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J A Jonasson

Publications and source records attributed to J A Jonasson.

10 recordsLinked to original sources

Structural studies of lipooligosaccharides from Haemophilus ducreyi ITM 5535, ITM 3147, and a fresh clinical isolate, ACY1: evidence for intrastrain heterogeneity with the production of mutually exclusive sialylated or elongated glycoforms.

The structures of the lipooligosaccharides (LOSs) from Haemophilus ducreyi ITM 5535 and ITM 3147 and a fresh clinical isolate, ACY1, have been investigated. Oligosaccharides were obtained from phenol-water-extracted LOS by mild acid hydrolysis and were studied by methylation analysis, fast atom bombardment and electrospray ionization mass spectrometry, and nuclear magnetic resonance spectroscopy. The major oligosaccharide obtained from all strains was a nonasaccharide with the structure beta-D-Galp-(1-->4)-beta-D-GlcNAcp-(1-->3)-beta-D-Galp-(1-->4)-D-a lpha-D-Hepp- (1-->6)-beta-D-Glcp-(1-->[L-alpha-D-Hepp-(1-->2)-L-alpha-D-Hepp - (1-->3)]4)-L-alpha-D-Hepp-Kdo (Kdo stands for 3-deoxy-D-manno-octulosonic acid) and is thus identical to that identified as the major oligosaccharide in H. ducreyi ITM 2665 (E. K. H. Schweda, A. C. Sundström, L. M. Eriksson, J.A. Jonasson, and A. A. Lindberg, J. Biol. Chem. 269:12040-12048, 1994). Electrospray ionization mass spectrometry on O-deacylated LOS from H. ducreyi ITM 5535 obtained after treatment with anhydrous hydrazine gave evidence for the presence of a sialylated major compound, Neu5Ac alpha(2-->3)-beta-D-Galp-(1-->4)-beta-D-GlcNAcp-(1-->3)-beta-D-Gal p- (1-->4)-D-alpha-D-Hepp-(1-->6)-beta-D-Glcp-(1-->[L-alpha-D-Hepp -(1-->2)-L- alpha-D-Hepp-(1-->3)]4)-L-alpha-D-Hepp-Kdo(P)-O-deacylated lipid A (Neu5Ac stands for N-acetylneuraminic acid). However, an even larger oligosaccharide could be isolated from all strains as a minor component, viz., the undecasaccharide beta-D-Galp-(1-->4)-beta-D-GlcNAcp-(1-->3)-beta-d-Galp-(1-->4)-beta-D-glcNAcp-(1-->3)-beta-D-Galp-(1-->4)-D-alpha-D-Hepp-(1-->6)-beta-D-Glcp-(1-->[L-alpha-D-Hepp-(1-->2)-L-alpha-D-Hepp-(1-->3)]4-L-alpha-D-Hepp-Kdo, which represents an N-acetyl lactosamine disaccharide unit elongation of the LOS outer core. No Sialylation of this latter minor component undecasaccharide was detected.

Carbohydrate Conformation↗

Structural studies of the cell envelope lipopolysaccharides from Haemophilus ducreyi strains ITM 2665 and ITM 4747.

The structures of the lipopolysaccharides from Haemophilus ducreyi strains ITM 2665 and ITM 4747 have been investigated. Oligosaccharides were obtained from phenol/water-extracted lipopolysaccharide by mild acid hydrolysis and were studied with methylation analysis, fast atom bombardment-mass spectrometry, and NMR spectroscopy. The major oligosaccharide obtained from strain 2665 is a nonasaccharide with the following structure: beta-D-Galp-1-->4-beta-D-GlcNAcp-1-->3-beta-D-Galp-1-->4-D-alpha-D -Hepp- 1-->6-beta-D-Glcp-1-->(L-alpha-D-Hepp-1-->2-L-alpha-D-Hepp-1 -->3)-4-L-alpha- D-Hepp-Kdo, where the reducing terminal 3-deoxy-D-manno-octulosonic acid (or Kdo) exists in reduced anhydro forms. The proposed structure complements the preliminary structure described for Haemophilus ducreyi strain 35000 (Melaugh, W., Phillips, N. J., Campagnari, A. A., Karalus, R., and Gibson, B. W. (1992) J. Biol. Chem. 267, 13434-13439) with the missing anomeric configurations. The saccharide isolated from strain 4747 is a markedly simpler hexasaccharide with the following structure: beta-D-Galp-1-->4-beta-D-Glcp-1-->(L-alpha-D-Hepp-1-->2-L-alpha-D- Hepp- 1-->3)4-L-alpha-D-Hepp-Kdo. Apart from a different phosphorylation of the inner core region the proposed structure is identical to the structure of lipopolysaccharide from an only distantly related bacterium, viz. Haemophilus influenzae nontypable strain 2019 (Phillips, N. J., Apicella, M. A., Griffiss, J. M., and Gibson, B.W. (1992) Biochemistry 31, 4515-4526). The implications of these findings as regards the role of lipopolysaccharide as a virulence factor are discussed.

