Biomedical subjects

C Jansson

Publications and source records attributed to C Jansson.

At least 19 recordsLinked to original sources

PubMed2001

D1' centers are less efficient than normal photosystem II centers.

One prominent difference between the photosystem II (PSII) reaction center protein D1' in Synechocystis 6803 and normal D1 is the replacement of Phe-186 in D1 with leucine in D1'. Mutants of Synechocystis 6803 producing only D1', or containing engineered D1 proteins with Phe-186 substitutions, were analyzed by 77 K fluorescence emission spectra, chlorophyll a fluorescence induction yield and decay kinetics, and flash-induced oxygen evolution. Compared to D1-containing PSII centers, D1' centers exhibited a 50% reduction in variable chlorophyll a fluorescence yield, while the flash-induced O(2) evolution pattern was unaffected. In the F186 mutants, both the P680(+)/Q(A)(-) recombination and O(2) oscillation pattern were noticeably perturbed.

Amino Acid Substitution↗

PubMed2001

Engineering of the protein environment around the redox-active TyrZ in photosystem II. The role of F186 and P162 in the D1 protein of Synechocystis 6803.

The photosystem II reaction centre protein D1 is encoded by the psbA gene. By activation of the silent and divergent psbA1 gene in the cyanobacterium Synechocystis 6803, a novel D1 protein, D1', was produced [Salih, G. & Jansson, C. (1997) Plant Cell 9, 869-878]. The D1' protein was found to be fully operational although it deviates from the normal D1 protein in 54 out of 360 amino acids. Two notable amino-acid substitutions in D1' are the replacements of F186 by a leucine and P162 by a serine. The F186 and P162 positions are located in the vicinity of the reaction centre chlorophyll dimer P680 and the redox-active Y161 (TyrZ), and F186 has been implicated in the electron transfer between Y161 and P680. The importance of F186 was addressed by construction of engineered D1 proteins in Synechocystis 6803. F186 was replaced by leucine, serine, alanine, tyrosine or tryptophan. Only the leucine replacement yielded a functional D1 protein. Other substitutions did not support photoautotrophic growth and the corresponding mutants showed no or very poor oxygen evolving activity. In the F186Y and F186W mutants, the D1 protein failed to accumulate to appreciable levels in the thylakoid membrane. The F186S mutation severely increased the light sensitivity of the D1 protein, as indicated by the presence of a 16-kDa proteolytic degradation product. We conclude that the hydrophobicity and van der Waals volume are the most important features of the residue at position 186. Exchanging P162 for a serine yielded no observable phenotype.

Amino Acid Substitution↗

PubMed2001

Transformation and regeneration capacities for five Nordic barley elite cultivars--evaluation of tissue culture response and transient expression.

Using both biolistic and Agrobacterium-mediated DNA delivery, we have investigated the transformation and regeneration capacity for five Nordic elite cultivars of barley. Transformation was followed as transient expression of the uidA or gfp gene in barley callus. Callus formation and regeneration of transformed callus were evaluated based on callus induction frequency, growth rate, callus appearance, and shoot formation frequency. From the accumulated results, one of the elite cultivars has been selected for our ongoing work in molecular breeding of barley.

Agrobacterium tumefaciens↗

PubMed2000

Deletion mutagenesis of the 5' psbA2 region in Synechocystis 6803: identification of a putative cis element involved in photoregulation.

Three Synechocystis 6803 mutants, with deletions in the upstream region of the light-regulated psbA2 gene, were constructed. All three mutants grew photoautotrophically but they were affected in psbA2 expression. In one of the mutants, DeltaU80, psbA2 transcription levels were not suppressed under low light conditions and production of psbA2 transcripts occurred also in darkness, in contrast to the situation in the control strain. Comparative sequence analyses of the mutants identified a hexanucleotide repeat as possible cis element in psbA2 light regulation. Electrophoresis mobility shift assays showed that protein binding to the hexanucleotide repeat decreased with light intensity, consistent with a hypothesis that the repeat harbors a binding site for a repressor protein. Repeats similar to that in psbA2 were found also in the upstream region of other light-regulated genes in Synechocystsis 6803. We refer to these hexanucleotide repeats as High Light Regulatory 1 (HLR1) sequences.

Base Sequence↗

PubMed1999

Analyses of isoamylase gene activity in wild-type barley indicate its involvement in starch synthesis.

