Showing papers by "Vrije Universiteit Brussel published in 1997"

Occurrence of multiple genomovars of Burkholderia cepacia in cystic fibrosis patients and proposal of Burkholderia multivorans sp. nov.

Abstract: We performed an integrated genotypic and phenotypic analysis of 128 strains of the genera Burkholderia, Ralstonia, and Pseudomonas in order to study the taxonomic structure of Burkholderia cepacia and its relationships with other Burkholderia species. Our data show that presumed B. cepacia strains isolated from cystic fibrosis patients belong to at least five distinct genomic species, one of which was identified as Burkholderia vietnamiensis. This group of five phenotypically similar species is referred to as the B. cepacia complex. The name Burkholderia multivorans is proposed for one of these genomic species, which was formerly referred to as B. cepacia genomovar II; the remaining B. cepacia groups are referred to as genomovars I, III, and IV, pending additional differential phenotypic tests. The role and pathogenic potential of each of these taxa, particularly in view of the potentially fatal infections in cystic fibrosis patients, need further evaluation. The data presented also demonstrate that Pseudomonas glathei and Pseudomonas pyrrocinia should be reclassified as Burkholderia species.

The nucleotide sequence of Saccharomyces cerevisiae chromosome VII.

Abstract: Here we report the sequence of 569,202 base pairs of Saccharomyces cerevisiae chromosome V. Analysis of the sequence revealed a centromere, two telomeres and 271 open reading frames (ORFs) plus 13 tRNAs and four small nuclear RNAs. There are two Ty1 transposable elements, each of which contains an ORF (included in the count of 271). Of the ORFs, 78 (29%) are new, 81 (30%) have potential homologues in the public databases, and 112 (41%) are previously characterized yeast genes.

The synthetic modeling of language origins

Abstract: This paper surveys work on the computational modeling of the origins and evolution of language. The main approaches are described and some example experiments from the domains of the evolution of communication, phonetics, lexicon formation, and syntax are discussed.

The in vitro micronucleus test: a multi-endpoint assay to detect simultaneously mitotic delay, apoptosis, chromosome breakage, chromosome loss and non-disjunction.

Abstract: Genotoxicity testing aims to detect a large range of genetic damage endpoints and evaluate such results in context of cell survival. The cytokinesis block micronucleus test offers the advantage to provide simultaneously information on both cell cycle progression and chromosome/genome mutations. Indeed, 1. frequencies of cytokinesis-blocked binucleated cells (and polynucleated) are good estimators of the mitotic rate; 2. frequencies of apoptotic figures in mononucleated and binucleated cells provide a measure for cell death before or after cell division; 3. combination of fluorescence in situ hybridization (FISH) for centromere/telomeres and micronucleus scoring allows discrimination between clastogenic and aneugenic events; 4. detection of FISH signals for chromosome specific sequences in both macronuclei and micronuclei, discriminates between aneuploidy due to chromosome non-disjunction or to chromosome loss. The cytokinesis block in vitro micronucleus test is thus a cytogenetic multi-test providing mechanistic information with a simple, rapid, objective, microscopical analysis.

Secular changes in growth and maturation: an update.

Abstract: Secular changes in growth and maturation in recent decades have been reviewed for various populations. The secular increase in attained height during the growth period is continuing in most countries, but has slowed down. The increase in adult stature over the past decades has varied between 0.3 and 3.0 cm/decade. The secular trend in the tempo of growth (earlier menarche and peak height velocity, and shortening of the growth cycle) has come to a halt in some populations, but is continuing or has been reversed in others. The secular trend in attained height and in the tempo of growth is usually more pronounced in children from low socioeconomic backgrounds, in those with poorly educated parents or in those from rural areas. It is concluded that updates of growth standards are required in all populations. More marked secular changes appear to occur in the lower height centiles, which may have direct implications on the future definition of 'short stature' in a population.

