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1.
Phys Rev Lett ; 132(22): 223402, 2024 May 31.
Article in English | MEDLINE | ID: mdl-38877935

ABSTRACT

Symmetry-breaking phase transitions are central to our understanding of states of matter. When a continuous symmetry is spontaneously broken, new excitations appear that are tied to fluctuations of the order parameter. In superconductors and fermionic superfluids, the phase and amplitude can fluctuate independently, giving rise to two distinct collective branches. However, amplitude fluctuations are difficult to both generate and measure, as they do not couple directly to the density of fermions and have only been observed indirectly to date. Here, we excite amplitude oscillations in an atomic Fermi gas with resonant interactions by an interaction quench. Exploiting the sensitivity of Bragg spectroscopy to the amplitude of the order parameter, we measure the time-resolved response of the atom cloud, directly revealing amplitude oscillations at twice the frequency of the gap. The magnitude of the oscillatory response shows a strong temperature dependence, and the oscillations appear to decay faster than predicted by time-dependent Bardeen-Cooper-Schrieffer theory applied to our experimental setup.

2.
J Biomed Mater Res A ; 87(4): 921-32, 2008 Dec 15.
Article in English | MEDLINE | ID: mdl-18228268

ABSTRACT

Porous scaffolds have been made from two polyurethanes based on thermally induced phase separation of polymer dissolved in a DMSO/water mixture in combination with salt leaching. It is possible to obtain very porous foams with a very high interconnectivity. A major advantage of this method is that variables like porosity, pore size, and interconnectivity can be independently adjusted with the absence of toxic materials in the production process. The obtained compression moduli were between 200 kPa and 1 MPa with a variation in porosity between 76 and 84%. Currently the biological and medical aspects are under evaluation.


Subject(s)
Polyurethanes/chemistry , Salts/chemistry , Tissue Scaffolds , Animals , Biocompatible Materials/chemistry , Dimethyl Sulfoxide/chemistry , Materials Testing , Porosity , Solvents/chemistry , Spectroscopy, Fourier Transform Infrared , Surface Properties , Temperature , Tissue Engineering/methods
3.
J Biomed Mater Res B Appl Biomater ; 76(2): 389-96, 2006 Feb.
Article in English | MEDLINE | ID: mdl-16211565

ABSTRACT

In earlier studies, meniscal replacement with a porous polymer implant led to regeneration of neo-meniscal tissue. To evaluate the influence of the chemical properties on the tissue regeneration in the implant, in the present study, the meniscus in the dog's knee was replaced with either an aromatic 4,4-diphenylmethanediisocyanate based polyesterurethane implant (Estane) (n = 6) or with an aliphatic 1,4-butanediisocyanate based polyesterurethane implant (PCLPU) (n = 6). After 6 months, the knee joints were resected and the tissue behavior in the two different prostheses was evaluated microscopically. In both prostheses, a meniscus-like distribution of the tissue phenotype was found with collagen type I in the peripheral fibrous zones and collagen type II in the central, more cartilaginous zones. The compression-stress behavior of the implant-tissue construct remained in between the stiffness of the polymer material and that of the native meniscus. The PCLPU implant seemed to provoke less synovial tissue reaction. After meniscectomy solely, in 5 out of 6 cases, a meniscus-like regenerate was formed. Furthermore, the articular cartilage degeneration after placing a PCLPU implant did also not exceed the degeneration after the Estane implant or after meniscectomy. The differences between these two implants did not seem to influence the tissue regeneration in the implant. However, PCLPU seemed to evoke less tissue reaction and, therefore, is thought to be less or even nontoxic as compared with the Estane implant. Therefore, for studies in the future, the authors prefer the PCLPU prostheses for replacement of the meniscus.


Subject(s)
Implants, Experimental , Menisci, Tibial , Polyesters , Polyurethanes , Regeneration , Animals , Biocompatible Materials/chemistry , Biocompatible Materials/metabolism , Biomedical Engineering , Collagen Type I/metabolism , Collagen Type II/metabolism , Dogs , Female , Male , Materials Testing , Menisci, Tibial/anatomy & histology , Menisci, Tibial/surgery , Molecular Structure , Polyesters/chemistry , Polyesters/metabolism , Polyurethanes/chemistry , Polyurethanes/metabolism , Surface Properties
4.
Infect Immun ; 73(10): 6812-21, 2005 Oct.
Article in English | MEDLINE | ID: mdl-16177359

