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PubMed2006

Regulation of Sam68 activity by small heat shock protein 22.

Sam68 associates with c-Src kinase during mitosis. We previously demonstrated that Sam68 functionally replaces and/or synergizes with HIV-1 Rev in rev response element (RRE)-mediated gene expression and virus production. Furthermore, we reported that knockdown of Sam68 inhibited Rev-mediated RNA export and it is absolutely required for HIV-1 production. In the present study, we identified small heat shock protein, hsp22, as a novel interacting partner of Sam68. Hsp22 binds to Sam68 in vitro and in vivo. Overexpression of hsp22 significantly inhibits Sam68-mediated RRE- as well as CTE (constitutive transport element)-dependent reporter gene expression. Furthermore, exposing 293T cells to heat shock inhibits Sam68/RRE function by virtue of elevating hsp22. The critical domain of hsp22 that interacts with Sam68 resides between amino acids 62 and 133. Our studies provide evidence for the first time that hsp22 specifically binds to Sam68 and modulates its activity, thus playing a role in the post-transcriptional regulation of gene expression.

Adaptor Proteins, Signal Transducing↗
PubMed2002

A comparison of electrochemical and electrokinetic parameters determined for cellophane membranes in contact with NaCl and NaNO3 solutions.

Electrochemical and electrokinetic characterizations of cellophane membrane samples have been carried out by measuring membrane potential, salt diffusion, and tangential streaming potential, which allow the determination of different characteristic membrane parameters. Experiments were made with the membrane samples in contact with NaCl and NaNO(3) solutions at different concentrations and under different external conditions (concentration gradients), in order to obtain differences in transport and membrane characteristic parameters, depending on the electrolyte considered. Salt permeability across the membrane, which was obtained from diffusion measurements, is about twice as high for NaCl solutions as for NaNO(3) solutions, which is attributed to the different sizes of the electrolytes. Membrane potential measurements keeping the concentration ratio constant (C(1)/C(2)=2) were used to determine both the effective fixed charge concentration in the membrane, X(f), and the average value of transport numbers, t(i); taking into account these values, concentration dependence of membrane potential under a different external condition (C(1)=cte=0.01 M, 5 x 10(-3)< or =C(M)< or =5 x 10(-2)) was predicted. Results show that cellophane membrane behaves as a weak cation-exchange membrane and its permselectivity to cations is practically independent of the electrolyte considered. From electrokinetic results, assuming a Langmuir-type adsorption of anions on the cellophane surface, the number of accessible sites per surface unit was obtained, which is higher for Cl(-) than for NO(3)(-), in agreement with the small radii of chlorine ions; however, no significant differences in the specific adsorption free energy were found (DeltaG(Nacl)=-22.0 x 10(3) J/mol) and (DeltaG(NaNO(3))=-23.2 x 10(3) J/mol).

Journal Article↗
PubMed1984

Antibodies to a nuclear/nucleolar antigen in patients with polymyositis overlap syndromes.

A precipitating antigen-antibody system has been characterized that occurs in patients with polymyositis. At least half of the patients not only have polymyositis but also have scleroderma. The proposed name for this antigen found in calf thymus extract (CTE) is PM-Scl, to indicate the almost universal presence of polymyositis and the frequent occurrence of scleroderma in the patients who make antibodies to this antigen. The antigen is probably nucleolar since all sera which precipitate with the PM-Scl antigen stain the nucleoli of Hep2 cells by indirect immunofluorescence. The PM-Scl immune system is a distinctive one different from the other known precipitins that occur in patients with polymyositis and dermatomyositis including Jo1, nRNP, and Mi. This PM-Scl antigen and its antibody represent one system which constitutes part of the reactions previously designated as PM1. Interlaboratory exchange of sera and extracts have established the unique nature of this reaction which occurs in patients with inflammatory myopathy.

Antigen-Antibody Reactions↗
PubMed1983

Steroid economising effects of a calf thymus extract in three patients with juvenile chronic arthritis.

Recent therapeutic trials in rheumatology using different immunomodulating agents have given encouraging results. In this study an aqueous calf thymus extract (CTE) was administered to three patients, two with systemic juvenile chronic arthritis (JCA), Still's disease, who could not be weaned from steroids during 2 years of conventional therapy, and one girl with a chronic juvenile monarthritis who had responded unsatisfactorily to nonsteroidal antirheumatics for 19 months. A striking clinical improvement was observed in all three patients. Prednisone (PRED) was discontinued in one case with systemic (JCA) and 0.25 mg/kg body weight/day is presently being given to the other patient. The girl is doing well on 4 mg chloroquin kg body weight/day; indomethacin (IND) was discontinued. Laboratory data including cellular immunoreactivity normalized in all three patients.

