Significance of alteration in biodistribution of labeled lymphocytes exposed to stannous ion

The biodistribution patterns of ^99mTc (^99mTc-lymph) and ¹¹¹In-lymphocytes with [¹¹¹In-(Sn)-lymph] or without (¹¹¹In-lymph) stannous ion treatment was compared in Lewis rats. Syngeneic lymphocytes were labeled with either 125 Ci (4.63 MBq) ^99mTc or 5 uCi (1985 kBq) ¹¹¹In per 2×10⁷ cells. Mean labeling efficiency for ^99mTc and ¹¹¹In was 68.61%±3.90% (SEM) and 87.22%±2.01% (SEM) respectively. ^99mTc-lymph (n=4), ¹¹¹In-lymph (n=6) and ¹¹¹In-(Sn)-lymph (n=6) rats received 2x10⁷ cells and were killed 18 h later. While ^99mTc-lymph demonstrated significantly less localization in spleen, lymph nodes, and blood (P(F)0.01) as compared with ¹¹¹In-lymph, ¹¹¹In-(Sn)lymph also demonstrated a significant difference (P[F]=0.0001) in lymph node accumulation when compared to ¹¹¹In-lymph. As the activity levels utilized are not associated with cell radiation damage, these alterations in biodistribution do not reflect viability or chromosomal damage, but appear related to stannous ion exposure.Support from the American Heart Association-Virginia Affiliate     

99Tcm标记双半胱氨酸-脱氧葡萄糖的小鼠实验研究

目的研究^99Tc^m标记双半胱氨酸一脱氧葡萄糖(EGDG)在正常小鼠及荷S180肉瘤小鼠体内的分布规律。方法正常小鼠及荷S180肉瘤小鼠各铝只，分8组，实验前禁食。尾静脉注射^99Tc^m-EC-DG后计算不同时间在小鼠体内的分布及肿瘤与血液、肌肉、肺、肝的放射性比值。同时，取荷瘤小鼠5只，在注射后不同时间进行显像，计算T／NT比值。结果肿瘤组织与血液、肌肉、肺的放射性比值随时间延长逐渐上升，在4h都超过1．0。显像示随时间延长瘤体周围组织放射性逐渐下降，瘤体在4h时显影清晰。结论^99Tc^m-EC-DG可用于肿瘤的葡萄糖代谢显像。     

Deuterioglucose: alteration of biodistribution by an isotope effect

Carbon-14 glucose, in which all carbon-hydrogen bonds were replaced by carbon-deuterium bonds (deuterioglucose), was extracted from algae growing in heavy water and exposed to [14C]carbon dioxide. The identity of [14C]deuterioglucose was confirmed by comparison with authentic material on two high performance liquid chromatography and two thin layer chromatography systems. Fermentation to lactate followed by oxidative decarboxylation demonstrated that 35% of the 14C was on carbons 3 and 4 for deuterioglucose isolated from a 24-hr algal incubation, and 61% for a 20-min incubation. Mice were injected intravenously with either (24-hr) deuterioglucose or with 14C-labeled (protio)glucose labeled uniformly. The deuterioglucose cleared more slowly from the blood, while heart and brain accumulated label more slowly. Tissue concentrations peaked at later times for deuterioglucose. Deuteration may be a useful feature of radiopharmaceutical design.     

Pharmacokinetics and biodistribution of a small radioiodine labeled nerve growth factor fragment

Nerve growth factor (NGF) exerts various actions on neuronal and non-neuronal tissues and has potential therapeutic utility, but difficulties in using the whole protein have stimulated interest in small NGF fragments. We radioiodinated a small cyclic peptide derived from NGF using the Bolton-Hunter method [125I-C(92-96)], and confirmed binding to high affinity NGF receptors by cross-linkage analysis. Pharmacokinetic characteristics in intravenously injected mice were T 1/2 alpha 5.2 min, T 1/2beta 121.3 min, clearance 11.8+/-0.5 ml/min, and volume of distribution 69.7+/-4.6 ml. Dose-proportionate increases in areas-under-curve and peak-concentrations indicated linear pharmacokinetics. Biodistribution data revealed that clinically relevant doses allowed C(92-96) accumulation sufficient to elicit biological responses in receptor expressing organs including the lungs, liver, spleen, and pancreas.     

