In vivo distributions of 111In and/or 3H labeled lymphocyte in C3H/He mouse (author's transl)

Although 99mTc and 51Cr have been used for lymphocyte labeling, these radionuclides have several disadvantages for study on immunological behaviour of lymphocyte; very high rate elution and low labeling efficiency for both radionuclides, and short half life for 99mTc. Indium-111 has quite suitable physical properties for clinical nuclear medicine, i.e. desirable photon energy (247,173 keV) and 2.8 day half life. 111In-oxine is lipid soluble and is known to pass through the cell membrane and attaches firmly to cytoplasmic component of the cell. On the other hand, 3H-thymidine is well known substance which incorporated to nucleic acid in the cell. In this study, distribution patterns of 111In-oxine and/or 3H-thymidine labeled lymphocyte in C3H/He mice were examined and the suitability of 111In-oxine labeled lymphocyte for radionuclide imaging in vivo was discussed. Thirty minutes after intravenous injection of 3H and/or 111In labeled lymphocyte, about 12% of lymphocyte were found in the lungs and rest of them were distributed mainly in the blood, kidneys and liver. After 24 hours the activity in the lung decreased markedly and the activity in the liver and kidneys increased with time. Between lymphocyte labeled with 111In-oxine and 3H-thymidine, there is not so much differences in terms of distribution patterns. From this study, it is concluded that the 111In-oxine labeled lymphocyte distributes in the same way as 3H labeled one, in spite of different labeling sites. This 111In-oxine labeling method can be used as a useful tool of radionuclide imaging in kinetic studies of lymphocyte in vivo.     

The study of a two-dimensional polar representation of the cardiac SPECT image--a new method of displaying left ventricular wall motion

A new method was developed to display left ventricular wall motion using two dimensional polar representation of cardiac SPECT image. After intravenous administration of 740 MBq (20 mCi) 99mTc (in vivo labeling of red blood cell), ECG-gated SPECT image of cardiac pool was recorded (32 directions, 180 degrees). The short axis images of left ventricle were constructed at end-diastole (ED) and end-systole (ES). Then the functional images of (ED-ES)/ED were calculated at each compatible slices and displayed in color according the degree of wall motion. In 8 cases with cardiac diseases this method was applied and clinically useful functional images could be obtained. The area of akinesia or hypokinesia are successfully demonstrated clearly separated from the area of normokinesia. We conclude that this method is useful to show left ventricular wall motion and to evaluate the segment and grade of abnormal wall motion of left ventricle.     

Characterization of technetium-99m complexes of pentane-2,4-dione bis(N-methylthiosemicarbazone)

Further characterization of the two neutral technetium-99m (99mTc) complexes of pentane-2,4-dione bis-(N-methylthiosemicarbazone) (PETS) was carried out using a new dianionic PETS derivative, 3,3-dimethyl-pentane-2,4-dione bis(N-methylthiosemicarbazone) (DM-PETS), and the well characterized 99mTc complex of 2,2,9,9-tetramethyl-4,7-diaza-1,10-decanedithiol (DADT) as references. While PETS generated two neutral 99mTc complexes, 99mTc-PETS-L1 and 99mTc-PETS-L2, by both the stannous reduction method and the ligand exchange reaction with six-coordinated 99mTc(V) complex of N,N'-ethylenebis(acetylacetone imine), DM-PETS formed only one neutral 99mTc complex. 99mTc-PETS-L2, the more lipophilic complex of the two 99mTc-PETS, was obtained with a much higher yield than 99mTc-PETS-L1 by the ligand exchange reaction of PETS with the five-coordinated 99mTc(V) complex of glucoheptonate. In addition, while 99mTc-PETS-L2 and 99mTc-DADT remained unchanged in the presence of CN- anions, a breakdown of the original complexes was observed in 99mTc-PETS-L1 and 99mTc-DM-PETS. All four 99mTc complexes exhibited similar brain, heart and pancreas extraction when injected into mice. These cumulative results imply that 99mTc-PETS-L1 and 99mTc-DM-PETS are six-coordinated mononuclear 99mTc(V) complexes and that 99mTc-PETS-L2 is a five-coordinated mononuclear 99mTc(V) complex. These results also suggest that while the chelate ring structure of the 99mTc-dithiosemicarbazone (DTS) chelate played a significant role in its stability, ionization of the third proton of the PETS molecule and the subsequent resonating structure afforded further stability to the 99mTc-PETS complex. Markedly high lipophilicity of the 99mTc-PETS-L2 may also be explained by assuming that 99mTc-PETS-L2 is the five-coordinated resonating structure.     

