Adduct of 2-[18F]FDG and 2-nitroimidazole as a putative radiotracer for the detection of hypoxia with PET: synthesis, in vitro- and in vivo-characterization.

A new sugar-coupled 2-nitroimidazole derivative ([18F](see structure in text)) has been prepared in good radiochemical yields starting from peracetylated 2-[18F]FDG obtained from an automated 2-[18F]FDG production module. The corresponding glucose derivative (see structure in text) has proved to be able to inhibit 2-[18F]FDG uptake into tumor cells in a concentration dependent way. However, [18F](see structure in text) failed to show a retention in hypoxic tumor tissue thus excluding itself from further investigations.     

The synthesis and radiolabeling of 2-nitroimidazole derivatives of cyclam and their preclinical evaluation as positive markers of tumor hypoxia.

The cyclam ligand (1,4,8,11-tetraazacyclotetradecane) was condensed with various azomycin-containing synthons to produce chemical compounds that could chelate radioactive metals. It was expected that these radiolabeled markers would become bound selectively to hypoxic cells on the bioreduction of their azomycin substituent. METHODS: The markers were radiolabeled with (99m)Tc, (67)Cu, or (64)Cu. Their uptake and binding to tumor cells in vitro was characterized as a function of time and oxygen concentration. These data defined the hypoxia-specific factor, the ratio of the initial rate of marker binding to severely hypoxic relative to aerobic cells. In addition, the concentration of oxygen (in the equilibrium gas phase) that inhibited binding to 50% of the maximum rate was determined. The in vivo biodistribution and clearance kinetics of the favorable markers were investigated with severe combined immune deficiency mice bearing EMT-6 tumors whose radiobiologic hypoxic fraction (RHF) was approximately 40%. The specific activity (percentage injected dose per gram [%ID/g]) in normal and tumor tissue and the tumor-to-blood and tumor-to-muscle ratios of the optimal markers were also measured for Dunning prostate carcinomas of anaplastic (RHF = 15%-20%) and well-differentiated (RHF < 1%) histology growing in Fischer X Copenhagen rats. Planar images were acquired with some markers from these tumor-bearing rats. RESULTS: The tumor uptake of these cyclam-based markers is approximately 10 times higher when they are labeled with copper isotopes than when labeled with (99m)Tc. FC-327 and FC-334, di-azomycin-substituted cyclams, exhibited hypoxia-specific factors > or = 7.0. The oxygen concentration that inhibited their binding to 50% of the maximal rate was approximately 0.5% O(2), similar to that of the radiobiologic oxygen effect. The %ID/g of (64)Cu-FC-334 retained in EMT-6 tumors in mice and in the anaplastic and well-differentiated prostate tumors in rats 6 h after administration was approximately 6.5, 0.4, and 0.1, respectively. Marker activity in tumor was always less than that in liver and kidney. The tumor-to-blood and tumor-to-muscle ratios of (64)Cu-FC-327 and (64)Cu-FC-334 activity in R3327-AT tumor-bearing rats are higher than those observed for (64)Cu-di-acetyl-bis (N(4)-methylthiosemicarbazone) and approach those of beta-D-(125)I-iodinated azomycin galactopyranoside, the optimal hypoxia marker of the azomycin-nucleoside class. CONCLUSION: These data suggest that some azomycin-cyclams exhibit good hypoxia-marking potential to tumor cells in vitro and to animal tumors of known RHF. Both PET and SPECT could be used to image tumor hypoxia with markers labeled with (64)Cu and (67)Cu, respectively.     

Investigation into 64Cu-labeled Bis(selenosemicarbazone) and Bis(thiosemicarbazone) complexes as hypoxia imaging agents

