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Biomedical subjects

W Janssens

Publications and source records attributed to W Janssens.

At least 73 records · Page 4Linked to original sources

Genetic and antigenic variability of HIV type 1 in Brazil.

Six Brazilian strains of human immunodeficiency virus type 1 (HIV-1) were isolated from infected individuals residing in different regions of Brazil between 1987 and 1989. Phylogenetic analysis based on an 860-base pair env fragment, including V3, V4, V5, and the beginning of gp41, classified the Brazilian strains significantly in genotype B, with interhost distances between 5.9 and 13.1% (mean value, 10%). Amino acid sequence analysis of the V3 loop revealed that three strains contained the North American/European GPGR motif as the tip of the loop whereas in the other three strains proline (P) was substituted by tryptophan (W), methionine (M), or phenylalanine (F). A consensus peptide, Bra-cons, was designed containing GWGR as the tip of the loop. Serological reactivity to the Bra-cons peptide and other V3 peptides (MN, SF2, HBX2, RF, MAL, ELI, Z6, and a Côte d'Ivoire peptide, CI-cons) was compared for 114 HIV-1-positive sera from Rio de Janeiro. Sixty-nine sera (60.5%) reacted with peptides belonging to genotype B, of which 10 sera also reacted with peptides belonging to genotype A (n = 7) and D (n = 3). Eighteen sera (15.8%) had binding antibodies to the Bra-cons peptide. A high number of sera (n = 43; 37.7%) had no antibodies to any of the V3 peptides tested. This result suggests that HIV-1 variants with aberrant V3 loops may circulate in Rio de Janeiro.

Acquired Immunodeficiency Syndrome↗

Isolation of simian immunodeficiency viruses from two sooty mangabeys in Côte d'Ivoire: virological and genetic characterization and relationship to other HIV type 2 and SIVsm/mac strains.

OBJECTIVE: To search for the presence of SIV in sooty mangabeys and other monkey species in Côte d'Ivoire, West Africa, and to compare viral isolates with HIV-2 strains from the same region. METHODS: Forty-three captive housed monkeys (28 African green monkeys, 6 sooty mangabeys, 6 baboons, and 6 patas monkeys) were tested for the presence of HIV and SIV antibodies. Virus was isolated from the peripheral blood lymphocytes of seropositive animals and from HIV-2 antibody-positive patients originating from Côte d'Ivoire, Ghana, Senegal, and Belgium. Viruses were characterized by Western blot and radioimmunoprecipitation assay. Proviral DNA was amplified by PCR, cloned, and sequenced to construct a phylogenetic tree. RESULTS: One African green monkey and three sooty mangabeys had antibodies that cross-reacted with HIV-2. From two mangabeys lentiviruses were isolated and designated as SIVsmCI2 and SIVsmCI8. Serological, virological, and sequence data showed that these isolates are members of the HIV-2/SIVsm/SIVmac group of primate lentiviruses. Furthermore, in the phylogenetic tree, these two new viruses form a distinct subgroup that is equidistant to the HIV-2 strains and the previously described SIVsm/SIVmac viruses. CONCLUSION: This study provides additional evidence that sooty mangabey monkeys can be infected with a lentivirus in their natural habitat. Within the SIVsm and SIVmac viruses extensive genetic variation is observed.

Acquired Immunodeficiency Syndrome↗

Molecular phylogeny of part of the env gene of HIV-1 strains isolated in Côte d'Ivoire.