Animals↗
PubMed2002

Adhesion promotion of Cu on C by Cr intermediate layers investigated by the SIMS method.

Copper-carbon composites are candidate materials for heat sinks for high speed/high-performance electronic components. They combine high thermal conductivity with low density and a tailorable coefficient of thermal expansion (CTE). Because of the low wettability of carbon by copper, a thin layer of chromium can be deposited to promote both the adhesion and the thermal contact of copper with the carbon fibers. Therefore, in a first step layers of Cr and Cu were deposited by magnetron sputtering on plane vitreous carbon substrates (Sigradur G), which serve as a model for carbon fibers. From pull-off-adhesion measurements an interlayer thickness of Cr in the range of 2-10 nm was found to provide the optimal adhesion for 1 micro m thick copper overlayers. To model the later serial fabrication of the composite that involves a hot pressing step following the deposition, the C/Cr/Cu samples were heat treated at 800 degrees C under vacuum for 1 h. Adhesion on the heat-treated samples was superior in comparison to the untreated ones. To obtain information about the adhesion mechanism secondary ion mass spectrometry (SIMS) investigations were done on the depth distribution of the main elements copper, chromium, and carbon. Two samples, one as deposited and one subjected to heat treatment after deposition, were compared in this investigation. We found that heat treatment mainly modifies the distribution of Cr in the C/Cr/Cu system.

Journal Article↗
PubMed1988

Comparison between required clothing insulation and that actually worn by workers exposed to artificial cold.

Required Clothing Insulation (IREQ) is a new thermal index submitted to the International Organisation for Standardisation (ISO) for discussion. It is designed to prevent general body cooling and is based on an analysis of heat exchanges. The thermal clothing insulation actually worn (lcl) is estimated using a new method, also submitted to ISO. IREQ of 54 workers exposed to artificial cold (air temperature between -30 degrees C and +10 degrees C) was compared with lcl actually worn by these workers. The results of the present study show that, on average, the workers choose accurately lcl they need if their IREQ is below and up to 1.5 clo. Moreover, these workers prefer to wear garments which provide them with thermal comfort. If IREQ of workers is higher than 1.5-2 clo (i e, workers exposed to -20 degrees C), it is difficult for them to increase their thermal insulation with additional garments. Although their lcl is not sufficient, there is no risk of gradual body cooling because of their continuous time exposure (CTE) which is shorter than the calculated Duration Limited Exposure (DLE). On the other hand, Wind Chill Index (WCI), which is proposed to prevent local cooling, is better adapted to prevent cold injuries than physiological thermal strain; for example, impairment of manual dexterity cannot be prevented with this index.

Journal Article↗
PubMed1991

Evidence for the stability and cytochrome P450 specificity of the phenobarbital-induced reductive halothane-cytochrome P450 complex formed in rat hepatic microsomes.

The hypothesis that the reduced spectral halothane-cytochrome P450 complex formed in rat hepatic microsomes is a stable cytochrome P450 specific species was examined. Comparisons of the cytochrome P450 inducers, phenobarbital (PB), pregnenolone-16 alpha-carbonitrile (PCN) and beta-naphthoflavone (beta-NF) showed that PB was the most effective inducer of the halothane-cytochrome P450 complex and the cytochrome P450 which liberates the halothane metabolites, 2-chloro-1,1-difluoroethene (CDE) and 2-chloro-1,1,1-trifluoroethane (CTE). However, the ratio of CDE produced to quantity of complex was found to be reduced 70-77% in these microsomes. A large portion of total microsomal cytochrome P450 was destroyed upon halothane reduction (up to 39%), yet the complexed cytochrome P450, particularly in microsomes from PB-treated animals, was resistant to the irreversible inactivation mechanisms of halothane reduction. The effects of reductive halothane metabolism on subsequent warfarin metabolism showed that 7-hydroxywarfarin formation from either (R)- or (S)-warfarin in microsomes from PCN-treated, PB-treated or untreated rats was highly susceptible to irreversible inhibition. In microsomes from PB-treated, but not PCN or untreated rats, the formation of one warfarin metabolite, 4'-hydroxywarfarin from (R)-warfarin, could be shown to be increased when complex was eliminated by photodissociation. These results suggest that PB-B is preferentially bound as complex and resistant to inactivation because of complex stability, and that halothane reduction readily destroys the cytochrome P450 form, PB-C.