Carbohydrate Conformation↗

High resolution gene mapping of the human alpha globin locus.

A combination of polymorphic DNA markers, cytogenetic analysis, and in situ hybridisation has been used for the high resolution assignment of the human alpha globin gene cluster on chromosome 16. Multiallelic DNA probes from within the alpha globin cluster were used to determine the number of copies of this locus in three cell lines containing trisomies of the short arm of chromosome 16 and one with a familial inversion, inv(16). The breakpoints in these rearrangements flank the alpha globin locus and locate a shortest region of overlap to 16p13.1. A meiotic crossover was also localised to this band. In situ hybridisation of biotinylated DNA probes to normal and inverted chromosomes 16 [inv(16)(p13.1;q22)] showed hybridisation sites at opposite ends of the chromosomes, consistent with this regional localisation.

Chromosome Inversion↗

Chromosomal localization of genes by scanning electron microscopy using in situ hybridization with biotinylated probes: Y chromosome repetitive sequences.

The feasibility of using scanning electron microscopy (SEM) to identify the position of specific DNA sequences was examined using a Y chromosome 'specific' probe (pHY2.1). Tests were carried out on chromosome spreads hybridized in situ with biotinylated pHY2.1. Chromosomal sites of hybridization of the probe were localized by an indirect immunohistochemical procedure which resulted in a gold product which could be amplified by silver precipitation. In the SEM, the specific location of the probe was easily identified due to the enhanced signal produced by the gold-silver complex. The probe was localized both on the long arm of the Y chromosome and within interphase nuclei. It was found that SEM was more sensitive than light microscopy since the probe could be identified without silver amplification. With refinements to the technique, SEM could provide a useful method for high resolution localizing of unique DNA sequences (i.e. single copy genes).

Biotin↗

Sensitive system for visualising biotinylated DNA probes hybridised in situ: rapid sex determination of intact cells.

A fast and sensitive method for detecting biotinylated deoxyribonucleic acid (DNA) probes was used for sex determination of cells and tissues by in situ hybridisation of a probe "specific" for the Y chromosome (pHY 2.1). Within 24 hours this procedure visualizes the Y chromosome in fetal and adult cells and tissue, without background noise. This procedure should facilitate antenatal determination of sex on small numbers of uncultured cells. The sensitivity of the procedure also permits the chromosomal assignment of genes present in low copy number.

Adult↗

Chromosome maps of man and mouse II.

Chromosome displays and listings are presented showing loci whose position is known in both man and mouse, in similar manner to our previous report (Dalton et al. 1981). There is now evidence for at least 27 conserved autosomal segments with two or more loci in the two species. The human and mouse chromosome maps show the location of homologous genes. The mouse map also shows the positions of translocations used in gene location and of some other genes used in linkage studies on them.

Animals↗

Haemophilus ducreyi.

Genital ulcer disease as a risk factor for HIV transmission has become apparent in the fight against AIDS. In poor communities in developing countries with people living under low hygienic conditions chancroid is often reported to be the most common form of genital ulcer. It is caused by Haemophilus ducreyi; a fastidious bacterium, notoriously difficult to grow in the laboratory. Apart from a few small micro-epidemics from time to time the disease, which has interesting immunological aspects, is now rare and almost forgotten in most industrialized countries. This may be part of the reason why there is no simple and inexpensive diagnostic test available yet that would be useful in the low-resource settings where chancroid is prevalent. The present review focuses on the diagnosis and pathogenesis of chancroid.

Chancroid↗