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PubMedDate not supplied

Generating renal cancer-reactive T cells using dendritic cells (DCs) to present autologous tumor.

Dendritic cells (DCs) have been used as professional antigen-presenting cells in vitro to prime T-cell responses. In this study, we generated both CD8+ and CD4+ renal cell carcinoma (RCC)-reactive T cells using a completely autologous system of DCs presenting engulfed whole-tumor cells. We compared DCs presenting RCC tumor cells in different preparations and found ultraviolet-irradiated apoptotic tumor cells to be more immunogenic than necrotic tumor cells or live untreated tumor cells in generating tumor-reactive T cells. In analyzing the T cells generated in this fashion, a CD8+ RCC-reactive T-cell clone generated from a patient recognized an epitope derived from fibroblast growth factor 5 in the context of human leukocyte antigen (HLA) B44*02. CD4+ T cells generated from another patient recognized multiple allogeneic RCC lines expressing HLA-DRbeta1*04, indicating a common shared tumor antigen presented by HLA-DRbeta1*04. The technique of using DCs to present whole-tumor cells can consistently generate both CD4+ and CD8+ RCC-reactive T cells for use in both antigen identification and therapeutic protocols.

Animals↗
PubMed2006

The effect of different anticoagulants on thrombin generation.

Decrease in thrombin generation is the key effect in anticoagulation. The aim of the present study was to investigate the effect of anticoagulants on thrombin generation and the relation to platelet count. Plasma samples from 10 healthy volunteers (mean age 43.0 +/- 9 years) were incubated at preset platelet counts with different doses of the anticoagulants lepirudin, fondaparinux and low molecular weight heparins. Thrombin generation was measured in a tissue factor-mediated assay using a fluorometer and a slow-reacting fluorogenic substrate. The endogenous thrombin potential, the lag phase, the maximum reaction velocity (Vmax) and the concentration of a given anticoagulant required for 50% inhibition of thrombin generation (IC50) are presented. All three anticoagulants decreased endogenous thrombin potential and prolonged the lag phase in a dose-dependent manner. Fondaparinux and low molecular weight heparins, but not hirudin, decreased Vmax in a concentration-dependent manner. With increasing platelet count, the IC50 increased but the extent of this increase was not uniform for the three anticoagulants and the three variables investigated. The influence of anticoagulants on thrombin generation is variable, depending on their basic mechanism of action. In defining and comparing their effects, the endogenous thrombin potential, the lag phase and the maximum reaction velocity should be considered together. Platelets have a considerable influence on the magnitude of thrombin generation.

Adult↗
PubMed2003

New CNS-active drugs which are second-generation valproic acid: can they lead to the development of a magic bullet?

PURPOSE OF REVIEW: Valproic acid (VPA) is one of the four first line antiepileptic drugs (AEDs). VPA is also an effective drug in migraine prophylaxis and in treatment of bipolar disorders. The use of VPA is limited by its two rare but potentially life-threatening side effects, teratogenicity and hepatotoxicity, and it is the least potent of the established AEDs. Consequently, there is an incentive to develop a second-generation VPA. A successful, second-generation VPA would need to possess the following characteristics: broad-spectrum antiepileptic activity; better potency than VPA; and lack of teratogenicity and hepatotoxicity. These characteristics would give such a drug the potential to be utilized in epilepsy and other CNS disorders. RECENT FINDINGS: Intensive research has been carried out in order to develop a second-generation VPA that would be more potent and safer than VPA. Amide derivatives of VPA have shown particular value as potential follow-up compounds and have better in-vivo performance than VPA. Several CNS-active valproylamides are more potent as antiepileptics than VPA, they possess broad-spectrum antiepileptic activity, and have been found to be nonteratogenic in animal models. The amide analogues of VPA that emerged from structure-pharmacokinetic-pharmacodynamic relationship studies as promising second-generation compounds are: N-methyl-tetramethylcyclopropane carboxamide, (2S,3S)-valnoctamide, (R)-propylisopropyl acetamide and valproyl glycinamide. SUMMARY: At present there are three compounds in clinical trials in patients with epilepsy that can be regarded as second-generation VPA: valproyl glycinamide, 3-methylbutanamide or isovaleramide and SPD421 (DP-VPA). For any one of these second-generation valproic acids to become a successful follow-up compound to VPA, it has to fulfil the above criteria and also possess favorable pharmacokinetics.

