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Biomedical subjects

H Nilsen

Publications and source records attributed to H Nilsen.

At least 19 recordsLinked to original sources
PubMed2001

Excision of deaminated cytosine from the vertebrate genome: role of the SMUG1 uracil-DNA glycosylase.

Gene-targeted mice deficient in the evolutionarily conserved uracil-DNA glycosylase encoded by the UNG gene surprisingly lack the mutator phenotype characteristic of bacterial and yeast ung(-) mutants. A complementary uracil-DNA glycosylase activity detected in ung(-/-) murine cells and tissues may be responsible for the repair of deaminated cytosine residues in vivo. Here, specific neutralizing antibodies were used to identify the SMUG1 enzyme as the major uracil-DNA glycosylase in UNG-deficient mice. SMUG1 is present at similar levels in cell nuclei of non-proliferating and proliferating tissues, indicating a replication- independent role in DNA repair. The SMUG1 enzyme is found in vertebrates and insects, whereas it is absent in nematodes, plants and fungi. We propose a model in which SMUG1 has evolved in higher eukaryotes as an anti-mutator distinct from the UNG enzyme, the latter being largely localized to replication foci in mammalian cells to counteract de novo dUMP incorporation into DNA.

Animals↗
PubMed2001

Sequence variation in the human uracil-DNA glycosylase (UNG) gene.

Spontaneous deamination of cytosine results in a premutagenic G:U mismatch that may result in a GC-->AT transition during replication. The human UNG-gene encodes the major uracil-DNA glycosylase (UDG or UNG) which releases uracil from DNA, thus, initiating base excision repair to restore the correct DNA sequence. Bacterial and yeast mutants lacking the homologous UDG exhibit elevated spontaneous mutation frequencies. Hence, mutations in the human UNG gene could presumably result in a mutator phenotype. We screened all seven exons including exon-intron boundaries, both promoters, and one intron of the UNG gene and identified considerable sequence variation in cell lines derived from normal fibroblasts and tumour tissue. None of the sequence variants was accompanied by significantly reduced UDG activity. In the UNG gene from 62 sources, we identified 12 different variant alleles, with allele frequencies ranging from 0.01 to 0.23. We identified one variant allele per 3.8kb in non-coding regions, but none in the coding region of the gene. In promoter B we identified four different variants. A substitution within an AP2 element was observed in tumour cell lines only and had an allele frequency of 0.10. Introduction of this substitution into chimaeric promoter-luciferase constructs affected transcription from the promoter. UDG-activity varied little in fibroblasts, but widely between tumour cell lines. This variation did not however correlate with the presence of any of the variant alleles. In conclusion, mutations affecting the function of human UNG gene are seemingly infrequent in human tumour cell lines.

Cells, Cultured↗
PubMed2001

Properties and functions of human uracil-DNA glycosylase from the UNG gene.

The human UNG-gene at position 12q24.1 encodes nuclear (UNG2) and mitochondrial (UNG1) forms of uracil-DNA glycosylase using differentially regulated promoters, PA and PB, and alternative splicing to produce two proteins with unique N-terminal sorting sequences. PCNA and RPA co-localize with UNG2 in replication foci and interact with N-terminal sequences in UNG2. Mitochondrial UNG1 is processed to shorter forms by mitochondrial processing peptidase (MPP) and an unidentified mitochondrial protease. The common core catalytic domain in UNG1 and UNG2 contains a conserved DNA binding groove and a tight-fitting uracil-binding pocket that binds uracil only when the uracil-containing nucleotide is flipped out. Certain single amino acid substitutions in the active site of the enzyme generate DNA glycosylases that remove either thymine or cytosine. These enzymes induce cytotoxic and mutagenic abasic (AP) sites in the E. coli chromosome and were used to examine biological consequences of AP sites. It has been assumed that a major role of the UNG gene product(s) is to repair mutagenic U:G mispairs caused by cytosine deamination. However, one major role of UNG2 is to remove misincorporated dUMP residues. Thus, knockout mice deficient in Ung activity (Ung-/- mice) have only small increases in GC-->AT transition mutations, but Ung-/- cells are deficient in removal of misincorporated dUMP and accumulate approximately 2000 uracil residues per cell. We propose that BER is important both in the prevention of cancer and for preserving the integrity of germ cell DNA during evolution.

