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Electron microscopic studies on the sinusoidal cells in the monkey liver.

The sinusoidal wall was observed by transmission electron microscopy in crab-eating monkey livers. The perikarya of sinusoidal endothelial cells were characterized by numerous macropinocytotic vacuoles and curved smooth-surfaced tubules of high electron density. Size and spacing of fenestrae in endothelial sieve plates corresponded to essentially those in other mammalian species including the human. It was verified that the high concentration of microfilaments was responsible for the electron dense appearance of the sieve plate in tangential sections. Besides occasional overlapping of endothelial sheets, complicated interdigitations of several short lamellae originating from endothelial processes occasionally caused a layered structure of the endothelial lining. Kupffer cells were strikingly rich in lysosomes and contained large mitochondria and phagosomes. They were fixed to the endothelial lining by patches of junctional complexes identical with those between endothelial cells. Ito cells of the monkey liver demonstrated, like those of the human liver, many smooth-surfaced caveolae and vesicles along their perisinusoidal surface, suggesting their micropinocytotic activity. They also contained glycogen beta-particles which were partly gathered around lipid vacuoles. The electron dense droplets enclosed by glycogen particles as revealed in human Ito cells and regarded as immature lipid droplets retaining the chemical properties of glycogen, could rarely be confirmed in the present study. Between the Ito cell and hepatocyte there occurred many junctional complexes. The space of Disse contained, besides abundant collagen fibrils, numerous fine filaments forming irregular meshworks or bundles which resembled fibrillar material (precursor of collagen) in appearance in the dilated cisternae of the rough endoplasmic reticulum (RER) of the Ito cell. These filaments often entwined the collagen fibrils in the Disse's space as if to participate by apposition in their development. The question, whether pinocytosis-like structures of the Ito cell might be involved in the precursor transport from the cisternae of the RER to the Disse's space, remained unanswered.

Adipose Tissue↗

Therapeutic radiopharmaceuticals.

A variety of radiopharmaceuticals have been introduced for the internal therapy of malignant and inflammatory lesions in nuclear medicine. In order to destroy the diseased tissues radionuclides with high linear energy transfer (LET) such as beta, alpha, Auger or low energy conversion electron emitters are needed. The range of beta particles is in mm's so they are effective for large tumors. The range of alpha particles is short, only a few cell diameters. Thus, they are effective in treating circulating malignant cells and micrometastases. The range of Auger and conversion electrons is <0.1 micro m. They are very effective in cell killing when they are carried across cell membrane into the nucleus to damage DNA. Appropriate ions, molecules and particles are labelled with such radionuclides and used as radiopharmaceuticals in many different applications. For an effective dose to be delivered high target to non target ratios must be attained. Monoclonal antibodies to specific antigens expressed on tumor cells have been developed to increase the uptake by malignant tissues by specific accumulation. Radiolabelled peptides such as somatostatin, small molecules such as metaiodo-benzylguanidine (MIBG) and many different nano-and micro-particles have been investigated. The effectiveness of therapy can be increased by direct locoregional administration of the radiopharmaceutical. This way the radiation effects are confined locally and the normal tissues are spared from radiation effects. In this review article selection criteria and characteristics of radionuclides and carrier ions, molecules and particles for various therapeutic applications will be discussed, including mainly the recent developments.

Animals↗

Treatment of acute leukaemias with monoclonal antibodies: current status and future prospects.