甘草次酸修饰海藻酸钠纳米给药系统的肝靶向性评估

通过EDC/NHS偶联反应将疏水性肝靶向小分子甘草次酸(GA)连接到天然多糖海藻酸钠(ALG)上,制备了具有双亲性肝靶向药物载体材料(GA-ALG).采用乳化法对广谱抗癌药物阿霉素(DOX)进行包载,得到肝靶向载药纳米粒子( DOX/GA-ALG NPs).利用单光子发射型计算机断层成像技术(SPECT)和药物体内分布...     

Short-cut method for the correction of light attenuation influences in the experimental data obtained from confocal laser scanning microscopy

Based on Lambert-Beer law and the light attenuation model, a new method will be introduced in this work in order to eliminate the disturbances caused by signal attenuation in the experimental data measured by confocal laser scanning microscopy (CLSM). This new method considers the attenuation effects which depend on concentration of fluorophore-labelled protein as well as attenuation effects which are independent from protein concentration. Furthermore, no solvent additive is required in order to match the refraction index of solvent to bead material. The determination of correction factors is, thus, easily done using the currently investigated chromatographic phase system, so that the validity of the correction factors in the current system can be guaranteed. The introduced correction method has been applied for the investigation of intraparticle protein distribution inside an HIC (hydrophobic interaction chromatography) particle.     

Evaluation of 99mTc-HIDA complex as a cholescintigraphic agent (author's transl)

In the reaction labeling N-(2,6-dimethylphenylcarbamoylmethyl) iminodiacetic acid (HIDA) with 99mTc, several complexes with different chemical characteristics were observed to occur with slight changes in the labeling conditions. Among these complexes, a complex detected in the bile of rats was limited to one complex, named as complex II. The preparation method of 99mTc-HIDA complex II and the exchange reaction between this complex and penicillamine indicate that 99mTc is coordinated with HIDA as low-hydrolyzed 99mTc in this complex. This complex is excreted rapidly through the bile and within 1 hr, about 65% of the total activity injected is recovered from bile in rats. The organ distribution of this complex was studied in mice by radioassay and in rabbits by scintillation camera and, in both cases, the radioactivity was accumulated in the gallbladder. These results suggest that the 99mTc chemical state, low-hydrolyzed state, relates to the bile excretion behavior of this complex, a potentially useful cholescintigraphic agent.     

Determination of residual activities in 99mTc generators and 81mKr generators

The residual activities present in spent 99Mo-99mTc generators and 81Rb-81mKr generators were determined from a view point of the disposal of radiopharmaceuticals. The results showed that seven kinds of residual nuclides such as 95Zr-95Nb, 103Ru, 90Sr, etc. were found in 99mTc generators with maximum level of 740 Bq/3.7 GBq (0.02 muCi/100 mCi) at one year after the reference date. In 81mKr generators one year old, six nuclides such as 57Co, 83Rb, 84Rb and others remained and these activities were less than 3.7 kBq/370 MBq (0.1 muCi/10 mCi). On the basis of these data, the disposal of 99mTc generators and 81mKr generators is discussed.     