The notion of debranching enzyme activity as a participant in starch synthesis is gaining acceptance. Inconsistent reports from mutant analyses implicate either isoamylase or pullulanase as a determinant in amylopectin formation and whether wild-type plants utilize one or the other, or both, of these debranching enzymes in starch synthesis is unclear. Recent results on the sul mutant in maize suggest that both forms of debranching enzymes might be involved in amylopectin formation. We wished to find out if isoamylase takes part in starch synthesis by comparing isoamylase gene activity under three conditions: (1) during starch accumulation in developing sink tissues; (2) during starch degradation in germinating seeds; (3) in ectopic expression after applying sucrose, a starch precursor. We isolated the gene for barley isoamylase, isol, and analysed its expression and regulation in germinating seeds, developing endosperm and vegetative tissues, and compared the isoamylase gene expression in sink tissues from three different species. Our results indicate that isoamylase gene activity is involved in starch synthesis in wild-type plants and is modulated by sucrose.

Amino Acid Sequence↗

PubMed1999

Transformation of nuclear and plastomic plant genomes by biolistic particle bombardment.

Microprojectile bombardment is a powerful method for the transformation of various organisms and tissues. For plants, the biolistic approach is primarily used for transformation of cereals and other monocotyledons, as well as for dicotyledonous plants shown to be recalcitrant to Agrobacterium-based transformation of organellar genomes, and transformation of plant and algal chloroplasts has recently been reported. In this protocol paper we provide methods for nuclear and plastomic transformation of plants using the biolistic technique.

Biolistics↗

PubMed1998

Engineering of N-terminal threonines in the D1 protein impairs photosystem II energy transfer in Synechocystis 6803.

Mutants of the cyanobacterium Synechocystis sp. PCC 6803 with N-terminal changes in the photosystem (PSII) II D1 protein were analysed by flash-induced oxygen evolution, chlorophyll a fluorescence decay kinetics and 77 K fluorescence emission spectra. The data presented here show that mutations of the Thr-2, Thr-3 and Thr-4 in D1 do not influence the oxygen evolution. A perturbation on the acceptor side was observed and the importance of the N-terminal threonines for an efficient energy transfer between the phycobilisome and PSII and for stability of the PSII complex was demonstrated.

Amino Acid Substitution↗

PubMed1998

Prevalence of bronchial hyper-responsiveness in the southern, central and northern parts of Sweden.

Studies have suggested that there is a higher prevalence of asthma in northern Sweden than in southern Sweden. Bronchial hyper-responsiveness (BHR) has been shown to be associated with asthma. The aim of this study was to explore the prevalence of bronchical hyper-responsiveness in different parts of Sweden. As part of the European Community Respiratory Health Survey (ECRHS), interviews, skin prick tests, lung function tests and methacholine provocation tests of the airways were performed in 1448 randomly selected subjects in southern, central and northern Sweden. The Mefar dosimeter was used according to the ECRHS protocol. The responsiveness was calculated both as the PD20 and as the dose response slope (DRS). BHR was defined as a PD20 of < or = 1.6 mg. Atopy was defined as at least one skin prick test of > or = 3 mm. The prevalence of BHR was 12.7%, 10.6% in men and 15.0% in women. No difference in prevalence was found between the three different regions of Sweden. The prevalence of BHR was higher in women than in men and higher in smokers than in non-smokers. Using multiple logistic regression, with BHR as the dependent variable, atopy, being female, having a low FEV1 (% predicted) and smoking (both own and passive) increased the odds of having BHR, while age and the region of Sweden did not influence BHR. Defining BHR as a PD20 of < or = 1.0 mg or a PD20 of < or = 2.0 mg did not change this. Multiple regression using log DRS as the dependent variable produced the same result. Both BHR and increasing DRS were associated with self-reported wheezing, attacks of shortness of breath during the daytime at rest or after strenuous activity, being awakened by a feeling of tightness in the chest or an attack of shortness of breath. In subjects without self-reported asthma, BHR was associated with self-reported wheezing and attacks of shortness of breath after strenuous activity. In conclusion, we found that the prevalence of BHR in the three investigated areas was 12.7%. We found a trend towards a higher prevalence of BHR in the most northerly of the study areas, but the difference between the areas was not statistically significant. BHR and DRS were associated with atopy, smoking, female sex and FEV1 (% predicted). The reporting of symptoms from the airways was associated with the degree of bronchical responsiveness.