Amides↗
PubMed2000

The origin of alternation of generations in land plants: a focus on matrotrophy and hexose transport.

A life history involving alternation of two developmentally associated, multicellular generations (sporophyte and gametophyte) is an autapomorphy of embryophytes (bryophytesphytes + vascular plants). Microfossil data indicate that Mid Late Ordovician land plants possessed such a life cycle, and that the origin of alternation of generations preceded this date. Molecular phylogenetic data unambiguously relate charophycean green algae to the ancestry of monophyletic embryophytes, and identify bryophytes as early-divergent land plants. Comparison of reproduction in charophyceans and bryophytes suggests that the following stages occurred during evolutionary origin of embryophytic alternation of generations: (i) origin of oogamy; (ii) retention of eggs and zygotes on the parental thallus; (iii) origin of matrotrophy (regulated transfer of nutritional and morphogenetic solutes from parental cells to the next generation); (iv) origin of a multicellular sporophyte generation; and (v) origin of non-flagellate, walled spores. Oogamy, egg/zygote retention and matrotrophy characterize at least some modern charophvceans, and are postulated to represent pre-adaptative features inherited by embryophytes from ancestral charophyceans. Matrotrophy is hypothesized to have preceded origin of the multicellular sporophytes of' plants, and to represent a critical innovation. Molecular approaches to the study of the origins of matrotrophy include assessment of hexose transporter genes and protein family members and their expression patterns. The occurrence in modern charophyceans and bryophytes of chemically resistant tissues that exhibit distinctive morphology correlated with matrotrophy suggests that Early-Mid Ordovician or older microfossils relevant to the origin of land plant alternation of generations may be found.

Animals↗
PubMed1992

Oxygen radical generation by isolated microsomes from soybean seedlings.

The generation of active oxygen species by microsomes isolated from soybean seedlings was studied. NADPH-dependent superoxide anion production was 5.0 +/- 0.4 nmol . min(-1) mg(-1) of microsomal protein. Hydrogen peroxide generation by microsomes was 1.40 +/- 0.05 nmol . min(-1) mg(-1) of protein. Hydroxyl radical production, in the presence of ferric EDTA, evaluated through the generation of formaldehyde from dimethyl sulfoxide or tert-butyl alcohol was 0.50 +/- 0.04 and 0.44 +/- 0.03 nmol . min(-1) mg(-1), respectively. NADH proved to be suitable as cofactor for oxygen radical generation by microsomes from soybean seedlings. Because transition metals are implicated in radical generation by biological systems, the ability of microsomal membranes to reduce iron complexes was studied. Ferric ATP, ferric citrate, ferric ADP, ferric diethylenetriamine pentaacetic acid, and ferric EDTA were efficiently reduced in the presence of either NADPH or NADH as cofactor. The pattern of effectiveness of the different ferric complexes, on superoxide anion, hydrogen peroxide, and hydroxyl radical production, was similar to that found with animal microsomes. The data presented here indicate that microsomal ability to catalyze oxygen radical generation must be considered as an important contribution to cellular radical steady-state concentrations in cells from soybean seedlings.

Journal Article↗
PubMed2000

MAP kinase and protein kinase A-dependent mobilization of triacylglycerol and glycogen during appressorium turgor generation by Magnaporthe grisea.

Magnaporthe grisea produces an infection structure called an appressorium, which is used to breach the plant cuticle by mechanical force. Appressoria generate hydrostatic turgor by accumulating molar concentrations of glycerol. To investigate the genetic control and biochemical mechanism for turgor generation, we assayed glycerol biosynthetic enzymes during appressorium development, and the movement of storage reserves was monitored in developmental mutants. Enzymatic activities for glycerol generation from carbohydrate sources were present in appressoria but did not increase during development. In contrast, triacylglycerol lipase activity increased during appressorium maturation. Rapid glycogen degradation occurred during conidial germination, followed by accumulation in incipient appressoria and dissolution before turgor generation. Lipid droplets also moved to the incipient appressorium and coalesced into a central vacuole before degrading at the onset of turgor generation. Glycogen and lipid mobilization did not occur in a Deltapmk1 mutant, which lacked the mitogen-activated protein kinase (MAPK) required for appressorium differentiation, and was retarded markedly in a DeltacpkA mutant, which lacks the catalytic subunit of cAMP-dependent protein kinase A (PKA). Glycogen and lipid degradation were very rapid in a Deltamac1 sum1-99 mutant, which carries a mutation in the regulatory subunit of PKA, occurring before appressorium morphogenesis was complete. Mass transfer of storage carbohydrate and lipid reserves to the appressorium therefore occurs under control of the PMK1 MAPK pathway. Turgor generation then proceeds by compartmentalization and rapid degradation of lipid and glycogen reserves under control of the CPKA/SUM1-encoded PKA holoenzyme.