Monoclonal antibodies are a new class of agents targeted at specific receptors on cancer cells. In addition to direct cellular effects, antibodies can carry substances to the targeted cells, such as radioactive isotopes, toxins, and antineoplastic agents. At present monoclonal antibodies, directed against both lymphoid antigens (CD 20 and CD 52) and a myeloid antigen (CD33) are available for clinical use. In ALL, rituximab, a humanized anti CD20 antibody, has been combined to chemotherapy mainly in mature B-ALL and Burkitt's lymphoma and preliminary results are promising. Alemtuzumab is an anti-CD52 humanized antibody, which showed anti-tumour activity in CLL; clinical effects were observed in some patients with relapsed adult ALL. Monoclonal antibodies against myeloid antigens have been prevalently used in acute myeloid leukaemias (AML), where the most utilised immunological target is CD33. The CD33 molecule is expressed by approximately 90% of AMLs but not on CD34(+) bone marrow-resident hematopoietic stem cells. The humanized anti-CD33 monoclonal antibody HuM195 has only modest activity against overt AML, but it can eliminate minimal residual disease. Radioimmunotherapy with beta-particle-emitting isotopes targeting CD33 shows a major efficacy. Targeted chemotherapy with the anti-CD33-calicheamicin construct gemtuzumab ozogamicin (GO) has produced remissions as a single agent in patients with relapsed AML and appears promising when used in combination with standard chemotherapy. GO is composed of a humanized immunoglobulin G(4) (IgG(4)) monoclonal antibody (mAb), targeting the CD33 antigen and linked to a calicheamicin derivative, a cytotoxic anthracycline antibiotic. GO has been approved by FDA as second-line therapy in older patients with AML. The most common adverse effects of monoclonal antibodies are myelosuppression, infusion-related reactions, and hypersensitivity reactions. Rituximab may cause tumour lysis syndrome. Alemtuzumab causes immunosuppression, increasing the risk of infection. GO may cause hepatotoxicity.

Acute Disease↗

Vacuolated glycogen-laden leukemic cells in a case of crisis type chronic adult T-cell leukemia.

We present a unique case of crisis type chronic adult T-cell leukemia (ATL), in which the majority of leukemic cells had abundant periodic acid-Schiff (PAS)-positive cytoplasmic inclusions. These inclusions were found to be composed of glycogen because the PAS-positivity completely disappeared after digestion with amylase or human saliva. Electron microscopy also revealed that the inclusions consisted of aggregated beta particles of glycogen. The mechanism of glycogen accumulation in leukemic cells remains unknown; however, the presence of such inclusions in leukemic cells may be helpful diagnostically in T-lymphocyte malignancies.

Glycogen↗

[A twin type heat flow microcalorimeter for radioactivity measurement].

A microcalorimeter is described which was developed and tested for measuring the activity of pure beta particle emitters. Two identically made calorimeter cells are used, each 46.5 mm i.d. and 54.0 mm deep and each fitted with thermomodules generating an emf of about 29 mV degrees C-1. The thermal energy from the cells flows into a surrounding mass of aluminium (15 kg) embedded in 30 mm thick styrofoam. The entire assembly is then immersed into an open thermostat bath controlled by electric heaters an arrangement which facilitates access for sample changing. The outputs of the modules (sensors) are connected in opposing polarities, the net output being amplified and this voltage signal is used to estimate the radioactivity in the samples. In order to demonstrate usefulness of this calorimeter, approximately 3.7 GBq(100 mCi) of tritiated water was measured. The result was compared with that of liquid scintilation counting method and found to be agreed within 5%.

Calorimetry↗

[Palliative treatment of bone metastases with bone-seeking radionuclides].

The skeleton is a common site for metastases in patients with prostate and breast cancer. Beside analgesic therapy and external beam radiotherapy, the use of bone-seeking radiopharmaceuticals gives pain relief. Analogues of these pharmaceuticals are also applied in skeletal scintigraphy. They accumulate at the site of high osteoblast activity and in this way they exert a local favourable influence on metastases through their radiation (beta particles or conversion electrons). The bone-seeking radiopharmaceuticals include strontium-89 chloride and rhenium-186 hydroxyethylidenediphosphonate. The main adverse reaction that has been observed is myelosuppression.

Bone Marrow↗

[A radiation hygiene assessment of the use of the deicing preparation Kama-M in Moscow].

The content of radionuclides and metals in the agent to control the ice-crusted ground Kama-M was determined. An investigation has shown that the radioactive background of the agent is caused only by the presence of 40K as part of KCl. Small quantities of 226Ra and 232Th are present in the insoluble sediment. The concentrations of metals are very low and produce no environmental effects. The levels of superficial pollutions vary from 1.3 to 4.8 beta-particles per cm2/min and the power of an exposure dose is at the level of background values (9-20 microR/hour), which is no greater than the permissible ones.

Freezing↗

211At- and 131I-labeled bisphosphonates with high in vivo stability and bone accumulation.

