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Biomedical subjects

L Andersson

Publications and source records attributed to L Andersson.

At least 163 records · Page 9Linked to original sources

Impact of plasmid presence and induction on cellular responses in fed batch cultures of Escherichia coli.

Fed batch cultivations of plasmid-free and recombinant Escherichia coli were employed in order to determine cellular responses and effects of plasmid presence and induction on the host cell physiology. While plasmid presence was shown to have minor influence on overall biomass yield, induction with 0.1 mM IPTG led to a marked reduction. The number of dividing cells, measured as colony forming ability, was influenced by plasmid presence and to a larger extent by induction. The latter caused a decline in the number of dividing cells to less than 10% of the population within 10 h. However, this cell segregation did not affect the specific rate of product formation, which was approximately constant throughout the cultivations. Analysis of the in vivo degradation rate of the product indicated that it was proteolytically stable. The cellular content of the stringent response signal substance, ppGpp, peaked immediately after transition from batch to fed batch mode to stabilise at a higher value than in the batch phase. When the specific growth rate declined below 0.06 h-1 an additional rise in ppGpp concentration was observed.

Cell Division↗

Purification of cysteine-containing synthetic peptides via selective binding of the alpha-amino group to immobilised Cu2+ and Ni2+ ions.

Peptides containing a cysteine residue but lacking histidine and tryptophan were synthesised by the solid-phase method. Their retention behaviour on Cu2+ - and Ni2+ -loaded immobilised metal ion affinity chromatography (IMAC) supports at pH 5-11 was studied and compared with that observed for the corresponding compounds without the free alpha-amino group and/or the thiol function. Unexpectedly, it was found that neither a cysteine side-chain nor a cysteine disulphide affects the retention of the peptides. A free alpha-amino group is required for binding; no retention is observed in its absence. At pH 9 substantial amounts of metal ions were transferred from the chromatographic support to an alpha-amino-protected cysteine-containing peptide. However, at pH 7 no such transfer occurred. Therefore, the lack of retention observed for peptides with a blocked alpha-amino function over the entire pH range is not solely caused by metal ion scavenging by the thiol group. Partial dimerisation may occur upon chromatography; the dimers formed are retained strongly due to the presence of two free alpha-amino groups. It seems that IMAC on a Cu2+ - or Ni2+ -loaded support can be used for the purification of cysteine-containing peptides synthesised by the solid-phase method. Inclusion of a capping protocol in the synthesis ensures that a free alpha-amino group, which can be used as an affinity handle, will be present only on the target peptide.

Amino Acid Sequence↗

Structure and organization of pig MHC class II DRB genes: evidence for genetic exchange between loci.

The pig major histocompatibility complex DRB genes were studied by polymerase chain reaction (PCR) amplification of exon 2 from eight domestic pigs and two European wild boars. Sequence comparisons together with a phylogenetic analysis showed the existence of at least three DRB genes of which only one appears to be expressed. The two putative DRB pseudogenes contained deletions in exon 2, making it possible to confirm the presence of three non-allelic DRB genes by analyzing the length polymorphism of the amplified PCR products. The expressed gene shows allelic polymorphism at the same positions as in the human DRB1 gene. In addition, this pig gene shows extensive allelic polymorphism at positions 84-88, whereas, e.g., human DRB genes do not. Surprisingly, the two putative DRB pseudogenes also display a considerable amount of allelic polymorphism, albeit of a different character as compared with the expressed DRB gene. Short stretches of sequences are shared between individual alleles at different loci. These sequence similarities cannot be due to natural selection, since two of the three DRB genes involved are polymorphic pseudogenes constituting allelic series that have diverged after the inactivation event. Instead, the results indicate that the sequences have been exchanged between the DRB genes by intergenic recombination.

Animals↗

A major locus (RN) affecting muscle glycogen content is located on pig chromosome 15.