Biological Transport↗
PubMed1996

Automated hexahedral mesh generation from biomedical image data: applications in limb prosthetics.

A general method to generate hexahedral meshes for finite element analysis of residual limbs and similar biomedical geometries is presented. The method utilizes skeleton-based subdivision of cross-sectional domains to produce simple subdomains in which structured meshes are easily generated. Application to a below-knee residual limb and external prosthetic socket is described. The residual limb was modeled as consisting of bones, soft tissue, and skin. The prosthetic socket model comprised a socket wall with an inner liner. The geometries of these structures were defined using axial cross-sectional contour data from X-ray computed tomography, optical scanning, and mechanical surface digitization. A tubular surface representation, using B-splines to define the directrix and generator, is shown to be convenient for definition of the structure geometries. Conversion of cross-sectional data to the compact tubular surface representation is direct, and the analytical representation simplifies geometric querying and numerical optimization within the mesh generation algorithms. The element meshes remain geometrically accurate since boundary nodes are constrained to lie on the tubular surfaces. Several element meshes of increasing mesh density were generated for two residual limbs and prosthetic sockets. Convergence testing demonstrated that approximately 19 elements are required along a circumference of the residual limb surface for a simple linear elastic model. A model with the fibula absent compared with the same geometry with the fibula present showed differences suggesting higher distal stresses in the absence of the fibula. Automated hexahedral mesh generation algorithms for sliced data represent an advancement in prosthetic stress analysis since they allow rapid modeling of any given residual limb and optimization of mesh parameters.

Artificial Limbs↗
PubMed2002

Damping, amplitude, aging tests of stacked transducers for shock wave generation.

New clinical concepts in lithotripsy demand small shock heads. Reducing the size of piezoelectric shock heads will be possible only if the pressure generated at the surface of each transducer can be increased so that the total pressure at the focus remains the same. To solve this problem, different solutions were proposed. For example, it has been demonstrated that piezocomposite material, as opposed to piezoceramic material, allows the generation of a higher surface pressure before breaking, mainly because radial modes are dramatically reduced. In addition, in a previous paper, we showed the feasibility of generating high-pressure pulse waves without increasing the transducer voltage by using sandwiched transducers, which are a stack of two or more transducers. Some discrepancies appeared, however, between the pressure measured at the surface of the front transducer and the arithmetic sum of the pressures generated by each transducer constituting the stack. In fact, development of such stacked transducers capable of generating surface pressures in the range of 2 to 5 MPa is very complex, which may explain why no aging tests have been reported in the literature thus far. In the first part of this paper, we theoretically determine the importance of the electroacoustical coupling between the two transducers on the generated surface pressure. We show that pressure losses due to these electroacoustical couplings are less than 5%. Experimental measurements done on a stacked transducer assembled and tightened in a castor oil-filled tank are in excellent accordance with the theoretical measurements. Using this assembly technique, it was possible to obtain, on average, out of four elements, a pressure of 7.5 MPa for the duration of 4 million shocks, which would allow the treatment of approximately 1000 patients.

Computer Simulation↗
PubMed2003

Sensitivity of point- and line-source laser-generated acoustic wave to surface flaws.

Laser generation and air-coupled detection were combined as a hybrid ultrasonic technique for the inspection of surface flaws in rail. Narrowband acoustic signals were generated using a formed laser source by focusing the laser light to a point and to a line on the surface of the rail. The pulse energy, and therefore the intensity of the laser source, varied such that the generated signal transitioned from the weak thermoelastic to the strong ablative regime. The detection of flaws using laser-generated surface acoustic wave, to the presence of surface flaws, was compared between both point and line laser sources operating under different pulse energy levels. The line source was found to be more sensitive to the presence of surface flaws than a point source. The sensitivity of the laser-generated acoustic signal appeared to be independent of the severity of the flaw and, within the ablative regime, independent of the laser-pulse energy. Theoretical analysis is provided to explain the underlying cause that influences the interaction of a formed laser-generated surface acoustic wave to surface flaws and how this sensitivity may vary between the thermoelastic and ablative regimes.