UNLABELLED: Bisphosphonates were synthesized for use as carriers for astatine and iodine radioisotopes to target bone neoplasms. METHODS: Radiohalogenated activated esters were coupled to the amino group in the side chain of the bisphosphonate. The bisphosphonate 3-amino-1-hydroxypropylidene bisphosphonate was combined with four different acylation agents: N-succinimidyl 3-[211At]astatobenzoate, N-succinimidyl 3-[131I]iodobenzoate, N-succinimidyl-5-[211At]astato-3-pyridinecarboxylate and N-succinimidyl-5-[131I]iodo-5-pyridinecarboxylate. The products, 3-[131I]iodobenzamide-N-3-hydroxypropylidene-3,3-bisphosphonate (IBPB), 3-[211At]astato-benzamide-N-3-hydroxypropylidene-3,3-bisphosphonat e (ABPB), 5-[131I]iodopyridine-3-amide-N-3-hydroxypropylidene-3,3-bisphospho nate (IPPB) and 5-[211At]astatopyridine-3-amide-N-3-hydroxypropylidene-3,3-bisphos phonate (APPB), were injected intravenously into Balb/c mice. MIRD and Monte Carlo methods were used on the basis of cumulated activity calculated from biodistribution data to estimate dose to organs and bone segments. RESULTS: All 131I- and 211At-labeled analogs were strongly incorporated into osseous tissue and retained there at stable levels, while a rapid clearance from blood was observed. The bone uptake was found to be similar for 211At- and 131I-labeled bisphosphonate when compared in paired label experiments. Bone uptake and bone-to-tissue ratios were better for IBPB compared with IPPB, and ABPB compared with APPB. All four compounds appeared to be highly resistant to in vivo dehalogenation as indicated by low uptake of 131I/211At in the thyroid gland and stomach. According to dosimetric estimates, the bone surface-to-bone marrow ratio was three times higher with 211At than with 131I. CONCLUSION: Both the beta-particle- and alpha-particle-emitting compounds showed high in vivo stability and excellent affinity for osseous tissue. Further preclinical evaluation is therefore warranted.

Animals↗

S values for radionuclides localized within the skeleton.

UNLABELLED: Calculations of radiation absorbed dose to the active marrow are important to radionuclide therapies such as radioimmunotherapy and bone pain palliation. In diagnostic nuclear medicine, calculations of the effective dose for radiopharmaceutical procedures also require the assessment of radiation dose to the skeletal endosteum. We have previously reported the development of 2 3-dimensional electron transport models for assessing absorbed fractions to both marrow and endosteum in trabecular and cortical bone, respectively. Here, we extend these calculations to the assignment of radionuclide S values. METHODS: Data published in International Commission on Radiological Protection Publication 70 were used to develop tables of masses for total marrow space, active and inactive marrow, endosteum, and bone matrix within 22 skeletal sites in the adult. Using our site-specific tissue masses, along with electron absorbed fractions given by our 3-dimensional transport models, radionuclide S values (electron and beta particle components only) were subsequently calculated using the MIRD schema for 32P, 33P, 89Sr, 90Sr, 90Y, 117mSn, 153Sm, 169Er, 177Lu, and 186Re. Specific consideration was given to the trabecular active marrow as both a source and a target region. RESULTS: Site-specific radionuclide S values are reported for 22 skeletal sites, for 9 source-target tissue combinations within trabecular bone, and for 6 source-target tissue combinations within cortical bone. Skeletal-averaged S values are also provided. CONCLUSION: A fully documented model is presented for the adult for use in radionuclide dosimetry of the skeleton. The model is based on both the latest international recommendations for skeletal tissue masses and results from three-dimensional electron transport calculations within the skeleton. Comparisons are additionally made against the radionuclide S values published in MIRD Pamphlet No. 11 and those calculated using the MIRDOSE2 and MIRDOSE3 computer codes. Differences in these datasets vary with the source-target combination considered and may be attributed to 1 of 3 causes: (a) assumptions on reference target masses, (b) transport models used to assign absorbed fractions, and (c) implicit assumptions made in considering the trabecular active marrow as both a source and a target tissue.

Adult↗

Peptide receptor imaging and therapy.