OBJECTIVES: To examine the genetic variation of HIV-1 isolates in Abidjan, Côte d'Ivoire, and to determine the extent to which phylogenetic trees based on sequence information of part of the env gene containing the principal neutralizing domain are representative for documenting genetic variability. DESIGN: Phylogenetic comparison of 13 HIV-1 strains isolated from patients in Abidjan with previously documented HIV-1 strains of different geographic origin. METHODS: To sequence a 900 base-pair fragment of the env gene containing V3, V4, V5 and the beginning of gp41 of three to four clones per isolate. Phylogenetic tree analysis was performed with the software package TREECON. RESULTS: Eleven HIV-1 isolates of Abidjan were classified as genotype A, while two were classed as genotypes B and D. Intra-genotype A distances at the nucleotide level were a maximum of 14.1%. Inter-genotype distances between genotype A and genotypes B, C, and D varied from 16.0 to 22.6%. Phylogenetic trees, based on sequence data of a 300 base-pair fragment containing the V3 loop, showed significant differences in tree topology and statistical confidence with phylogenetic trees based on sequence data of the 900 base-pair env fragment. CONCLUSIONS: Genotype A Côte d'Ivoire HIV-1 strains, which comprise 11 out of 13 isolates, predominate in Abidjan, which may indicate a local burst of particular variants. Phylogenetic trees should be interpreted with caution when based on a more limited number of nucleotides, such as the V3 region.

Base Sequence↗

Cross-neutralizing antibodies to HIV-1ANT70 and HIV-1IIIB in sera of African and Belgian HIV-1-infected individuals.

OBJECTIVES: To determine the neutralizing antibody patterns to HIV-1ANT70 (ANT70) and HIV-1IIIB (IIIB) in human sera obtained from HIV-1-infected individuals from different African countries and Belgium. Second, to correlate the presence of neutralizing antibodies in sera and their ability to bind to synthetic peptides derived from eight different HIV-1 V3 loop sequences. DESIGN AND METHODS: Forty sera from Belgium and 88 obtained from seven countries in Africa were tested for their ability to neutralize ANT70 (one of the most genetically divergent HIV-1 isolates documented), and IIIB. Sera found to cross-neutralize both viruses were further challenged with four HIV-1 field isolates. All sera were tested on a panel of V3 loop peptides obtained from different HIV-1 genotypes. RESULTS: Four patterns of sera were identified, including 33 (26%) sera not neutralizing any of the isolates, seven (5%) sera neutralizing only ANT70, 45 (35%) sera neutralizing only IIIB, and 43 (34%) sera cross-neutralizing both isolates. Sera capable of cross-neutralizing both ANT70 and IIIB consistently neutralized other field isolates tested, with a remarkable similarity in neutralizing antibody titre. A significantly higher number of sera cross-neutralizing both ANT70 and IIIB compared with sera lacking neutralizing antibodies, reacted simultaneously in enzyme-linked immunosorbent assays (ELISA) with three or more V3 loop peptides belonging to HIV-1 strains of different genotypes. However, none of the sera cross-neutralizing ANT70 and IIIB were reactive in ELISA with the ANT70 V3 loop peptide. CONCLUSION: These results suggest that despite pronounced genomic variation of the HIV-1ANT70 isolate, there are strongly conserved neutralizing epitopes situated outside the V3 loop that are shared by other HIV-1 isolates. These findings suggest that genetic variation might be surmountable in the design of a polyvalent HIV vaccine, if neutralizing antibodies are found to be correlates of protection in HIV infection.

Acquired Immunodeficiency Syndrome↗

Genotypic subtypes of HIV-1 in Cameroon.

OBJECTIVE: The only two HIV-1 strains (ANT70 and MVP5180) reported to date from Cameroon are members of the outlier clade (group O). In this study, we assessed the prevalence of group O viruses and other HIV-1 subtypes in Cameroon. DESIGN: A phylogenetic analysis of 18 HIV-1 strains isolated from seropositive individuals from Yaoundé and Douala, Cameroon. METHODS: A 900 base-pair fragment of the env gene coding for V3, V4, V5, and the beginning of gp41 of 17 out of 18 HIV-1 isolates from Cameroon was amplified, cloned and sequenced using polymerase chain reaction. A phylogenetic tree was constructed. RESULTS: The overall env nucleotide sequence divergence among the Cameroon isolates ranged from 6.1 to 27.5%. In a phylogenetic tree, six subtypes were identified when compared with 23 reference strains of different geographic origin. Of these 17 Cameroonian strains, 11 (61%) were of subtype A of which the interpatient distances at the sequence level varied from 6.1% to 18.3% (average, 11.9%). Three (17%) strains were of subtype F, and the other three strains (6% each) belonged to subtypes B, E and H, respectively. The remaining isolate was classified as belonging to group O, on the basis of the sequence of part of the pol gene. A very broad spectrum of different tetrameric amino-acid sequences was observed at the apex of the V3 loop. Eleven strains contained the tetrameric globally predominant GPGQ sequence at the tip of the V3 motif. Two strains had the GPGR sequence typical of the American and European HIV-1 strains. The remaining tetrameric sequences included GPGS, GSGQ, GRGQ, and GLGR. CONCLUSION: These findings on a limited number of viruses suggest extensive env gene diversity of HIV-1 strains from Cameroon, and could have implications for vaccine development in Africa.

