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PubMed2006

Novel approach in radionuclide tumor therapy: dose enhancement by high Z-element contrast agents.

PURPOSE: The aim of this study was to first calculate the dose-enhancement effect from internalized radiation by the presence of exogenous contrast media using Monte Carlo simulations, and then provide in vitro proof-of-concept for this novel method of radiation-dose enhancement. MATERIALS AND METHODS: The Monte Carlo program EGSnrc (Electron Gamma Shower) was used to simulate the interaction of internalizing radiation with iodine (I) or gadolinium (Gd) containing contrast media. Experimentally, the doseenhancement effect induced by I or Gd was evaluated in cell culture assays using internalizing peptides chelated with beta- emitting radionuclides and clinically available contrast media. RESULTS: Monte Carlo simulations predicted significant enhancement (approximately 70-340%) of radiation dose in the presence of high Zelement contrast media. This enhancement is radiation and Z-element dependent. Calculations showed that in the presence of contrast media, low-energy radionuclides favor localization of secondary particles, whereas higher energy beta- emitters localize radiation by reducing the pathway of the primary beta-particle. The dose enhancement was verified in vitro in two cell lines. CONCLUSIONS: Monte Carlo simulations in parallel with in vitro studies provide proof-of-principle for dose enhancement that occurs when utilizing an internalized source of radiation followed by the addition of exogenous contrast media. This dose enhancement is both radiation and Z-element dependent.

Cell Line, Tumor↗
PubMed2003

Alterations in high-density lipoprotein metabolism and reverse cholesterol transport in insulin resistance and type 2 diabetes mellitus: role of lipolytic enzymes, lecithin:cholesterol acyltransferase and lipid transfer proteins.

Insulin resistance and type 2 diabetes mellitus are generally accompanied by low HDL cholesterol and high plasma triglycerides, which are major cardiovascular risk factors. This review describes abnormalities in HDL metabolism and reverse cholesterol transport, i.e. the transport of cholesterol from peripheral cells back to the liver for metabolism and biliary excretion, in insulin resistance and type 2 diabetes mellitus. Several enzymes including lipoprotein lipase (LPL), hepatic lipase (HL) and lecithin: cholesterol acyltransferase (LCAT), as well as cholesteryl ester transfer protein (CETP) and phospholipid transfer protein (PLTP), participate in HDL metabolism and remodelling. Lipoprotein lipase hydrolyses lipoprotein triglycerides, thus providing lipids for HDL formation. Hepatic lipase reduces HDL particle size by hydrolysing its triglycerides and phospholipids. A decreased postheparin plasma LPL/HL ratio is a determinant of low HDL2 cholesterol in insulin resistance. The esterification of free cholesterol by LCAT increases HDL particle size. Plasma cholesterol esterification is unaltered or increased in type 2 diabetes mellitus, probably depending on the extent of triglyceride elevation. Subsequent CETP action results in transfer of cholesteryl esters from HDL towards triglyceride-rich lipoproteins, and is involved in decreasing HDL size. An increased plasma cholesteryl ester transfer is frequently observed in insulin-resistant conditions, and is considered to be a determinant of low HDL cholesterol. Phospholipid transfer protein generates small pre beta-HDL particles that are initial acceptors of cell-derived cholesterol. Its activity in plasma is elevated in insulin resistance and type 2 diabetes mellitus in association with high plasma triglycerides and obesity. In insulin resistance, the ability of plasma to promote cellular cholesterol efflux may be maintained consequent to increases in PLTP activity and pre beta-HDL. However, cellular cholesterol efflux to diabetic plasma is probably impaired. Besides, cellular abnormalities that are in part related to impaired actions of ATP binding cassette transporter 1 and scavenger receptor class B type I are likely to result in diminished cellular cholesterol efflux in the diabetic state. Whether hepatic metabolism of HDL-derived cholesterol and subsequent hepatobiliary transport is altered in insulin resistance and type 2 diabetes mellitus is unknown. Specific CETP inhibitors have been developed that exert major HDL cholesterol-raising effects in humans and retard atherosclerosis in animals. As an increased CETP-mediated cholesteryl ester transfer represents a plausible metabolic intermediate between high triglycerides and low HDL cholesterol, studies are warranted to evaluate the effects of these agents in insulin resistance- and diabetes-associated dyslipidaemia.

Biological Transport↗
PubMed1998

Interactions of griseofulvin with cyclodextrins in solid binary systems.