ABSTRACT

Ornithobacterium rhinotracheale is a bacterial pathogen known for causing respiratory disease in poultry. In this study, we demonstrate for the first time that cross-protective immunity against different O. rhinotracheale serotypes can be induced by live vaccination. Sera from these live-vaccinated and cross-protected birds were used to identify new vaccine targets by screening an O. rhinotracheale expression library. Out of 20,000 screened plaques, a total of 30 cross-reactive clones were selected for further analysis. Western blot analysis and DNA sequencing identified eight different open reading frames. The genes encoding the eight cross-reactive antigens were amplified, cloned in an expression vector, and expressed in Escherichia coli. Purified recombinant proteins with a molecular mass ranging from 35.9 kDa to 62.9 kDa were mixed and tested as a subunit vaccine for (cross-)protection against challenge with homologous and heterologous O. rhinotracheale serotypes in chickens. Subunit vaccination resulted in the production of antibodies reactive to the recombinant proteins on Western blot, and this eight-valent vaccine conferred both homologous and heterologous protection against O. rhinotracheale challenge in chickens.


Subject(s)
Antigens, Bacterial/immunology , Bacterial Vaccines , Flavobacteriaceae Infections/veterinary , Ornithobacterium/immunology , Poultry Diseases/prevention & control , Animals , Antibodies, Bacterial/immunology , Antigens, Bacterial/genetics , Bacterial Vaccines/immunology , Base Sequence , Cloning, Molecular , Cross Reactions , Flavobacteriaceae Infections/immunology , Flavobacteriaceae Infections/prevention & control , Genomic Library , Molecular Sequence Data , Open Reading Frames/genetics , Poultry Diseases/immunology , Protein Subunits/genetics , Protein Subunits/immunology , Vaccination
5.
Vaccine ; 23(26): 3404-11, 2005 May 16.
Article in English | MEDLINE | ID: mdl-15837364

ABSTRACT

Unravelling of the protective immunity acquired during a natural infection may contribute to vaccine development. To assess the role of antibody-mediated immunity in protection against Ornithobacterium rhinotracheale infection in chickens, a novel experimental method was applied that combined immune depletion and passive transfer of immunity within the same host. Administration of cyclophosphamide (CY) to broiler chickens successfully suppressed B lymphocyte development, and therefore humoral immunity, as confirmed by histological and serological analysis. Challenge of CY-treated birds with O. rhinotracheale revealed a significantly higher pathology score in comparison to immune-competent birds that received the same bacterial challenge. Measurement of serum immunoglobulin levels of immune-competent birds revealed a positive correlation between IgA and/or IgG production and protection against infection. Passive transfer of O. rhinotracheale-specific antiserum to the immune-suppressed birds prior to pathogen challenge significantly decreased morbidity. This protective effect was not observed after administration of control sera containing similar concentrations of immunoglobulins. Together, these results provide firm evidence that chicken humoral immunity to O. rhinotracheale is a key component in protection against infection. Our data confirm that the applied immune depletion and reconstitution approach is an attractive tool to analyse the nature of the protective immune response.


Subject(s)
Antibodies, Bacterial/administration & dosage , Flavobacteriaceae Infections/prevention & control , Immunization, Passive , Ornithobacterium , Animals , Antibodies, Bacterial/immunology , Chickens , Flavobacteriaceae Infections/veterinary , Immunity, Cellular , Immunization
6.
J Mater Sci Mater Med ; 15(4): 423-7, 2004 Apr.
Article in English | MEDLINE | ID: mdl-15332611

ABSTRACT

Longitudinal lesions in menisci are among the most frequent orthopedic problems of the knee. Repair by simple techniques is only limited to the vascular part of the meniscus. For repair of the avascular part of the meniscus a scaffold, which will assist the body in the formation of new meniscus cell tissue, might be applicable. In this study a biomedical segmented polyurethane with poly(epsilon-caprolactone) as soft segment and 1,4-butanediisocyanate and 1,4-butanediol as uniform hard segments has been synthesised. The material has a micro phase separated morphology and excellent mechanical properties. A porous scaffold was prepared via a combination of liquid-liquid phase separation and salt leaching. The foams prepared combined a very high interconnectivity and porosity with the desired compression modulus. After six months of implantation in the knees of beagles full ingrowth with cells was obtained and it was found that meniscus like tissue had been formed in the scaffold. Moreover, compression behaviour appeared to be comparable to native meniscus tissue.