Animals↗
PubMed2000

Base excision repair of DNA in mammalian cells.

Base excision repair (BER) of DNA corrects a number of spontaneous and environmentally induced genotoxic or miscoding base lesions in a process initiated by DNA glycosylases. An AP endonuclease cleaves at the 5' side of the abasic site and the repair process is subsequently completed via either short patch repair or long patch repair, which largely require different proteins. As one example, the UNG gene encodes both nuclear (UNG2) and mitochondrial (UNG1) uracil DNA glycosylase and prevents accumulation of uracil in the genome. BER is likely to have a major role in preserving the integrity of DNA during evolution and may prevent cancer.

Amino Acid Sequence↗
PubMed2000

Analysis of uracil-DNA glycosylases from the murine Ung gene reveals differential expression in tissues and in embryonic development and a subcellular sorting pattern that differs from the human homologues.

The murine UNG: gene encodes both mitochondrial (Ung1) and nuclear (Ung2) forms of uracil-DNA glyco-sylase. The gene contains seven exons organised like the human counterpart. While the putative Ung1 promoter (P(B)) and the human P(B) contain essentially the same, although differently organised, transcription factor binding elements, the Ung2 promoter (P(A)) shows limited homology to the human counterpart. Transient transfection of chimaeric promoter-luciferase constructs demonstrated that both promoters are functional and that P(B) drives transcription more efficiently than P(A). mRNAs for Ung1 and Ung2 are found in all adult tissues analysed, but they are differentially expressed. Furthermore, transcription of both mRNA forms, particularly Ung2, is induced in mid-gestation embryos. Except for a strong conservation of the 26 N-terminal residues in Ung2, the subcellular targeting sequences in the encoded proteins have limited homology. Ung2 is transported exclusively to the nucleus in NIH 3T3 cells as expected. In contrast, Ung1 was sorted both to nuclei and mitochondria. These results demonstrate that although the catalytic domain of uracil-DNA glycosylase is highly conserved in mouse and man, regulatory elements in the gene and subcellular sorting sequences in the proteins differ both structurally and functionally, resulting in altered contribution of the isoforms to total uracil-DNA glycosylase activity.

3T3 Cells↗
PubMed2000

Uracil-DNA glycosylase (UNG)-deficient mice reveal a primary role of the enzyme during DNA replication.

Gene-targeted knockout mice have been generated lacking the major uracil-DNA glycosylase, UNG. In contrast to ung- mutants of bacteria and yeast, such mice do not exhibit a greatly increased spontaneous mutation frequency. However, there is only slow removal of uracil from misincorporated dUMP in isolated ung-/- nuclei and an elevated steady-state level of uracil in DNA in dividing ung-/- cells. A backup uracil-excising activity in tissue extracts from ung null mice, with properties indistinguishable from the mammalian SMUG1 DNA glycosylase, may account for the repair of premutagenic U:G mispairs resulting from cytosine deamination in vivo. The nuclear UNG protein has apparently evolved a specialized role in mammalian cells counteracting U:A base pairs formed by use of dUTP during DNA synthesis.

Animals↗
PubMed2000

Essential fatty acids in the nutrition of severely neurologically disabled children.