Sodium pertechnetate (Na99mTcO4) biodistribution in mice exposed to cigarette smoke

BACKGROUND: The biological effects of cigarette smoke are not fully known. To improve our understanding of the action of various chemical agents, we investigated the biodistribution of sodium pertechnetate (Na99mTcO4) in mice exposed to cigarette smoke. METHODS: Fifteen BALB/c male mice were exposed to the smoke of nine whole commercial cigarettes per day, 3 times/day, for up to 10 days to whole body exposure in a chamber. A control group of 5 BALB/c male mice was sham-smoked. One day later, the exposed and control groups of mice received (7.4 MBq/0.3 ml) of Na99mTcO4 before being killed at 30 min. Bones, brain, heart, intestine, kidney, liver, lungs, muscle, pancreas, spleen, stomach, testis and thyroid were weighed and these organs and blood radioactivity recorded with a gamma counter. The percentage per gram of tissue of injected dose (%ID/g) was determined for each organ. RESULTS: Cigarette smoke significantly decreased (p < 0.05) the %ID/g in red blood cells, bone, kidney, lung, spleen, stomach, testis and thyroid of the exposed mice. CONCLUSION: The toxic effects of cigarette smoke reduced the Na99mTcO4 biodistribution.     

Synthesis and biodistribution of radioarsenic labeled dimethylarsinothiols: derivatives of penicillamine and mercaptoethanol

Arsenic analogs of sulfhydryl containing biomolecules can be derived from dimethylchloroarsine as a precursor. Arsenic-76 labeled dimethylarsinothiols (dimethylarsinopenicillamine and dimethylarsinomercaptoethanol) were synthesized, purified by chromatography, and their biodistributions obtained in mice. The present study demonstrates the possibility of developing a group of radioarsenicals from SH-containing biomolecules.     

Preparation and biological distribution of technetium diphosphonate radiotracers synthesized without stannous ion

Two HEDP complexes of technetium (either Tc-99 or a mixture of Tc-99 and Tc-99m) have been prepared without the use of stannous ion. The first, Tc(NaBH4)-HEDP, is synthesized by reduction of TcO4- with NaBH4 in the presence of excess HEDP; this is analogous to the preparation of Tc(Sn)-HEDP in commercial kits wherein SN(II) functions as the reductant. The second, Tc-HEDP, is prepared by substitution of HEDP onto the pre-formed, pre-reduced, technetium center TcBr62-. The HEDP-to-Tc ratio in Tc-HEDP was found to be 1.0 by double-labeling procedures (Tc-99 and [3H]HEDP), implying that in solution this material is polymeric or at least dimeric. Preparations of Tc(NaBH4)-HEDP and Tc-HEDP with Tc-99m are excellent bone-imaging agents in both rats and dogs. Tissue distribution studies in rats show that uptake of Tc(NaBH4)-HEDP and Tc-HEDP by the bone is at least equivalent to that achieved by Tc(Sn)-HEDP prepared in commercial kits with Sn(II) as the reductant. Tin is therefore not necessary for the bone-seeking properties of Tc(Sn)-HEDP, and the in vivo distribution of a given HEDP radiotracer seems to depend primarily on the presence of the HEDP ligand and not on the exact nature of the technetium complex itself. Synthesis of technetium radiotracers by a substitution route, rather than by redox, is practicable; this route has the potential of introducing hitherto unattainable flexibility and subtlety into the preparation of technetium radiotracers.     