The RN locus in pigs has a major effect on the amount of stored glycogen in white muscle and affects meat quality. The fully dominant RN- allele, associated with high glycogen content, occurs in the Hampshire breed. We have mapped the RN locus using a large half-sib family comprising one heterozygous RN-/rn+ Hampshire boar mated to homozygous rn+/rn+ Swedish Landrace x Swedish Yorkshire sows. The segregation at the RN locus was inferred from data on glycolytic potential and residual glycogen in white muscle which both showed clear bimodal distributions. Highly significant evidence for genetic linkage was obtained against microsatellite markers on Chromosome (Chr) 15. Multipoint analysis revealed the order Sw1111-8.0-S0088-10.6-RN-4.8-Sw936,Sw906 (recombination estimates are given as Kosambi cM). Comparative mapping data imply that the human homolog of RN is located on Chr 2q.

Animals↗

Pigs with the dominant white coat color phenotype carry a duplication of the KIT gene encoding the mast/stem cell growth factor receptor.

Comparative mapping data suggested that the dominant white coat color in pigs may be due to a mutation in KIT which encodes the mast/stem cell growth factor receptor. We report here that dominant white pigs lack melanocytes in the skin, as would be anticipated for a KIT mutation. We found a complete association between the dominant white mutation and a duplication of the KIT gene, or part of it, in samples of unrelated pigs representing six different breeds. The duplication was revealed by single strand conformation polymorphism (SSCP) analysis and subsequent sequence analysis showing that white pigs transmitted two nonallelic KIT sequences. Quantitative Southern blot and quantitative PCR analysis, as well as fluorescence in situ hybridization (FISH) analysis, confirmed the presence of a gene duplication in white pigs. FISH analyses showed that KIT and the very closely linked gene encoding the platelet-derived growth factor receptor (PDGFRA) are both located on the short arm of Chromosome (Chr) 8 at band 8p12. The result revealed an extremely low rate of recombination in the centromeric region of this chromosome, since the closely linked (0.5 cM) serum albumin (ALB) locus has previously been in situ mapped to the long arm (8q12). Pig Chr 8 shares extensive conserved synteny with human Chr 4, but the gene order is rearranged.

Animals↗

A missense mutation in the gene for melanocyte-stimulating hormone receptor (MC1R) is associated with the chestnut coat color in horses.

The melanocyte-stimulating hormone receptor gene (MC1R) is the major candidate gene for the chestnut coat color in horses since it is assumed to be controlled by an allele at the extension locus. MC1R sequences were PCR amplified from chestnut (e/e) and non-chestnut (E/-) horses. A single-strand conformation polymorphism was found that showed a complete association to the chestnut coat color among 144 horses representing 12 breeds. Sequence analysis revealed a single missense mutation (83Ser-->Phe) in the MC1R allele associated with the chestnut color. The substitution occurs in the second transmembrane region, which apparently plays a key role in the molecule since substitutions associated with coat color variants in mice and cattle as well as red hair and fair skin in humans are found in this part of the molecule. We propose that the now reported mutation is likely to be the causative mutation for the chestnut coat color. The polymorphism can be detected with a simple PCR-RFLP test, since the mutation creates a TaqI restriction site in the chestnut allele.

Alleles↗

Lung cancer, smoking and diet among Swedish men.

In a prospective case-control lung cancer study in the west of Sweden, the relationship between lung cancer, smoking and dietary factors was investigated. Suspected cases were collected from pulmonary units at two central hospitals in the area investigated and population controls of the same age and sex were selected from registers. The majority of cases and controls were interviewed by specially trained nurses, using a food frequency questionnaire. The lung cancer diagnosis (ICD 7, 162.1) was made using data from the local cancer register. In an analysis based on 308 cases and 504 controls, a dose-related increase in lung cancer risk for smokers was found, although no significant risk was found for males smoking 1-10 cigarettes/day for less than 20 years. A lower consumption of vegetables was related to a higher risk, both for smokers and nonsmokers. A higher consumption of milk was related to an increased risk.

Adult↗

The PCR typing of MHC-DRB genes in the sheep using primers for an intronic microsatellite: application to nematode parasite resistance.