Subject(s)
Absorbable Implants , Menisci, Tibial/cytology , Menisci, Tibial/physiology , Polyesters/chemistry , Polyurethanes/chemistry , Regeneration/physiology , Tissue Engineering/methods , Animals , Biocompatible Materials/chemical synthesis , Butylene Glycols/chemistry , Cell Division , Collagen/metabolism , Compressive Strength , Dogs , Elasticity , Equipment Design , Isocyanates/chemistry , Materials Testing , Membranes, Artificial , Menisci, Tibial/surgery , Porosity , Proteoglycans/metabolism , Surface Properties , Tissue Engineering/instrumentation , Transition Temperature , Treatment Outcome
7.
Biomaterials ; 24(14): 2541-8, 2003 Jun.
Article in English | MEDLINE | ID: mdl-12695081

ABSTRACT

Meniscal lesions often occur in the avascular area of the meniscus with little chance of spontaneous repair. An access channel in the meniscal tissue can function as an entrance for ingrowing repair tissue from the vascular periphery of the meniscus to the lesion in the avascular zone which again induced healing of the lesion. Implantation of a porous polymer in a full-thickness access channel induced healing. However, a better integration between meniscal tissue and the implant might be achieved with the combination of the newly developed porous polymers and a modified surgical technique. This might improve meniscal lesion healing and the repair of the access channel with neo-meniscal tissue. Longitudinal lesions were created in the avascular part of 24 canine lateral menisci and a partial-thickness access channel was formed to connect the lesion with the meniscal periphery. In 12 menisci, the access channel was left empty (control group), while in the remaining 12 menisci the polymer implant was sutured into the access channel. Repair of the longitudinal lesions was achieved with and without polymer implantation in the partial-thickness access channel. Polymer implants induced fibrous ingrowth with cartilaginous areas, which resembled neo-meniscal tissue. Implantation did not prevent articular cartilage degeneration.


Subject(s)
Menisci, Tibial/pathology , Menisci, Tibial/surgery , Plastic Surgery Procedures/methods , Polyesters/chemistry , Prostheses and Implants , Tibial Meniscus Injuries , Animals , Culture Techniques , Dogs , Female , Male , Treatment Outcome
8.
Osteoarthritis Cartilage ; 11(1): 78-84, 2003 Jan.
Article in English | MEDLINE | ID: mdl-12505490

ABSTRACT

OBJECTIVE: Partial meniscectomy is the golden standard for treating a bucket-handle tear in the meniscus of the knee, but it inevitably leads to articular cartilage degeneration. Surgical creation of an access channel between the lesion and the vascularized synovial lining is intended to induce ingrowth of repair tissue and thus avoid degeneration of articular cartilage. DESIGN: The presence and mechanism of cartilage degeneration were evaluated in 24 canine menisci after a longitudinal lesion and access channel had been created in the avascular part of the meniscus. In 12 menisci the channel was implanted with a porous polymer scaffold, while the remaining 12 were left empty. Evaluation was performed using routine histology and antibodies directed against denatured type II collagen (Col2-3/4M). RESULTS: Articular degeneration was apparent in the polymer implant group and the empty channel group. This consisted of fibrillation, loss of chondrocytes and decreased proteoglycan content. Areas of fibrillated cartilage always showed positive labeling with the collagen degradation antibody Col2-3/4M. Collagen degradation was also visible in non-fibrillated areas. The upper zone of the cartilage showed swelling especially in the implant group, with empty cell lacunae and moderate levels of Col2-3/4M antibody labeling. DISCUSSION: This reconstruction technique cannot be considered superior to partial meniscectomy. We propose that degradation of the collagen type II network is a result of cartilage fibrillation and vice versa.


Subject(s)
Cartilage Diseases/metabolism , Cartilage, Articular/metabolism , Collagen Type II/metabolism , Orthopedic Procedures/adverse effects , Animals , Cartilage Diseases/etiology , Dogs , Hindlimb/surgery , Menisci, Tibial/surgery
9.
Leukemia ; 16(1): 13-21, 2002 Jan.
Article in English | MEDLINE | ID: mdl-11840258