Animals↗
PubMed2006

Elasticity reconstruction for ultrasound elastography using a radial compression: an inverse approach.

To reduce the inherent mechanical artifacts in the strain images, many groups have investigated solutions to the inverse problem in elastography. However, in prostate elastography or intravascular elastography where the compression direction is radial, the inverse problem has not been studied thoroughly. In this paper, an iterative approach is proposed to reconstruct tissue elasticity for ultrasound elastography using a radial compression. The method is based upon the stress-strain relations in the polar coordinates. Computer simulations in an intravascular model are performed to illustrate the feasibility of this method in reducing the mechanical artifacts of the strain images. The reconstructed elasticity error and the contrast-transfer efficiency (CTE) as a function of the iteration number show that the inverse approach converges with a few iterations.

Algorithms↗
PubMed2004

Immunization with the hybrid protein vaccine, consisting of Leishmania major cysteine proteinases Type I (CPB) and Type II (CPA), partially protects against leishmaniasis.

Cysteine proteinases (CPs) are enzymes that belong to the papain superfamily, which are found in a number of organisms from prokaryotes to mammals. On the parasitic protozoan Leishmania, extensive studies have shown that CPs are involved in parasite survival, replication and the onset of disease, and have, therefore, been considered as attractive drugs and/or vaccine targets for the control of leishmaniasis. We have previously shown that cysteine proteinases, Type I (CPB) and Type II (CPA), in Leishmania major (L. major), delivered as recombinant proteins or in plasmid DNA, induce partial protection against infection with the parasite in BALB/c mice. We had shown that the level of protection was greater if a cocktail of cpa and cpb containing DNA constructs was used. Therefore, to reduce the costs associated with the production of these vaccine candidates, a construct was developed, whereby the cpa and cpb genes were fused together to give rise to a single hybrid protein. The genes were fused in tandem where the C-terminal extension (CTE), encoding region of CPB, was located at the 3' of the fused genes, and ultimately expressed in the bacterial expression construct pET-23a. The expression of the CPA/B hybrid protein (60 kDa) was verified using rabbit anti-CPA and anti-CPB antibodies by SDS-PAGE and immunoblotting. The protective potential of the CPA/B hybrid protein against the infection with Leishmania was then assessed in BALB/c mice. The animals were vaccinated with CPA/B, challenged with live L. major promastigotes, and the degree of protection was examined by measuring footpad lesion sizes. It was found that there was a delay in the expansion of lesions size compared to control groups. Furthermore, an immunological analysis of antibody isotypes, before and after infection, showed high levels of IgG2a compared to IgG1 (more than five-fold) in the CPA/B hybrid protein vaccinated group. In addition, a predominant Th1 immune response characterized by in vitro IFN-gamma production was observed, along with little, if any, IL-5 production. This finding indicates that the hybrid CPA/B is able to elicit a protective immune response against L. major in the mice model. In addition, 54% of individuals tested, who had recovered from cutaneous leishmaniasis, produced more than 50 pg/ml IFN-gamma, in response to the CPA/B hybrid protein in an in vitro assay, demonstrating the importance of cysteine proteinases as targets of immune response in humans.

Animals↗
PubMed2002

Gaucher and Niemann-Pick diseases--enzymatic diagnosis in dried blood spots on filter paper: retrospective diagnoses in newborn-screening cards.

BACKGROUND: Gaucher disease (GD) and Niemann-Pick (NP) disease are caused by deficient activity of the lysosomal enzymes acid beta-D-glucosidase (ABG) and acid sphingomyelinase (ASM), respectively. For diagnosis, these enzymes are usually measured in the extracts of leukocytes or cultured fibroblasts. Chitotriosidase (CTE), a chitinolytic enzyme, is markedly increased in the plasma of Gaucher patients. We describe methods for the assay of acid beta-D-glucosidase, acid sphingomyelinase, chitotriosidase, and alpha-N-acetyl-galactosaminidase (NAGA) as a control enzyme in blood spots that were dried onto filter paper. METHODS: To tubes containing a 3 mm-diameter blood spot, we added elution liquid and substrate solution. After incubation at 37 degrees C, the amount of hydrolyzed product was compared with a calibrator to allow the quantification of enzyme activity. We examined 80 healthy controls, 54 Gaucher patients, 8 Niemann-Pick patients, 27 obligate carriers, and the newborn-screening cards (NSC) from a case of Gaucher and a case of Niemann-Pick disease. RESULTS AND CONCLUSION: The described methodology is useful to identify Gaucher and Niemann-Pick patients and controls, using samples that are sufficiently stable to be transported to the testing laboratory by mail. The diagnosis of both diseases on a newborn-screening card was clearly established. The newborn-screening card has been added to the biological materials that allow the identification of patients with Gaucher and Niemann-Pick diseases.