Amino Acid Sequence↗

Identification of an env G subtype and heterogeneity of HIV-1 strains in the Russian Federation and Belarus.

OBJECTIVE: To identify HIV-1 envelope sequence subtypes in infected individuals from the Russian Federation and Belarus. PATIENTS: A cohort of children infected after exposure to non-sterile needles during the 1988-1989 HIV-1 epidemic in southern Russia (n = 20) and HIV-1-seropositive individuals from Russia (n = 1) and Belarus (n = 7) infected via sexual transmission. METHODS: DNA samples derived from peripheral blood mononuclear cells were analysed for their HIV-1 genotypes by the heteroduplex mobility assay (HMA). The 1.3 kilobase-pair env gene fragments encoding a portion of gp120 were amplified by nested polymerase chain reaction, cloned and sequenced. The env sequences derived from these patients were aligned and phylogenetic neighbour-joining and maximum parsimony-derived trees generated. RESULTS: The env sequences derived from eight individuals infected in Russia and Belarus belong to subtype A (one), B (four), C (two), and D (one). Sequences derived from children, infected during parenteral manipulations in southern Russia, and one mother were closely related, but highly divergent, as a group, from all prototypic strains (genetic divergence, 17.2-22.9%). However, they clustered together with env sequences of the V1525 and LBV21-7 isolates from Gabon, recently described to be members of a new HIV-1 env subtype G. CONCLUSION: Extensive heterogeneity of HIV-1 subtypes was evident in the Russian Federation and Belarus. Our data also support the existence of an HIV-1 env genetic subtype G, and such isolates are now apparently present on both the African and European continents. These variants were identified through V3 peptide enzyme-linked immunosorbent assay screening and subsequent HMA analysis. The combination of these techniques represents a model for screening HIV variants within a large population.

Amino Acid Sequence↗

Study of the hepatic growth hormone (GH) receptor at different ages in chickens selected for a good feed conversion (FC) and a fast weight gain (GL).

The influence of genetic selection for improved feed conversion (FC) and fast growth (GL) in broiler chickens, on the hepatic growth hormone receptor was investigated as a function of age. Selection for improved feed conversion resulted in lean chickens, whereas selection for fast growth resulted in fat chickens. Growth hormone receptor characteristics were determined in liver microsomal fractions and plasma GH concentrations were measured by radioimmunoassay. Embryos from the FC line showed higher specific GH binding than GL embryos at day 16 of embryonic development only (P < 0.05). On day 18 of embryonic development, the FC line had slightly more GH receptors than the GL line (P < 0.05), whereas simultaneous plasma GH concentrations were greater in the GL line. The effect of selection on hepatic growth hormone binding was greatest in 4 week old growing chicks at which age the GL line showed a 2- to 3-fold higher % of specific binding than the FC line (P < 0.001). The difference at this stage is probably due to down regulation of the receptor by the significantly higher GH concentrations in the FC line. The adult GL chickens also showed higher specific binding than FC, but the difference was less pronounced than in the growing stage (P < 0.05). This was due to a higher number of GH receptors, while receptor affinity was unchanged. Because no line difference in plasma GH concentrations was found in adult hens, other unknown mechanisms probably play a role in determining differences in GH receptor binding between these selected lines at older ages.

Aging↗

Relation between slow-wave frequency and spiking activity during the migrating myoelectric complex in dogs.

