to separate any 99mTc-pertechnetate impurity from 99mTc-ATC with physiologic saline as the eluent. Quality control results were compared with the results from the manufacturer's recommended method, which uses an instant thin-layer chromatography (ITLC) strip with saline as the migrating solution.The quality control results obtained with a cartridge method in 2-3 min compared favorably with those obtained with the ITLC method with saline when the tracer was prepared by heating at 115 degrees C in a dry block heater for 35 min (RCPs, 99.4%+/-0.3% [mean+/-SD] and 99.2%+/-0.3%, respectively; n=25). The cartridge and ITLC quality control results also were in excellent agreement (correlation coefficient, 0.99) over a range of RCPs (80%-100%). An alternative anion-exchange cartridge that was tested in this study was not suitable for assaying the RCP of 99mTc-ATC because of the complete retention of 99mTc-pertechnetate on the sorbent.Compared with the established ITLC method, the cartridge quality control method tested in this study is rapid and provides a reliable assessment of the RCP of 99mTc-ATC. For the preparation of 99mTc-ATC, a dry block heater can be successfully substituted for a boiling water bath and is recommended for heating at high altitudes.

99m Tc-aprotinin scintigraphy has been demonstrated to be a useful noninvasive imaging technique for amyloid deposits located in extraabdominal regions of patients. The aim of this study was to develop an improved aprotinin cold kit formulation, to validate the kit for long-term stability, as well as to assess the radiotracer stability by novel quality control methods. The aprotinin cold kit formulation of Trasylol, pyrophosphate (PYP)-chelated stannous reductant and an alkaline buffer, was dispensed into nitrogen-filled vials and aliquots frozen at −20°C. After 0, 1, 2, 3 and 6 months of storage, three samples were reconstituted with 750–850 MBq of 99m Tc-pertechnetate, followed by quality control analyses by paper chromatography methods at 25, 85 and 265 min postreconstitution (pr). Cation-exchange cartridge quality control methods were also investigated. The cold kits proved to be stable to long-term storage for up to 6 months, and the radiotracer was stable for at least 4 h pr. 99m Tc-aprotinin was formed at greater than 95% efficiency at all time points tested with 99m TcO 2 present as the major impurity (1–4%) and 99m Tc-pertechnetate and 99m Tc-PYP present in trace amounts. An alternative, rapid, safe and reliable method was found in Oasis MCX–BSA-treated cartridges using saline as the eluting solution to assay for 99m Tc-aprotinin.

The effects of fat on gastric emptying, gastrointestinal hormone release and energy intake are dependent on the digestion of fat to free fatty acids (FFAs). There is limited information about the comparative effects of FFAs and triglycerides (TGs) on these factors. In animals, oleic acid inhibits energy intake more potently than an isocaloric infusion of its TG, triolein. Nine healthy males (23±2 years, BMI 22±1 kg/m 2 ) were studied on three occasions to evaluate the effects of (i) 40 g of oleic acid (“FFA”) (1830 kJ), (ii) 40 g of macadamia oil (“TG”) (1856 kJ) (both 600 ml oil-in-water emulsions stabilized with 4% milk protein and labelled with 15 MBq 123 I), or (iii) 600 ml 4% milk protein (“control”, 352 kJ), administered intragastrically, on gastric emptying, plasma CCK and PYY, appetite and energy intake. Gastric emptying of FFA was much slower than of TG ( P P P P P P Conclusion Free fatty acids empty from the stomach more slowly, but stimulate CCK and PYY release and suppress appetite and energy intake much more potently, than triglycerides.