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Biomedical subjects

M Cornelissen

Publications and source records attributed to M Cornelissen.

At least 73 records · Page 4Linked to original sources
PubMed1995

cry IA(b) transcript formation in tobacco is inefficient.

Chimaeric PCaMV35Scry genes direct in tobacco mesophyll protoplasts mRNA levels of less than one transcript per cell. We provide evidence that this low cytoplasmic cry IA(b) mRNA level is not due to a rapid turnover but rather results from a marginal import flow of cry messenger into the cytoplasm. Run-on assays indicate that the frequency of transcription initiation is not limiting. However, the cry precursor mRNA carries at least three regions that are recognized as introns. The absence of high cytoplasmic levels of spliced cry mRNAs suggests that these mRNAs are unstable and/or not efficiently made. Point mutations in the 5' splice site of the most distal intron allows high accumulation levels of the full-length mRNA. This implies that the inefficient formation of full-size mRNA is a major cause of the low expression level of chimaeric cry IA(b) genes in tobacco.

Bacillus thuringiensis Toxins↗
PubMed1995

Maintenance of syncytium-inducing phenotype of HIV type 1 is associated with positively charged residues in the HIV type 1 gp120 V2 domain without fixed positions, elongation, or relocated N-linked glycosylation sites.

The prevalence of HIV-1 sequences of the envelope domains V1V2 and V3 was analyzed by RT-PCR amplification. Two distinct biological phenotypes of HIV-1 have been described: the nonsyncytium-inducing (NSI) phenotype, best characterized by the inability to infect MT-2 cells, and the syncytium-inducing phenotype (SI), with the ability to infect MT-2 cells. Viral phenotype SI has been associated with HIV pathogenesis. The presence of positively charged amino acids at position 306 and 320 in the V3 domain of gp120 has been shown to be required for the support of the SI phenotype. In addition, V2 elongation and relocation of N-glycosylation sites were postulated to herald an NSI to SI phenotype switch. The present study was designed to assess the stability of an elongated V2 region with relocated N-glycosylation sites observed in SI isolates compared to NSI isolates. Eleven isolates with the SI phenotype and 19 isolates with the NSI phenotype were included in the study. Nine of the SI and 1 of the NSI isolates had a positively charged residue at position 306 or 320 (p < 0.001) in the V3 domain as assessed by direct sequencing of the viral RNA. In contrast, elongation and/or relocation of N-glycosylation sites of the V2 variable region were not found to be a consistent genetic feature of the SI phenotype. However, SI isolates had more positively charged amino acid residues in the hypervariable V2 region compared with NSI isolates. In one of the two SI isolates lacking positively charged amino acids at positions 306 or 320 in the V3 loop an elongation of 26 amino acids with 4 additional N-linked glycosylation sites was observed in the V2 region. This is consistent with the theory that elongation of V2 may be transiently required for SI conversion. These results suggest that maintenance of the SI phenotype requires positively charged amino acids in V3 in the majority of the virus population, but not an elongated V2 region with added or relocated N-linked glycosylation sites. Although increased charged residues in the V2 region may contribute.

Amino Acid Sequence↗
PubMed1995

Transmission of zidovudine-resistant human immunodeficiency virus type 1 variants following deliberate injection of blood from a patient with AIDS: characteristics and natural history of the virus.

We describe the development and persistence of severe immunodeficiency in a previously healthy young woman shortly after she was deliberately injected with blood that was drawn from a patient with AIDS. The heterogenous populations of human immunodeficiency virus (HIV) in donor and recipient were as closely related as those reported in previous cases of proven transmission. The relatively large proportion of non-syncytium-inducing (NSI) clones in the recipient suggests a selective but not complete suppression of syncytium-inducing (SI) viruses. The continuous presence of SI viruses might explain the severe immunosuppression that persisted once the recipient seroconverted. A codon 215 mutation (indicative of zidovudine resistance) was present in SI and NSI clones of the donor and in NSI clones of the recipient. The relative increase in codon 215 resistance mutation in the absence of zidovudine therapy was secondary to the increase in NSI clones. Findings in this case suggest that qualities of an inoculum and/or the route of transmission are important determinants in the subsequent clinical course of HIV disease.

Adult↗
PubMed1995

Differences in human immunodeficiency virus type 1 V3 sequences from patients with and without AIDS dementia complex.