The quantitative relation between slow-wave periods and spiking activity was evaluated in vivo in canine small intestine during the fasted state. Experiments were performed in three conscious dogs with three bipolar electrodes, implanted respectively 10, 25 and 40 cm beyond the ligament of Treitz. Digitized electrical recordings were automatically processed for the individual slow-wave periods and spike-burst intensities using a set of computer programs developed in our laboratory. A linear correlation existed between the degree of spiking activity and the average length of the preceding slow-wave period. The slopes of the regression lines were less steep for more distal electrodes. A second series of experiments showed that an increase in the slow-wave period precedes the onset of phase 3 of the migrating myoelectric complex and that a fall in slow-wave period precedes the end of phase 3. These data show that a low slow-wave frequency is accompanied by a facilitation of spiking activity, whereas shortening of the slow-wave period is accompanied by a decrease in spike burst intensity. This relation between slow-wave period and spiking activity shows an aboral trend that may be related to intrinsic slow-wave frequency.

Animals↗

Variation of slow-wave frequency and locking during the migrating myoelectric complex in dogs.

Slow waves determine rhythm and polarity of spike bursts. We measured the variation of slow-wave frequency (swf) and locking (swl) in the canine jejunum during the various phases of the migrating myoelectric complex (MMC) and during induced phase III (erythromycin 125 micrograms/kg iv bolus or somatostatin 2.5 micrograms.kg-1.h-1 iv infusion), blocked phase III (atropine 20 micrograms/kg iv bolus), and so-called stationary phase III activity (cisapride 150 micrograms/kg iv bolus). The EMG of 4 dogs, implanted with 10 bipolar electrodes, was recorded on a polygraph. Our results indicate that swf and swl change during the MMC from a stepwise swf gradient with slow waves locked in plateaus during phase I to a continuous swf gradient without or with significantly reduced phase locking during phase III. The length of the first swf plateau decreases significantly from 42 +/- 12 cm post Treitz during phase I to 11 +/- 4 cm during spontaneous phase III. Atropine block of phase III activity prevents phase unlocking and development of a continuous swf gradient. Our hypothesis is that phase unlocking may be one of the induction mechanisms of spike-burst activity.

Animals↗

Heterologous gene expression by filamentous fungi: secretion of human interleukin-6 by Aspergillus nidulans.

Expression vectors for human interleukin-6 (hIL6) contain an expression cassette consisting of the Aspergillus niger glaA promoter and the Aspergillus nidulans argB terminator. The secretion signals were either those of glaA or that of the authentic hIL6 peptide. The constructs under study were introduced into A. nidulans and A. niger by means of cotransformation. No IL6 activity could be detected in the medium of a cotransformed A. niger strain, although transcripts corresponding with the IL6 cDNA were present. Evidence is presented that this apparent lack of IL6 expression is due to extracellular proteolytic activity. In the media of a cotransformed A. nidulans strain grown on starch, IL6 activity was detected by means of a bioassay. Up to 25 ng/ml of biologically active hIL6 could be secreted by A. nidulans transformed with the plasmid containing the mature hIL6-encoding gene fused to the glaA signal peptide nucleotide sequences. hIL6 of the expected 23-kDa size was also observed by Western-blot analysis of the medium. There was no evidence for glycosylation of the protein.

Amino Acid Sequence↗

Tissue characterization of atheromatous plaques: correlation between ultrasound image and histological findings.

Fifty-four atheromatous plaques of carotid arteries, obtained at autopsy, were scanned with a high-resolution real-time instrument, then histologically examined. Comparison between atherogenesis assessed first by histology then by ultrasonography allowed us to differentiate young from old plaques. An old plaque is characterized by a dense heterogeneous echo pattern and irregular borders or ulceration. Previous studies have shown a correlation between the presence of hemorrhage and ulceration of atheromatous plaques in the carotid, and the frequency of cerebral complications. Since these characteristics were mainly found in older plaques, we consider them to be the most important ones clinically.

Arteriosclerosis↗

Pharmacological and hemodynamic profile of nebivolol, a chemically novel, potent, and selective beta 1-adrenergic antagonist.