Essential fatty acids (EFA) are important for the normal development and functioning of the brain, retina and immune competent cells. Severely neurologically handicapped children often have feeding difficulties, and the composition of the diet may be critical with respect to an optimal nutrient content. The aim of the present investigation was to evaluate if the dietary intakes and serum phospholipid concentrations of EFA were adequate in a group of severely neurologically impaired children in an institution. To achieve this, a prospective study was done. The investigation showed low dietary intakes of both n-6 fatty acids (FA) and n-3 FA. The serum concentrations of total n-6 FA, linoleic acid and 22:6n-3 (docosahexaenoic acid) as proportions of the total serum phospholipid FA concentration were initially low. The serum concentrations of 20:3n-9 and 22:5n-6 cholesterol, triacylglycerol, total saturated FA, total monounsaturated FA and apolipoproteins A-I and B were high compared with levels in a reference group of healthy children. Following supplementation with fish oil and soyabean oil, the serum lipid profile approached normal. We conclude that the study children had suboptimal intakes of EFA and that elevated serum concentrations of 20:3n-9 and 22:5n6 were useful serological markers of suboptimal EFA status. Recommended dietary allowances for EFA given as a percentage of energy underestimate EFA requirements in children with a low energy intake. Severely disabled children with feeding difficulties should probably be monitored with serum phospholipid FA measurements or calculation of dietary absolute intakes of EFA.

Adolescent↗
PubMed2000

Effect of omega-3 fatty acids and simvastatin on hemostatic risk factors and postprandial hyperlipemia in patients with combined hyperlipemia.

Patients with combined hyperlipemia have lipid abnormalities associated with an increased tendency to develop atherosclerosis and thrombosis. This tendency may be accelerated during postprandial hyperlipemia. In the present double-blind parallel study, 41 patients with combined hyperlipemia and serum triacylglycerols between 2.0 and 15.0 mmol/L and serum total cholesterol >5.3 mmol/L at the end of a 3-month dietary run-in period were treated with simvastatin at 20 mg/d for at least 10 weeks; patients were then randomized into 2 groups receiving simvastatin+omega-3 fatty acids at 3.36 g/d or placebo (corn oil) for an additional 5 weeks. Hemostatic variables that have been associated with increased thrombotic tendency were evaluated with subjects in the fasting state and during postprandial hyperlipemia before and after combined treatment. Supplementation of omega-3 fatty acid reduced tissue factor pathway inhibitor antigen (P<0.05) in the fasting state, reduced the degree of postprandial hyperlipemia (P<0.005), and reduced activated factor VII concentration appearing during postprandial hyperlipemia. In conclusion, omega-3 fatty acids given in addition to simvastatin to patients with combined hyperlipemia reduced the free tissue factor pathway inhibitor fraction in the fasting state and inhibited the activation of factor VII occurring during postprandial lipemia, thus representing a potential beneficial effect on the hemostatic risk profile in this patient group.

Adult↗
PubMed2000

Relationship between apolipoprotein E polymorphism, postprandial hyperlipemia and hemostatic variables in patients with combined hyperlipemia.

BACKGROUND AND AIM: Combined hyperlipemia is a common lipid abnormality associated with coronary heart disease (CHD). Aim of this study was to investigate the relationship between this abnormality and hemostatic risk factors related to thrombosis. METHODS AND RESULTS: Forty-one patients were examined in the fasting state and during postprandial hyperlipemia. They had high levels of factor VIIc in the fasting state. During postprandial hyperlipemia a highly significant (p < 0.001) increase of activated factor (VIIa) occurred. This activation was not correlated to the increase in free fatty acids, but to the degree of postprandial hypertriglyceridemia. The absolute postprandial triglyceridemia was highest in subjects with the apolipoprotein (apo) E2 allele. Tissue factor pathway inhibitor activity (TFPIa) was correlated to LDL cholesterol and apo B concentrations and was highest in subjects with genotypes containing the E4 allele. CONCLUSIONS: Subjects with combined hyperlipemia may have an increased thrombotic risk related to activation of factor VII during postprandial hyperlipemia. Subjects with genotypes including the apoE4 allele may have a high lipid risk profile associated with high levels of LDL cholesterol.

Adult↗
PubMed1999

Dietary and nutrient intake of a Sami population living in traditional reindeer herding areas in north Norway: comparisons with a group of Norwegians.