Interleukin-10 prevents the generation of dendritic cells from human peripheral blood mononuclear cells cultured with interleukin-4 and granulocyte/macrophage-colony-stimulating factor

Abstract: We evaluated the effects of interleukin (IL)-10 on the differentiation of dendritic cells (DC) obtained by culturing plastic-adherent peripheral blood mononuclear cells for 7 days in presence of granulocyte/macrophage-colony-stimulating factor (GM-CSF) + IL-4. The addition of IL-10 at the initiation of culture resulted in the generation of macrophage-like cells with expressing high levels of CD14 and decreased levels of CD1a and CD1c. Furthermore, cells generated in presence of IL-10 secreted lower levels of IL-12, but higher levels of IL-8 compared with DC generated in absence of IL-10, both spontaneously and after CD40 engagement. Finally, cells generated in presence of IL-10 were less efficient than DC in stimulating the production of IL-2, interferon-gamma, and IL-4 by allogeneic T cells. We conclude that IL-10 prevents the generation of DC induced by GM-CSF + IL-4 and favors the development of macrophages with a lower T cell stimulatory potential, but secreting higher levels of IL-8 than DC.

Grounding adaptive language games in robotic agents.

Abstract: Trabajo presentado a la 4th European Conference on Artificial Life, celebrada en Cambridge, Massachusetts (US).

Use of siderophores to type pseudomonads: the three Pseudomonas aeruginosa pyoverdine systems.

Abstract: Eighty-eight Pseudomonas aeruginosa isolates, most of them from the Collection of Bacterial Strains of the Institut Pasteur, Paris, were analysed for their pyoverdine-mediated iron incorporation system by different methods, including pyoverdine isoelectrofocusing analysis, pyoverdine-mediated growth stimulation, immunoblot detection of (ferri)pyoverdine outer-membrane receptor and pyoverdine-facilitated iron uptake. The same grouping of the strains was reached by each of these methods, resulting in the classification of the P. aeruginosa isolates, even those which were devoid of pyoverdine production, into three different siderophore types. Forty-two percent of the strains were identified with the type-strain P. aeruginosa ATCC 15692 (group I). 42% were identical with the second type-strain P. aeruginosa ATCC 27853 (group II) and 16% reacted identically with the clinical isolate P. aeruginosa Pa6, whose pyoverdine was recognized in this study to be identical in structure to the pyoverdine produced by a natural isolate, P. aeruginosa strain R. No new pyoverdine species was detected among these strains.

Direct detection and identification of Pseudomonas aeruginosa in clinical samples such as skin biopsy specimens and expectorations by multiplex PCR based on two outer membrane lipoprotein genes, oprI and oprL.

Abstract: A multiplex PCR test based on the simultaneous amplification of two lipoprotein genes, oprI and oprL, was designed and evaluated for its ability to directly detect fluorescent pseudomonads (amplification of oprI open reading frame, 249 bp) and Pseudomonas aeruginosa (amplification of oprL open reading frame, 504 bp) in clinical material. A collection of reference strains including 20 different species of fluorescent pseudomonads was tested. Positive PCR results for both genes were observed only for P. aeruginosa isolates (n = 150), including strains of clinical and environmental origin, while only one gene, oprI, was amplified from the other fluorescent pseudomonads. All other bacteria tested (n = 15) were negative by the amplification test. The lower detection level for P. aeruginosa was estimated to be 10(2) cells/ml. Preliminary evaluation on testing skin biopsy specimens from patients with burns (n = 14) and sputum samples from cystic fibrosis patients (n = 49) and other patients (n = 19) showed 100% sensitivity and 74% specificity in comparison with culture. This multiplex PCR assay appears promising for the rapid and sensitive detection of P. aeruginosa in clinical specimens. Further evaluation of its specificity in longitudinal clinical studies is warranted.

Human dendritic cell responses to lipopolysaccharide and CD40 ligation are differentially regulated by interleukin-10

Abstract: We evaluated the effects of interleukin (IL)-10 on the maturation of human dendritic cells (DC) induced either by lipopolysaccharide (LPS) or CD40 engagement. For this purpose, DC generated by culturing plastic-adherent peripheral blood mononuclear cells for 7 days with granulocyte/macrophage-colony-stimulating factor and IL-4 were incubated for 3 days with either LPS (10 ng/ml) or 3T6 fibroblasts transfected with the gene encoding CD40 ligand, in absence or presence of IL-10. First we found that the membrane expression of CD83, a marker of mature DC, was inhibited by IL-10 when induced by LPS but not by CD40 engagement. Likewise, IL-10 inhibited LPS-induced but not CD40-dependent CD86 (B7.2) up-regulation on DC. Furthermore, IL-10 inhibited the production of IL-8 and tumor necrosis factor-alpha by DC when activated by LPS but not by CD40. In contrast, IL-10 inhibited IL-12 production in both activation systems. We conclude that IL-10 differentially influences LPS-dependent and CD40-dependent pathways of DC maturation.