The rabbit biodistribution of a therapeutic dose of zoledronic acid labeled with Tc-99m

The aim of the present study was to label a therapeutic dose of zoledronic acid (ZOL) with Tc-99m, evaluate its in vitro stability and compare its biodistribution to ^99mTc-methylene biphosphonate (^99mTc-MDP) in normal rabbits. Preparation of 0.50 mg of ^99mTc-ZOL was carried out by the reduction of ^99mTc-pertechnetate in the presence of stannous chloride. The radiolabeling efficiency was found to be greater than 99%. The labeled complex was stable at least up to 6 h at room temperature determined by paper chromatography. ^99mTc-ZOL and ^99mTc-MDP were administered intravenously to the rabbits for scintigraphic studies. Between ^99mTc-ZOL and ^99mTc-MDP, there were no significant differences in the ratios of femur/BG and lumbar vertebrae/BG, whereas epiphysis/BG and the kidney/BG ratios of ^99mTc-MDP were higher than ^99mTc-ZOL at the static studies.     

mBAND analysis of chromosome aberrations in lymphocytes exposed in vitro to alpha-particles and gamma-rays

PURPOSE: To investigate the profiles of chromosome damage induced in vitro by exposure to alpha-particles and gamma-rays. MATERIALS AND METHODS: Human peripheral blood lymphocytes were exposed to three dose regimes: alpha-particle doses of 0.2 and 0.5 Gy and a gamma-ray dose of 1.5 Gy. After culturing for 47 hours, chromosome aberrations involving the number 5 chromosomes were identified using a multi-coloured banding (mBAND) technique. RESULTS: Analysis of the frequencies of chromosome 5 breaks within aberrant cells and within aberrant number 5 chromosomes demonstrated that alpha-particle irradiation is more likely to result in multiple breaks in a chromosome than gamma-irradiation. Additionally, overdispersion was observed for all doses for the distribution of breaks amongst all cells analysed and breaks amongst total number 5 chromosomes, with this being greatest for the 0.2 Gy alpha-particle dose. The ratio of interchanges to intrachanges (F ratio) was 1.4 and 2.4 for 0.2 and 0.5 Gy alpha-particles respectively and 5.5 for 1.5 Gy gamma-rays. Evaluation of simple versus complex exchanges indicated ratios of 1.9 and 2.7 for 0.2 and 0.5 Gy alpha-particles respectively and 10.6 for 1.5 Gy gamma-rays. The majority of the intrachanges involving chromosomes 5 induced by alpha-particle radiation were associated with more complex exchanges. CONCLUSIONS: This study has confirmed that exchanges induced by exposure to high linear energy transfer (LET) alpha-particle radiation comprise a greater proportion of intrachanges than those induced by exposure to low LET gamma-rays. However, since the majority of these are associated with complex rearrangements and likely to be non-transmissible, this limits their applicability as a marker of past in vivo exposure.     

Electrolytically labeled [99mTc]MDP: chromatographic pattern, stability and biodistribution in rats

The labeling of methylene diphosphonate with 99mTc is possible after reduction of pertechnetate by means of controlled potential electrolysis. This results in a [99mTc]MDP complex which differs slightly from 99mTc(Sn)-MDP in paper and gel chromatography. The scintigraphic images of both preparations are comparable in quality. Biodistribution in rats shows a higher bone uptake for the 99mTc(Sn)-MDP complex, whereas [99mTc]MDP shows a higher uptake in the gastric wall.     

Primary lung cancer: biodistribution and dosimetry of two In-111- labeled monoclonal antibodies

This study was undertaken to measure the biokinetics and organ dosimetry of indium-111-labeled monoclonal antibodies (MoAbs) with a whole-body gamma camera imaging technique. Twenty patients with primary lung cancer were studied with two different MoAb agents (anti-carcinoembryonic antigen ZCEO25 and antiadenocarcinoma LA20207). Imaging was performed at 1, 24, 72, and 144 hours after injection. Scintigraphic whole-body retention was verified by means of comparison with the results from in vitro counting of excreta. Organ retention was verified in an abdominal phantom. The MoAb cleared slowly from the heart and lungs, the brain and spleen showed no clearance, and the liver showed increased activity over the 6-day period. Dosimetry for ZCE025 showed a dose to the liver of 1.3 rad/mCi (0.36 mGy/MBq); heart, 1.5 rad/mCi (0.40 mGy/MBq); spleen, 1.1 rad/mCi (0.29 mGy/MBq); total body, 0.49 rad/mCi (0.13 mGy/MBq); and testes, 0.39 rad/mCi (0.11 mGy/MBq). The dosimetry for LA20207 was similar.     

