Synthesis and evaluation of a novel 99m Tc nitrido radiopharmaceutical with alendronate dithiocarbamate as a potential bone-imaging agent.

Currently, a popular strategy for designing novel radioprobes as bone-imaging agents is based on the concept of bifunctional radiopharmaceuticals. Considering the dithiocarbamate ligand can act as a suitable bifunctional linking agent to attach technetium-99m (99m Tc) to corresponding target molecules, in this study, alendronate dithiocarbamate (ALNDTC) was synthesized and radiolabeled with [99m Tc≡N]2+ core by ligand exchange reaction to produce 99m TcN-ALNDTC complex, for the potential use as a novel probe for bone imaging. The radiochemical purity of the complex was over 90%. The complex was stable in vitro and could bind to hydroxyapatite. The partition coefficient result indicated it was hydrophilic, and an evaluation of biodistribution in mice indicated that the complex exhibited a higher bone uptake than did 99m Tc-labeled methylenediphosphonate (99m Tc-MDP). Further, single photon emission computed tomography imaging study indicated clear accumulation in bone, suggesting that 99m TcN-ALNDTC would be a promising candidate for bone imaging.

Development of the full-length cDNA clones of two porcine epidemic diarrhea disease virus isolates with different virulence.

The recently emerged highly virulent variants of porcine epidemic and diarrhea virus (PEDV) remain a huge threat to the worldwide swine industry. Here, we describe the development of a bacterial artificial chromosome (BAC) reverse genetics system for PEDV based on two recent Chinese field isolates, namely CHM2013 and BJ2011C. Phylogenetically, CHM2013 is closely related to the vaccine strain SM98 whereas the isolate BJ2011C belongs to the GIIb group, a cluster that contains many recent pandemic strains. The full-length cDNA clones of the two isolates were constructed into BAC under the control of CMV promoter. The rescued viruses rBJ2011C and rCHM2013 were found to replicate at the kinetics similar to their respective parental viruses in cell culture. When tested in the 2-day-old pig model, rBJ2011C caused severe diarrhea of piglets with extensive damages to the intestinal epithelium, leading to 100% fatality within 48 hours. In contrast, the rCHM2013-inoculated piglets all survived with only very minor tissue damage observed. Thus, we have successfully established a convenient platform for PEDV genome manipulation. This study also represents the first description of a DNA-launched reverse genetics system for the highly virulent PEDV.

Kit formulation for preparation and biological evaluation of a novel (99m)Tc-oxo complex with metronidazole xanthate for imaging tumor hypoxia.

INTRODUCTION: Achieving an ideal (99m)Tc labeled nitroimidazole hypoxia marker is still considered to be of great interest. Metronidazole xanthate (MNXT) ligand was synthesized and radiolabeled with (99m)Tc-glucoheptonate (GH) to form the (99m)TcO-MNXT complex, for the potential use as a novel probe for imaging tumor hypoxia. METHODS: For labeling, (99m)TcO-MNXT was prepared by ligand-exchange reaction with (99m)Tc-GH. The radiochemical purity of the (99m)TcO-MNXT complex was measured by thin layer chromatography (TLC) and high performance liquid chromatography (HPLC). The distribution coefficient and stability of the complex was investigated. The structure of the (99m)TcO-MNXT complex was verified by preparation and characterization of the corresponding stable rhenium complex. The cellular uptake of the (99m)TcO-MNXT complex was determined in murine sarcoma S180 cell lines under hypoxic and aerobic conditions. The biodistribution and single photon emission computed tomography (SPECT) image studies of the (99m)TcO-MNXT complex were performed in mice bearing S 180 tumor. RESULTS: The radiochemical purity of the (99m)TcO-MNXT complex was over 90%. It had good in vitro stability and its distribution coefficient indicated that it was a hydrophilic complex. When (99m)Tc and Re complexes were coinjected in HPLC, both radioactivity (for (99m)Tc complex) and UV detectors (for Re complex) showed nearly identical HPLC profiles, suggesting their structures are similar. The tumor cell experiment and the biodistribution in mice bearing S 180 tumor showed that the (99m)TcO-MNXT complex had a good hypoxic selectivity and accumulated in the tumor with high uptake and good retention. Single photon emission computed tomography (SPECT) image studies showed that the tumor detection was observable. CONCLUSIONS: (99m)TcO-MNXT is prepared from a kit without the need for purification and shows high tumor uptake, tumor/blood and tumor/muscle ratios, suggesting that it would be a promising candidate for imaging tumor hypoxia.