The distribution of Fe in the antarctic circumpolar current

Abstract: The large-scale distributions of dissolved and total Fe in surface and deep waters of the Antarctic Circumpolar Current exhibit strong relationships with hydrography and biological processes. The mean dissolved Fe concentrations are low in surface waters of the Antarctic Circumpolar Current (0.31–0.49 nM, with a minimum of 0.17 nM) and higher (averaging 1.11.9 nM) in the Polar Frontal region. Enhanced dissolved surface water concentrations in the Polar Frontal region are attributed to input from the continental shelf and coincide with phytoplankton spring blooms of large diatoms. The effects of sea-ice melting and iceberg melting on the Fe concentrations were relatively small. Dissolved deep-water concentrations ( > 400 m) in the Antarctic Circumpolar Current ranged from 0.4 to 2.8 nM. Circumpolar Deep Water has relatively high dissolved Fe concentrations in the Polar Frontal region (0.4–2.8 nM) compared with deep waters further to the south (0.6–1.1 nM). Similarly, total dissolvable (unfiltered) Fe concentrations in the Upper Circumpolar Deep Water tend to decrease southward from the Polar Frontal region. In the Lower Circumpolar Deep Water total dissolvable Fe concentrations are higher than in the Upper Circumpolar Deep Water due to the existing nepheloid layer and sources on the Mid-Atlantic Ridge. Dissolved and total dissolvable Fe concentrations in the Antarctic Bottom Water are higher than those of other water masses in the Antarctic Circumpolar Current, consistent with the nepheloid layer as well as diagenetic input from shelf sediments. The High-Nutrient/Low-Chlorophyll areas of the Antarctic Ocean and northeast Pacific Ocean have different major Fe input sources of similar magnitude. In the Antarctic Circumpolar Current upward transport of Fe is the main input source, whereas in the North Pacific Ocean, aerosols are the dominant source.

Endurance training effects on neurotransmitter release in rat striatum: an in vivo microdialysis study.

Abstract: In the present study we use the in vivo microdialysis sampling technique to register extracellular levels of neurotransmitters in the striatum of trained and untrained rats. We further evaluate the influence of 1 h of exercise on the striatal release of dopamine (DA), noradrenaline (NA), glutamate (GLU) and gamma-aminobutyric acid (GABA) in trained and untrained rats. Male Wistars were randomly assigned to a training or control group. The exercise training consisted of running on a treadmill for 6 weeks, 5 days week-1, with running time and speed gradually increased from 30 min at 19 m min-1 during the first week to 80 min at 26 m min-1 during the final training week. The animals of the control group were placed on the treadmill twice a week, and received a total of four 'adaptation sessions', in which they exercised 15-45 min at 26 m min-1. Brain dialysates were analysed with microbore liquid chromatography (LC), with electrochemical detection (monoamines and GABA) and fluorescence detection (GLU). Soleus citrate synthase and basal striatal concentrations of DA, NA and GLU were significantly different between the trained and control animals. Sixty minutes of exercise significantly increased extracellular DA, NA and GLU levels in both groups, but there was no statistically significant difference in the exercise-induced increase between trained and control animals. There was no statistical difference in basal or exercise-induced GABA levels between trained and control animals. The results indicate that exercise training appears to result in diminished basal activity of striatal neurotransmitters, while maintaining the necessary sensitivity for responses to acute exercise.