ABSTRACT

Using red cell phenotyping (RCP) and/or cytogenetics (CYT) we identified 19 patients with persisting mixed chimerism (MC) among 231 patients transplanted with partially T cell-depleted stem cell grafts from HLA-identical siblings. Persisting MC is defined as MC for more than 2 years in patients without any evidence of relapse. Median leukemia-free survival in these patients was 150 (range, 50-218) months. Diagnoses were ALL (n= 10); AML (n = 2); CML (n = 2); NHL (n = 2); MDS (n= 1); MM (n = 1) and SAA (n = 1). Purpose of this study was the long-term follow-up of MC and definition of patterns of chimerism in the various subsets of PBMCs and granulocytes. Using a PCR-STR technique CD3(+)/CD4(+) (T4 lymphocytes), CD3(+)/CD8(+) (T8 lymphocytes), CD45(+)/CD19(+) (B lymphocytes), CD45(+)/CD14(+) (monocytes), CD45(+)/CD15(+) (granulocytes) and CD3(-)/CD56(+) (NK-cells) were analyzed. The majority of patients with persisting MC were conditioned with a less intensive conditioning regimen and had little GVHD. Sequential monitoring of the chimerism resulted in a group of patients (n = 7) with very slow transient mixed chimerism that resulted in complete DC after median 7 years. Another nine patients had a relatively high percentage of persisting autologous cells for a median of 12 years and in three patients we observed a stable low percentage of autologous cells. Only two out of 19 patients (AML-CR1, CML-CP1) relapsed during follow-up. Both patients had a relatively high percentage of autologous cells. Chimerism in granulocytes and PBMC subsets was analyzed at a median of 8 years after SCT in nine patients. In five patients mixed chimerism simultaneously detected by RCP and CYT was associated with MC in all subsets. Within each individual patient the percentages of donor and recipient cells were very different between the different subsets. Two CML-CP1 patients were mixed chimera in only two subsets and in one patient these subsets represented pending relapse. In another two patients mixed chimerism with a very low number of autologous red cells was not found in the PBMCs because of the different sensitivity level of the RCP and the PCR-STR technique. We conclude that in patients with persisting mixed chimerism after partially T cell-depleted SCT a remarkable number of patients had lymphoid malignancies, the majority of the patients were conditioned with less intensive conditioning regimens and the mixed chimerism was not correlated with relapse. Chimerism in granulocytes and PBMC subsets did show great intra-individual differences in the subsets and these data correlated well with RCP and CYT data with the exception of the NK cells.


Subject(s)
Hematologic Neoplasms/therapy , Hematopoietic Stem Cell Transplantation , Adolescent , Adult , Cell Survival , Disease-Free Survival , Female , Follow-Up Studies , Graft Survival , Graft vs Host Disease/epidemiology , Graft vs Host Disease/etiology , Graft vs Host Disease/prevention & control , Hematologic Neoplasms/mortality , Hematopoietic Stem Cell Transplantation/methods , Humans , Immunophenotyping , Lymphocyte Count , Lymphocyte Depletion , Lymphocyte Subsets , Male , Middle Aged , Myeloid Cells , Polymerase Chain Reaction , Sensitivity and Specificity , T-Lymphocytes , Transplantation Conditioning , Transplantation, Homologous
10.
Cytometry ; 47(1): 24-31, 2002 Jan 01.
Article in English | MEDLINE | ID: mdl-11774346

ABSTRACT

BACKGROUND: To study the apoptotic process in time, we used the following flow cytometric (FCM) techniques: phosphatidylserine (PS) translocation by Annexin-V (AnV), DNA fragmentation by in situ end labeling (ISEL), and propidium iodide (PI) staining. Because PS translocation is assumed to be an early feature of programmed cell death (PCD), we questioned if AnV positivity implies inevitable cell death. METHODS: Apoptosis was induced in Jurkat cells by gamma-irradiation, incubation with camptothecin (CPT), or cytosine beta-D-arabinofuranoside (Ara-C). At different time intervals, PCD was quantified by AnV/PI and ISEL. To analyze the influence of cell handling procedures on PCD, we applied these three FCM techniques on CD34+ bone marrow (BM) stem cells after selection and after a freeze-thaw procedure. Various AnV/PI- CD34+ fractions were cultured in a single-cell single-well (SCSW) assay. RESULTS: Jurkat cells under three different detrimental conditions showed essentially the same pattern of apoptosis in time. Initially developed AnV+/PI- cells subsequently (within 1 h) showed ISEL positivity, after which they turned into AnV+/PI++ cells with even higher levels of ISEL positivity (80-90%). Eventually, they lost some of their PI and ISEL positivity and formed the AnV+/PI+ fraction. Cell handling of CD34+ cells caused high and variable AnV+/PI- fractions (overall range 23-62%). Within total AnV+ and AnV+/PI- populations, only a minority of CD34+ cells showed ISEL positivity (range 4-8% and 0.8-6%, respectively). Different fractions of AnV+/PI- CD34+ cells did have clonogenic capacity. CONCLUSIONS: PCD of cell suspensions in vitro can be followed accurately in time by these three FCM techniques. PS translocation is followed rapidly (within 1 h) by oligo-nucleosomal DNA fragmentation, after which cell (and nuclear) membrane leakage occurs. Detection of PS asymmetry by AnV-fluorescein isothiocyanate (FITC) is not always associated with (inevitable) apoptosis, as can be concluded from the proliferative capacity of AnV+ /PI- CD34+ cells in the SCSW assay.