Preparation and biodistribution of a (99m)Tc tricarbonyl complex with deoxyglucose dithiocarbamate as a tumor imaging agent for SPECT.

The deoxyglucose dithiocarbamate (DGDTC) was successfully labeled with the (99m)Tc(CO)3 core to provide the corresponding (99m)Tc(CO)3-DGDTC complex in good yields. The radiochemical purity of the (99m)Tc(CO)3-DGDTC complex was over 90%, as measured by high performance liquid chromatography (HPLC). The complex possessed good stability in saline at room temperature and in mouse plasma at 37 °C. Its partition coefficient result indicated that it was a hydrophilic complex. The electrophoresis results showed the complex was neutral. The biodistribution of (99m)Tc(CO)3-DGDTC in mice bearing S 180 tumor showed that the complex clearly accumulated in tumor, exhibiting high tumor/blood and tumor/muscle ratios and good tumor retention. Single photon emission computed tomography (SPECT) image studies showed there was a visible uptake in tumor sites, suggesting (99m)Tc(CO)3-DGDTC could be considered as a potential tumor imaging agent.

Synthesis and biodistribution of a new (99m) Tc-oxo complex with deoxyglucose dithiocarbamate for tumor imaging.

The deoxyglucose dithiocarbamate (DGDTC) was radiolabeled with (99m) Tc(V)-glucoheptonate (GH), for the potential use as radiopharmaceuticals for tumor imaging. For labeling, (99m) TcO-DGDTC was prepared by ligand-exchange reaction with (99m) Tc-GH. The radiochemical purity of the (99m) TcO-DGDTC complex was over 90% by thin-layer chromatography and high-performance liquid chromatography, without any notable decomposition at room temperature over a period of 6 h. Its partition coefficient indicated that it was a hydrophilic complex. The ligand-exchange reaction occured at neutral condition and under 100 °C for 15 min to achieve high radiochemical purity. In vitro cell studies showed there was an increase in the uptake of (99m) TcO-DGDTC as a function of incubation time and the cellular uptake of (99m) TcO-DGDTC was possibly mediated by way of a d-glucose mechanism. The biodistribution of (99m) TcO-DGDTC in mice bearing S 180 tumor showed that the complex accumulated in the tumor with good uptake and excellent retention. As compared with other reported (99m) Tc radiolabeled glucose derivatives, (99m) TcO-DGDTC showed the highest tumor uptake and good tumor/muscle ratios. The tumor/muscle ratio of (99m) TcO-DGDTC uptake was higher than that of [(18) F] FDG uptake. Single photon emission computed tomography (SPECT) image studies showed there was a visible accumulation in tumor sites, suggesting (99m) TcO-DGDTC would be a promising candidate for tumor imaging.

Synthesis and biodistribution of a novel (99m)TcN complex of norfloxacin dithiocarbamate as a potential agent for bacterial infection imaging.