BACKGROUND: Cu-diacetyl-bis(N4-methylthiosemicarbazone) [Cu-ATSM], although excellent for oncology applications, may not be suitable for delineating cardiovascular or neurological hypoxia. For this reason, new Cu hypoxia positron emission tomography (PET) imaging agents are being examined to search for a higher selectivity for hypoxic or ischemic tissue at higher oxygen concentrations found in these tissues. Two approaches are to increase alkylation or to replace the sulfur atoms with selenium, resulting in the formation of selenosemicarbazones. METHODS: Three 64Cu-labeled selenosemicarbazone complexes were synthesized and one was screened for hypoxia selectivity in vitro using EMT-6 mouse mammary carcinoma cells. Rodent biodistribution and small animal PET images were obtained from BALB/c mice implanted with EMT-6 tumors. One alkylated thiosemicarbazone was synthesized and examined. RESULTS: Of the three bis(selenosemicarbazone) ligands synthesized and examined, only 64Cu-diacetyl-bis(selenosemicarbazone) [64Cu-ASSM] was isolated in high-enough radiochemical purity to undertake cell uptake experiments where uptake was shown to be independent of oxygen concentration. The bis(thiosemicarbazone) complex synthesized, 64Cu-diacetyl-bis(N4-ethylthiosemicarbazone) [64Cu-ATSE], showed hypoxia selectivity similar to 64Cu-ATSM although at a higher oxygen concentration. Biodistribution studies for 64Cu-ASSM and 64Cu-ATSE showed high tumor uptake at 20 min (64Cu-ASSM, 10.33+/-0.78% ID/g; 64Cu-ATSE, 7.71+/-0.46% ID/g). PET images of EMT-6 tumor-bearing mice visualized the tumor with 64Cu-ATSE and revealed hypoxia selectivity consistent with the in vitro data. CONCLUSION: Of the compounds synthesized, only 64Cu-ASSM and 64Cu-ATSE could be examined in vitro and in vivo. Although the stability of bis(selenosemicarbazone) complexes increased upon addition of methyl groups to the diimine backbone, the fully alkylated species, 64Cu-ASSM, demonstrated no hypoxia selectivity. However, the additional alkylation present in Cu-ATSE modifies the hypoxia selectivity and in vivo properties when compared with Cu-ATSM.     

Evaluation of Tc-labeled modified serum albumin for tumor detection

Serum albumin (SA) modified and labeled with ¹³¹l-tyramine N-1′-desoxysorbitol (¹³¹I-TDS) has been shown to localize in tumors [Sinn et al., (1990) Nucl. Med. Biol. Part B 17, 819–827]. We prepared similar TDS complexes labeled with ^99mTc and evaluated their potential for tumor imaging. Derivatization of SA with TDS was optimized using cyanuric chloride or 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDAC) as coupling agents. A high TDS loading yield of 38 mol/mol SA was obtained with the latter reagent. Modified SA (8 and 38 mol TDS/mol SA) were labeled with ^99mTc via the stannous reduction method and injected i.v. into EMT-6 tumor bearing mice. ¹²⁵I-TDS-SA (8 mol ¹²⁵I-TDS/mol SA) revealed a high tumor uptake of 10% ID/g at 3 h post-injection. The ^99mTc-labeled SA and TDS-SA complexes lacked tumor specificity, instead TDS loading of SA resulted in increased liver/spleen uptake, suggesting colloid formation. This study confirms the potential of modified SA for tumor imaging but highlights the importance of choice of radioisotope, as well as site of attachment of the radiolabel to the modified SA for optimal tumor localization.     

Radioiodinated 1-(5-iodo-5-deoxy-beta-D-arabinofuranosyl)-2-nitroimidazole (iodoazomycin arabinoside: IAZA): a novel marker of tissue hypoxia

1-(5-Iodo-5-deoxy-beta-D-arabinofuranosyl)-2-nitroimidazole (IAZA) has been synthesised and labeled with 125I. Radioiodinated IAZA was shown to undergo hypoxia-dependent binding in EMT-6 cells in vitro and to have an initial binding rate of 284 pmole/10(6) cells/hr at a substrate concentration of 30 microM. This binding rate is more than three times that of the reference compound, misonidazole (89 pmole/10(6) cells/hr). The elevated binding rate was accompanied by in vitro cytotoxicity 30-40 times greater than that observed for misonidazole. Whole-body elimination and biodistribution studies in BALB/c mice bearing implanted, subcutaneous EMT-6 tumors showed a rapid excretion (greater than 98% in 24 hr) with moderate tissue levels which, in general, declined as a function of blood clearance. Tumor-to-blood ratios of 4.6 (4 hr) and 8.7 (8 hr), with respective tumor uptake values of 2.08% and 1.22% ID/g of tissue, form a rational basis for evaluation of this and related 2-nitroimidazole analogs as radiopharmaceuticals suitable for scintigraphic evaluation of tissue (tumor) hypoxia.     