Comparison of SPECT images with four kinds of 99mTc collimators

Performance of SPECT imaging systems which use a rotating gamma camera, are affected by characteristics of the detector-collimator assembly, the data acquisition method, and the filter used in image reconstruction. The purpose of this study is to examine image qualities of SPECT with different types of low energy collimators. The SPECT imaging system in this study is a rotating gamma camera ZLC-7500 (Siemens) and a data processing unit Scintipac-700 (Shimadzu). The four types of collimators compared are UHR (ultra high resolution), HR (high resolution), AP (general all purpose), and HS (high sensitivity), with 0.27, 0.66, 1.00, and 2.06 relative sensitivity, respectively. In the case of the same collimator, the spatial resolutions measured in the slice plane showed a slight difference in the FWHM values (mean values of UHR, HR, AP, and HS were 11.3 mm, 13.6 mm, 15.8 mm, and 20.4 mm, respectively.) between the center and the circumference of the field of view, in the radial direction, but a large difference in the tangential direction, with lower FWHM values (values of UHR, HR, AP, and HS were 8.4 mm, 8.7 mm, 9.3 mm, and 10.8 mm at 12 cm from the center, respectively.). In comparison of SPECT images with the four types of collimators, except for the HS collimator, image qualities of UHR, HR, and AP collimators showed only a slight difference. From the pont of view of sensitivity and spatial resolution of the collimator, it is expected that the AP collimator would be suitable for SPeCT imaging with 99mTc.     

Evaluation of additional absorbed radiation dose from radioactive impurities in radiopharmaceuticals

The additional absorbed radiation doses to total body from radioactive impurities in radiopharmaceuticals were estimated for a total of 10 samples (i.e., 99mTc generator eluates, 99mTc solutions, 67Ga, 81mKr generator eluates, 111In, 123I, 201Tl, 75Se, 197Hg, 198Au) selected from commonly used radiopharmaceuticals. The radioactive impurities in radiopharmaceuticals used in this estimation were previously identified. The radiation doses were calculated according to the MIRD procedures, assuming each radionuclide to be uniformly distributed throughout the body. In the case of the cyclotron-produced radioisotopes and 198Au, the additional total body absorbed doses from impurities were found to be rather high: 280% for 123I, 55% for 81mKr, 5% for 198Au, 4% for 111In and 2% for 201Tl. The absorbed doses due to impurities in the other radioisotopes were less than 1%.     

Indium-111 labeled bleomycin for targeting diagnosis and therapy of liver tumor: optimized preparation,biodistribution and SPECT imaging with xenograft models

In this work, the preparation of ¹¹¹In radiolabeled bleomycin (¹¹¹In-BLM) was optimized systematically and used for SPECT imaging of liver cancer xenograft models. ¹¹¹In-BLM with a high radiochemical yield (>?99%) could be obtained at pH 0.5–1 at a mixture ratio of 9:1 (¹¹¹In/BLM, mCi/mg). The radiochemical purity of ¹¹¹In-BLM retained?>?99% in either buffers or serum for 3 days. Biodistribution of ¹¹¹In-BLM revealed its excellent stability in vivo. The SPECT imaging studies showed ¹¹¹In-BLM has great specificity to liver tumor xenograft. All the results implied ¹¹¹In-BLM is a potential radiopharmaceutical for targeting diagnosis and therapy of liver tumor.

     

Technical study for SPECT of the head using slant hole collimator

The performance of a 160 keV slant hole collimator (160 keV SH) was compared with that of a 140 keV high resolution collimator (140 keV HR) for SPECT imaging of the head using 99mTc. The minimum radius of revolution was reduced from about 23 cm to 13 cm using 160 keV SH. Relative sensitivity of the 160 keV SH was 108% of that of the 140 keV HR. Compared with SPECT using 140 keV HR, 160 keV SH SPECT yielded approximately a 15% increase in resolution. The 160 keV SH SPECT images of phantom were superior in resolution and contrast to that of 140 keV HR SPECT. We concluded that high quality SPECT images of the head can be obtained from rotating gamma cameras equipped with slant hole collimators.     

High performance liquid chromatography of 99mTc labeled human serum albumin using an N-methylpyridinium polymer column.