Low-temperature synthesized aluminosilicate glasses. Part III Influence of the composition of the silicate solution on production, structure and properties

Abstract: The low-temperature reaction between an aqueous sodium or potassium silicate solution and metakaolinite yields a solid aluminosilicate. The influence of the molar ratios H2O/R2O (between 6.6 and 21.0) and SiO2/R2O (between 0.0 and 2.3) of the silicate solution (R=Na or K) on the aluminosilicate's production, on the reaction stoichiometry and on the aluminosilicate's molecular structure is studied with differential scanning calorimetry, 27Al and 29Si magic angle spinning nuclear magnetic resonance (MAS NMR), cross-polarization MAS NMR, Fourier transform infrared spectroscopy and X-ray diffractometry. The reaction stoichiometry is determined by a one to one ratio for R/Al. H2O/R2O has no influence on the molecular structure of the aluminosilicate. Aluminium in the aluminosilicate is four-fold coordinated for the whole range of silicate solutions investigated. Moreover, Si and Al are homogeneously distributed and the ratio Al/Si in the aluminosilicate is the same as in the reaction mixture if the stoichiometric one-to-one ratio for R/Al is used. If SiO2/R2O in the Na-silicate solution is equal to or higher than 0.8, the low-temperature reaction yields an amorphous aluminosilicate or “inorganic polymer glass”. For smaller values of SiO2/R2O the Na-aluminosilicates are partially crystalline. Thermomechanical analysis and dynamic mechanical analysis indicate that a variation in the composition of the amorphous aluminosilicates can shift the glass transition over a few hundreds of degrees, with a minimum value of 650°C.

The role of vascular endothelial growth factor and interleukins in the pathogenesis of severe ovarian hyperstimulation syndrome

Abstract: Ovarian hyperstimulation syndrome (OHSS) is a dramatic complication of ovulation induction. In its most severe form, OHSS is characterized by massive cystic enlargement of the ovaries associated with third space fluid shift, resulting in the formation of ascites and pleural effusion. Ascites developes because of increased peritoneal capillary permeability. In this study we examined the role of vascular endothelial growth factor (VEGF) and interleukins in the pathogenesis of increased capillary permeability. VEGF is a member of the family of heparin binding proteins that act directly on endothelial cells to induce proliferation and angiogenesis. VEGF mRNA and protein are expressed by human ovarian granulosa and theca cells late in follicular development and subsequent to ovulation by granulosa and theca cells. Therefore, VEGF is ideally positioned to provoke the increased permeability of theca blood vessels that occurs shortly before ovulation. Hybridization studies in the rat and primate ovary have demonstrated VEGF mRNA expression predominantly after the luteinizing hormone (LH) surge known to be essential for OHSS. The gonadotrophin-releasing hormone antagonist results in a decreased mRNA expression, implying such expression is dependent on LH. The expression of VEGF mRNA has been recently shown to be enhanced by human chorionic gonadotrophin (HCG) in a dose- and time-dependent fashion. These studies confirm the timely association between VEGF and HCG that has been clinically known for many years to be integral in the development of OHSS. VEGF concentrations in serum, peritoneal fluid and follicular fluid of patients at risk for OHSS have been shown to be significantly related to the development of the syndrome. Furthermore, the kinetics of VEGF in the plasma of patients who actually develop severe OHSS are closely correlated with the clinical course of the syndrome and with certain biological characteristics of OHSS and of capillary leakage, such as leukocytosis and increased hematocrit. Studies on ascitic fluid from patients with sever OHSS have proved that VEGF is the major capillary permeability agent. Incubation with VEGF antiserum decreased the vascular permeability activity by 70%. Interleukin-2 (IL-2) is the first of a series of lymphocytotrophic hormones to be recognized as pivotal for the regulation of immune response. However, hard data to confirm its central role in the pathogenesis of OHSS are still lacking, despite the fact that some preliminary studies suggest a positive association between the pooled follicular fluid IL-2 concentration and the development of OHSS. IL-6 is a mediator of the acute phase response to injury, a systemic reaction characterized by leukocytosis, increased vascular permeability and increased synthesis of acute phase proteins by the liver. Significantly higher serum and ascites IL-6 concentrations were seen in OHSS patients. The immunohistochemical localization pattern suggested that IL-6 is LH or HCG dependent. However, the use of IL-6 as a predictor for the occurrence of OHSS has not been successful. The kinetics of IL-6 in patients with severe OHSS are correlated with the clinical symptoms and the biochemical parameters known to be associated with the severity of the syndrome, suggesting a possible role for IL-6. Further molecular biology studies similar to those performed on VEGF are needed to confirm if this interleukin is central in the cascade of events. IL-8 is a chemoattractant, activating cytokine and a potent angiogenic agent. The peritoneal fluid levels is increased in patients with severe OHSS; its concentration in peritoneal fluid is increased inpatients with severe OHSS. The place of this interleukin in the cascade of events is as yet undetermined and further studies are needed. In conclusion, molecular biology and clinical studies strongly suggest that VEGF is the principal mediator by which HCG might increase capillary permeability in OHSS.