Caffeine stimulates ventilation in athletes with exercise-induced hypoxemia

INTRODUCTION/PURPOSE: Many athletes with exercise-induced hypoxemia (EIH) show an insufficient ventilatory response to exercise and low resting ventilatory responsiveness. The purpose of this project was to determine whether a moderate dosage of caffeine, a common ventilatory stimulant, could augment resting ventilatory responsiveness, exercise ventilation (V E), end-tidal O2 partial pressure (PetO2), and arterial oxyhemoglobin saturation (HbSaO2) in athletes with EIH. METHODS: Eight highly trained males ([formula: see text], 69.2 +/- 4.0 mL.[kg.min]) who demonstrated EIH at [formula: see text] (HbSaO2, 88.0 +/- 1.7%), ingested in a randomized design a placebo or caffeine (CAF, 8 mg.kg body wt) 1 h before testing. Ventilatory responsiveness at rest was assessed via the isocapnic hypoxic and hyperoxic hypercapnic ventilatory responses (HVR and HCVR, respectively). Dependent measures of metabolic variables, ventilation, and saturation were determined during progressive treadmill exercise to exhaustion. RESULTS: V E was higher at 75%, 80%, and 100% of [formula: see text] with CAF (P < 0.05). V E/V O2, PetO2, and HbSaO2 were increased at 75%, 80%, and 90% of [formula: see text] with CAF but were not different at [formula: see text] despite an increase in V e. No change in [formula: see text] was observed between treatments. HVR and HCVR were not different between the two conditions, indicating that the increased V E likely came from central stimulation or secondary effects of CAF. CONCLUSION: The failure of HbSaO2 to increase at [formula: see text] despite an increase in V E suggests that mechanisms influencing HbSaO2 other than an inadequate hyperventilatory response may operate to different degrees across individuals as [formula: see text] is approached.     

Technetium-99m-cyclam AK 2123: a novel marker for tumor hypoxia.

Technetium-99m labeled cyclam N-2'-methoxyethyl-2-(3'-nitro-1'-triazole) acetamide (cyclam AK 2123) has been synthesized, radiolabeled and characterized as a hypoxic tumor imaging agent. Radiochemical purity was greater than 95%. Marker biodistribution was measured in normal Wistar strain rats at different time intervals after intra venous (i.v.) administration. In vivo distribution and scintigraphic imaging studies were performed after i.v. injection into mammary tumor-bearing rats using a gamma camera and associated computer. Intratumor partial oxygen pressure (pO2) and oxygen saturation measurements were performed to estimate the oxygenation status of the tumors. Tumor to muscle ratio (T/M) of 99mTc-cyclam AK 2123 was 8.5 which was compared with other tumor seeking radiopharmaceuticals, viz. 99mTc-(V) DMSA (3.07), 99mTc-citrate (5.29) and 201T1C1 (3.29). T/M ratios were also evaluated in comparison with radioiodinated iodoazomycin galactopyronoside (125I-IAZG). The ratio obtained was 18 for 99mTc-cyclam AK 2123 and 20 for 125I-IAZG, respectively. The increased concentration of radioactivity in these tumors suggests that this agent could be labelling hypoxic cells and have utility as an imaging agent.     

乏氧组织显像剂99Tcm-DTPA-甲硝唑的制备和荷瘤动物实验研究

目的　进行新型乏氧组织显像剂DTPA 甲硝唑的化学合成、药盒制备、99Tcm 标记、质量控制以及药理研究。方法　合成的DTPA 甲硝唑用氯化亚锡作还原剂 ,99Tcm 标记 ,用多元正交法确立99Tcm 标记一步法药盒的最佳配方。纸层析法鉴定99Tcm DTPA 甲硝唑的标记率、标记稳定性 ,完成了H2 2肝癌小鼠体内生物分布实验及显像研究。结果　DTPA 甲硝唑一步法药盒与99TcmO-4 混合 ,沸水浴反应 10min ,放化纯度大于 99%。H2 2肝癌小鼠生物分布显示 ,肿瘤 /血液比值 1、2h分别为3 43、6 40 ,肿瘤 /肌肉比值 1、2h分别为 5 2 4、7 42 ;注射血管舒张药肼苯哒嗪组肿瘤组织摄取明显高于对照组 (P <0 0 1)。结论　研制的99Tcm DTPA 甲硝唑有望成为一种新型的肿瘤乏氧组织显像剂。     

Preparation, biodistribution, and small animal PET of 45Ti-transferrin.