Subject(s)
Apoptosis , Annexin A5 , Antigens, CD34 , Camptothecin/pharmacology , Cells, Cultured , Coloring Agents , Enzyme Inhibitors/pharmacology , Flow Cytometry/methods , Humans , In Situ Nick-End Labeling/methods , Jurkat Cells , Kinetics , Propidium , Time Factors
11.
Biomaterials ; 21(23): 2433-42, 2000 Dec.
Article in English | MEDLINE | ID: mdl-11055291

ABSTRACT

The cytotoxicity of poly(96L/4D-lactide) (PLA96), and of its accumulated degradation products, was investigated following different sterilization methods and pre-determined heat-accelerated degradation intervals. PLA96 samples sterilized by either steam, ethylene oxide, or gamma irradiation were left untreated (S0 samples), or were degraded for 30 h or 60 h (S30 and S60 samples) at 90 degrees C in water. Extracts of the samples and of the remaining degradation fluids (F30 and F60) were prepared. The toxicity of both unfiltered and filtered extracts was analyzed in a cell growth inhibition (CGI) assay and a lactate dehydrogenase (LDH) leakage assay. Physical analysis of the extracted samples and of the degradation fluids also was performed. The S0 extracts demonstrated no significant CGI. The CGI of the S30 extracts ranged from 37 to 78%, whereas the CGI of the S60 extracts ranged from 6 to 33%. The CGI of the F30 extracts ranged from 19 to 38% and the CGI of the F60 extracts was 98 to 123%. The LDH leakage assay only showed a high response to the unfiltered F60 extracts. Neither sterilization nor filtration appeared to influence the cytotoxicity of the extracts. Particle accumulation, however, might affect cell membrane permeability resulting in LDH leakage. The results of this study suggest that the cytotoxicity of PLA96 is related to the pH and possibly the osmolarity of the tested extracts. The pH and osmolarity, in turn, may depend on variations in the amounts of solubilized lactic acid and oligomers. These variations appear to result from degradation stage-dependent differences in crystallinity, molecular weight and molecular weight distribution of the PLA96 samples.


Subject(s)
Cell Survival/drug effects , Polyesters/pharmacology , Animals , Cell Division/drug effects , Cell Line , Chromatography, Gel , Fibroblasts/cytology , Fibroblasts/drug effects , Fibroblasts/enzymology , L-Lactate Dehydrogenase/metabolism , Mice , Polyesters/chemistry , Sterilization
12.
Biomaterials ; 21(23): 2453-60, 2000 Dec.
Article in English | MEDLINE | ID: mdl-11055293

ABSTRACT

New porous polyurethane urea and polyurethane amide scaffolds for meniscal reconstruction have been developed in a solvent-free process. As soft segments, copolymers of 50/50 L-lactide/epsilon-caprolactone have been used. After terminating the soft segment with diisocyanates, chain extension was performed with adipic acid and water. Reaction between the isocyanate groups and adipic acid or water provides carbon dioxide and results in a porous polymer. Extra hydroxyl-terminated prepolymer was added in order to regulate the amount of carbon dioxide formed in the foaming reaction. Furthermore, salt crystals ranging in size from 150 to 355 microm were added in order to induce macroporosity. The pore size was regulated by addition of surfactant and by the use of ultrasonic waves. The resulting porous polymer scaffolds exhibit good mechanical properties like a high-compression modulus of 150 kPa. Chain extension with adipic acid results in better mechanical properties due to better defined hard segments. This results from the lower nucleophilicity of carboxylic acids compared to water and alcohols. By adjusting the reaction conditions, materials in which macropores are interconnected by micropores can be obtained. On degradation only non-toxic products will be released; importantly, the materials were obtained by a simple, reproducible and solvent-free procedure.