Adolescent↗
PubMed1999

Substrate and binding specificities of bacterial polyhydroxybutyrate depolymerases.

The substrate specificities of three extracellular polyhydroxybutyrate (PHB) depolymerases from Alcaligenes faecalis (PhaZ Afa), Pseudomonas stutzeri (PhaZ Pst), and Comamonas acidovorans (PhaZ Cac), which are grouped into types A and B based on the position of a lipase box sequence in the catalytic domain, were examined for films of 12 different aliphatic polyesters. Each of these PHB depolymerases used was capable of hydrolyzing poly(3-hydroxybutyrate) (P(3HB)), poly(3-hydroxypropionate) (P(3HP)), poly(4-hydroxybutyrate) (P(4HB)), poly(ethylene succinate) (PESU), and poly(ethylene adipate) (PEA) but could not hydrolyze another seven polyesters. In addition, the binding characteristics of substrate binding domains from PhaZ Afa, PhaZ Cac, and PHB depolymerase from Comamonas testosteroni (PhaZ Cte) were studied by using fusions with glutathione S-transferase (GST). All of fusion proteins adsorbed strongly on the surfaces of polyester granules of P(3HB), P(3HP), and poly(2-hydroxypropionate) (P(2HP)) which was not hydrolyzed by the PHB depolymerases used in this study, while they did not bind on Avicel and chitin granules. The adsorption kinetics of the fusion proteins to the surface of P(3HB) and P(2HP) granules were found to obey the Langmuir isotherm. The cross-area per molecule of fusion protein bound to P(3HB) granules was estimated to be 12+/-4 nm2/molecule. It has been suggested that the active sites in catalytic domains of PHB depolymerases have a similar conformational structure, and that several amino acids in substrate-binding domains of PHB depolymerases interact specifically with the surface of polyesters.

Adsorption↗
PubMed2001

Thermo-debonding mechanisms in dentin bonding systems using finite element analysis.

The finite element method (FEM) has been extensively used in evaluating the interfacial status of biomaterials. We used FEM to explore the microscopic debonding mechanism of the dentin/hybrid layer/resin adhesive interface. The stress status of the local material was used as an index to judge whether the adhesive interface would develop a debonding mechanism. To generate the local stress concentration, the thermal boundary condition was applied to the model which has the phenomenon of the coefficient of thermal expansion (CTE) mismatch. The thermal boundary condition was used to emulute a previous study conducted with a laser thermoacoustic technique (LTAT). The materials, Scotchbond MP, Optibond, and Tenure bonding systems, used in the previous experiment were also tested in this study. The results show that interfacial debonding in the finite element model occurred through the hybrid layer for both the Scotchbond MP and Tenure systems, as well as within the adhesive layer itself for the Optibond system. These findings are compatible with observations by SEM obtained by LTAT. Another transformed model was created to test the "elastic cavity wall" concept. The result also confirms the importance of the elastic cavity wall concept. These compatible results between FEM and LTAT indicate that FEM can be a very useful supplement to thermoacoustic testing.

Biocompatible Materials↗
PubMed2000

Finite element analysis of thermo-debonding mechanism in dental composites.

Finite element method (FEM) has been extensively used for evaluating interfacial status inside biomaterials. This study using FEM was designed to evaluate the thermal stress behavior of a filler-matrix interface. The results were then compared to those of a previous study obtained by a laser thermoacoustic technique (LTAT). The experimental systems (75/25 Bis-GMA/TEGDMA resin reinforced with 0, 25, 50, and 75 wt% 8-microm silanized/unsilanized BaSiO6) as used in the previous study were modeled in this study. The established finite element models were based on coefficient of thermal expansion (CTE) Mismatch Phenomenon. The mechanical properties of the silane coupling agent, such as elastic modulus and thermal expansion coefficient used in the silanized model, were assumed to have optimal heat flux transfer. A third (imaginary) material was proposed to block the transfer of thermal stress between the filler and matrix in the unsilanized model. The thermal load simulation was based on steady-state thermal analysis. The results showed that: (1) The strain energy and interfacial shearing stress calculated from FEM validate the results from the previous LTAT study. (2) Comparing the stress distribution of silanized and unsilanized FEM models, the acoustic signals in LTAT study are mainly derived from debonding of the filler-matrix interface of silanized specimens, and from the matrix area of unsilanized specimens. Based on results to date, we conclude that the finite element method may be a powerful tool for exploring thermoacoustic mechanisms of dental composites.