Human serum albumin (HSA) labeled with 99mTc (99mTc-HSA) was analyzed by high performance liquid chromatography using a 4-vinylpyridinium polymer column which specifically resolved albumin components such as human mercaptalbumin (HMA), human non-mercaptalbumins (HNA), etc. The 99mTc-HSA radiochromatogram revealed that 99mTc-HSA consisted of several components. The radiochromatographic profile was similar to that of 99mTc-HMA prepared with 99mTc and separated HMA. This suggested that HMA participated mainly in 99mTc-labeling of HSA. When HSA was labeled with a stoichiometric concentration of 99Tc, the HMA peak was significantly decreased and new peaks were revealed by absorbance at 280 nm. From these results, the role of HMA in labeling HSA with 99mTc was elucidated.     

99mTc-2-mercaptopropionylglycine: different biliary excretion behavior in different animal species (author's transl)

Labeling of 2-mercaptopropionylglycine (2-MPG) with 99mTc, was studied and its chemical characteristics were examined. Further, biliary excretion behavior of this complex was comparatively estimated in mice, rats, and rabbits. 99mTc-2-MPG was rapidly excreted in large quantities into the bile in mice and rats: within 1 hr after injection, 51% of the injected dose was recovered from the bile in rats. On the other hand, the ligand exchange reaction between this complex and penicillamine indicates that a low hydrolyzed 99mTc species is coordinated with 2-MPG. These results suggest that a low hydrolyzed 99mTc is an effective feature in biliary excretion behavior of 99mTc compounds. Another interesting in vivo behavior of 99mTc-2-MPG is the difference observed in mice and rabbits: in mice, very high 99mTc activity is concentrated in the gallbladder and the clearance from tissues other than the gallbladder is rapid, whereas in rabbits, although a rapid and high excretion into the gallbladder is observed, a considerable high 99mTc activity is retained in the liver and the kidney. One reasons for this different in vivo behavior is the low stability of this complex at high dilution: a big animal has the large dilution volume which lead to higher decomposition estimated by the higher liver and kidney retention or the lower bile excretion. In conclusion, studies carried on 99mTc-2-MPG showed a good biliary excretion behavior but an in vivo unstableness in big animals.     

"Click-to-chelate": design and incorporation of triazole-containing metal-chelating systems into biomolecules of diagnostic and therapeutic interest

The site-specific conjugation of metal chelating systems to biologically relevant molecules is an important contemporary topic in bioinorganic and bioorganometallic chemistry. In this work, we have used the CuI-catalyzed cycloaddition of azides and terminal alkynes to synthesise novel ligand systems, in which the 1,2,3-triazole is an integral part of the metal chelating system. A diverse set of bidentate alkyne building blocks with different aliphatic and aromatic backbones and various donor groups were prepared. The bidentate alkynes were reacted with benzyl azide in the presence of a catalytic amount of CuI to form tridentate model ligands. The chelators were reacted with [ReBr3(CO)3]2- to form well-defined and stable complexes with different overall charges, structures and hydrophilicities. In all cases tridentate coordination of the ligands, including through N3 of the 1,2,3-triazole ring, was observed. The ligand systems could also be quantitatively radiolabelled with the precursor [99 mTc (H2O)3(CO)3]+ at low ligand concentrations. Similarly the alkynes were reacted with an azido thymidine derivative to form a series of compounds, which could be radiolabelled in situ to form single products. Subsequent incubation of the neutral and cationic organometallic 99 mTc thymidine derivatives with human cytosolic thymidine kinase, a key enzyme in tumour proliferation, revealed that only the neutral compounds maintained substrate activity towards the enzyme. Bioconjugation, radiolabelling and enzymatic reactions were successfully performed in a matter of hours. Thus, click chemistry provides an elegant method for rapidly functionalising a biologically relevant molecule with a variety of efficient metal chelators suitable for (radio)labelling with the M(CO)3 core (M=99 mTc, Re), to offer new potential for technetium-99 m in clinical and preclinical tracer development.     