The pharmacological profile of nebivolol (N), a chemically novel beta-adrenergic antagonist, was assessed in investigations on isolated tissues, awake spontaneously hypertensive rats (SHR), closed-chest anesthetized dogs, and humans. In vitro, N was found to be a potent antagonist of beta 1-adrenergic receptors (A2 value, 5.8 X 10(-9) M) and only a weak beta 2-adrenergic antagonist (A2 value, 1.7 X 10(-6) M). The selectivity for the beta 1-adrenergic receptor was higher for N than for any of the reference compounds. In dogs--similarly with atenolol--N was more potent in blocking the isoprenaline (I)-induced increases in left ventricular performance than the I-induced decrease in arterial pressure. In dogs, as compared with propranolol, N (0.025 and 0.01 mg.kg-1 i.v.) increased cardiac output and stroke volume, lowered systemic vascular resistance, and had no significant effect on the variables related to left ventricular contraction. In contrast to other beta-adrenergic antagonists, N acutely lowered arterial blood pressure in SHR (1.25 mg.kg-1 i.p.) and in hypertensive patients (1 oral dose of 5 mg) for several hours. In healthy volunteers N (5 mg) lowered systemic vascular resistance during daily oral treatment and did not negatively affect left ventricular function. In conclusion, N is a potent and selective beta 1-adrenergic blocking agent with an interesting hemodynamic profile. In hypertensive subjects and SHR, a single dose lowers arterial blood pressure for substantial periods of time.

Adrenergic alpha-Antagonists↗

BM 13.177, a selective blocker of platelet and vessel wall thromboxane receptors, is active in man.

BM 13.177, a sulphonamide derivative, prevented platelet aggregation by thromboxane A2 in vitro and selectively inhibited contraction of isolated rabbit femoral arteries induced by two stable endoperoxide analogues. In a double-blind placebo-controlled study, oral BM 13.177 inhibited platelet aggregation induced by arachidonic acid, low dose collagen, and the two stable endoperoxide analogues, and slightly prolonged the bleeding time. Generation of thromboxane or of other prostaglandins was not affected. No side-effects were seen. BM 13.177 appears selectively and safely to block platelet and vessel wall thromboxane receptors and should be useful in elucidating the role of thromboxane A2 in disease.

15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5↗

Sources of calcium used during alpha 1- and alpha 2-adrenergic contractions in canine saphenous veins.

Experiments were performed to determine the role of intra- and extracellular Ca2+ during contractions of canine saphenous veins evoked with 'alpha 1'- and 'alpha 2'-adrenergic agonists. After incubation in Ca2+-free medium for 45 min, noradrenaline, clonidine and phenylephrine caused contractions, reaching a maximal amplitude rapidly and then declining to a lower stable level within 6 min. The maximal amplitude amounted to about 10% of the response to the agonists in physiological saline containing Ca2+ ions. After a previous exposure to one of the alpha-adrenergic agonists in Ca2+-free medium, no peak response was observed when the preparations were exposed to any of the agonists a second time. The stable smaller contractions which followed the peak response could however be repeated after a previous exposure to one of the agonists. In normal physiological salt solution (PSS), iproveratril depressed the maximal response to all three agonists to about 60% of the control value. However, the shift of the median effective dose (ED50) to the right was larger for clonidine than for the other two agonists. Noradrenaline, clonidine and phenylephrine (all at 10(-5) M) increased the uptake of 45Ca. Iproveratril reduced this increased uptake of 45Ca. Noradrenaline, clonidine, guanfacine and phenylephrine augmented the efflux rate of 45Ca. After exposure to one of the agonists, noradrenaline caused no or only a moderate increase of the efflux rate of 45Ca. The present experiments indicate that in the dog's saphenous vein: (1) alpha 1- and alpha 2-adrenergic activation cause both influx of Ca2+ ions and release of intracellular Ca2+; (2) the source of the intracellular Ca2+ is the same; (3) the mechanism underlying the influx of extracellular Ca2+ ions into the smooth muscle cells, to judge from the inhibiting effect of iproveratril, is most likely to be the same.

Adrenergic alpha-Agonists↗