Achieving a (99m)Tc-labeled fluoroquinolone derivative as a single photon emission computed tomography (SPECT) tracer is considered to be of great interest. The norfloxacin dithiocarbamate (NFXDTC) was synthesized and radiolabeled with a [(99m)TcN]²(+) intermediate to form the (99m)TcN-NFXDTC complex in high yield. The radiochemical purity of (99m)TcN-NFXDTC was over 90%, as measured by thin layer chromatography (TLC) and high performance liquid chromatography (HPLC), without any notable decomposition at room temperature over a period of 6 h. The partition coefficient and electrophoresis results indicated that (99m)TcN-NFXDTC was lipophilic and neutral. The bacterial binding assay studies showed tht (99m)TcN-NFXDTC had a good binding affinity. Biodistribution results in bacterial infected mice showed that (99m)TcN-NFXDTC had a higher uptake at the sites of infection and better abscess/blood and abscess/muscle ratios than those of (99m)Tc-ciprofloxacin and (99m)TcN-CPFXDTC (CPFXDTC = ciprofloxacin dithiocarbamate). The biodistribution results of (99m)TcN-NFXDTC in bacterially infected mice and in mice with turpentine-induced abscesses indicated that (99m)TcN-NFXDTC was suited to be a bacteria-specific infection imaging agent. Single photon emission computed tomography (SPECT) image studies showed there was a visible accumulation in infection sites, suggesting that it would be a promising candidate for bacterial infection imaging.

Increased liver and spleen accumulation of Tc-99m methylene diphosphonate associated with intravenous injection of MRI contrast gadolinium-diethylenetriaminepentaacetic acid.

Several conditions that can cause diffuse hepatic or splenic uptake of Tc-99m methylene diphosphonate (Tc-99m MDP) have been previously reported. Nevertheless, diffuse abnormal liver and spleen uptake of Tc-99m MDP associated with intravenous injection of magnetic resonance imaging contrast gadolinium-diethylenetriaminepentaacetic acid is not previously known. In our series, we reported diffuse increased Tc-99m MDP activity in the liver and spleen in bone scans in patients who received Tc-99m MDP injection shortly after contrast-enhanced magnetic resonance imaging.

An anti-DLK1 monoclonal antibody produced using ELISA and hybridoma techniques.

DLK1 is a newly identified prognostic factor associated with liver cancer survival. To prepare specific monoclonal antibody (MAb) against DLK1, cDNA of DLK1 was cloned by RT-PCR and inserted into prokaryotic expression vector pGEX-4T1, respectively. The fusion proteins were expressed in Escherichia coli. Monoclonal antibody against DLK1 was obtained with hybridoma technique and specific ELISA screening. Western blotting and immunohistochemistry assays showed that MAb 6D6 had specific binding ability with DLK1 protein in eukaryotic cells and cancer tissues. This MAb will be a helpful tool for the detection of DLK1 protein in the tissues and serum of liver cancer and other cancer patients.

Progenitor gene DLK1 might be an independent prognostic factor of liver cancer.

BACKGROUND: Delta-like 1 homolog (DLK1) is a marker for progenitor cells of the liver. The gene encoding DLK1 is expressed early during embryonic development but, importantly, it is also expressed in some human liver cancers. However, the prognostic value of the DLK1 gene has not been investigated. OBJECTIVES: To examine the association between the DLK1 gene and survival time and whether high levels of expression of DLK1 are a prognostic factor for liver cancer. METHODS: We evaluated 60 cases of primary liver cancer, and investigated the link between the expression of DLK1 and patient survival. Clinical characteristics of the cases used for our study, such as tumor size, differentiation and staging, are statistically evenly distributed. Using RT-PCR, western blotting and immunohistochemistry, we analyzed the expression of DLK1 in the tumor samples and evaluated the results statistically. RESULTS: DLK1 was expressed in 22 of the 60 cases (36.7%), and analysis of the survival of the patients revealed that DLK1-positive patients had a shorter survival time than DLK1-negative patients. Cox regression analysis also showed that DLK1 is a risk factor. However, DLK1 expression does not seem to correlate with other classic prognostic factors such as alpha-fetoprotein (AFP), tumor-node-metastasis (TNM) and vascular invasion, which implies that it is an independent prognostic factor.