This article reviews the results of somatostatin receptor imaging (SRI) in patients with somatostatin receptor-positive neuroendocrine tumors, such as pituitary tumors, endocrine pancreatic tumors, carcinoids, gastrinomas, and paragangliomas, or other diseases in which somatostatin receptors may also be expressed, like sarcoidosis and autoimmune diseases. [(111)In-DTPA0]octreotide is a radiopharmaceutical that has great potential for helping visualize whether somatostatin receptor-positive tumors have recurred. The overall sensitivity of SRI to localize neuroendocrine tumors is high. In several neuroendocrine tumor types, inclusion of SRI in the localization or staging procedure may be very rewarding in terms of cost effectiveness, patient management, or quality of life. The value of SRI in patients with other tumors, such as breast cancer or malignant lymphomas, or in patients with granulomatous diseases has to be established. The application of radiolabeled peptides may be clinically useful in another way: after the injection of [(111)In-DTPA0]octreotide, surgeons can detect tumor localizations by a probe that is used during the operation. This may be of particular value if small tumors with a high receptor density are present (e.g., gastrinomas). As the success of peptide receptor scintigraphy for tumor visualization became clear, the next logical step was to try to label these peptides with radionuclides emitting alpha or beta particles, or Auger or conversion electrons, and to perform radiotherapy with these radiolabeled peptides. The results of the described studies with 90Y- and (111)In-labeled octreotide show that peptide receptor radionuclide therapy using radionuclides with appropriate particle ranges may become a new treatment modality. One might consider the use of radiolabeled somatostatin analogs first in an adjuvant setting after surgery of somatostatin receptor-positive tumors to eradicate occult metastases and second for cancer treatment at a later stage.

Humans↗

Two-step targeting of xenografted colon carcinoma using a bispecific antibody and 188Re-labeled bivalent hapten: biodistribution and dosimetry studies.

UNLABELLED: Radioimmunotherapy (RIT) is currently being considered for the treatment of solid tumors. Although results have been encouraging for pretargeted 131I RIT with the affinity enhancement system (AES), the radionuclide used is not optimal because of its long half-life, strong gamma emission, poor specific activity, and low beta particle energy. 188Re, though unsuitable for direct antibody labeling, could be used with the AES two-step targeting technique. The purpose of this study was to compare the distribution and dosimetry of a bivalent hapten labeled with 188Re or 125I. For dosimetry calculations and biodistribution data, 125I was substituted for 131I. METHODS: After preliminary injection of a bispecific anticarcinoembryonic antigen (CEA) or antihapten antibody (Bs-mAb F6-679), AG 8.1 or AG 8.0 hapten radiolabeled with 188Re or 125I was injected into a nude mouse model grafted subcutaneously with a human colon carcinoma cell line (LS-174-T) expressing CEA. A dosimetry study was performed for each animal from the concentration of radioactivity in tumor and different tissues. RESULTS: Radiolabeling of AG 8.1 with 125I afforded a 40% yield with a specific activity of 11.1 MBq/nmol after purification. Radiolabeling of AG 8.0 with 188Re afforded a 72% yield with a specific activity of 31.82 MBq/nmol. In all experiments, the percentage of tumor uptake of 125I-AG 8.1 was always significantly greater than that of 188Re-AG 8.0. The corresponding tumor-to-tissue ratios reflected uptake values. The least favorable tumor-to-normal tissue ratios in the dosimetry study were 8.1 and 8.5 for 131I (tumor-to-blood ratio and tumor-to-kidney ratio, respectively) and 2.3 for 188Re (tumor-to-intestine ratio). CONCLUSION: This study indicates that 188Re can be used for radiolabeling of hapten in two-step radioimmunotherapy protocols with the AES technique. 188Re has a greater range than 131I, which should allow the treatment of solid tumors around 1 cm in diameter. Although the method used for hapten radiolabeling did not provide optimal tumor uptake, the use of a bifunctional chelating agent associated with AG 8.1 should solve this problem.

Animals↗

GPI-specific phospholipase D associates with an apoA-I- and apoA-IV-containing complex.

