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Biomedical subjects

A Forsberg

Publications and source records attributed to A Forsberg.

At least 55 records · Page 3Linked to original sources

Genetic analysis of the yopE region of Yersinia spp.: identification of a novel conserved locus, yerA, regulating yopE expression.

The yopE gene of Yersinia pseudotuberculosis was recently sequenced, and YopE was identified as an indispensable virulence determinant when tested in a mouse model (A. Forsberg and H. Wolf-Watz, Mol. Microbiol. 2:121-133, 1988). In the study described here, the DNA sequences of the yopE genes of Yersinia pestis EV76 and Yersinia enterocolitica 8081 were determined and compared with that of the Y. pseudotuberculosis gene. Only two codons were found to differ, both leading to amino acid replacements, when the gene from Y. pestis was compared. These two replacements were also present in the gene from Y. enterocolitica; in addition, 18 other codons were found to differ. Thirteen of these substitutions led to amino acid replacements. Downstream of the yopE gene, the plasmid partition locus par was found to be conserved in all three species. In Y. enterocolitica 8081, the sequence homology was interrupted by a putative insertion sequence element inserted between the yopE gene and the par region at a position only 5 base pairs downstream of the yopE stop codon. Upstream of the yopE gene, 620 base pairs were conserved in the three species. This region contained a 130-amino-acid-long open reading frame reading in the opposite direction to the yopE gene and expressed a 14-kilodalton protein in minicells. An insertion mutation in this region constructed in Y. pseudotuberculosis expressed significantly lower amounts of YopE protein in vitro than did the corresponding wild type. The expression level could be restored by transcomplementation. This new locus was designated yerA, for yopE-regulating gene A. The yerA mutant was avirulent when mice were challenged by oral infection.

Amino Acid Sequence↗

Studies on metal content in the brown seaweed, Fucus vesiculosus, from the Archipelago of Stockholm.

Concentrations of eleven metals (Al, Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb, V and Zn) were determined in the brown seaweed Fucus vesiculosus collected from the Archipelago of Stockholm. Several factors which influence the metal content in the seaweed have been studied, including errors caused by epiphytes, sea exposure and differences depending on which part of the seaweed is analysed. It is concluded that, if all these factors are considered, Fucus vesiculosus plants are excellent bio-indicators of metal pollution. This is also demonstrated by a significant increase in metal content in transplanted Fucus vesiculosus near the city of Stockholm. The results from this investigation also indicate increasing metal concentrations, especially Cd, in samples from the northern parts of the Archipelago and the reason for this is discussed.

Journal Article↗

Concentrations of cadmium and other metals in Fucus vesiculosus L. and Fontinalis dalecarlica Br. Eur. from the northern Baltic Sea and the southern Bothnian Sea.

Concentrations of Cd and ten other metals (Al, Co, Cr, Cu, Fe, Mn, Ni, Pb, V and Zn) were determined in the brown seaweed Fucus vesiculosus L. and the aquatic moss, Fontinalis dalecarlica Br. Eur. from the northern Baltic Sea and the southern Bothnian Sea. Elevated concentrations of metals were found in samples taken close to densely populated areas, such as Stockholm and Nynäshamn. Very high concentrations of especially Zn were found in both Fucus and Fontinalis samples taken from the area south of the Gulf of Gävle. The results indicate that mining and industrial activities along the river Dalälven are the main sources of Zn and several other metals. Cd concentrations in Fucus plants reached maximum values (24.5 mg kg(-1)) at the northern site. The gradual increase of Cd concentrations in Fucus plants northward could not be totally explained by the salinity gradient in the Baltic Sea; reasons for this are discussed in this paper.

Journal Article↗

Identification and mapping of the temperature-inducible, plasmid-encoded proteins of Yersinia spp.