The Samis are an ethnic minority living in the Northern region of Norway, Sweden, Finland and Russia. Traditionally the Samis made their living from reindeer herding with some fishing and agriculture. Earlier studies have shown that their diet consisted of large amounts of reindeer meat, some fish and wild berries with low intakes of other fruits, vegetables and dairy products. Due to the introduction of technical improvements like snowmobiles and terrain vehicles which makes moving with the herd less necessary, their lifestyle has changed. There is little documentation how this has affected their dietary habits. In this study, the dietary pattern and nutrient intake of a group of Samis (n = 75) living in traditional reindeer herding areas of North Norway were investigated and compared with that of a group of Norwegians (n = 65). Dietary information was obtained through an interview by a nutritionist using the dietary history method. The findings indicate that nutrient content of the Sami diet is adequate except for folic acid. Calcium and iron intake was slightly below recommended levels for Sami females. There seems to be some difference between the diet of the Samis and Norwegians. The Samis consume more meat, fat, table sugar and coffee and less fruits and vegetables. The dietary pattern of the Samis seems, however, to be changing toward a more typically Norwegian diet.

Adult↗
PubMed1998

Antibodies to orthopoxvirus in domestic cats in Norway.

The prevalence of antibodies to orthopoxvirus in 217 sera collected from domestic cats in the western part of Norway was 10.1 per cent as measured by a competitive ELISA. In one of the seropositive cats antibodies were also detected by an immunofluorescence assay. The average age of the cats sampled was 4.9 years, but the average age of the seropositive individuals was 7.3 years, higher than the average age of clinical cowpox virus cases in Britain (4.2 years), and in Germany (3.9 years). Antibodies against feline immunodeficiency virus (FIV) were detected in nine of 30 (30 per cent) of the seropositive cats, and in five of 30 (17 per cent) of the seronegative cats, which suggests that FIV infection may influence the susceptibility of domestic cats to orthopoxvirus, or vice versa. Orthopoxvirus infections, have recently been detected in rodent populations in several areas of Norway, and the infection may therefore be present in cats all over the country; cat owners and animal handlers should be aware of this (re)emerging zoonosis.

Animals↗
PubMed1998

Effects of highly purified eicosapentaenoic acid and docosahexaenoic acid on fatty acid absorption, incorporation into serum phospholipids and postprandial triglyceridemia.

Fourteen healthy volunteers were randomly allocated to receive 4 g highly purified ethyl esters of eicosapentaenoic acid (EPA) (95% pure, n = 7) or docosahexaenoic acid (DHA) (90% pure, n = 7) daily for 5 wk in supplement to their ordinary diet. The n-3 fatty acids were given with a standard high-fat meal at the beginning and the end of the supplementation period. EPA and DHA induced a similar incorporation into chylomicrons which peaked 6 h after the meal. The relative uptake of EPA and DHA from the meal was > 90% compared with the uptake of oleic acid. During absorption, there was no significant elongation or retroconversion of EPA or DHA in total chylomicron fatty acids. The concentration of EPA decreased by 13% and DHA by 62% (P < 0.001) between 6 and 8 h after the meal. During the 5-wk supplementation period, EPA showed a more rapid and comprehensive increase in serum phospholipids than did DHA. DHA was retroconverted to EPA, whereas EPA was elongated to docosapentaenoic acid (DPA). The postprandial triglyceridemia was suppressed by 19 and 49% after prolonged intake of EPA and DHA, respectively, indicating that prolonged intake of DHA is equivalent to or even more efficient than that of EPA in lowering postprandial triglyceridemia. This study indicates that there are metabolic differences between EPA and DHA which may have implications for the use of n-3 fatty acids in preventive and clinical medicine.

Adult↗
PubMed1998

Effects of Simvastatin and omega-3 fatty acids on plasma lipoproteins and lipid peroxidation in patients with combined hyperlipidaemia.