The strong association between polymorphisms in an intronic microsatellite and the coding sequences for (BoLA)-DRB3 genes, previously described for demonstrating alleles of class II major histocompatibility complex (MHC) in the cow, was examined in sheep to see if similar polymorphisms could be demonstrated in the DRB region of the MHC. The bovine primes LA53 and LA54, previously used to amplify the bovine DRB3 microsatellites, were used with DNA from Australian sheep, eight DRB alleles were identified by length polymorphisms of polymerase chain reaction (PCR) products amplified from the DRB microsatellite region. Incomplete amplification of both alleles was sometimes found for sheep DNA samples using bovine primers, so a modified primer (LA53b) was used, and found to amplify the microsatellite next to intron 2 of the MHC more reliably than the LA53 primer. Two additional primers (LA31 and LA32), used in amplification of the exon 2 region of bovine DRB3, were used in the sheep, and the PCR products were analysed by single-stranded conformation polymorphism (SSCP). These primers successfully amplified the variable region of the ovine DRB region coded by exon 2, and the SSCP technique demonstrated polymorphisms with sheep DNA. Family studies demonstrated the segregation of alleles, by amplification both of intronic microsatellites and of the exon 2 variable region. Close correspondence was found between the two regions for several alleles, suggesting that the intronic microsatellites were closely linked to DRB-variable region alleles. Three families of Merino sheep with different antibody responses to intestinal nematode parasites were examined. The sire group with the highest antibody levels possessed two microsatellite alleles of closely similar length (alleles 3 and 4) inherited from the sire and present in high frequency in the lambs. In contrast, the other two sires did not possess these two alleles and the alleles were in low frequency in their progeny. Further studies are required in unrelated sheep to confirm whether these two alleles are associated with resistance to nematode parasites.

Alleles↗

Dietary factors and lung cancer among men in west Sweden.

BACKGROUND: Previous studies have reported an association between tea drinking and lung cancer. In view of these data, the relationship between tea drinking as well as other dietary factors and lung cancer was investigated in a case-control study in the west of Sweden. METHODS: Patients with suspected lung cancer were collected from pulmonary units at central hospitals in the area investigated, and population controls were matched for age. The material reported here comprises 308 male cases with a confirmed diagnosis of lung cancer and 504 controls. The participants were interviewed by specially trained nurses, using a questionnaire to assess smoking, dietary habits, occupational exposures and conditions in the residential area (local air pollution). This paper reports the results from dietary factors studied with a food frequency technique. RESULTS: The results demonstrated a strong protective effect of vegetables (odds ratio [OR] = 0.69, 95% confidence interval [CI]: 0.46-1.05, and OR = 0.37, 95% CI: 0.23-0.61 for intermediate and high consumption classes respectively). A low OR was consistent for all histological types of lung cancer. High consumption of fruits did not show any similar protective effect. Drinking milk was associated with a dose-response related risk increase after adjustment for smoking and vegetable consumption (P for trend = 0.07). Odds ratio was 1.73, 95% CI: 1.00-3.01 for high consumption of milk. CONCLUSIONS: High intake of vegetables had a strong protective effect among males. Diet is thus a potential confounding factor in studies on lung cancer and environmental factors and should thus be taken into consideration in the planning of such studies.

Adenocarcinoma↗

Report of the first workshop on the genetic map of bovine chromosome 23.

A report of the first workshop on the genetic map of bovine chromosome 23 (BTA23) is given. Five laboratories contributed data from 29 loci, including a total 11586 informative genotypes. The combined pedigrees represented 1930 potentially informative meioses. Eighteen of the 29 loci were common to two or more data sets and were used to construct a framework linkage map of BTA23. Twelve of the 18 could be ordered on the linkage map with a likelihood ratio of greater than 1000:1. Thus, a low resolution consensus map was constructed with a high level of support for order. The sex-averaged, female and male maps span 54.5, 52.7 and 55.8 cM, respectively. Sex-specific differences in recombination frequency were identified for eight pairs of framework loci. Average genetic distance between framework loci on the sex-averaged map is 5.0 cM.

Animals↗

A comprehensive linkage map of the pig based on a wild pig-Large White intercross.

A comprehensive linkage map, including 236 linked markers with a total sex-average map length of about 2300 cM, covering nearly all parts of the pig genome has been established. Linkage groups were assigned to approximately all 18 autosomes, the X chromosome and the X/Y pseudoautosomal region. Several new gene assignments were made including the assignment of linkage group U1 (EAK-HPX) to chromosome 9. The linkage map includes 77 types I loci informative for comparative mapping and 72 in situ mapped markers physically anchoring the linkage groups on chromosomes. A highly significant heterogeneity in recombination rates between sexes was observed with a general tendency towards an excess of female recombination. The average ratio of female to male recombination was estimated at 1.4:1 but this parameter varied between chromosomes as well as between regions within chromosomes. An intriguing finding was that blood group loci were overrepresented at the distal ends of linkage groups.