The structural genes of the outer membrane polypeptides of Yersinia spp. (YOPs) and the V antigen of plasmid pIB1 of Yersinia pseudotuberculosis were recently cloned and mapped (A. Forsberg, I. Bölin, L. Norlander, and H. Wolf-Watz, Microb. Pathogen. 2:123-137, 1987). The corresponding genes were localized on pYV019 and pYV8081 of Yersinia pestis and Yersinia enterocolitica, respectively. No obvious differences were observed on comparison of pIB1 and pYV019, whereas pYV8081 showed intragenic as well as extragenic changes. However, one region of plasmid pYV8081, which coded for the V antigen, YOP3, and YOP4a, was essentially conserved among the three plasmids. Since this region is connected with the Ca2+ region, we suggest that the conserved region of the virulence plasmids of Yersinia spp. be extended to include both of these regions. Low amounts of the YOPs were detected in the membrane fraction at 37 degrees C in the presence of 2.5 mM calcium. Only minor differences were noticed when the individual YOPs of Y. pestis and Y. pseudotuberculosis were compared. Several differences were observed when the YOPs of Y. enterocolitica were included for comparison. All Y. enterocolitica proteins, except YOP1, YOP4b, and the V antigen, exhibited changes in their characteristic molecular sizes. Although these differences were within a range of +/- 2 kilodaltons, the isoelectric point was retained for each protein type.

Antigens, Bacterial↗

The virulence protein Yop5 of Yersinia pseudotuberculosis is regulated at transcriptional level by plasmid-plB1-encoded trans-acting elements controlled by temperature and calcium.

The DNA sequence of the structural gene (yopE) of one of the Yersinia pseudotuberculosis virulence plasmid-plB1-encoded proteins, Yop5, is presented. The deduced protein showed a molecular weight of 22,971 Daltons. A specific mutant, having a kanamycin-resistance fragment inserted within the yopE gene was no longer virulent for mice. The expression of the Yop5 protein is regulated at the level of transcription by temperature as well as by the Ca2+-concentration of the medium. A significant increase in the level of transcription was not detected until 45 min after a temperature shift from 26 degrees C to 37 degrees C in the absence of calcium; addition of Ca2+ inhibited the expression. The yopE promoter is under positive, as well as negative, plB1-encoded control. The positive function is solely regulated by temperature, while the regulation of the negative function involves at least five different plasmid-encoded gene loci; one of these genes encodes the V-antigen.

Amino Acid Sequence↗

Molecular cloning and expression of calcium-regulated, plasmid-coded proteins of Y. pseudotuberculosis.

A number of plasmid-associated proteins (YOPs) of Y. pseudotuberculosis are induced and expressed at high levels when the pathogen is grown at 37 degrees C in absence of Ca2+ ions. These proteins were recovered both from the outer membrane fraction and the culture supernatant. Two hours after a temperature-shift the YOPs were only found in the culture supernatant, amounting to about 5% of the total cell protein. After 4 h of incubation they were also detected in the outer membrane fraction. Separation by 2-D gel electrophoresis revealed that the YOPs could be separated into 6 different polypeptides; YOP2a (45 kDa), YOP2b (45 kDa), YOP3 (41-42 kDa), YOP4a (34 kDa), YOP4b (34 kDa) and YOP5 (26 kDa). The structural genes of all of these YOPs, except the YOP2a gene, were cloned to pBR322 and their respective genetic localization was established. It was found that the genes were not part of a common operon but scattered around plasmid plB1. Only the YOP4b protein was found to map within the Ca2+ region. The hybrid plasmid plB572 coded for a number of plasmid plB1 specific proteins, one of which showed a molecular weight of 38 kDa. This polypeptide could be precipitated by monospecific V-antiserum, showing that this protein is the V-antigen.

Antigens, Bacterial↗

Immunologic aberrations, HIV seropositivity and seroconversion rates in patients with hemophilia B.

Because there have been reports that factor IX concentrate is less immunosuppressive and therefore factor IX users have less immunologic aberrations, we have studied a group of 22 patients with hemophilia B and six patients with factor VIII deficiency and high titer inhibitors with respect to lymphocyte numbers and function, human immunodeficiency virus (HIV) serology, and factor usage. This group was compared to 111 patients with hemophilia A and a group of 28 healthy male volunteer controls. When the study began in 1983, the majority of patients with hemophilia B and with higher titer factor VIII inhibitors were seronegative, 77% and 83% respectively, as compared to only 30% of patients with hemophilia A. At that time the factor IX users also had milder immune aberrations than the hemophilia A group. However, with time and increasing clotting factor concentrate usage, seroconversion and more striking abnormalities in immune function have occurred in the hemophilia B group. In a subgroup of 16 patients with hemophilia B studied twice, the incidence of seropositivity increased from 31% in 1983 to 69% in 1985. We thus conclude that factor IX concentrate in itself is not less immunosuppressive than factor VIII concentrate. Seroconversion in factor IX concentrate users appears to be lagging behind seroconversion in factor VIII concentrate users, perhaps secondary to the lower cumulative dosage of concentrate that patients with hemophilia B utilize.