Investigation of 45Ti-transferrin was pursued to provide insight into the mechanism of action of titanocene dichloride, a chemotherapeutic agent currently in clinical trials. METHODS: Plasma protein-binding studies of processed 45Ti were performed by solubilizing the 45Ti residue in 0.05N HCl, of which 1.22 MBq (33 microCi) in 10 microL were added to 250 microL of dog plasma. 45Ti-Transferrin was prepared by redissolving the processed 45Ti in 25 micromol/L apotransferrin or by in vivo incorporation through preparation and introduction of 45Ti-citrate. Biodistribution studies were performed on normal Sprague-Dawley rats and EMT-6 tumor-bearing BALB/c mice with 45Ti-transferrin coinjected with 67Ga-citrate for direct comparison. microPET was performed on mice bearing EMT-6 tumors and the images were analyzed for tumor-to-muscle uptake ratios. RESULTS: Direct labeling of apotransferrin in situ with 45Ti was achieved as well as in vivo incorporation by 2 h after injection with 45Ti-citrate. The biodistribution of 45Ti-transferrin and 67Ga-citrate showed similar trends. In Sprague-Dawley rats, initial blood uptake was higher for the 45Ti-transferrin, whereas bone uptake increased more for the 67Ga-citrate. EMT-6 tumor uptake in both cases was relatively high (14.6 +/- 1.83 %ID/g for 45Ti and 8.72 +/- 0.98 %ID/g for 67Ga [%ID/g = percentage injected dose per gram]) and remained elevated even out to 24 h after injection. The tumor-to-muscle ratio of the 67Ga-citrate reached 6.7 at 24 h, whereas the ratio of the 45Ti-transferrin increased to 4.3 at this time point. Uptake of 45Ti-transferrin was visualized in the EMT-6 murine mammary carcinoma tumor with microPET. In all cases, the tumor was clearly delineated from the surrounding tissue with tumor-to-muscle ratios on the order of 1.6. CONCLUSION: 45Ti forms a complex with apotransferrin that remains intact in vivo. Results of the biodistribution in mice showed that the tumor had increased uptake compared with nontarget organs (e.g., muscle). The microPET images of tumor-bearing mice clearly delineate the tumors from the surrounding tissue. Comparison of the data suggests that tissue uptake is similar whether injecting 45Ti-transferrin directly or as 45Ti-citrate, which transchelates to transferrin before the time of imaging.     

131I]Iodoazomycin arabinoside for low-dose-rate isotope radiotherapy: radiolabeling, stability, long-term whole-body clearance and radiation dosimetry estimates in mice

BACKGROUND: The preliminary characterization of [(131)I]iodoazomycin arabinoside ([(131)I]IAZA) as a potential radiotherapeutic radiopharmaceutical is described. METHODS: High-specific-activity [(131)I]IAZA was prepared in therapeutic doses (up to 3 GBq per batch) by isotope exchange in pivalic acid melt and was purified on Sep-Pak cartridges. Stability in 15% ethanol in saline at 4 degrees C was determined by high-performance liquid chromatography. IAZA cytotoxicity (IC(50), approximately 0.1 mM) against both murine (EMT-6) and human (143B, 143B-LTK) tumor cells determined by MTT test was in the range previously reported for EMT-6 cells using a clonogenic assay. Tissue radioactivity levels were measured in a murine tumor model for the 24- to 168-h postinjection period. Radiation dose estimates obtained from the tissue activity levels for this period were calculated from pharmacokinetic (WinNonlin) and dosimetry (MIRD and RAdiation Dose Assessment Resource) parameters. RESULTS: The radioiodination efficiency was >90%, but with systematic losses during Sep-Pak purification, the recovered yields of [(131)I]IAZA were approximately 75%. The product (specific activity, 4.6-6.4 GBq/micromol) was stable for at least 2 weeks, with only approximately 6% degradation over this storage period. Extended biodistribution studies in Balb/c mice bearing implanted EMT-6 tumors showed that the highest tumor/blood radioactivity ratio (T/B; 4.8) occurred 24 h after dosing; the T/B ratio was approximately 1.5 at the end of the 7-day study. The 24- to 168-h tissue radioactivity data fit a one-compartment model except for liver data, which best fit a two-compartment model. Dosimetry estimates showed a tumor self-dose of 7.4 mGy/MBq, which is several-fold higher than for the liver or the kidney. CONCLUSIONS: [(131)I]IAZA can be efficiently radiolabeled at high specific activity, purified by a simple Sep-Pak technique and stored with little radiolysis or chemical decomposition at these specific activities. Based on measured radioactivity burdens during the week following injection and on published animal ([(125)I]IAZA) and clinical ([(123)I]IAZA) dosimetry data, the current dose estimates point to selective tumor irradiation at low dose rates.     