Griseofulvin/cyclodextrin interactions were investigated in aqueous environment and in solid state. Two cyclodextrin derivatives (beta-cyclodextrin and 2-hydroxypropyl beta-cyclodextrin) were used to prepare different physical mixtures and kneaded systems, and the drug/cyclodextrin ratios were 1:1 and 1:2 mol/mol. Scanning electron microscopy (SEM), hot-stage microscopy (HSM), differential scanning calorimetry (DSC), thermogravimetry (TG), and X-ray powder diffractometry were employed to characterize pure substances and their kneaded counterparts and all of the binary systems. The solubility of griseofulvin was increased in accord with the quantity of cyclodextrin added. HSM examination revealed that 2-hydroxypropyl-beta-cyclodextrin was dissolved in the droplets of melted griseofulvin, but did not show any interactions between melted griseofulvin and beta-cyclodextrin particles. The presence of griseofulvin endothermic peak in the DSC curves of all binary systems suggests the absence of any griseofulvin/cylcodextrin inclusion compound in the solid state. In TG, data of weight loss owing to the dehydration of cyclodextrins was similar for both kneaded systems and physical mixtures. X-ray diffraction patterns exhibited the amorphous nature of 2-hydroxypropyl-beta-cyclodextrin and the crystalline nature of griseofulvin and binary systems. Griseofulvin dissolution profiles from all binary systems showed an improvement in drug dissolution, which indicates that an "in situ" drug/cyclodextrin inclusion compound was formed in the aqueous dissolution medium.

2-Hydroxypropyl-beta-cyclodextrin↗
PubMed1995

A generic model for estimating the risk of deterministic effects of partial organ irradiation by hot particles.

A generic (Weibull-type) model is presented for evaluating the risk of a specified deterministic effect of hot-particle (i.e., highly radioactive particle) exposure of the skin, eye, ear, respiratory tract, or gastrointestinal tract. The generic model is a composite of the two-parameter Weibull function used for evaluating the risk of deterministic effects of total-organ irradiation and an isoeffect equation for partial-volume (i.e., fractional-volume) irradiation. Isoeffect refers to a constant level of risk rather than a constant level of severity. The generic model provides a way to evaluate the uniform-exposure, isoeffect equivalent dose, H'T,V, to a partial volume V that incurs the same risk as is associated with a nonuniform exposure of volumes within V. When V = 1, the volume is called a reference volume. The reference volume can be associated with a specific organ or mass of tissue. The reference-volume-specific, uniform-exposure, isoeffect equivalent dose, H'T,1, can be used to limit the risk of deterministic effects of hot-particle exposure by requiring that H'T,1 remain below the equivalent dose limit for any reference volume of tissue at risk. Local doses H'T,V, can be converted to isoeffect doses H'T,1 by multiplying H'T,V by a volume weighting factor Wv, where Wv equals Vn. For deterministic effects, n < 1. For stochastic effects, n = 1. For stochastic effects, H'T,1 can be converted to an effective dose by multiplying H'T,1 by an appropriate tissue-weighting factor WT. Results presented apply to circumstances where, at most, only a few hot particles are involved. In such circumstances, it is highly unlikely that local tissue will be irradiated by more than one hot particle. Although the focus of this paper is on hot-particle effects, results presented also apply to partial-organ or partial-tissue irradiation by any radiation source including external beams (e.g., electrons, neutrons, protons, heavy ions, and X-ray and gamma-ray photons).

Animals↗
PubMed2006

Preparation of a radionuclide/gel formulation for localised radiotherapy to a wide range of organs and tissues.

Localised radiotherapy by instillation of radiolabeled particles is being used to treat rheumatoid arthritis and certain tumors. Such therapy is limited to organs and tissues capable of retaining the radioactive compound until the radioactivity is sufficiently low, and the leakage to other parts of the body is no longer unacceptable. In this study, radiolabeled particles, i.e. 90Y-silica colloid particles, were encapsulated in calcium alginate gels, and the leakage of radioactivity from the gels was monitored. The purpose of the study was to develop a formulation suitable for the localised delivery of radiation therapy to a wide range of organs and tissues. An injectable gel formulation was developed, liberating only small amounts of radioactivity into the surrounding medium. The formulation is a viscous liquid at room temperature and forms a gel on heating to normal body temperature. Thus, it should be suitable for the localised delivery of radiolabeled particles to a wide range of organs and tissues. The study also includes a formulation exhibiting time-dependent gelation. However, this formulation was not found to be suitable for the purpose.