Journal Article↗
PubMed1987

Mitomycin C-treated 3T3 fibroblasts used as feeder layers for human keratinocyte culture retain the capacity to generate eicosanoids.

Murine 3T3 fibroblasts, treated with mitomycin C in order to inhibit their proliferation, have been extensively used as feeder layers to enhance the cultivation of human keratinocytes in vitro. In order to identify possible factors responsible for this enhancing effect, studies were undertaken to determine whether eicosanoid generation by 3T3 cells was preserved following mitomycin C treatment. Accordingly, both untreated and mitomycin C-treated 3T3 cells were incubated with [1-14C]arachidonic acid and their qualitative generation of [1-14C]labeled eicosanoids assessed by thin layer chromatography. Levels of endogenously generated eicosanoids were quantitatively determined by specific radioimmunoassays performed on culture supernatants obtained from incubations of both treated and untreated cells. Results of these studies demonstrate that mitomycin C treatment, although preventing the proliferation of 3T3 cells, failed to alter their generation of prostaglandin E2 or 6-keto-prostaglandin F1 alpha. Furthermore, in parallel cultures, the untreated 3T3 cells stopped generating eicosanoids upon reaching confluence, whereas the mitomycin C-treated 3T3 cells that did not proliferate continued to generate eicosanoids.

Animals↗
PubMed1991

Kinin-generating cascade in advanced cancer patients and in vitro study.

The role of the bradykinin-generating system in the pathogenesis of cancer was explored by simultaneously measuring plasma prekallikrein (PK), the precursor of kallikrein, which is the major enzyme responsible for kinin generation, and plasma kininogens (KNG), which are precursors of kinin, in patients with various cancers. The mean value of plasma PK in healthy volunteers was 2.5 +/- 0.5 (mean +/- SD) units/mg plasma protein and that in cancer patients (all stage IV) was 1.7 +/- 0.7 units/mg plasma protein. The mean value of plasma KNG in healthy volunteers was 12.5 +/- 2.0 ng kinin equivalents/mg plasma protein and that in cancer patients was 10.9 +/- 2.8 ng. These data showed that plasma PK and plasma KNG values were significantly lower in cancer patients compared with healthy volunteers (P less than or equal to 0.005 for PK; 0.0005 less than P less than or equal to 0.005 for KNG; n = 28 for healthy subjects; n = 29 for cancer patients). These data appear to indicate that conversion of PK to kallikrein would probably occur with concomitant consumption of KNG by newly generated kallikrein for kinin generation in cancer patients. Early stage cancer patients showed little difference from healthy volunteers. For the in vitro study, activation of purified Hageman factor (HF) and PK was examined by using cancer cell lines and virus-transformed cells that produced plasminogen activator (PA) at a high rate. Both HF and PK were activated in the presence of plasminogen. Diploid cell lines and primary fibroblasts, which did not produce PA, activated neither HF nor PK. Taking all these data together, we conclude that kinin generation does occur in the plasma of patients with advanced cancer, and that one of the initiation mechanisms of the kinin-generating cascade appears to be mediated by plasmin and to depend on cancer cell-derived PA activity.

Adult↗
PubMed1981

Determination of steady state generation time distributions from labelled mitosis experimental data.

The proper application of detailed deterministic cell kinetic models depends on the way in which cells are assigned their generation times. A method is presented for the determination of population generation time distributions from labelled mitoses experiments. The model assumes that the generation time of each new cell is a function of both the steady-state generation time distribution function of the population, and also the generation time frequency-function of the previous generation of cells. This approach is applied to two different cell types to successfully simulate extended labelled mitoses curves using a population balance model with constant maturation rates.

Animals↗
PubMed2005

Influence of interleukin-4 on the phenotype and function of bone marrow-derived murine dendritic cells generated under serum-free conditions.