[<Superscript>99m</Superscript>Tc]Demotensin 5 and 6 in the NTS1-R-targeted imaging of tumours: synthesis and preclinical results

Purpose The aim of this study was to evaluate the applicability of [^99mTc]Demotensin 5 and 6 in the targeted diagnostic imaging of neurotensin subtype 1 receptor (NTS1-R)-expressing tumours. Methods Labelling of Demotensin 5 and 6 with ^99mTc was conducted by brief incubation with ^99mTcO₄ ⁻, SnCl₂ and citrate anions in alkaline medium at ambient temperature. Affinities of conjugates for the NTS1-R were determined by competition binding experiments in WiDr cell membranes using [¹²⁵I-Tyr³]NT as the radioligand. Saturation binding assays were conducted for [^99mTc/^99gTc]Demotensin 6 in WiDr cell membranes. Internalisation of [^99mTc]Demotensin 5 and 6 was studied at 37°C in WiDr cells. Biodistribution of [^99mTc]Demotensin 5 and 6 was performed in female Swiss nu/nu mice bearing human WiDr xenografts. Results Unlabelled conjugates showed a high affinity for the human NTS1-R (Demotensin 5 IC₅₀=0.03±0.01 nM; Demotensin 6 IC₅₀=0.08±0.02 nM), while high affinity was also exhibited by (radio)metallated [^99mTc/^99gTc]Demotensin 6 (K _d=0.13±0.01 nM). [^99mTc]Demotensin 5 and 6 internalised rapidly and specifically in WiDr cells. After injection in WiDr tumour-bearing mice, radiopeptides, and especially the doubly stabilised [^99mTc]Demotensin 6, showed NTS1-R-mediated uptake in the intestines and in the implanted tumour (4.30±0.45%ID/g at 1 h post injection) and rapid renal excretion from non-target tissues into the urine. Conclusion [^99mTc]Demotensin 6 shows a favourable preclinical profile and further testing in patients is warranted to monitor its eventual applicability as a radiotracer in the diagnostic imaging of NTS1-R-positive tumours.     

Biodistribution of hypoxic marker, 99mTc-HL91 (4,9-diaza-3,3,10,10-tetramethyldodecan-2,11-dione dioxime)

99mTc-HL91 (4,9-diaza-3,3,10,10-tetramethyldodecan-2,11-dione dioxime) was developed as a hypoxic marker. Athymic mice bearing human tumors were administered with 99mTc-HL91 to evaluate it from the clinical point view. The tumor was visualized clearly 4 hours after injection. The biodistribution study revealed that 99mTc-HL91 was accumulated in the liver (tissue-to-blood ratio (T/B) = 11.5) and kidney (2.25) with higher than in the tumor (1.01). Oxygen condition of the tumor and muscle was measured by using the probe. 99mTc-HL91 uptake of tumors with PO2 (tumor-to-muscle ratio) under 0.55 was higher than that with 0.55 or higher PO2. Oxygen condition of tumors with 1.0 or higher uptake (tumor-to-blood ratio) was lower than that with the lower uptake. Autoradiography of tumor sections indicated that 99mTc-HL91 was scarcely accumulated in the necrotic and viable areas while strong radioactivity was observed in the border zone (speculated to be hypoxic condition). Our experimental results suggest that the 99mTc-HL91 scintigraphy may be useful for evaluating oxygenation status of some tumors in non-abdominal region.     