Experimental considerations of myocardial single photon emission CT image with a cardiac phantom

We studied the quantity of thallium-201 myocardial single photon emission CT (SPECT) image with a cardiac phantom. The myocardial SPECT image is influenced by several causes, i.e., the absorption of gamma rays, collimator aperture, statistical noise and limited angle of rotation in data acquisition. The attenuation correction under the assumption of uniform distribution is not sufficient, so we have to consider the actual distribution of absorption. The effects of the nonuniform attenuation are most prominently appeared when we evaluate the quantity by the integral method. The aperture characteristics of a collimator are decided by the distance between the rotational center of the gamma camera and the collimator surface. Simultaneously scattered gamma rays affect the reconstructed image in the low frequency component, so the effective attenuation coefficient varies gradually. The statistical noise intermixed in the projection data generate artifacts like as lump-shaped pattern. The noise are clearly appeared in the spatial frequency upper than 0.25 cycle, so the low-pass filter are required with that of cut-off frequency. Moreover to enhance the details of the radionuclide distribution of cardiac muscle, the Wiener filter should be applied. The scan area also affects the appearance of the artifact in the myocardial SPECT image. The 180-degree scan usually generates false defects just nearby the center of reconstructed cross section. And the positions are changed with the conditions of gamma ray absorber and its relative position to the myocardial area. The 360-degree scan should be used from the standpoint of reconstructing more quantitative image.     

Rhenium analogues of promising renal imaging agents with a [99mTc(CO)3]+ core bound to cysteine-derived dipeptides, including lanthionine

The coordination chemistry of lanthionine (LANH2) and cystathionine (CSTH2) dipeptides, which respectively consist of two cysteines and one cysteine and one homocysteine linked by a thioether bridge, is almost unstudied. Recently for fac-[99mTc(CO)3(LAN)]- isomers, the first small 99mTc(CO)3 agents evaluated in humans were found to give excellent renal images and to have a high specificity for renal excretion. Herein we report the synthesis and characterization of Re complexes useful for interpreting the nature of tracer 99mTc radiopharmaceuticals. Treatment of [Re(CO)3(H2O)3]OTf with commercially available LANH2 (a mixture of meso (d,l) and chiral (dd,ll) isomers) gave three HPLC peaks, 1A, 1B, and 1C, but treatment with CSTH2 (l,l isomer) gave one major product, Re(CO)3(CSTH) (2). Crystalline Re(CO)3(LANH) products were best obtained with synthetic LANH2, richer in meso or chiral isomers. X-ray crystallography showed that these dipeptides coordinate as tridentate N2S-bound ligands with two dangling carboxyls. The LANH ligand is meso in 1A and 1C and chiral in 1B. While 1A (kinetically favored) is stable at ambient temperature for days, it converted into 1C (thermodynamically favored) at 100 degrees C; after 6 h, equilibrium was reached at a 1A:1C ratio of 1:2 at pH 8. The structures provide a rationale for this behavior and for the fact that 1A and 1C have simple NMR spectra. This simplicity results from fluxional interchange between an enantiomer with both chelate rings having the same delta pucker and an enantiomer with both chelate rings having the same lambda pucker. Agents with the [99mTc(CO)3]+ core and N2S ligands show promise of becoming an important class of 99mTc radiopharmaceuticals. The chemistry of Re analogues with these ligands, such as the LAN2- complexes reported here, provides a useful background for designing new small agents and also tagged large agents because two uncoordinated carboxyl groups are available for conjugation with biological molecules such as proteins.     

An efficient Monte Carlo algorithm for the fast equilibration and atomistic simulation of alkanethiol self-assembled monolayers on a Au(111) substrate