Acoustics↗
PubMed1998

An analysis of elastographic contrast-to-noise ratio.

We present a theoretical formalism and simulation results that allow the incorporation of the elastic contrast properties of tissues with simple geometries into the elastographic noise models developed previously. This analysis results in the computation of the elastographic contrast-to-noise ratio (CNRe). The CNRe in elastography is an important quantity that is related to the detectability of a lesion or inhomogeneity. In this paper, the upper bound on the elastographic CNRe is derived for both a one-dimensional (1-D) and 2-D analytic plane-strain tissue model. The CNRe in the elastogram depends on the contrast-transfer efficiency (CTE) for both the 1-D and 2-D geometries discussed in this paper. The 1-D model is used to characterize layered structures and the 2-D model is derived for circular inclusion within a background of uniform elasticity. A previously derived classical analytic solution of the elasticity equations, for a circular inclusion embedded in an infinite medium and subjected to a uniaxial compression, is used to compute the upper bound of the CNRe. Monte Carlo simulations illustrate the close correspondence between the theoretical and simulation results.

Elasticity↗
PubMed2002

Induction of sister chromatid exchanges and chromosome aberrations in cultured mammalian cells treated with aminophenylnorharman formed by norharman with aniline.

Aminophenylnorharman (APNH) is a newly identified mutagenic heterocyclic amine formed by coupling of norharman with aniline in the presence of S9 mix. Furthermore, mutagenic amino-3'-methylphenylnorharman (AMPNH) and aminophenylharman (APH) have been identified from a reaction mixture of norharman and o-toluidine and that of harman and aniline, respectively, with S9 mix. Among these three heterocyclic amines, APNH shows most potent mutagenic activity towards Salmonella typhimurium TA98 and YG1024 with S9 mix. In the present study, the induction of sister chromatid exchanges (SCEs) by APNH was examined in Chinese hamster lung (CHL) cells in vitro, comparing it to those of AMPNH and APH. On incubation with rat S9 for 6h, followed by a recovery culture period of 18h, a dose-dependent effect was found at concentrations between 0.00125 and 0.01 microg/ml for APNH and between 0.3125 and 5 microg/ml for AMPNH and APH. The approximate chemical concentrations leading to a three-fold of control SCE levels calculated from slopes of the linear regressions of induced SCEs were 0.005 for APNH, 0.51 for AMPNH and 1.7 microg/ml for APH. Because of the very strong SCE-causing ability of APNH, we further explored its genotoxicity by examining the induction of chromosome aberrations in CHL cells. A dose-dependent effect was found for chromosome aberrations at concentrations between 0.00125 and 0.04 microg/ml of APNH. The aberrations observed were primarily chromatid exchanges (cte) and breaks (ctb). In conclusion, the potency of SCE induction and clastogenic activity induced by APNH is stronger than Actinomycin D, Mitomycin C (MMC) or 1,8-dinitropyrene which are considered to be the potent clastogens in the literature. Further studies are needed for elucidating mechanisms of the genotoxic actions of these compounds and for evaluating their potential hazards to human health.

Aniline Compounds↗
PubMed2002

Relativistic spin-orbit coupling effects on secondary isotope shifts of (13)C nuclear shielding in CX(2) (X = O, S, Se, Te).

Rovibrational corrections, temperature dependence, and secondary isotope shifts of the (13)C nuclear shielding in CX(2) (X = O, S, Se, Te) are calculated taking into account the relativistic spin-orbit (SO) interaction. The SO effect is considered for the first time for the secondary isotope shifts. The nuclear shielding hypersurface in terms of nuclear displacements is calculated by using a density-functional theory method. Ab initio multiconfiguration self-consistent field calculations are done at the equilibrium geometry for comparison. (13)C NMR measurements are carried out for CS(2). The calculated results are compared with both present and earlier experimental data on CO(2), CS(2), and CSe(2). The heavy-atom SO effects on the rovibrational corrections of (13)C shielding are shown to be significant. For CSe(2) and CTe(2), reliable prediction of secondary isotope effects and their temperature dependence requires the inclusion of the SO corrections. In particular, earlier discrepancies of theory and experiment for CSe(2) are fully resolved by taking the SO interactions into account.