Administered dose and tumor dose of bleomycin labeled with cobalt-57 in mice and men.

Tumor concentrations of the chemotherapeutic drug, bleomycin, labeled with cobalt-57 (Co-bleo) were compared in mouse tumor models and in human lung tumors using quantitative single-photon emission computed tomography. Drug concentrations in histologically similar human tumors showed marked variability for the same injected dose (ID). Small cell carcinomas showed concentrations between 1.09 and 8.85 %ID/cc x 10(-3) while non-small cell lung tumors showed a concentration variation between 0.36 and 6.75 %ID/cc x 10(-3). In contrast to the situation in human tumors, uptake in mouse tumors showed only slight variability in animals with the same tumor model. EMT-6 tumors in mice showed at 6 hr significantly higher uptake of Co-bleo (p less than 0.001) and significantly higher tumor-to-lung ratio (p less than 0.001) when compared to murine fibrosarcomas. The EMT-6 tumors in contrast to the fibrosarcomas responded to bleomycin treatment in a dose dependent manner. The results indicate that while in mice the tumor dose closely follows the administered dose, in humans, the tumor dose and the tumor-to-lung ratio in the individual patient cannot be predicted from the administered dose.     

99Tcm-HL91和18F-FDG显像与放疗关系的实验研究

目的 用99Tcm-4,9-二氮-2,3,10,10-四甲基十二烷-2,11-二酮肟(99Tcm-HL91)和18F-FDG为显像剂,探讨肿瘤放疗前、后的乏氧状态与放疗疗效之间的关系.方法 ①动物:选取20只昆明种、清洁级健康成年雄性小鼠供实验用(另备5只用于腹水模型),采用完全随机方法分为2组(对照组和实验组),每组...     

99Tcm-HL91乏氧显像在甲状腺结节良恶性鉴别诊断中的应用

目的 评价99Tcm4,9-二氮-3,3,10,10-四甲基十二烷-2,11-二酮肟(HL91)对甲状腺结节良恶性鉴别诊断的临床价值.方法 对58例经触诊或超声检查存在甲状腺结节的患者进行99Tcm-HL91甲状腺早期(10 min)及延迟(4 h)平面显像,应用感兴趣区(ROI)技术计算靶/非靶(T/N)比值并进行比较.组间比较采用成组设计两样本均数t检验,率的比较用χ2检验.结果 患者均经手术或细针穿刺病理检查,其中21例为甲状腺癌,37例为甲状腺良性结节.99Tcm-HL91显像诊断甲状腺癌的灵敏度、特异性和准确性分别为85.7%,94.6%和91.4%,不同甲状腺癌病理类型的显像灵敏度差异无统计学意义(χ2=0.778,P＞0.05),但结节越大灵敏度越高.甲状腺癌与甲状腺良性结节组早期及延迟相T/N比值分别为1.07±0.04,1.25±0.03和0.92±0.10,0.91±0.12,组间在10 min时T/N比值差异无统计学意义(t=1.900,P＞0.05),4 h时T/N比值差异有统计学意义(t=3.885,P＜0.001).结论 用99Tcm-HL91乏氧显像判断甲状腺结节的良恶性有一定的临床价值,显像时间以4 h为佳.     

A transferrin-mediated uptake of gallium-67 by EMT-6 sarcoma. II. Studies in vivo (BALB/c mice): concise communication

The EMT-6 sarcoma-like tumor of BALB/c mice can be grown as a solid subcutaneous transplantable tumor in vivo or as a monolayer culture in vitro. We have studied the uptake of gallium-67 by this tumor growing subcutaneously on the backs of 6-week-old BALB/c mice. After i.v. administration of Ga-67 citrate, tumor uptakes were as high as any others reported for mouse tumors. Also, for unknown reasons, there was appreciable reduction in tumor uptake with increasing amounts of Ga-67 citrate, even in the microcurie range. Furthermore, when mouse serum is prelabeled with Ga-67 and then injected, the EMT-6 uptake is greater than with Ga-67 administered as citrate (p less than 0.02). We believe that the finding of avid Ga-67 uptake in vivo helps to establish this unique in vivo/in vitro tumor system as a valid experimental model for studies regarding the mechanism of Ga-67 accumulation by neoplastic tissue.     