Subject(s)
Knee Injuries/therapy , Knee Joint , Polyurethanes , Microscopy, Electron
13.
Blood ; 94(7): 2414-23, 1999 Oct 01.
Article in English | MEDLINE | ID: mdl-10498614

ABSTRACT

Expression of the multidrug resistance (MDR) mechanisms P-glycoprotein (Pgp) and MDR-related protein (MRP) decrease cellular retention and consequently cytotoxicity of anthracyclines. MDR is expressed on normal human hematopoietic progenitors and leukemic blasts. Normal CD34(+) progenitors showed rhodamine efflux in 20% to 30% of the cells, which could be blocked by verapamil. These cells appeared noncycling, in contrast to the proliferating rhodamine bright (RhoB) cells. We postulated that MDR expression can be downregulated by proliferation induction. Triggering rhodamine dull (RhoD) CD34(+) cells to proliferate indeed resulted in a higher rhodamine retention and significantly decreased efflux modulation by verapamil (P =.04). Also in acute myeloid leukemia (AML), the proliferation rate (percentage S/G(2)+M and Iododeoxyuridine labelings index) was significantly less in the RhoD blasts (P

Subject(s)
ATP Binding Cassette Transporter, Subfamily B, Member 1/genetics , Antibiotics, Antineoplastic/toxicity , Cell Cycle , Drug Resistance, Multiple , Hematopoietic Stem Cells/cytology , Hematopoietic Stem Cells/pathology , Leukemia, Myeloid, Acute/pathology , Antigens, CD34 , Blast Crisis/pathology , Bone Marrow Cells/cytology , Bone Marrow Cells/pathology , Cell Cycle/drug effects , Cell Division/drug effects , Cell Survival/drug effects , Cells, Cultured , Daunorubicin/toxicity , Gene Expression Regulation , Hematopoietic Stem Cells/drug effects , Humans , Idarubicin/toxicity , Idoxuridine , Leukemia, Myeloid, Acute/genetics , Verapamil/pharmacology
14.
Leukemia ; 13(9): 1390-8, 1999 Sep.
Article in English | MEDLINE | ID: mdl-10482990

ABSTRACT

Currently available data regarding the substrate specificity of the multi-drug resistance (MDR) mechanisms P-glycoprotein (Pgp) and MDR-associated protein (MRP1) for idarubicin are inconclusive. A multiparameter flow cytometry method was developed which allows simultaneous quantitative measurement of total cellular fluorescence and the amount of anthracyclines intercalated into the DNA. Anthracycline DNA intercalation was measured by fluorescence resonance energy transfer (FRET) between Hoechst 33342 and anthracyclines. Daunorubicin and idarubicin accumulation were studied and compared in established cell lines expressing Pgp and MRP1. The data demonstrate that daunorubicin DNA intercalation is affected by both Pgp and MRP1 whereas idarubicin DNA intercalation is affected only by MRP1. MRP1 and Pgp function could be blocked completely by 5 microM PAK 104P, while higher concentrations of verapamil, PSC 833 and cyclosporin A were necessary to attain complete blocking of MRP1 compared to Pgp. Daunorubicin DNA intercalation correlates better with cell survival and is more sensitive at physiological MDR expression as observed in hematopoietic progenitors than daunorubicin levels measured by total cellular fluorescence. In conclusion, idarubicin DNA intercalation is reduced by MRP1 but not by Pgp. PAK-104P is an effective modulator for both Pgp and MRP1 and may further improve idarubicin efficacy.


Subject(s)
ATP Binding Cassette Transporter, Subfamily B, Member 1/physiology , Antibiotics, Antineoplastic/pharmacology , Base Pair Mismatch , DNA-Binding Proteins/physiology , Idarubicin/pharmacology , Intercalating Agents/pharmacology , Multidrug Resistance-Associated Proteins , Antibiotics, Antineoplastic/metabolism , Benzimidazoles , Daunorubicin/metabolism , Energy Transfer , Flow Cytometry , Fluorescent Dyes , Humans , Idarubicin/metabolism , MutS Homolog 3 Protein , Rhodamines , Sensitivity and Specificity , Spectrometry, Fluorescence/methods , Tumor Cells, Cultured
15.
Ann Rheum Dis ; 58(2): 122-5, 1999 Feb.
Article in English | MEDLINE | ID: mdl-10343529