Ventilation distribution during histamine provocation

Abstract: Verbanck, S., D. Schuermans, A. Van Muylem, M. Paiva, M. Noppen, and W. Vincken. Ventilation distribution during histamine provocation. J. Appl. Physiol. 83(6): 1907–1916, 1997.—We investigated ven...

A high proportion of novel mutations in BRCA1 with strong founder effects among Dutch and Belgian hereditary breast and ovarian cancer families

Abstract: We have identified 79 mutations in BRCA1 in a set of 643 Dutch and 23 Belgian hereditary breast and ovarian cancer families collected either for research or for clinical diagnostic purposes. Twenty-eight distinct mutations have been observed, 18 of them not previously reported and 12 of them occurring more than once. Most conspicuously, a 2804delAA mutation has been found 19 times and has never been reported outside the Netherlands. A common haplotype spanning > or = 375 kb could be identified for each of the nine examined recurrent mutations, indicating the presence of multiple BRCA1 founder mutations in the Dutch population. The 2804delAA mutation has been estimated to have originated approximately 32 generations ago. No specific breast or ovarian cancer phenotype could be assigned to any of the common mutations, and the ovarian cancer incidence among 18 families with the 2804delAA mutation was heterogeneous.

IA-2-autoantibodies complement GAD65-autoantibodies in new-onset IDDM patients and help predict impending diabetes in their siblings. The Belgian Diabetes Registry.

Abstract: IA-2 has been identified as an autoantigen that is recognized by immunoglobulins from insulin-dependent diabetic (IDDM) patients. Using a liquid phase radiobinding assay, we performed an IA-2-autoantibody (IA-2-Ab) assay in 474 IDDM patients and 482 non-diabetic control subjects aged 0–39 years. IA-2-Ab were detected in 58 % of the patients and 0.8 % of control subjects. Their prevalence in patients was lower than that of islet cell autoantibodies (ICA; 73 %) or glutamic acid decarboxylase (Mr 65 kDa)-autoantibodies (GAD65-Ab; 82 %) but higher than that of insulin autoantibodies (IAA; 42 %). IA-2-Ab were more frequent in patients under age 20 years (70 %) than between 20 and 40 years (45 %; p < 0.001). In the whole IDDM group, 92 % of patients were positive for at least one of the three molecular assays, which is higher than the positivity for the ICA assay (73 %). Only 1 % was negative in the molecular assays and positive in the ICA assay. IA-2-Ab levels were positively correlated with ICA titres (p < 0.001) and HLA DQ A1*0301 – DQ B1*0302 (p < 0.003) by multivariate analysis. In a group of 481 non-diabetic siblings (age 0–39 years) of IDDM patients only 7 were IA-2-Ab positive (1.5 %). All seven were under age 20 years and positive for at least two other autoantibodies and for DQ A1*0301 – DQB1*0302. Four of these seven developed IDDM during the 6–70-month follow-up period. The positive predictive value of IA-2-Ab (57 %) was higher than that of ICA, GAD65-Ab or IAA alone, or in combination (≤ 20 %) but these calculations are restricted by the relatively short observation period and the small number of cases. The only IA-2-Ab-negative case of pre-diabetes was also negative for IAA and GAD65-Ab, while it was strongly positive for ICA. In conclusion, IA-2-Ab show a high diagnostic specificity for IDDM and are predictive markers of impending diabetes in siblings of patients. In combination with other molecular antibody assays they may replace ICA testing in future. Our data also indicate that other autoantibodies than IA-2-Ab, GAD65-Ab and IAA contribute to ICA. [Diabetologia (1997) 40: 95–99]

Frequency domain system identification using arbitrary signals

Abstract: It is the common conviction that frequency domain system identification suffers from the drawback that it cannot handle arbitrary signals without introducing systematic errors. This paper shows that it is possible to deal with nonperiodic signals without any approximation and under the same assumptions as in the time domain, by estimating simultaneously some initial conditions and the system model parameters.