Animals↗

Limited polymorphism at major histocompatibility complex (MHC) loci in the Swedish moose A. alces.

The Swedish moose was analysed for genetic variability at major histocompatibility complex (MHC) class I and class II DQA, DQB and DRB loci using restriction fragment length polymorphism (RFLP) and single strand conformation polymorphism (SSCP) techniques. Both methods revealed limited amounts of polymorphism. Since the SSCP analysis concerned an expressed DRB gene it can be concluded that the level of functional MHC class II polymorphism, at least at the DRB locus, is low in Swedish moose. DNA fingerprinting was used to determine if the unusual pattern of low MHC variability could be explained by a low degree of genome-wide genetic diversity. Hybridizations with two minisatellite probes gave similarity indices somewhat higher than the average for other natural population, but the data suggest that the low MHC variability cannot be explained by a recent population bottleneck. However, since minisatellite sequences evolve more rapidly than MHC sequences, the low levels of MHC diversity may be attributed to a bottleneck of more ancient origin. The selection pressure for MHC variability in moose may also be reduced and we discuss the possibility that its solitary life style may reduce lateral transmission of pathogens in the population.

Animals↗

Incidence of traumatic tooth injuries in children and adolescents in the county of Västmanland, Sweden.

The study reports the total incidence as well as the incidence of different types of traumatic tooth injuries in a Swedish county in the age interval 0-19 years during 1989/90. The incidence of individuals experiencing tooth injuries was 13 per 1000 per year. Boys were more frequently injured in the age-groups 3-4 years and 7-9 years and girls, in the age-group 5-6 years. A method is presented defining uncomplicated and complicated tooth injuries as a basis for estimating the economic consequences of these injuries in the community. Boys more often suffered uncomplicated injuries to permanent teeth and girls, to primary teeth (p < 0.05). The same distribution was found for uncomplicated multiple tooth injuries. Boys sustained more uncomplicated hard tissue injuries and girls, more uncomplicated luxation injuries (p < 0.01). Using a classification according to the most serious tooth injury in each episode, 33% of the episodes had resulted in complicated injuries in which the pulpal tissue and/or periodontal membrane was severely damaged.

Adolescent↗

Variability of the blood/breath alcohol ratio in drinking drivers.

The ratio of blood-alcohol concentration (BAC) to breath-alcohol concentration (BrAC) was determined for 799 individuals apprehended for driving under the influence of alcohol (DUI) in Sweden. The BrAC was determined with an infrared analyzer (Intoxilyzer 5000S) and venous BAC was measured by headspace gas chromatography. The blood samples were always taken after the breath tests were made and the average time delay was 30 +/- 12 min (+/- SD), spanning from 6 to 60 min. The blood/breath ratios of alcohol decreased as the time between sampling blood and breath increased (F = 15.4, p < 0.001), being 2337 +/- 183 (6 to 15 min), 2302 +/- 202 (16 to 30 min), 2226 +/- 229 (31 to 45 min), and 2170 +/- 225 (46 to 60 min). When the BAC was corrected for the metabolism of alcohol at a rate of 0.019 g%/h, the mean blood/breath ratios were 2395 +/- 193 (6 to 15 min), 2416 +/- 211 (16 to 30 min), 2406 +/- 223 (31 to 45 min), and 2407 +/- 210 (45 to 60 min); no significant differences (F = 0.197, p > 0.05). The overall mean time-adjusted blood/breath ratio (+/- SD) was 2407 +/- 213 and the 95% limits of agreement (LOA) were 1981 and 2833. During 1992, 1993, and 1994, the mean blood/breath ratios of alcohol were remarkably constant, being 2409 +/- 288, 2407 +/- 206, and 2421 +/- 235, respectively, and the values were not significantly influenced by the person's age, gender, or blood-alcohol content. In 34 individuals (4.3%), the blood/breath ratio was less than 2100 after compensating for metabolism of alcohol between the times of sampling blood and breath. This compares with 156 individuals (19.6%) having a blood/breath ratio less than 2100:1 without making any correction for the metabolism of alcohol.

Adult↗