^99Tc^m标记RGD环肽在荷肺腺癌裸鼠中的显像研究

目的制备^99Tc^m标记的含RGD序列的^99Tc^m-联肼尼克酰胺（HYNIC）-c（RGDfK）环肽单体,评价其在整合素表达阳性的肺腺癌严重联合免疫缺陷（SCID）小鼠肿瘤模型中的生物学分布,并进行显像研究。方法（1）以HYNIC为双功能螯合剂,以三羟甲基甘氨酸（tricine）和乙二胺二乙酸为协同配体,采用二步法制备^99Tc^m标记HYNIC—c（RGDfK）,进行细胞结合实验,测定标记物生物学活性;（2）将荷A549肺腺癌模型小鼠分为7组[第7组作为竞争性抑制组,注射显像剂前0．5h先注射HYNIC-c（CRDGfk）100μg],每组5只,经尾静脉注射7．4MBq的^99Tc^m-HYNIC-c（RGDfK）,于注射后0．5,1,2,4,8,12h处死,计算荷A549肺腺癌小鼠模型各脏器％ID／g,同时采用ROI技术研究^99Tc^m-HYNIC—c（RGDfK）在小鼠体内的生物学分布,计算不同时间点的T／NT比值（NT选取肌肉）;（3）取6只荷瘤裸鼠,其中3只为竞争性抑制组,经尾静脉注射7．4MBq的^99Tc^m-HYNIC—c（RGDfK）,于注射后0．5,1,2,4,8,12h进行静态1显像。结果^99Tc^m-HYNIC—c（RGDfK）的标记率〉90％,放化纯〉95％。^99Tc^m-HYNIC—c（RGDfK）与A549肺腺癌细胞特异性结合率最高为36．14％,体内分布实验显示^99Tc^m-HYNIC—c（RGDfK）在肾的摄取率始终高于20％ID／g,注射后0．5h肿瘤％ID／g为10．52±1．48,8h为17．26±2．81,12h为8．93±0．90,竞争性抑制组注射后0．5h为2．29±0．85。通过ROI技术测得T／NT在8h达6．87。注射后1h肿瘤可显影,4～8h显影更清晰。结论^99Tc^m标记HYNIC—c（RGDfK）易于制备,具有良好的靶向性。     

Preparation and properties of 99Tcm-exametazime using stannous ion augmentation of fractionated cold kits

Fractionation of 99Tcm-exametazime enables several patients doses to be prepared from one vial. To overcome the disadvantages of storage at -70 degrees C, storage at higher temperatures was investigated with replacement of stannous ions before use. Kits were fractionated using a nitrogen-purged 0.9% w/v sodium chloride injection, sub-dispensed into five nitrogen-filled vials in an aseptic environment and stored at -20 degrees C. Under these conditions, exametazime vials failed quality control in approximately 1 week. However, the addition of stannous pyrophosphate solution (10 micrograms Sn(II) in 0.1 ml, prepared by aseptic dilution of a red blood cell labelling kit) immediately before 99Tcm-pertechnetate produced 90.1 +/- 1.6% (n = 8) lipophilic complex with a normal rate of decomposition. Biodistribution studies of this product in mice compared with the full kit showed identical brain and other organ uptakes. This product was used for cerebral imaging in patients with 90.5 +/- 3.3% lipophilic complex (n = 647) at approximately 3 min and produced excellent clinical results. It is concluded that stannous ion augmentation ('reboot') using approved Sn(II)-pyrophosphate kits is a viable option for institutions with facilities for aseptic preparation of fractionated exametazime kits, enabling substantial cost savings. Other advantages include removal of restrictions on generator eluate and 'rejuvenation' of expired kits.     

The stability of 99Tcm directly labelled to an Fab' antibody via stannous ion and mercaptoethanol reduction.