Alginates↗
PubMed1992

Effects of apolipoprotein E, beta-very low density lipoproteins, and cholesterol on the extension of neurites by rabbit dorsal root ganglion neurons in vitro.

Previous studies suggest that during nerve regeneration apoE acts as a lipid transport protein that assists in the rapid initial extension of axons and then in their myelination. To determine whether apoE and/or apoE-containing lipoproteins can modulate axon growth, we assessed their effect on the out-growth of neurites from neurons in mixed cultures of fetal rabbit dorsal root ganglion cells in vitro. Incubation with beta-very low density lipoprotein (beta-VLDL) particles, which are rich in apoE and cholesterol, increased neurite outgrowth and branching. Unesterified cholesterol added to the cultures had a similar, but less pronounced, effect. These data suggest that cholesterol might be the component responsible for the enhanced neurite growth. In contrast, purified, lipid-free apoE added to the cultures reduced neurite branching. Neurite branching was also reduced when purified apoE was added along with beta-VLDL or cholesterol; however, the striking finding was that under these conditions the neurites extended farther from the neuronal cell body. Dorsal root ganglion cells were examined for the presence of receptors for native and apoE-enriched beta-VLDL. Immunocytochemistry, ligand blots, 45Ca2+ blots, and studies of the interaction of the cells with fluorescent lipoproteins provided evidence of two types of receptors for apoE-containing lipoproteins on neurons: the low density lipoprotein (LDL) receptor, which binds native beta-VLDL, and the LDL receptor-related protein, which binds apoE-enriched beta-VLDL. These findings indicate that apoE may play two complementary roles in neurite outgrowth. When complexed with lipoproteins, apoE stimulates neurite growth by the receptor-mediated delivery of cholesterol and perhaps other components necessary for neurite outgrowth. When apoE as a free protein is added together with apoE-containing lipoproteins, apoE decreases neurite branching and promotes neurite extension away from the cell body. These actions, which would be complementary in promoting target-directed nerve growth in vivo, provide the first direct evidence that apoE and apoE-containing lipoproteins can modulate the outgrowth of neuronal processes.

Animals↗
PubMed1998

Variation in cytokines induced by particles from different prosthetic materials.

Particles of prosthetic material stimulate macrophages to release cytokines, which may cause bone loss and loosening of the prosthesis. This study investigates the possibility that particles of different prosthetic materials may induce different cytokines and thus have different effects on bone remodeling. The in vitro response of human monocytes to particles of cast and forged cobalt chrome alloy, stainless steel, and titanium aluminum vanadium alloy were compared. There was no difference in the biologic response to cobalt-chrome particles derived from cast or forged material. Cobalt-chrome particles were toxic to the cells, but titanium aluminum vanadium particles did not affect cell viability. Stainless steel particles were approximately 10 times more toxic than were cobalt-chrome particles. All particles induced the release of tumor necrosis factor and interleukin 1 beta; stainless steel particles were the most potent stimulators of interleukin 1 beta; titanium aluminum vanadium particles were the strongest stimulators of interleukin 6 and prostaglandin 2. The study showed that particles derived from prosthetic materials of different metal compositions can elicit significantly different biologic responses. Understanding these different responses may help identify materials better suited for prostheses.

Alloys↗
PubMed1998

Mechanisms of high density lipoprotein-mediated efflux of cholesterol from cell plasma membranes.

The participation of HDL in the reverse cholesterol transport (RCT) from peripheral cells to the liver is critical for the antiatherogenic properties of this lipoprotein. Experimental results showing that efflux of cholesterol from cells growing in culture is mediated by HDL and lipoprotein particles containing apo A-I, in particular, support this conclusion. A bidirectional flux of unesterified cholesterol molecules between the plasma membrane of cells and HDL particles in the extracellular medium occurs. Net efflux of cholesterol mass from the cells involves passive diffusion of cholesterol molecules through the aqueous phase and down their concentration gradient between the membrane and HDL; the concentration gradient is maintained by LCAT-mediated esterification of cholesterol molecules in the HDL particles. Fully lipidated apo A-I is important in promoting this aqueous diffusion mechanism because it: (1) acts as a cofactor for LCAT; and (2) solubilizes phospholipid into small HDL-sized particles that are efficient at absorbing cholesterol molecules diffusing away from the cell surface. Apo A-I also exists in an incompletely lipidated state in plasma. Apo A-I molecules in this state are able to solubilize phospholipid and cholesterol from the plasma membrane of cells. This membrane-microsolubilization process is enhanced by enrichment of the plasma membrane with cholesterol and is the mechanism by which pre-beta-HDL particles in the extracellular medium remove cholesterol and phospholipid from cells. The relative contributions in vivo of the aqueous diffusion and membrane-microsolubilization mechanisms of apo A-I-mediated cell cholesterol efflux are not predicted readily from cell culture experiments. Confounding issues are the variations with cell type and the dependence on the degree of cholesterol loading of the cell plasma membrane.