ABSTRACT

OBJECTIVE: To investigate purine metabolism in patients with systemic lupus erythematosus (SLE) for possible abnormalities that might be related to their overall impaired immune function. METHODS: This pilot study included 17 patients with SLE (2 men, 15 women). Enzyme activities of the purine enzymes 5'-nucleotidase (5'NT), purine nucleoside phosphorylase (PNP), and hypoxanthine-guanine-phosphoribosyltransferase (HGPRT) were measured in peripheral blood mononuclear cells (PBMC) and also in fractions of T cells (differentiation antigen CD3+) (n = 12) and B cells (CD19+) (n = 9). The activity of the thiopurine enzyme thiopurine-methyltransferase (TPMT) was measured in red cell lysate. Routine blood tests and indices of disease activity were measured as well. Results were compared with those of healthy volunteers. RESULTS: Compared with their controls, the female SLE patients had a more than 50% reduced activity of 5'NT in the T cell fraction (p = 0.001) and in PBMC (p < 0.000). 5'NT activity was also lower in B cells, but this was not statistically significant. Enzyme activities did not correlate with indices of disease activity, disease duration or the B cell/T cell ratio and no influence of medication was found. CONCLUSION: Reduced lymphocyte 5'NT activity is a novel finding in SLE. These results indicate that purine metabolism in SLE may be disturbed. Consequences of a low 5'NT activity may be an intracellular accumulation of (deoxy)purine nucleotides and a reduction of adenosine production. It is hypothesised that these factors may play a part in the overall impaired immune function and in the chronicity of inflammation in SLE.


Subject(s)
Leukocytes, Mononuclear/enzymology , Lupus Erythematosus, Systemic/enzymology , Nucleotidases/metabolism , Adult , Aged , Antigens, CD19 , B-Lymphocytes/enzymology , B-Lymphocytes/immunology , CD3 Complex , Case-Control Studies , Female , Humans , Lupus Erythematosus, Systemic/immunology , Male , Middle Aged , Pilot Projects , T-Lymphocytes/enzymology , T-Lymphocytes/immunology
16.
Leukemia ; 12(4): 571-7, 1998 Apr.
Article in English | MEDLINE | ID: mdl-9557616

ABSTRACT

Mobilized peripheral blood progenitor cells (PBPC) have been shown to differ qualitatively from bone marrow (BM) progenitors. The released progenitor cells are predominantly in G0/G1 and show a relatively high percentage of rhodamine dull cells. Within the BM these last two features are characteristic of the more primitive progenitors. Although the mobilized PB cells can give rise to long-term repopulation and thus contain stem cells, the frequency of stem cells is not much higher if long-term initiating cell (LTC-IC) assays are used. To determine whether quiescent stem cells are selectively released or the low-cycle status of PB progenitors is related to the release from the BM microenvironment, the cell cycle status and rhodamine content in the PB and BM during mobilization were studied and compared with steady-state BM. More differentiated and more primitive progenitors were separated based on differentiation markers and cloned in single cell assay. In mobilized PB 54% of the CD34+ cells (n=5) were rhodamine dull compared to 22% in steady-state BM (P=0.014) [n=6]. The percentage of CD34+ cells in the S/G2M phases of the cell cycle was 2.1% in the mobilized PB (n=11), and 18% in steady-state BM (n=11) [P=0.002]. During mobilization the fraction of cells in the S/G2M phase of the cell cycle was 16% in BM (n=7), similar to steady-state BM (P=0.34). The released progenitors represented a selection of BM progenitors, with significantly more primitive progenitors (CD34+/13+/33dim) and less lymphoid precursors (CD34+/19+). Within the more differentiated CD34+113+/33bright, myelomonocytic precursors, both in PB as well as in BM, the percentage S/G2M was relatively higher than in the CD34+/13+/33dim subfraction: in normal BM: median 18% vs 8% (P=0.006) [n=8]; in mobilized PB 3% vs 2% (P=0.03) [n=10]; and in BM during mobilization 24% vs 7% (P=0.01) [n=6]. The cycle status of mobilized PB progenitors was low both in the primitive and more differentiated subfractions. During the mobilization period the BM progenitors are cycling as in steady-state BM. The low-cycle status of the mobilized PB progenitors may be related to the loss of contact with the micro-environment.