NMDA receptor‐mediated pilocarpine‐induced seizures: characterization in freely moving rats by microdialysis

Abstract: Pilocarpine administration has been used as an animal model for temporal lobe epilepsy since it produces several morphological and synaptic features in common with human complex partial seizures. Little is known about changes in extracellular neurotransmitter concentrations during the seizures provoked by pilocarpine, a non-selective muscarinic agonist. Focally evoked pilocarpine-induced seizures in freely moving rats were provoked by intrahippocampal pilocarpine (10mm for 40min at a flow rate of 2μl min−1) administration via a microdialysis probe. Concomitant changes in extracellular hippocampal glutamate, γ-aminobutyric acid (GABA) and dopamine levels were monitored and simultaneous electrocorticography was performed. The animal model was characterized by intrahippocampal perfusion with the muscarinic receptor antagonist atropine (20mm), the sodium channel blocker tetrodotoxin (1μm) and the N-methyl-d-aspartate (NMDA) receptor antagonist MK-801 (dizocilpine maleate, 100μm). The effectiveness of locally (600μm) or systemically (10mg kg−1 day−1) applied lamotrigine against the pilocarpine-induced convulsions was evaluated. Pilocarpine initially decreased extracellular hippocampal glutamate and GABA levels. During the subsequent pilocarpine-induced limbic convulsions extracellular glutamate, GABA and dopamine concentrations in hippocampus were significantly increased. Atropine blocked all changes in extracellular transmitter levels during and after co-administration of pilocarpine. All pilocarpine-induced increases were completely prevented by simultaneous tetrodotoxin perfusion. Intrahippocampal administration of MK-801 and lamotrigine resulted in an elevation of hippocampal dopamine levels and protected the rats from the pilocarpine-induced seizures. Pilocarpine-induced convulsions developed in the rats which received lamotrigine perorally. Pilocarpine-induced seizures are initiated via muscarinic receptors and further mediated via NMDA receptors. Sustained increases in extracellular glutamate levels after pilocarpine perfusion are related to the limbic seizures. These are arguments in favour of earlier described NMDA receptor-mediated excitotoxicity. Hippocampal dopamine release may be functionally important in epileptogenesis and may participate in the anticonvulsant effects of MK-801 and lamotrigine. The pilocarpine-stimulated hippocampal GABA, glutamate and dopamine levels reflect neuronal vesicular release.

Assessment of the need for follicle stimulating hormone in early preantral mouse follicle culture in vitro.

Abstract: In two consecutive controlled experiments 160 early preantral follicles were cultured in order to evaluate effects of recombinant follicle stimulating hormone (r-FSH) on survival, differentiation, oestradiol and inhibin secretion, cumulus mucification and cumulus-corona-oocyte detachment by human chorionic gonadotrophin (HCG) stimulation. Nuclear maturation in oocytes was also assessed following addition of HCG. A histological analysis of cultured follicles was carried out on semi-thin sections at various culture stages. Addition of r-FSH was essential for follicle survival for 16 days: without r-FSH only 11% of the follicles survived for 12 days (with r-FSH: 79%) and none of these mucified after the HCG stimulus. r-FSH promoted granulosa cell proliferation and antral-like cavity formation. Without r-FSH, histology of the cultures demonstrated degeneration and reduced granulosa cell proliferation; oestradiol and inhibin production were reduced. This study illustrates the essential role of FSH in promoting the in-vitro growth of early preantral mouse ovarian follicles and in maintaining the oocyte under meiotic arrest.

The chromosomal constitution of embryos developing from abnormally fertilized oocytes after intracytoplasmic sperm injection and conventional in-vitro fertilization.