Animals↗
PubMed1999

Association behavior of native beta-lactoglobulin.

The association behavior of beta-lactoglobulin has been studied by small-angle neutron scattering as a function of protein concentration, temperature, pH, and NaCl concentration of the solution. By indirect Fourier transformation of the spectra, pair-distance distribution functions for the various samples were obtained. These functions provided information on the maximum size, the weight-averaged molecular mass, and the z-averaged radius of gyration of the beta-lactoglobulin particles. At room temperature and pH values below 4 and above 5.2 the protein consists predominantly of monomers and dimers, consistent with literature. In these pH regimes the formation of dimers is favored upon increasing ionic strength and decreasing protein charge (pH values closer to the isoelectric point of the protein). Around pH 4.7, larger oligomeric structures are formed, enhanced by a decrease in temperature and a decrease in ionic strength. beta-Lactoglobulin A associates more strongly than beta-lactoglobulin B. Surprisingly, at pH 6.9 larger structures than dimers seem to be formed at high protein concentrations (> 30 mg mL-1).

Animals↗
PubMed2003

Autoradiographical methods for the assessment of radionuclides in hot particles on filter samples.

Three different autoradiographical methods were tested for detection, localisation and quantification of radionuclides in hot particles deposited on air filters. With all methods, hot particles could successfully be localised on air filters. Two methods, a X-ray film technique and a digital autoradiography technique using a microchannel array detector, have been examined with particles of known activity and with particles on air filter samples of the Chernobyl area to be able to quantify the 90Sr activity in the hot particles consistently. The results obtained in this study suggest that the digital autoradiography system is a very useful tool having a high efficiency with a low measurement uncertainty and allowing short exposure times for extensive measurement tasks.

Air Pollution, Radioactive↗
PubMed2004

Ionizing radiation attracts soil fungi.

During the last 15 years, about 2000 strains of 200 species of 98 genera of fungi have been isolated from around the Chernobyl Atomic Energy Station. Many of these microfungi are capable of growing into and decomposing 'hot particles'; carbon based radioactive graphite from the reactor and there are suggestions that some fungi actively direct their growth toward sources of radioactivity, possibly attracted to the carbon skeleton of these structures. In our experiments, we eliminated the confounding effects of carbon as a fungal resource, by developing experimental protocols that expose fungal spores and their germinating hyphae to directional sources of ionizing radiation allowing us to measure fungal response to ionizing radiation per se. We show that both beta and gamma radiation promote directional growth of hyphae towards the source of ionizing radiation.

Beta Particles↗
PubMed2001

A wide dynamic range, high-spatial-resolution scanning system for radiochromic dye films.

This paper describes the evaluation of an inexpensive, commercially available 35 mm transparency slide scanner as a potential alternative scanning device for GafChromic HD-810 radiochromic dye film. Besides its low cost, the principal advantages of this type of scanner are high spatial resolution and high speed (a typical scan taking less than 1 min). With broad-band illumination the useful dose range using grey-scale imaging of GafChromic HD-810 is limited to about 50-800 Gy. By using the colour-scale imaging capability of the scanner we have been able to achieve a significant extension covering a similar range (15-2000 Gy) to that attainable using monochromatic illumination. The short-term reproducibility of the system is good, with a coefficient of variation of doses estimated from repeat scanning of uniformly exposed calibration films of less than 2%. Long-term stability is ensured by the scanning of a manufacturer-supplied test slide. The slide scanner system has been used in the determination of depth dose distributions from a model 'hot particle' source containing 106Ru/Rh. GafChromic dye film stacks irradiated by the source were read out on both the slide scanner and a conventional Joyce Loebl MDM6 scanning stage microdensitometer. The overall agreement between the dose estimates provided by the two systems was within 10%.