Paired serum and cerebrospinal fluid (CSF) samples from 10 AIDS patients with and 10 without AIDS dementia complex (ADC) were studied, in an attempt to uncover ADC-associated variation in V3 sequences. Sequences were obtained from four of the patients with and eight of those without ADC. Comparison of the sequences using a resampling technique revealed a significant ADC-associated difference occurring at several amino acid positions. Results from serum and CSF sequences were comparable. These differences may indicate that the virus found in ADC and that in non-ADC patients have different biological properties. Comparison of serum versus CSF sequences within samples from both ADC and non-ADC patients, using the same resampling technique, revealed no clear distinctions. In some patients, the sequence populations in serum and CSF were completely distinct, while in others, there was no difference in distribution. These patterns were not associated with ADC.

AIDS Dementia Complex↗
PubMed1995

Virus load in chimpanzees infected with human immunodeficiency virus type 1: effect of pre-exposure vaccination.

Many reports indicate that a long-term asymptomatic state following human immunodeficiency virus type 1 (HIV-1) infection is associated with a low amount of circulating virus. To evaluate the possible effect of stabilizing a low virus load by non-sterilizing pre-exposure vaccination, a quantitative virus isolation method was developed and evaluated in four chronically infected chimpanzees infected with a variety of HIV-1 related isolates. This assay was then used to monitor a group of chimpanzees (n = 6) challenged with HIV-1 following vaccination with gp120 or gp160. Data indicated that of the three vaccinated animals which became infected after challenge, the animal with the lowest neutralizing titre at the time of challenge acquired a virus load similar to the control animals, whereas the two other chimpanzees had reduced numbers of virus producing cells in their peripheral circulation. One animal became virus isolation negative, developed an indeterminant PCR signal on lymph node DNA and subsequently became negative for HIV-1 DNA as determined by PCR on PBMC (peripheral blood mononuclear cells) and bone marrow DNA. Recently, the second animal has also become PCR negative. To confirm observations from quantitative virus isolations, quantification of HIV-1 DNA in PBMC and virus RNA in serum was performed by PCR on serially diluted samples at two different time points. Comparison of virus load as determined by these three methods confirmed that there was an effect of vaccination in reducing virus load and demonstrated a correlation between decreased numbers of virus producing cells, HIV-1 DNA containing cells and virus RNA molecules in serum.

Animals↗
PubMed1995

Post-transcriptional cosuppression of beta-1,3-glucanase genes does not affect accumulation of transgene nuclear mRNA.

Silencing of a Nicotiana plumbaginifolia beta-1,3-glucanase (gn1) transgene in tobacco line T17 occurs in homozygous and in haploid plants with one transgene locus dosage per chromosome set. We have previously shown that the silent state is manifested by a reduced gn1 steady state mRNA level and results from a post-transcriptional process that is under developmental control in homozygous T17 plants. In this study, we show that specific endogenous beta-1,3-glucanase genes are cosuppressed with gn1 in homozygous T17 plants. We also demonstrate that the developmental timing of cosuppression depends on environmental conditions and that once silencing is established it is stably maintained during plant development. Analyses of additional transgenic tobacco lines revealed that gn1 silencing is not restricted to the T17 line and showed that silencing can also take place in R0 plants containing independent loci in hemizygous states. Furthermore, silencing can also be obtained in progeny plants in which expressing loci have been combined. Importantly, cosuppression occurs via a post-transcriptional mechanism that does not interfere with the accumulation of transgene nuclear mRNA. These results strongly suggest that the silencing mechanism operates at RNA transport and/or RNA stability levels.

Base Sequence↗
PubMed1995

Syncytium-inducing (SI) phenotype suppression at seroconversion after intramuscular inoculation of a non-syncytium-inducing/SI phenotypically mixed human immunodeficiency virus population.

Two distinct biological phenotypes of human immunodeficiency virus (HIV) have been described: the non-syncytium-inducing (NSI) phenotype, best characterized by the inability to infect MT-2 cells, and the syncytium-inducing (SI) phenotype, with the ability to infect MT-2 cells. The earliest virus population observed following HIV transmission is generally of the NSI phenotype, even after exposure to inocula of mixed NSI/SI phenotype. In this study, the issue of intrapatient selection of virus phenotype following transmission was addressed by studying two cases of accidental transmission. A comparison of the sequences of the V1-V2 and the V3 coding regions of the envelope gene and the p17 region of the gag gene showed that the donor-recipient pairs were tightly clustered in all gene segments, but away from local and published transmission controls. The intrasample variation of the p17 sequence was greater in the recipients and smaller in the donors than that of the V3 region sequence, indicating selection of V3 at transmission. In these transmission cases, the effects of an intravenous inoculation of a small quantity of blood containing predominantly SI V3 sequences (6 of 8 clonal sequences) were compared with those of an intramuscular inoculation of a large quantity of blood containing predominantly NSI viruses (14 of 16 clonal sequences). Both SI and NSI V3 regions were demonstrated to be phenotypic expressions of genetically related viral strains. The inoculation of the predominantly SI virus population resulted in the persistence of an SI virus population in the recipient and a rapid CD4+ T-cell decline. The inoculation of the predominantly NSI population resulted in a selective amplification of SI viruses before seroconversion, followed by a suppression of SI viruses at seroconversion and a rapid decline of CD4+ T-cell numbers. These data suggest that the suppression of SI viruses can be accomplished following the development of HIV-specific immunity and that the ability to suppress SI viruses does not prevent the development of immunodeficiency.