Adolescent↗

High doses of fluoride do not affect cyclic AMP levels in human and rat plasma or urine.

The effect of fluoride ingestion on plasma and urinary cyclic AMP levels was studied in healthy volunteers, children undergoing prophylactic fluoride treatment and in rats. In the first series of experiments fluoride ingestion led to a 20-fold increase in plasma fluoride concentration, which was unrelated to changes in plasma cyclic AMP. The only significant effect on plasma cyclic AMP occurred prior to fluoride, an effect attributed to stress. In the second series performed in children increases in urinary F- of up to 200-fold were associated with slight but nonsignificant changes in cyclic AMP excretion. In rat experiments the blood sampling procedure was associated with a rise in plasma cyclic AMP. When this was prevented by pretreatment with propranolol (3 mg/kg), the effect of fluoride (50 ppm in the drinking water) was tested. A fall in urine production was not associated with any change in cyclic AMP excretion or in nephrogenic cyclic AMP. It is concluded that if fluoride alters cyclic AMP in rats and man the effect is small and easily masked by other factors such as catecholamine release following stress.

Adult↗

Kinetics for the inhibition of acetylcholinesterase from the electric eel by some organophosphates and carbamates.

The inhibition kinetics for some organophosphates (paroxon, diisopropylfluorophosphate, sarin, VX, soman and soman isomers) and carbamates (physostigmine, neostigmine, pyridostigmine and carbaryl) in the reaction with acetylcholinesterase from electric eel have been studied. Dissociation constants and rate constants for the irreversible step were determined. The great differences in inhibitory power of the organophosphates were almost entirely due to differences in affinity. A possible correlation between affinity and bonding rate is discussed.

Animals↗

Platelet activation and mitral valve prolapse.

Mitral valve prolapse (MVP) is a predisposing factor for cerebral ischemia, especially in young adults. Cerebral embolization of intracardiac thrombi is the probable mechanism in many cases. Platelets play a key role in the development of thrombi. We found that platelet factor 4, a marker protein of platelet activation, was elevated in 12 of 33 MVP patients (36%) without a history of stroke. This finding indicates that platelets are frequently activated in asymptomatic MVP patients and may allow identification of a subgroup of MVP patients with activated platelets who are at increased risk for emboli.

Adolescent↗

Marker proteins of platelet activation in patients with cerebrovascular disease.

Platelet activation is assumed to occur in many patients with strokes of transient ischemic attacks (TIAs). Determination of this activity can now be measured by assaying the platelet-specific proteins, platelet factor 4 and beta-thromboglobulin. These proteins are platelet specific and are released during irreversible aggregation. The plasma level of these proteins was measured in patients with strokes and TIAs and both groups had significantly greater values than healthy control subjects. These markers of platelet activation may provide further understanding of the role of platelets in the etiology, diagnosis, and treatment of cerebrovascular disease.

Adolescent↗

Morphological muscle characteristics in rowers.

Biopsies were taken from the vastus lateralis and deltoid muscle in twelve major senior rowers. Histochemical analyses showed a predominance of the oxidative type I fiber, a low content of the highly glycolytic type IIB fiber, large fiber areas, and a large number of capillaries surrounding the fibers. All these trends were more pronounced in the internationally competitive compared to the nationally competitive rowers. Exceptionally high aerobic capacities and muscular strengths, in competitive rowers, have been reported in previous papers as well as confirmed in the present study. The muscle morphology reported here is in line with the high aerobic demand in race rowing. However, strength has been shown to be more favoured by a high content of type II fibers which may appear contradictory to the present results. This question together with possible effects of physical training on the muscle morphology is discussed.

Adult↗