Cellular accumulation and retention of the technetium-99m-labelled hypoxia markers BRU59-21 and butylene amine oxime

BRU59-21 and ^99mTc-butylene amine oxime (BnAO, HL91) are being evaluated for imaging hypoxia in tumors. Both tracers: 1) rapidly reached a plateau in aerobic Chinese hamster ovary cells in vitro but continuously accumulated in hypoxic cells; 2) ceased to accumulate when hypoxic cells were exposed to air; 3) showed 40% retention upon washing the cells; 4) showed selective hypoxic accumulation only at 37°C; 5) accumulation could be modulated by addition of electron-affinic compounds; and 6) exhibited higher accumulation in cells which overexpress cytochrome P450 reductase. Both BRU59-21 and ^99mTc-BnAO share properties making them suitable for hypoxia imaging.     

An improved (99m)Tc-aprotinin kit formulation: quality control analysis of radiotracer stability and cold kit shelf life

^99mTc-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 ^99mTc-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. ^99mTc-aprotinin was formed at greater than 95% efficiency at all time points tested with ^99mTcO₂ present as the major impurity (1–4%) and ^99mTc-pertechnetate and ^99mTc-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 ^99mTc-aprotinin.     

A transferrin-mediated uptake of gallium-67 by EMT-6 sarcoma. I. Studies in tissue culture

We have studied the in vitro uptake of gallium-67 by exponentially growing EMT-6 sarcoma cells in long-term tissue culture. In this system, the addition of transferrin to the medium was required before an appreciable cellular uptake of Ga-67 occurred. The transferrin effect was complex, with an initial stimulation to a peak cell-to-medium ratio of 8--10:1 at low concentrations of transferrin (0.2 mg/ml), followed by a gradual decline in uptake as transferrin in the medium was increased further. EMT-6 tumor-cell uptake of Ga-67 was probably mediated by a specific cellular receptor for transferrin. Scatchard analysis of the EMT-6 cellular binding of human transferrin labeled with iodine-125 indicated a cellular receptor with affinity for transferrin of 5 X 10(6) l/mole and abundance of 500,000 receptors per cell. Over the experimental range of transferrin concentration in the medium, the observed uptake of Ga-67 was closely correlated with the degree of formation of Ga-67-labeled transferrin and the fraction of transferrin bound to the cellular receptor (N = 69, r = 0.86, p less than 0.0001).     

The effects of interval–exercise duration and intensity on oxygen consumption during treadmill running

The magnitude of improvement in peak oxygen uptake ( [Formula: see text] ) and performance to an exercise training regime is related to the [Formula: see text] of prior accumulated exercise training bouts. However, it is unclear whether constant rate training (CRT) or interval training (INT) preferentially alters the [Formula: see text] of running exercise. Therefore, the purpose of this study was to compare the acute [Formula: see text] response to constant, and interval training sessions. Consequently, this study compared the mean average [Formula: see text] of 17 moderately trained participants to a 20-min CRT and two different 20min INT treadmill runs. Participants completed three treatments (twice) in random order over 3 weeks. In 1min INT participants completed 10x1min efforts at the velocity corresponding to [Formula: see text] (V(peak)) interspersed with 10x1min efforts at 0.5V(peak). In the 2min INT, participants completed 5x2min efforts at the V(peak) interspersed with 5x2min efforts at 0.5 at V(peak). In CRT participants ran at a velocity equivalent to the mean velocity of INT (75% V(peak)). Mean average [Formula: see text] for 2min INT, 1min INT and CRT were, respectively, 3200+/-661; 3076+/-6041; 2909+/-584mlmin(-1). Both INT sessions resulted in a significantly higher mean average [Formula: see text] than CRT. Furthermore, 2min INT resulted in 90% of [Formula: see text] being exceeded more frequently than 1min INT. We conclude that INT serves as a more potent stimulus for improvement in [Formula: see text] and subsequent endurance performance than CRT.