Acquired Immunodeficiency Syndrome↗
PubMed1994

A 6' gentamicin acetyltransferase gene allows effective selection of tobacco transformants using kanamycin as a substrate.

6' gentamicin acetyltransferases detoxify aminoglycoside antibiotics containing a 6' amino group. We tested whether a 6' gentamicin acetyltransferase gene (6' gat) of Shigella sp. is suitable as selectable gene in plant transformation using kanamycin (Km) as a substrate. A comparative transformation experiment using Nicotiana tabacum SR1 protoplasts showed that 6' gat is as effective for selection of transformants as the commonly used neomycin phosphotransferase II (nptII). In stably transformed plants we detected moderate levels of the 6' gat mRNA. An enzymatic assay was developed with which the acetyltransferase activity of the protein is easily demonstrated.

Acetyltransferases↗
PubMed1994

Comparative and combined effects of interleukin 6, interleukin 1 beta, and tumor necrosis factor alpha on proteoglycan metabolism of human articular chondrocytes cultured in agarose.

OBJECTIVE: To study the effects of recombinant tumor necrosis factor alpha (TNF-alpha), interleukin 1 beta (IL-1 beta) and interleukin 6 (IL-6) on proteoglycan metabolism of isolated chondrocytes. METHODS: Human articular cartilage cells were cultured in agarose gel. In these culture conditions, chondrocytes keep their phenotypic stability. They release cartilage specific proteoglycans into the surrounding artificial matrix. Proteoglycan synthesis was measured by the incorporation of 35sulfate (35S). RESULTS: TNF-alpha and IL-1 beta depressed proteoglycan synthesis and induced proteoglycan degradation. The effects of both cytokines were additive, when used in submaximal doses. No mutual induction of TNF-alpha and IL-1 beta was shown, but both cytokines stimulated the chondrocytes to release IL-6, up to 100,000 pg/ml. Equal amounts of human recombinant IL-6 did not affect proteoglycan synthesis. IL-6 did not alter proteoglycan quality, nor did it modulate the IL-1 beta activities on proteoglycan metabolism. CONCLUSION: These findings illustrate the role of IL-1 beta and TNF-alpha in cartilage degradation and suggest that the role of the large amounts of IL-6 released in response to IL-1 in chronic arthritis is not directly protective with regard to proteoglycan metabolism.

Arthritis↗
PubMed1993

Radiation effects on the matrix synthesis in non-ossifying embryonic cartilage in vitro: a functional and morphological study.

Radiation effects on collagen and glycosaminoglycan (GAG) synthesis were studied in non-ossifying embryonic chick cartilage, in vitro. Fragments of the non-calcifying region of 15-day-old chick sterna were irradiated with 20, 50 and 100 Gy. The counterparts served as controls. Fragments were cultured up to 14 days. Pulse labeling experiments with 3H-proline and 3H-glucosamine started immediately after irradiation and on day 3, 6 and 13 post-irradiation. Fragments for morphological evaluation were fixed on day 1, 4, 7 and 14, simultaneously with the end of the labeling period. The same dose- and time-dependent inhibition was observed for the collagen and GAG synthesis. Radiation doses of 20, 50 and 100 Gy cause respectively and inhibition of about 20, 35 and 60% at the end of the culture period. Morphological alterations were observed after radiation doses of 50 and 100 Gy. Dose- and time-dependent necrosis all over the sternum was observed. A restricted number of cells showed an increased GAG synthesis after irradiation (50 Gy). This could explain the levelling off of the radiation effect on the incorporation of the labeled precursors towards the end of the culture period for radiation doses up to 50 Gy.

Animals↗
PubMed1993

Supplementation of vitamin K in pregnant women receiving anticonvulsant therapy prevents neonatal vitamin K deficiency.