Beta Particles↗
PubMed1981

Role of beta 1H for the binding of C3b-coated particles to human lymphoid and phagocytic cells.

Coating of EAC14oxy23b with highly purified human serum beta 1H globulin (beta 1H) led to acceleration of rosette formation with human peripheral blood lymphocytes (PBL), tonsil lymphocytes, B lymphoblastoid (Raji) cells, granulocytes and monocytes. This reaction was discernible from C3bi-dependent rosette formation. Enhancement of rosette formation of C3b cells by beta 1H was most effective at limiting amounts of C3 per EAC14oxy23b. The beta 1H effect was not due to trace contamination with C3b inactivator. beta 1H-dependent rosette formation with the various lymphoid and phagocytic cells could be suppressed by the F(ab')2 fragment of anti-beta 1H suggesting beta 1H-mediated binding of beta 1H-coated particles to complement receptor-positive (CR+) cells. In turn, binding of fluid-phase beta 1H to lymphoid and phagocytic cells could be demonstrated by fluorescence and by 14C-labeled beta 1H. In addition, the functional status of these cells with respect to their receptor reactivity was altered. Treatment of normal lymphocytes (PBL, tonsil lymphocytes) and of granulocytes with beta 1H improved their rosette formation with both EAC14oxy23b and EAC14oxy23b-beta 1H. The reaction of monocytes was hardly affected. The beta 1H effect on Raji cells resulted in reduced rosette formation with EAC14oxy23b-beta 1H, while binding of EAC14oxy23b remained unchanged. These results suggest the presence of sites on CR+ cells, to which soluble and particle-bound beta 1H can bind, leading to alteration of the functional status of the cells. In all likelihood, EAC14oxy23bi can attach to the beta 1H-binding sites on CR+ cells.

Animals↗
PubMed2003

Histologic effect of pure-phase beta-tricalcium phosphate on bone regeneration in human artificial jawbone defects.

The effect of the pure-phase beta-tricalcium phosphate (beta-TCP) Cerasorb on bone regeneration was evaluated in hollow titanium cylinders implanted in the posterior jaws of five volunteers. Beta-TCP particles were inserted inside the cylinders and harvested 6 months after placement. The density of the newly formed bone inside the bone-growing chambers measured 27.84% +/- 24.67% in test and 17.90% +/- 4.28% in control subjects, without a statistically significant difference. Analysis of the histologic specimens revealed that the density of the regenerated bone was related to the density of the surrounding bone. The present study demonstrates the spontaneous healing of infrabony artificial defects, 2.5 mm diameter, in the jaw. The pure beta-TCP was resorbed simultaneously with new bone formation, without interference with the bone matrix formation.

Absorbable Implants↗
PubMed1991

On the nature of spontaneous RNA synthesis by Q beta replicase.

Numerous RNA species of different length and nucleotide sequence grow spontaneously in vitro in Q beta replicase reactions where no RNA templates are added deliberately. Here, we show that this spontaneous RNA synthesis by Q beta replicase is template directed. The immediate source of template RNA can be the laboratory air, but there are ways to eliminate, or at least substantially reduce, the harmful effects of spontaneous synthesis. Solitary RNA molecules were detected in a thin layer of agarose gel containing Q beta replicase, where they grew to form colonies that became visible upon staining with ethidium bromide. This result provides a powerful tool for RNA cloning and selection in vitro. We also show that replicating RNAs similar to those growing spontaneously are incorporated into Q beta phage particles and can propagate in vivo for a number of phage generations. These RNAs are the smallest known molecular parasites, and in many aspects they resemble both the defective interfering genomes of animal and plant viruses and plant virus satellite RNAs.

Air↗
PubMed1993

Beta camera low activity tumor imaging.

A new technique, the beta camera, to complement film autoradiography, with fast quantitative imaging of beta particle-emitting radionuclides has been developed. It consists of a thin plastic scintillator and a light-sensitive microchannel plate detector. The thin tissue sample is mounted on the scintillator. Our first system had a high background and a moderate spatial resolution of 900 microns. We now report an improved system with a photomultiplier tube mounted on the scintillator of the microchannel plate detector. Only events registered by both detectors are accepted. A fast coincidence unit processes the signals, and if a time overlap exists, an event is generated in the beta camera. In the coincidence mode, images with low activity distribution of 201Tl (count rate 1 s-1) in 50 microns-thick slices of a human glioma tumor could be recorded with a spatial resolution of 500 microns.

