Synthesis and evaluation of gallium-68-labeled nitroimidazole-based imaging probes for PET diagnosis of tumor hypoxia.

OBJECTIVE: In this study, we designed and synthesized four novel 68Ga-radiolabeled compounds ([68Ga]DN-3, [68Ga]DN-4, [68Ga]NN-3, and [68Ga]NN-4) composed of a nitroimidazole and two types of bifunctional chelates (DOTA or NOTA) via several alkyl linkers of different length. Then, we evaluated their properties as hypoxia imaging probes for positron emission tomography (PET) compared with conventional compounds ([68Ga]DN-2 and [68Ga]NN-2). METHODS: The precursors of 68Ga-radiolabeled compounds were synthesized through a two-step reaction, and then reacted with 68GaCl3 to be 68Ga-radiolabeled compounds. FaDu cells were treated with 68Ga-radiolabeled compounds and then incubated under normoxic (21% O2) or hypoxic (1% O2) conditions. The radioactivity of these cells was measured 2 h after incubation. The biodistribution and PET/CT imaging of 68Ga-radiolabeled compounds in FaDu-bearing Balb/c nude mice were evaluated 2 h after intravenous injection. RESULTS: The 68Ga-radiolabeled compounds were synthesized with radiochemical purities over 95%. In the in vitro study, the levels of 68Ga-radiolabeled compounds were significantly higher in hypoxic cells than in normoxic cells. In hypoxic cells, the compounds we designed in this study demonstrated higher accumulation than the conventional compounds. In the in vivo biodistribution study, [68Ga]DN-3 exhibited the highest accumulation in tumor. In the in vivo PET/CT imaging study, the tumor tissues of the FaDu-xenografted mice were visualized at 2 h after intravenous administration of 68Ga-radiolabeled compounds. CONCLUSIONS: Our study suggested that the length of the linkers connecting nitroimidazole to a bifunctional chelate affect PET imaging of hypoxic tumors with 68Ga-radiolabeled compounds.

Increased [18F]FMISO accumulation under hypoxia by multidrug-resistant protein 1 inhibitors.

BACKGROUND: [18F]Fluoromisonidazole ([18F]FMISO) is a PET imaging probe widely used for the detection of hypoxia. We previously reported that [18F]FMISO is metabolized to the glutathione conjugate of the reduced form in hypoxic cells. In addition, we found that the [18F]FMISO uptake level varied depending on the cellular glutathione conjugation and excretion ability such as enzyme activity of glutathione-S-transferase and expression levels of multidrug resistance-associated protein 1 (MRP1, an efflux transporter), in addition to the cellular hypoxic state. In this study, we evaluated whether MRP1 activity affected [18F]FMISO PET imaging. METHODS: FaDu human pharyngeal squamous cell carcinoma cells were pretreated with MRP1 inhibitors (cyclosporine A, lapatinib, or MK-571) for 1 h, incubated with [18F]FMISO for 4 h under hypoxia, and their radioactivity was then measured. FaDu tumor-bearing mice were intravenously injected with [18F]FMISO, and PET/CT images were acquired at 4 h post-injection (1st PET scan). Two days later, the same mice were pretreated with MRP1 inhibitors (cyclosporine A, lapatinib, or MK-571) for 1 h, and PET/CT images were acquired (2nd PET scan). RESULTS: FaDu cells pretreated with MRP1 inhibitors exhibited significantly higher radioactivity than those without inhibitor treatment (cyclosporine A: 6.91 ± 0.27, lapatinib: 10.03 ± 0.47, MK-571: 10.15 ± 0.44%dose/mg protein, p < 0.01). In the in vivo PET study, the SUVmean ratio in tumors [calculated as after treatment (2nd PET scan)/before treatment of MRP1 inhibitors (1st PET scan)] of the mice treated with MRP1 inhibitors was significantly higher than those of control mice (cyclosporine A: 2.6 ± 0.7, lapatinib: 2.2 ± 0.7, MK-571: 2.2 ± 0.7, control: 1.2 ± 0.2, p < 0.05). CONCLUSION: In this study, we revealed that MRP1 inhibitors increase [18F]FMISO accumulation in hypoxic cells. This suggests that [18F]FMISO-PET imaging is affected by MRP1 inhibitors independent of the hypoxic state.

Composting cattle dung wastes by using a hyperthermophilic pre-treatment process: characterization by physicochemical and molecular biological analysis.

To solve malodorous odor problems by ammonia emission in composting of cattle dung wastes, we developed an alternative composting method consisting of a hyperthrmophilic pre-treatment reactor (HTPRT) (first step) combined with a general windrow post-treatment system (WPOT) (second step). In this study, physicochemical and microbiological differences in compost materials during the HTPRT-WPOT process and a simple windrow composing process (SWC) were investigated. The HTPRT-WPOT process removed excess ammonia in the compost materials by physical ammonia stripping, and controlled the malodorous ammonia emission. The organic matter evolution index showed that the HTPRT-WPOT process also contributed to accelerate formation of humic acids in composting. Quantitative real-time PCR analyses using Bacterial-, Archaeal- and fungal-protozoan-specific primer sets showed that small subunit ribosomal RNA (SSU rRNA) gene copy numbers differed much between composting materials of these two processes. Particularly, the SSU rRNA gene copy of eukaryotic microbes (fungi-protozoa) in the HTPRT-WPOT process was much higher than in the SWC process. From these results, we conclude that the HTPRT-WPOT process has great advantages for the control of malodorous odor problems caused by ammonia emission, and for high rate of composting evaluated by the humification rate and microbial characterization of the composting materials.