Glycosylphosphatidylinositol-specific phospholipase D (GPI-PLD) is abundant in serum and associates with high density lipoproteins (HDL). We have characterized the distribution of GPI-PLD among lipoproteins in human plasma. Apolipoprotein (apo)-specific lipoproteins containing apoB (Lp[B]), apoA-I and A-II (Lp[A-I, A-II]), or apoA-I only (Lp[A-I]) were isolated using dextran sulfate and immunoaffinity chromatography. In six human plasma samples with HDL cholesterol ranging from 39 to 129 mg/dl, 79 +/- 14% (mean +/- SD) of the total plasma GPI-PLD activity was associated with Lp[A-I], 9 +/- 12% with Lp[A-I, A-II], and 1 +/- 1% with Lp[B]; and 11 +/- 10% was present in plasma devoid of these lipoproteins. Further characterization of the GPI-PLD-containing lipoproteins by gel-filtration chromatography and nondenaturing polyacrylamide and agarose gel electrophoresis revealed that these apoA-I-containing particles/complexes were small (8 nm) and migrated with pre-beta particles on agarose electrophoresis. Immunoprecipitation of GPI-PLD with a monoclonal antibody to GPI-PLD co-precipitated apoA-I and apoA-IV but little or no apoA-II, apoC-II, apoC-III, apoD, or apoE. In vitro, apoA-I but not apoA-IV or bovine serum albumin interacted directly with GPI-PLD, but did not stimulate GPI-PLD-mediated cleavage of a cell surface GPI-anchored protein. Thus, the majority of plasma GPI-PLD appears to be specifically associated with a small, discrete, and minor fraction of lipoproteins containing apoA-I and apoA-IV. -- Deeg, M. A., E. L. Bierman, and M. C. Cheung. GPI-specific phospholipase D associates with an apoA-I- and apoA-IV-containing complex. J. Lipid Res. 2001. 42: 442--451.

Adult↗

Endocytic activity in the thrombocytes of the turtle Phrynopys hilarii (freshwater South American species).

The phagocytic process in cells depends on lysosomal enzymes, high-energy metabolism and cellular recognition. In this paper, we investigated the presence of energy and recognition factors in thrombocytes of turtle Phrynopys hilarii (a freshwater South American species). Turtle thrombocytes (P. hilarii) present glycogen - possibly beta particles - dispersed in their cytoplasm and glycoproteins in the cell surface, as well as a large number of enzymes involved in the endocytic process (Pellizzon, 1996). The activity of these enzymes depends on high-energy metabolism and on cellular recognition provided by specific glycoconjugates (Alberts et al., 1994). This metabolic characterization is demonstrated by the large amount of glycogen particles observed in the cytoplasm by Thiéry's method. Glycogen labeling was also observed when concanavalin A-peroxidase was used as a marker for thrombocytes and for endocyted charcoal particles. Our results show that these cells have phagocytic ability, suggesting that their function in blood circulation is not limited to aggregation but may also involve a great potential for phagocytosis.

Animals↗

Tumor response after [(90)Y-DOTA(0),Tyr(3)]octreotide radionuclide therapy in a transplantable rat tumor model is dependent on tumor size.

UNLABELLED: A promising application of radiolabeled somatostatin analogs is peptide receptor-targeted radionuclide therapy of somatostatin receptor-expressing tumors. A suitable radionuclide is (90)Y, which emits high-energy beta-particles with a pathlength of several millimeters in tissue, making it especially promising for treatment of large tumors. METHODS: We investigated the radiotherapeutic effect of different activities (111 and 370 MBq) of [(90)Y-1,4,7,10-tetraazacyclododecane-N,N',N",N"'-tetraacetic acid (DOTA)(0),Tyr(3)]octreotide in Lewis rats bearing somatostatin receptor-positive rat pancreatic CA20948 tumors of different size (0.08-15 cm(2)) in their flank. RESULTS: Dose-dependent radiotherapeutic effects of (90)Y-labeled octreotide in this rat tumor model were found. Tumor control (100% complete response) was found in animals bearing tumors of 3-9 cm(2) (mean, 7.8 cm(2)) after intravenous injection of the highest activity (370 MBq [(90)Y-DOTA(0),Tyr(3)]octreotide). In rats bearing tumors of < or =1 cm(2) or > or =14 cm(2), the effects were less pronounced (50% and 0% complete response, respectively). In tumors of < or =1 cm(2) the (90)Y radiation energy will not be absorbed completely in the tumor, whereas in tumors of > or =14 cm(2) the increased number of clonogenic and probably hypoxic tumor cells may explain the failure to reach a cure. CONCLUSION: This study shows the ability of [(90)Y-DOTA(0),Tyr(3)]octreotide to control tumor growth, especially in medium-sized tumors. The effect of radionuclide therapy appeared to be dependent on tumor size at the onset of therapy.