OBJECTIVES: Patients with combined hyperlipidemia are at increased risk for development of coronary heart disease. The purpose of this study was to evaluate the efficiency and the safety of treatment with Simvastatin and omega-3 fatty acids in patients with this lipid disorder. DESIGN: A double-blind placebo controlled, randomized study evaluating the effects of Simvastatin separately and in combination with omega-3 fatty acids in 41 healthy patients with defined hyperlipidemia. After a 16 weeks dietary run-in period the patients were treated in periods of 5 weeks. RESULTS: As expected Simvastatin (20 mg day[-1]) reduced serum total cholesterol, triacylglycerols, apolipoproteins B and E and increased HDL-cholesterol and apolipoprotein A1. Addition of omega-3 fatty acids (4 g day[-1]) further decreased serum triacylglycerols (P = 0.007), total cholesterol (P = 0.052) and apolipoprotein E (P = 0.035). No significant changes attributable to supplementation of polyunsaturated fatty acids on the content of lipid peroxides in plasma and in the various lipoprotein fractions were observed. CONCLUSIONS: The combined treatment of Simvastatin and omega-3 fatty acids seems to be an efficient and safe alternative for patients with combined hyperlipidemia.

Adult↗
PubMed1997

Nuclear and mitochondrial uracil-DNA glycosylases are generated by alternative splicing and transcription from different positions in the UNG gene.

A distinct nuclear form of human uracil-DNA glycosylase [UNG2, open reading frame (ORF) 313 amino acid residues] from the UNG gene has been identified. UNG2 differs from the previously known form (UNG1, ORF 304 amino acid residues) in the 44 amino acids of the N-terminal sequence, which is not necessary for catalytic activity. The rest of the sequence and the catalytic domain, altogether 269 amino acids, are identical. The alternative N-terminal sequence in UNG2 arises by splicing of a previously unrecognized exon (exon 1A) into a consensus splice site after codon 35 in exon 1B (previously designated exon 1). The UNG1 sequence starts at codon 1 in exon 1B and thus has 35 amino acids not present in UNG2. Coupled transcription/translation in rabbit reticulocyte lysates demonstrated that both proteins are catalytically active. Similar forms of UNG1 and UNG2 are expressed in mouse which has an identical organization of the homologous gene. Constructs that express fusion products of UNG1 or UNG2 and green fluorescent protein (EGFP) were used to study the significance of the N-terminal sequences in UNG1 and UNG2 for subcellular targeting. After transient transfection of HeLa cells, the pUNG1-EGFP-N1 product colocalizes with mitochondria, whereas the pUNG2-EGFP-N1 product is targeted exclusively to nuclei.

Alternative Splicing↗
PubMed1996

Studies on nutrition in severely neurologically disabled children in an institution.

Severe neurological handicaps in children are frequently accompanied by growth retardation. We have studied 13 severely neurologically impaired children in an institution to see if their poor growth was related to a low intake of energy and nutrients, if this was reflected in biochemical nutritional parameters, and to modify their diet according to the results. The investigation showed low dietary intakes of energy and of several of the nutrients, with corresponding low Hb values and serum values of ferritin, selenium and vitamins E and D in some of the children. All the children were initially light for age, with catch-up growth after intervention. We conclude that severely disabled children are at high risk for under- and malnutrition, and that this may partly explain the growth retardation in the study group. To avoid the potential detrimental effects of malnutrition, it is important to aim at providing an optimal diet.

Adolescent↗
PubMed1995

Sequence specificity for removal of uracil from U.A pairs and U.G mismatches by uracil-DNA glycosylase from Escherichia coli, and correlation with mutational hotspots.

The rate of removal of uracil from different positions in double-stranded DNA by uracil-DNA glycosylase from Escherichia coli varied more than 15-fold. Consensus sequences for good and poor removal were 5'-(A/T)UA(A/T)-3' and 5'-(G/C)U(T/G/C)-3', respectively. In general, the sequence context surrounding U was more important for the rate of removal than whether U was present in U.A pairs or U.G mispairs. Rates of removal of U from sites of amber mutations in the lacI gene, where mutation frequencies and deamination rates were known, indicated that the observed variation in removal is biologically significant.

Base Composition↗