Murine bone marrow-derived dendritic cells (DC) can be generated by culture in the presence of granulocyte/macrophage colony-stimulating factor (GM-CSF) alone or GM-CSF in conjunction with interleukin-4 (IL-4). However, these two culture methods result in the production of heterogeneous DC populations with distinct phenotypic and stimulatory properties. In this study, we investigated the properties of DC generated under serum-free conditions in the presence or absence of IL-4 and compared their yield and phenotype to that of DC generated in the presence of fetal calf serum (FCS) (+/-IL-4). We did not observe a significant difference in the total cell yield between these four culture conditions, although the proportion of CD11c+ DC in cultures that received FCS was higher than that of their counterparts generated under serum-free conditions. Also, the four culture conditions generated CD11c+ DC with comparable levels of major histocompatibility complex (MHC) class II, CD40, CD80 and CD86 expression, with the exception of cells cultured under serum-free conditions in the absence of IL-4, which displayed suboptimal levels of these markers. Moreover, we compared the functional and stimulatory properties of DC generated under serum-free conditions in the presence or absence of IL-4. DC cultured in the presence of IL-4 were stronger stimulators of allogeneic splenocytes in a primary mixed lymphocyte reaction (MLR) and of naive antigen-specific OT-II transgenic T cells when pulsed with the class II ovalbumin (OVA)323-339 peptide or whole OVA protein than DC cultured in the absence of IL-4. However, both DC populations displayed a similar capacity to take up fluorescein isothiocyanate (FITC)-albumin by macropinocytosis and FITC-Dextran by the mannose receptor and to secrete IL-12 in response to stimulation with lipopolysaccharide (LPS) or an agonistic anti-CD40 monoclonal antibody. Therefore, we conclude that although both DC culture methods result in the production of DC with similar functional abilities, under serum-free conditions, DC cultured in GM-CSF and IL-4 show an increased stimulatory potential over DC cultured in GM-CSF alone. This is an important consideration in the design of experiments where DC are being exploited as immunotherapeutic vaccines.

Animals↗
PubMed2005

Leukaemia-derived dendritic cells can be generated from blood or bone marrow cells from patients with myelodysplasia: a methodological approach under serum-free culture conditions.

Functional dendritic cells (DC) are professional antigen-presenting cells (APC) and can be generated in vitro from healthy as well as from leukaemic cells from AML patients giving rise to APC of leukaemic origin presenting leukaemic antigens. In a comparative methodological analysis of 50 AML samples, we could already show that leukaemia-derived DC can regularly be generated under serum-free culture conditions. In this study, we describe the generation and characterization of DC from different mononuclear cell (MNC) fractions from 24 myelodysplastic syndrome (MDS) patients under those different serum-free culture conditions, determine the optimal culture conditions and compare the results with that from 23 healthy donors. In parallel cultures, we compared DC harvests after 7- or 14-day culture, with total or adherent MNC or T-cell-depleted MNC or PB or BM-MNC, thawn or fresh MNC, in Xvivo or CellGro serum-free media, +/-10% autologous plasma or +/-FL. In detail, we could show that MDS-DC harvests compared to healthy DC were higher after 10- to 14-day culture; total or adherent PB or BM-MNC fractions yield comparable DC counts; however, from MACS-depleted MNC fractions or thawn MNC lower DC counts can be generated. Whereas the addition of FL increases the DC harvest, the addition of autologous plasma in many cases has inhibitory influence on DC maturation, CellGro and Xvivo media yield comparable DC counts. Optimal harvest of vital and mature DC from MDS samples was obtained with a GM-CSF, IL-4, FL and TNF-alpha containing serum-free Xvivo medium after 10-14 days of culture (18/26% DC; 54/64% vital DC; 59/51% mature DC were generated from MDS/healthy MNC samples). Surface marker profiles (e.g. costimulatory antigen expression) of DC obtained from MDS samples were comparable with that of healthy DC. The leukaemic derivation of MDS-DC was demonstrated by the persistence of the clonal cytogenetic aberration in the DC or by coexpression of leukaemic antigens on DC. Autologous T-cell activation of leukaemia-derived DC was demonstrated in cases with MDS. Autologous T cells proliferate and upregulate DC-contact-relevant antigens. We are the first who demonstrate that the generation of leukaemia-derived DC is feasible not only in AML but also in MDS under serum-free culture conditions giving rise to DC with comparable characteristics as healthy DC and offering an antileukaemia-directed immunotherapeutical vaccination strategy in AML and MDS.

Adult↗
PubMed2005

Signal pathway of cytokines produced by reactive oxygen species generated from phorbol myristate acetate-stimulated HMC-1 cells.