Animals↗

High-dose 166Ho-DOTMP in myeloablative treatment of multiple myeloma: pharmacokinetics, biodistribution, and absorbed dose estimation.

UNLABELLED: Thirty-two patients with multiple myeloma were treated with high doses of 166Ho-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetramethylene-phosphonic acid (DOTMP) and were a subset of patients enrolled in a multicenter phase I/II dose escalation myeloablative trial. 166Ho with beta-emission (half-life, 26.8 h; beta-particle energies, 1.85 MeV [51%] and 1.77 MeV [48%]; gamma-photons, 80.6 keV [6.6%] and 1.38 MeV [0.9%]) was complexed to DOTMP, a macrocyclic tetraphosphonate. Pharmacokinetics, dosimetry, and biodistribution were studied. METHODS: Patients were treated at escalating dose levels of 20, 30, and 40 Gy to the bone marrow in combination with high-dose melphalan, with or without total-body irradiation, to evaluate toxicity and efficacy. After infusion with 1,110 MBq (30 mCi) of 166Ho-DOTMP for evaluation of biodistribution and dosimetry calculation, patients received the calculated amount of radioactivity for therapy in a single administration based on estimated dose calculations. RESULTS: Thirty-two patients participated in the study and were then treated. The average amount of administered radioactivity was 74.3 GBq (2,007 mCi) (range, 21.5-147.5 GBq [581-3,987 mCi]) of 166Ho-DOTMP. CONCLUSION: 166Ho-DOTMP has physical and pharmacokinetic characteristics compatible with high-dose myeloablative treatment of multiple myeloma.

Antineoplastic Agents, Alkylating↗

[Demonstration, content and distribution of glycogen in rat ova during segmentation].

A large amount of glycogen can be detected in the cytoplasm of rat eggs; it is evenly distributed in all the cytoplasm of blastomeres and polar globules. During cleavage, eggs show non significative change in glycogen content. This polysaccharide was observed and identified in ultrathin sections. It exists in the form of beta particles.

Animals↗

A comparison of 4 radionuclides conjugated to antibodies for single-cell kill.

UNLABELLED: We previously found that (67)Ga was more potent and more specific in single-cell kill than other Auger electron emitters and beta-particle emitters, using an anti-CD74 antibody (Ab) (major histocompatibility complex [MHC] class II invariant chain). Because anti-CD74 Abs follow an unusual processing pathway, with rapid delivery in very large amounts to lysosomes, it was important to determine if similar results would be obtained with other, more typical Abs. METHODS: Target cells were Raji B-lymphoma cells, and the Abs tested were antimature MHC class II antigen (lacking the invariant chain) and anti-CD20, both of which react with high-density antigens. Labeling was with (125)I or (131)I, by conventional iodination; with (111)In using the chelator benzyl-diethylenetriaminepentaacetic acid; or with (67)Ga using the chelator 1,4,7-triazacyclononane-1,4,7-triacetic acid. Abs were incubated with the cells for 2 d, uptake of radioactivity was assayed at various times, and toxicity was assayed primarily by a clonogenic assay. The fraction surviving was plotted versus cumulative disintegrations per cell to determine relative potency. RESULTS: The ranking of the radionuclides for potency was (131)I > (67)Ga > (125)I > (111)In. (67)Ga was approximately 2- to 3-fold more potent than (111)In. This was very similar to previous results with anti-CD74. Dosimetry calculations were generally consistent with the level of toxicity observed. In previous studies of nonspecific toxicity, the order of ranking was the same. CONCLUSION: The subcellular location of the bound Ab (whether on the cell surface or the cytoplasm) does not appear to be an important variable in the choice of radionuclide for single-cell kill. (67)Ga is a promising radionuclide for killing micrometastases, for high-density target antigens, but methods for achieving higher specific activity are required to fully exploit this approach. Each of the 4 radionuclides tested has certain advantages, and further studies are required to select the optimal radionuclide for a particular purpose.

Antibodies↗