A new Monte Carlo algorithm is presented for the simulation of atomistically detailed alkanethiol self-assembled monolayers (R-SH) on a Au(111) surface. Built on a set of simpler but also more complex (sometimes nonphysical) moves, the new algorithm is capable of efficiently driving all alkanethiol molecules to the Au(111) surface, thereby leading to full surface coverage, irrespective of the initial setup of the system. This circumvents a significant limitation of previous methods in which the simulations typically started from optimally packed structures on the substrate close to thermal equilibrium. Further, by considering an extended ensemble of configurations each one of which corresponds to a different value of the sulfur-sulfur repulsive core potential, sigmass, and by allowing for configurations to swap between systems characterized by different sigmass values, the new algorithm can adequately simulate model R-SH/Au(111) systems for values of sigmass ranging from 4.25 A corresponding to the Hautman-Klein molecular model (J. Chem. Phys. 1989, 91, 4994; 1990, 93, 7483) to 4.97 A corresponding to the Siepmann-McDonald model (Langmuir 1993, 9, 2351), and practically any chain length. Detailed results are presented quantifying the efficiency and robustness of the new method. Representative simulation data for the dependence of the structural and conformational properties of the formed monolayer on the details of the employed molecular model are reported and discussed; an investigation of the variation of molecular organization and ordering on the Au(111) substrate for three CH3-(CH2)n-SH/Au(111) systems with n=9, 15, and 21 is also included.     

Scintigraphic evaluation of brain death with 99mTc-d,l-hexamethyl-propyleneamine oxime (HMPAO)

Lately, the criteria of brain death is being discussed. Cerebral scintigram, especially scintigraphic evaluation of brain death by dynamic study, has been previously reported. Cerebral imaging using radiolabeled amines such as 123I-IMP N-isopropyl-p-iodoamphetamin (IMP) or 99mTc d,l-hexamethyl-propyleneamine oxime (HMPAO) offers a significant information of brain death by the finding of "non visualized brain". However, the dynamic scintigram acquired during the bolus injection of 99mTc-HMPAO provides an additional information of brain death by classical "hot nose sign". 99mTc-HMPAO is able to be administered relatively in a large amount of dose. This cerebral perfusion tracer is lipophilic and remains in the central nervous system, which characterize its role as an reliable indicator of cerebral blood flow. As a result, this compound became suitable for the non-invasive study of brain circulation when the diagnosis of brain death is uncertain. We report a case of brain death in which diagnosis was made by the classical "hot nose sign" in dynamic scintigraphy performed when 99Tc-HMPAO was injected as well as the SPECT which showed a lack of cerebral visualization at the equilibrium state. As far as we are informed, this additional procedure used in the diagnosis of brain death has not reported before. The importance of performing a dynamic scintigram at the administration of 99mTc-HMPAO is also discussed in this report.     

Evaluation of the system performance and clinical images of the single photon emission computed tomography for head using ring arranged detector

To evaluate the system performance, several preoperational fundamental tests of single photon emission computed tomography (SPECT) were carried out. Spatial resolutions (FWHM) measured with the point-spread functions of a 99mTc line source were 12.5 mm with a high resolution (HR) collimator and 17.2 mm with a high sensitivity (HS) collimator respectively. Slice thicknesses (FWHM) obtained from the profile curves of slice images were 17.5 mm (HR) and 29.0 mm (HS) at the center of rotation. System sensitivities were 5.4 kcps/slice (HR) and 27.8 kcps/slice (HS). Uniformities calculated from the SPECT images of a pool phantom were 4.7% (HR) and 2.7% (HS) at the condition of 3,000 kcounts to be acquired. SPECT images of the HEADTOME SET-031 were considered very useful to diagnose the cerebrovascular disease.     

Stable and lipophilic technetium-99m dithiosemicarbazone complexes with 5-6-5 membered chelate ring structure

Modification of the chelate ring structure of technetium-99m (99mTc) dithiosemicarbazone (DTS) chelate was carried out in pursuit of a more stable and lipophilic compound. A new DTS chelating molecule, pentane-2,4-dione bis(N-methylthiosemicarbazone) (PETS), with a 5-6-5 membered chelate ring structure, was synthesized and labeled with 99mTc, PETS generated two 99mTc compounds as major products. Both had much higher stability and lipophilicity than a 5-5-5 membered 99mTc DTS compound, as well as great stability in plasma. Both 99mTc-PETS compounds were rapidly extracted by the brain and heart when injected into mice. Thus, the modified chelate ring structure afforded a preferable characteristics to DTS chelate as for the chelating site for technetium radiopharmaceuticals.