Journal Article↗
PubMed1998

High dose chemotherapy with autologous bone marrow rescue for children with diffuse pontine brain stem tumors. Children's Cancer Group.

PURPOSE: Diffuse pontine tumors are highly lethal, and there are few long-term survivors with the standard treatment of external beam irradiation. We investigated the effectiveness of high-dose thiotepa and etoposide-based chemotherapy regimens with autologous bone marrow rescue (ABMR) in children with pontine tumors. PATIENTS AND METHODS: Sixteen children with diffuse pontine tumors were treated. Ten had resistant or recurrent tumors. All ten had previously received irradiation; five had also received chemotherapy and one, beta-interferon. Three high-dose chemotherapy regimens were employed. Six patients received three days of thiotepa (300 mg/m2/day) and etoposide (250-500 mg/m2/day) (TE); two received three days of carmustine (BCNU) (200 mg/m2/day divided every 12 hours) followed by TE (BTE); and two received three days of carboplatin (500 mg/m2/day) followed by TE (CTE). Six other patients had newly-diagnosed tumors and had not received any prior treatment. They all received the BTE regimen and subsequently were treated with hyperfractionated irradiation (7200-7800 cGy) beginning approximately six weeks post-ABMR. RESULTS: There were two toxic deaths (13%), both in previously treated patients, due to multiorgan system failure and Candida septicemia in one case each. Median survival of the patients with resistant or recurrent disease was 4.7 months (range 0.1-18.7) from time of ABMR. Median survival of the newly-diagnosed patients was 11.4 months (range 7.6-17.1) from the time of ABMR. CONCLUSION: High-dose chemotherapy utilizing these regimens followed by ABMR did not appear to prolong survival compared to conventional therapy in these children with pontine tumors. Alternative strategies need to be developed for this highly lethal disease.

Adolescent↗
PubMed1998

High-dose thiotepa and etoposide-based regimens with autologous hematopoietic support for high-risk or recurrent CNS tumors in children and adults.

The prognosis in patients with primary brain tumors treated with surgery, radiotherapy and conventional chemotherapy remains poor. To improve outcome, combination high-dose chemotherapy (HDC) has been explored in children, but rarely in adults. This study was performed to determine the tolerability of three-drug combination high-dose thiotepa (T) and etoposide (E)-based regimens in pediatric and adult patients with high-risk or recurrent primary brain tumors. Thirty-one patients (13 children and 18 adults) with brain tumors were treated with high-dose chemotherapy: 19 with BCNU (B) and TE (BTE regimen), and 12 with carboplatin (C) and TE (CTE regimen). Patients received growth factors and hematopoietic support with marrow (n = 15), peripheral blood progenitor cells (PBPC) (n = 11) or both (n = 5). The 100 day toxic mortality rate was 3% (1/31). Grade III/IV toxicities included mucositis (58%), hepatitis (39%) and diarrhea (42%). Five patients had seizures and two had transient encephalopathy (23%). All patients had neutropenic fever and all pediatric patients required hyperalimentation. Median time to engraftment with absolute neutrophil count (ANC) >0.5 x 10(9)/l was 11 days (range 8-37 days). Time to ANC engraftment was significantly longer (P = 0.0001) in patients receiving marrow (median 14 days, range 10-37) than for PBPC (median 9.5 days, range 8-10). Platelet engraftment >50 x 10(9)/l was 24 days (range 14-53 days) in children. In adults, platelet engraftment >20 x 10(9)/l was 12 days (range 9-65 days). In 11 patients supported with PBPC, there was a significant inverse correlation between CD34+ dose and days to ANC (rho = -0.87, P = 0.009) and platelet engraftment (rho = -0.85, P = 0.005), with CD34+ dose predicting time to engraftment following HDC. Overall, 30% of evaluable patients (7/24) had a complete response (CR) (n = 3) or partial response (PR) (n = 4). Median time to tumor progression (TTP) was 7 months, with an overall median survival of 12 months. These TE-based BCNU or carboplatin three-drug combination HDC regimens are safe and tolerable with promising response rates in both children and older adults.

Adolescent↗