OBJECTIVE: The null hypothesis of this study is that extra vitamin K administered to pregnant women on a regimen of enzyme-inducing anticonvulsant therapy will not decrease the frequency of symptoms of vitamin K deficiency in their neonates. STUDY DESIGN: A multicenter case-control study was performed on 16 pregnant women on anticonvulsant therapy who received 10 mg of vitamin K1 daily from 36 weeks of pregnancy onward. Concentrations of PIVKA-II (protein induced by vitamin K absence for factor II) and of vitamin K1 were determined in cord blood and compared with those in 20 controls. RESULTS: In none of 17 cord samples was PIVKA-II detectable, compared with 13 of 20 in controls (chi 2, p < 0.001). Median cord vitamin K1 level was 530 pg/ml compared with below detection limit in most controls. CONCLUSIONS: Antenatal vitamin K1 treatment decreases the frequency of vitamin K deficiency in neonates of mothers on anticonvulsant therapy.

Anticonvulsants↗
PubMed1993

Increased incidence of neonatal vitamin K deficiency resulting from maternal anticonvulsant therapy.

OBJECTIVE: The null hypothesis of our study is that the incidence of vitamin K deficiency in mother-infant pairs exposed to anticonvulsant drugs is not higher than in controls. STUDY DESIGN: In this multicenter observational case-control study, 25 pregnant women receiving anticonvulsant therapy and 25 pregnant controls were studied for PIVKA-II (protein induced by vitamin K absence of factor II) and vitamin K1 concentrations at 32 weeks' gestation and at delivery. RESULTS: PIVKA-II was detectable in 54% of cord samples of the anticonvulsant group and in 20% of controls (chi 2, p = 0.01). In both groups vitamin K1 cord blood levels were predominantly below the detection limit. Maternal vitamin K1 concentrations were lower in women with epilepsy than in controls (Wilcoxon's rank sum test, p < 0.05), but PIVKA-II was rarely present. CONCLUSIONS: The incidence of vitamin K deficiency is increased in neonates exposed to anticonvulsant drugs prenatally. Their mothers, however, are rarely vitamin K deficient.

Anticonvulsants↗
PubMed1993

The 3' untranslated region of satellite tobacco necrosis virus RNA stimulates translation in vitro.

The RNA of satellite tobacco necrosis virus (STNV) is a monocistronic messenger that lacks both a 5' cap structure and a 3' poly(A) tail. We show that in a cell-free translation system derived from wheat germ, STNV RNA lacking the 600-nucleotide trailer is translated an order of magnitude less efficiently than full-size RNA. Deletion analyses positioned the translational enhancer domain (TED) within a conserved hairpin structure immediately downstream from the coat protein cistron. TED enhances translation when fused to a heterologous mRNA, but the level of enhancement depends on the nature of the 5' untranslated sequence and is maximal in combination with the STNV leader. The STNV leader and TED have two regions of complementarity. One of the complementary regions in TED resembles picornavirus box A, which is involved in cap-independent translation but which is located upstream of the coding region.

Base Sequence↗
PubMed1993

Effects of ionizing radiation on the size distribution of proteoglycan aggregates synthesized by chondrocytes in agarose.

Effects of ionizing radiation on the structure and size-distribution of newly synthesized proteoglycan aggregates are studied in-vitro. Chondrocytes, isolated from embryonic chick sterna, are cultured for 7 days in a tri-dimensional agarose system. Single radiation doses of 10, 20 and 50 Gy are given before starting the culture. Digestion of the artificial agarose matrix liberates the newly synthesized proteoglycans. Spreading with cytochrome C allows electron microscopic investigations of the individual, newly synthesized molecules. The structure of aggregates synthesized by control and irradiated chondrocytes is comparable. However, radiation causes alterations in the size-distributions of the aggregate-populations. For the control cultures, an average aggregate contains 27 aggrecans per aggregate. 34 pro mille of the molecules contain more than 100 aggrecans per aggregate. Irradiation with 10 Gy doesn't cause alterations. With radiation doses of 20 and 50 Gy, an average molecule contains 20 aggrecans. Only about 9 pro mille of the aggregates contain more than 100 aggrecans. Stimulation of lysosomal activity after irradiation could explain the observed alterations.

Animals↗
PubMed1992

Analysis of human immunodeficiency virus type 1 LTR-LTR junctions in peripheral blood mononuclear cells of infected individuals.

Circularized DNA species containing two long terminal repeat circle junctions were analysed in peripheral blood mononuclear cells of human immunodeficiency virus type 1 (HIV-1)-infected individuals. The circle junction fragments found could be classified into four groups: fragments containing a normal circle junction, fragments with deletions at the circle junction, fragments containing the primer binding site inserted at the circle junction, and fragments containing insertions at the circle junction derived from other regions of the HIV-1 genome.

Base Sequence↗