Brain Neoplasms↗
PubMed2002

Dose-dependent action of atorvastatin in type IIB hyperlipidemia: preferential and progressive reduction of atherogenic apoB-containing lipoprotein subclasses (VLDL-2, IDL, small dense LDL) and stimulation of cellular cholesterol efflux.

Type IIB hyperlipidemia is associated with premature vascular disease, an atherogenic lipoprotein phenotype characterised by elevated levels of triglyceride-rich VLDL and small dense LDL, together with subnormal levels of HDL. The dose-dependent and independent effects of a potent HMGCoA reductase inhibitor, Atorvastatin, at daily doses of 10 and 40 mg, were evaluated on triglyceride-rich lipoprotein subclasses (VLDL-1, VLDL-2 and IDL), on the major LDL subclasses (light LDL, LDL-1+LDL-2, D: 1.019-1.029 g/ml; intermediate LDL, LDL-3, D: 1.029-1.039 g/ml and small dense LDL, LDL-4+LDL+5, D: 1.039-1.063 g/ml), on CETP-mediated cholesteryl ester transfer from HDL to apoB-containing lipoproteins, on phospholipid transfer protein activity and on plasma-mediated cellular cholesterol efflux in patients (n=10) displaying type IIB hyperlipidemia. Plasma concentrations of triglyceride-rich lipoprotein subclasses (TRL: VLDL-1, Sf 60-400; VLDL-2, Sf 20-60 and IDL, Sf 12-20) and of LDL (D: 1.019-1.063 g/ml) were markedly diminished after 6 weeks of statin treatment at 10 mg per day (-31 and -36%, respectively; P<0.002) and by 42 and 51%, respectively at the 40 mg per day dose. Increasing doses of atorvastatin progressively normalised both the quantitative and qualitative features of the LDL subclass profile, in which dense LDL predominated at baseline. Indeed, dense LDL levels were reduced by up to 57% at the 40-mg dose, leading to a shift in the peak of the density profile towards larger, buoyant LDL particles typical of normolipidemic subjects. In addition, marked reduction in numbers of apoB100-containing particle acceptors led to a 30% decrease (P<0.02) in CETP-mediated CE transfer from HDL. Finally, a significant dose-dependent statin-mediated elevation (+15% at 10 mg; P=0.0003 and +35% at 40 mg; P<0.0001 compared to baseline) in the capacity of plasma from type IIB subjects to mediate free cholesterol efflux from Fu5AH hepatoma cells was observed. Moreover, atorvastatin (40 mg per day) significantly increased plasma apoAI levels (+24%; P<0.05), thereby suggesting that this statin enhances production of apoAI and with it, formation of nascent pre-beta HDL particles. Plasma PLTP activity was not affected by either dose of atorvastatin. We conclude that increasing the dose of atorvastatin leads to dose-dependent, preferential and progressive reduction in particle numbers of atherogenic VLDL-2, IDL and dense LDL, and concomitantly, to enhanced cellular cholesterol efflux in type IIB dyslipidemia, thereby diminishing the atherosclerotic burden in subjects characterised by high cardiovascular risk.

Adult↗
PubMed2000

Solubilized cell wall beta-glucan, CSBG, is an epitope of Candida immune mice.

Antibody to beta-glucan is generally difficult to produce in mice. We have recently developed a protocol to obtain a soluble Candida spp. beta-(1-->3)-D-glucan (CSBG) by sodium hypochlorite (NaClO) oxidation and subsequent dimethyl sulfoxide (Me2SO) extraction. CSBG is composed mainly of beta-(1-->3) and beta-(1-->6)-glucosidic linkages with a small amount of branch. In this paper, mice were immunized with Candida albicans and the specificity of the resulting sera to CSBG was examined by ELISA. Using CSBG coated plate, sera of the Candida immune mice showed higher reactivity than non-immune, normal mice and the reactivity was neutralized by adding soluble CSBG as a competitor. However, the reactivity could not be neutralized by a beta-(1-->6) branched beta-(1-->3)-glucan, grifolan. Similar specificity of the sera was obtained by commercially available beta-glucan particle, zymosan or zymocel, immune mice. These facts strongly suggested that CSBG included epitopes of the specific antibody in Candida immune mice.

Animals↗