The relationship of cytokine production and reactive oxygen species (ROS) generated in mast cells has not been reported yet. This study aimed to examine the signal pathway in the production of cytokines [interleukin-8 (IL-8) and tumour necrosis factor-alpha (TNF-alpha)] by ROS generated from phorbol myristate acetate (PMA)-stimulated human mast cell line-1 cells (HMC-1). HMC-1 cells were stimulated with 25 ng/ml of PMA. The ROS generation and production of cytokines (IL-8 and TNF-alpha) were measured by fluorescence-activated cell sorter and enzyme-linked immunosorbent assay method, respectively. Phosphorylation of mitogen-activated protein kinase family (extracellular signal-regulated kinase, p38 and c-Jun N-terminal kinase) was detected by the Western blotting method. The expression of cytokine's mRNA was measured by reverse transcriptase--polymerase chain reaction, and the DNA-binding activity of the transcription factors [nuclear factor-kappaB (NF-kappaB) and activator protein-1] was detected by electrophoretic mobility shift assay. PMA-stimulated HMC-1 cells immediately generated ROS, and the generated ROS was inhibited by 1,3-dimethyl-2-thiourea (DMTU), but partially inhibited by catalase or N-acetyl-L-cysteine. The production of cytokines in PMA-stimulated HMC-1 cells reached the maximum at 3-5 h and was inhibited by DMTU and specific kinase inhibitor for p38, SB203580. DMTU and SB203580 also inhibited the expressed cytokine's mRNA level and the increased DNA-binding activity of transcription factors, NF-kappaB in PMA-stimulated HMC-1 cells. These data suggest that intracellular ROS generated from PMA-stimulated HMC-1 cells contributes to the production of inflammatory cytokines via p38 kinase/NF-kappaB.

Antioxidants↗
PubMed1992

Specificity and sensitivity of first and second generation anti-HCV ELISA in a low prevalence population.

Specificity and sensitivity of the second generation enzyme immunosorbent assay for hepatitis C antibodies were calculated from data on 208,554 blood donors screened with either the first or the second generation tests (ELISA I, ELISA II) or both (4,639 donors). The second generation ELISA was no more specific than the first generation test in this low-prevalence blood donor population (specificity 99.7% vs 99.6%). Most of the donors (93.5%) who were found to be reactive using ELISA II were negative or indeterminate according to a supplemental test, the second generation recombinant immunoblot assay (RIBA II). The second generation ELISA was found to be more sensitive than the first. It identified all 16 donors who were positive according to RIBA II during screening with ELISA I. The latter found only 13 out of 30 donors (43%) who were positive according to RIBA II during screening with ELISA II. If the number of RIBA II positive donors found during the first year of screening with ELISA I (74, 0.024%), and the number of RIBA II positive but ELISA I negative donors found during screening with ELISA II, are taken into consideration it can be estimated that the added sensitivity due to using ELISA II is about 40%.

Blood Donors↗
PubMed2006

Generation of the B*41 group of alleles as indicated by intron sequences.

The generation of the B*41 alleles has been analysed using exon 1, intron 1, exon 2, intron 2 and exon 3 sequences. Results showed that B*4102 may have been generated as the first B*41 allele by a recombination mechanism between B*400102 and B*0801 or B*4201 involving intron 2. B*4101, B*4104 and B*4107 alleles could have been generated from B*4102 by a gene conversion event taking three different fragments from sequences belonging to intron 2/exon 3 of B*45, B*50 or B*49 alleles. B*4105 and B*4106 could be generated from B*4101 allele by point mutations, and B*4103 generation is unclear due to the lack of intron 2. The importance of introns in HLA-B allele polymorphism generation is stressed.

Alleles↗
PubMed1992

Second-generation anti-HCV tests predict infectivity.

Twenty-four blood donors found positive for the first-generation hepatitis C antibody (anti-HCV) test (Ortho EIA-I) and 88 of their recipients over the period from 1972 to 1990 were retrospectively investigated with different first- and second-generation anti-HCV tests. The aim of the study was to identify the infective donors and to evaluate the tests. Seven donors, who probably were infective carriers of HCV, were also second-generation test (EIA-II) positive, compared to only 3 out of 17 noninfective donors. Among the infected recipients, 14 out of 29 (48%) were positive for the second-generation test only. The second-generation test identified the infective donors in our study and was more sensitive than the first-generation test. We therefore recommend that blood donors are screened with EIA-II. Positive test results should be confirmed by the recombinant immunoblot assay (RIBA-II), and persons with positive or not conclusive RIBA-II should not be accepted as blood donors.

Blood↗