Subject(s)
Antigens, CD34/blood , Antigens, CD34/physiology , Bone Marrow Cells/cytology , Hematopoietic Stem Cells/cytology , Bone Marrow Cells/metabolism , Cell Cycle/physiology , Cell Differentiation/physiology , Cell Division/physiology , Cell Fractionation , Cell Movement/physiology , Flow Cytometry , Fluorescent Dyes/pharmacokinetics , G1 Phase/physiology , Hematopoietic Stem Cells/metabolism , Humans , Immunophenotyping , Indicators and Reagents , Propidium , Resting Phase, Cell Cycle/physiology , Rhodamine 123 , Rhodamines/pharmacokinetics , S Phase/physiology , Staining and Labeling/methods
17.
J Mater Sci Mater Med ; 9(10): 561-6, 1998 Oct.
Article in English | MEDLINE | ID: mdl-15348688

ABSTRACT

Surface oxidation of ultra-high molecular weight polyethylene (UHMWPE) powder has an influence on the mixing procedure of chopped fibres and UHMWPE powder. Due to this oxidation hydrogen bonds can be formed between the fibres and powder particles, leading to a more homogeneous fibre-powder mixture. This treatment improves the fibre-matrix interface and thus the physical properties of the composite. Chromic acid treatment also has an influence on the mechanical and tribological properties of the aramid-UHMWPE composite. Although only a relatively small improvement is observed in the modulus, yield stress and stress at break, of 33, 17 and 9%, respectively, a substantial enhancement in wear resistance of 117% is observed.

18.
J Mater Sci Mater Med ; 9(12): 675-8, 1998 Dec.
Article in English | MEDLINE | ID: mdl-15348920

ABSTRACT

A new approach to the synthesis of biomedical polyurethanes based on epsilon-caprolactone and 1,4-butanediisocyanate with a high modulus, has been developed. By chain extending an epsilon-caprolactone prepolymer with a long uniform-size diisocyanate block, a segmented polyurethane with uniform-size hard segments was obtained. It shows excellent mechanical properties; an extremely high modulus of 105 MPa and a tensile strength of 35 MPa. The polymer is soluble at high concentrations in various volatile solvents such as chloroform and 1,4-dioxane. By a combination of salt-leaching and freeze-drying, porous materials have been obtained in which macropores ranging in size from 150-300 microm are highly interconnected by micropores. The material shows a sufficiently high compression modulus of 200 kPa and appears to be suitable for biomedical applications such as meniscal prostheses.

19.
J Biomed Mater Res ; 36(3): 337-46, 1997 Sep 05.
Article in English | MEDLINE | ID: mdl-9260105

ABSTRACT

The aim of this study was to evaluate the degradation and foreign-body reaction of poly(DL-lactide-epsilon-caprolactone) (PLA85CL50) bars. This specific biomaterial is used for the construction of nerve guides, which can be used in the reconstruction of short nerve gaps. Subcutaneously implanted PLA85CL50 bars were harvested after implantation periods ranging from 3 to 12 months and evaluated for the rate of degradation and the degree of foreign-body reaction. It was observed that this copolymer degraded completely within 12 months and that no lactide or epsilon-caprolactone crystals were formed. Furthermore, we conclude that the foreign-body reaction of PLA85CL50 is very mild. These properties make the amorphous copolymer of DL-lactide and epsilon-caprolactone (50:50) suitable for the construction of nerve guides.


Subject(s)
Biocompatible Materials/adverse effects , Foreign-Body Reaction/chemically induced , Polyesters/adverse effects , Administration, Cutaneous , Animals , Biocompatible Materials/administration & dosage , Male , Polyesters/administration & dosage , Rats , Rats, Wistar
20.
Int J Artif Organs ; 20(8): 463-8, 1997 Aug.
Article in English | MEDLINE | ID: mdl-9323510

ABSTRACT

The aim of this study was to evaluate functional nerve recovery following reconstruction of a 1 cm gap in the sciatic nerve of a rat, using a new biodegradable p (DLLA-epsilon-CL) nerve guide. To evaluate both motor and sensory nerve recovery, walking track analysis and electrostimulation tests were carried out after implantation periods, ranging from 3 to 15 weeks post-operatively. The first signs of functional nerve recovery were observed after 3 weeks. After 15 weeks, 70% of the motor- and 90% of the sensory nerve function was re-established. Return of nerve function was better, in comparison with results from other studies. This study demonstrated successful functional nerve recovery after the reconstruction of a 1 cm nerve gap with a biodegradable p(DLLA-epsilon-CL) nerve guide.


Subject(s)
Biocompatible Materials , Implants, Experimental , Nerve Regeneration , Polyesters , Sciatic Nerve/surgery , Animals , Electric Stimulation , Male , Motor Activity/physiology , Motor Neurons/physiology , Neurons, Afferent/physiology , Rats , Rats, Wistar , Plastic Surgery Procedures , Sciatic Nerve/physiology
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