The anti-CEA FO23C5 F(ab')2 antibody was directly radiolabelled with 99mTcm by two methods (stannous ion and mercaptoethanol reduction) and compared in vitro and in vivo for label stability. By both methods, reduction of the F(ab')2 fragment produced primarily Fab' fragments. By both methods, the label was stable to 99Tcm-pertechnetate formation in vitro. Analysis by high performance liquid chromatography (HPLC) of serum, urine, kidney and liver homogenates from mice injected with 99Tcm-antibodies by both methods consistently showed a prominent radiolabelled peak with an estimated molecular weight of about 300 daltons. An identical peak was observed in the analysis of patient samples in a related investigation from this laboratory. Cysteine was radiolabelled with reduced 99Tcm and analysed by HPLC and thin layer chromatography (TLC); one of the 99Tcm-cysteine species so produced showed the same chromatographic behaviour as that of the 300 dalton species. In conclusion, the FO23C5 and other antibodies are stably labelled with 99Tcm via either stannous ion or mercaptoethanol reduction. In mice and in patients, the labelled proteins are either catabolized or, more likely, the 99Tcm label is transchelated such that the label is present on several low molecular weight species, the most prominent of which is postulated to be 99Tcm-cysteine.     

Synthesis, biodistribution and quantitative structure-activity relationship studies of new 99mTc labeled pseudo-peptide complexes.

Twelve new peptide or pseudo-peptide chelators have been synthesized in the course of our continuing investigation of 99mTc-labelled peptides for application for renal imaging agents. All compounds were characterized on the basis of, IR, 1HNMR, 13CNMR spectroscopy, as well as MS and elemental analysis. All peptides yield stable 99mTc complexes using Sn(II) reactive coupling and exhibit renal uptake. Linear regression analysis between Logarithm of renal uptake value (RU) and the parameters obtained by the ZINDO/1 method was performed. Some equations were obtained which showed that molecular polar and charge have some relationship with their renal uptake.     

Labeling of human lymphocytes with 99mTc by means of stannous pyrophosphate. Scintigraphic applications

An original ^99mTc-labeling method applied to human lymphocytes is described. This technique is based on the use of stannous pyrophosphate to reduce sodium pertechnetate. The proposed procedure has two main advantages: firstly, the labeling process takes place in a neutral and buffered medium which prevents any cellular aggregation; secondly, the labeling yield obtained is compatible with scintigraphic studies in man. The influence of each parameter on the labeling quality had been studied as well as the binding stability of the technetium fixed on cells. A systematic study of lymphocyte surface markers, before and after labeling, has led to the suggestion that the labeling process seems to affect cellular membranes, probably because of the technetium binding. Finally, scintigraphic results are presented, the study being performed on eight healthy volunteers. These results are compared with those published by other authors who used either a different radioisotope or a different labeling method.     

99Tcm标记反义肽核酸探针的制备及其荷瘤裸鼠体内分布

目的 探讨99Tcm标记反义肽核酸(PNA)探针的新方法及其在生物体内的分布.方法 合成12mer且5'端含有四肽G-(D)-A-G-G的c-myc mRNA反义、无义PNA片段,利用G-(D)-A-G-G形成的N4结构为螯合基团进行99Tcm标记,用聚酰胺薄膜层析法和高效液相色谱仪法(HPLC)测定其标记率和标记物的稳定性,并行人结肠癌荷瘤裸鼠体内分布[每克组织百分注射剂量率(%ID/g)]及显像研究.采用SAS 6.22软件对数据进行分析.结果 反义、无义PNA片段合成物的纯度>95%.99Tcm标记c-myc mRNA反义、无义PNA的标记率>95%,标记物室温放置18 h测定其标记率仍可达95%以上.c-myc mRNA反义、无义PNA片段4～8℃下放置3个月标记率仍>95%.HPLC测定标记物呈单峰.99Tcm标记c-myc mRNA反义PNA主要分布在荷瘤鼠肾、脾、肿瘤、肠道、肝组织中,99Tcm标记c-myc mRNA无义PNA在荷瘤鼠血液、脾、肾、肝及肺组织中分布较多;注射后4 h两者在荷瘤鼠肿瘤组织中的分布差异有统计学意义[(1.11±0.12)%ID/g和(0.14±0.02)%ID/g;t=14.75,P<0.01].99Tcm标记c-mycmRNA反义PNA在小鼠体内肿瘤/肌肉、肿瘤/肺的摄取比值较高,肿瘤显像明显.结论 用99Tcm标记c-myc mRNA反义、无义PNA的方法简单,标记率高,标记物稳定,前者有望成为一种新型肿瘤显像剂.