Abstract: The aim of this study was to analyse the chromosomal constitution of embryos developing from mono- (1PN) and tripronuclear (3PN) oocytes, after in-vitro fertilization (IVF) and after intracytoplasmic sperm injection (ICSI) into oocytes, by means of the fluorescent in-situ hybridization (FISH) technique with specific probes for the chromosomes X, Y and 18. FISH analysis was carried out on embryos from 3PN oocytes: 106 after ICSI and 71 after conventional IVF. In the 3PN embryos after ICSI, equal ratios of XXX and XXY were observed and no XYY embryos were present. This shows the digynic origin of such 3PN embryos. On the other hand, after conventional IVF, the XYY status indicative of dispermic fertilization was observed in some embryos. After IVF, only 12.7% of the 3PN oocytes developed into embryos with uniformly triploid blastomeres, compared with 55.7% after ICSI (P < 0.001). On the other hand, after ICSI only 16.0% of the embryos developing from 3PN oocytes were mosaic, compared with 42.3% after conventional IVF (P < 0.001). FISH was also carried out on embryos from 1PN oocytes: 61 after ICSI and 115 after conventional IVF. In 35.6% of IVF embryos developing from 1PN oocytes Y-specific hybridization signals were observed. This indicates that in 70-75% of such cases a spermatozoon had penetrated the oocyte and that only 25-30% of them were parthenogenetic. A significantly higher proportion (P < 0.001) of embryos developing from 1PN oocytes were diploid after IVF (48.7%) than after ICSI (27.9%); equal ratios of XX and XY embryos were observed in the two groups. Formation of a single pronucleus in an embryo subsequently shown to be diploid indicates that normal fertilization was followed by asynchronous formation of pronuclei. A significantly (P < 0.001) higher proportion of 1PN oocytes developed into haploid embryos after ICSI (31.2%) than after conventional IVF (13.1%). In both groups most of the haploid embryos were X-bearing (IVF, 93.3%; ICSI, 84.2%) and only a few were Y-bearing (IVF, 6.7%; ICSI, 15.8%). A contribution of normal fertilization and androgenetic activation thus led to 1PN oocytes. Gynogenetic and/or parthenogenetic activation, both leading to indistinguishable chromosomal distributions, also contributed to the formation of 1PN oocytes after ICSI and IVF.

Specific Uptake of Tumor Necrosis Factor-α Is Involved in Growth Control of Trypanosoma brucei

Abstract: Trypanosoma brucei is lysed by tumor necrosis factor-α (TNF-α) in a dose-dependent way, involving specific binding of the cytokine to a trypanosomal glycoprotein present in the flagellar pocket of the parasite. TNF-α–gold particles are endocytosed via coated pits and vesicles and are directed towards lysosome-like digestive organelles. The specific uptake of the cytokine by the parasite results in a developmentally regulated loss of osmoregulatory capacity. TNF-α specific lysis is prevented when lysis assays are performed at a temperature <26°C, despite uptake of the cytokine. Inhibition of lysis is also observed when a lysosomotropic agent is added during the first 2 h of incubation. Both monomorphic and pleomorphic trypanosomes are lysed but only when isolated during the peak of parasitaemia. Lysis is not observed with early infection stage parasites or procyclic (insect-specific) forms. Anti– TNF-α treatment of T. brucei -infected mice reveals a dramatic increase in parasitaemia in the blood circulation, the spleen, the lymph nodes, and the peritoneal cavity. These data suggest that in the mammalian host, TNF-α is involved in the growth control of T. brucei.

Calculation of ionization energies, electron affinities, electronegativities, and hardnesses using density functional methods

Abstract: The performance of two exact exchange methods is tested in the calculation of ionization energies, electron affinities, electronegativities, and hardnesses using Dunning’s correlation consistent basis sets. Comparison is made to experiment and other density functional methods, including the local density approximation and two gradient corrected functionals. The obtained electronegativities and hardnesses are also compared with high level coupled cluster results. Both the exact exchange methods show an excellent performance in the calculation of all four properties, yielding mean absolute deviations from experiment below 0.20 eV for all basis sets.