Adult↗

PubMed1998

Site-specific mutations of the N-terminal threonines in the D1 protein affects photoautotrophic growth but not D1 protein stability in Synechocystis 6803.

The cyanobacterium Synechocystis 6803 was engineered to produce a D1 protein where one or more of the N-terminal threonines at positions 2, 3 and 4 were replaced by other amino acid residues. No phenotypic effects were found for the T2S or T2L mutations, whereas the T2V, T2L;T4V and T2V;T3V;T4V mutations resulted in reduced photoautotrophic growth rate and oxygen evolving activity. The mutant strain T2V;T3V;T4V exhibited an oxygen evolution activity that was only half of that for the wild-type strain. Despite of that, both accumulation and stability of the D1 protein in the thylakoid membrane appeared unaffected in the mutant.

Cyanobacteria↗

PubMed1998

The two genes encoding starch-branching enzymes IIa and IIb are differentially expressed in barley.

The sbeIIa and sbeIIb genes, encoding starch-branching enzyme (SBE) IIa and SBEIIb in barley (Hordeum vulgare L.), have been isolated. The 5' portions of the two genes are strongly divergent, primarily due to the 2064-nucleotide-long intron 2 in sbeIIb. The sequence of this intron shows that it contains a retro-transposon-like element. Expression of sbeIIb but not sbeIIa was found to be endosperm specific. The temporal expression patterns for sbeIIa and sbeIIb were similar and peaked around 12 d after pollination. DNA gel-blot analysis demonstrated that sbeIIa and sbeIIb are both single-copy genes in the barley genome. By fluorescence in situ hybridization, the sbeIIa and sbeIIb genes were mapped to chromosomes 2 and 5, respectively. The cDNA clones for SBEIIa and SBEIIb were isolated and sequenced. The amino acid sequences of SBEIIa and SBEIIb were almost 80% identical. The major structural difference between the two enzymes was the presence of a 94-amino acid N-terminal extension in the SBEIIb precursor. The (beta/alpha)8-barrel topology of the alpha-amylase superfamily and the catalytic residues implicated in branching enzymes are conserved in both barley enzymes.

1,4-alpha-Glucan Branching Enzyme↗

PubMed1997

Increased LH and FSH secretion after cranial irradiation in boys.

The effect of high-dose cranial- and craniospinal irradiation and chemotherapy on the gonadotropin-sex steroid axis was studied during different stages of puberty by measuring pulsatile secretion of luteinizing hormone (LH), follicle-stimulating hormone (FSH) and testosterone. The patients were thirteen boys who had been treated for malignant brain tumor residing well away from the hypothalamo-pituitary region. The median time to follow-up was 9 (1-16) years. The onset of puberty was early in the patients, median 10.5 years, compared to the average age for Swedish boys, which is at median 12.4 years. There was, before puberty, no significant difference in LH and FSH secretion between patients and a control group of normal boys. In early, mid- and late stages of puberty, however, LH and FSH secretion was increased in the patients overall, whereas testosterone secretion was maintained within the normal range in spite of signs of gonadotoxocity with small testicular volumes. These results indicate that the vulnerable parts of the gonadotropin releasing hormone (GnRH)-gonadotropin (LH, FSH)-gonadal axis are the regulatory system that determines the timing of pubertal induction and the gonads. The GnRH-LH, FSH-releasing neurons appear relatively resistant to cranial irradiation as they are able to respond with supranormal LH and FSH levels for long periods of time after treatment.

Adolescent↗

PubMed1997

Activation of the silent psbA1 gene in the cyanobacterium Synechocystis sp strain 6803 produces a novel and functional D1 protein.

The photosystem II reaction center protein D1 in Synechocystis sp strain 6803 is encoded by the psbA2 and psbA3 genes of the three-membered psbA gene family. The silent and divergent psbA1 copy of the psbA gene family was activated by exchanging part of its upstream region with a corresponding fragment of the psbA2 copy. The light-regulated expression of the activated psbA1 gene showed that the inserted psbA2 segment contains the information necessary for light-dependent as well as high-light-stimulated transcription. The activated psbA1 gene expressed a novel D1 protein, D1'. A mutant strain containing psbA1 as the only active psbA gene grew photoautotrophically at a rate comparable to that of the wild type. This finding demonstrates that despite its unusual amino acid sequence, D1 is exchangeable for D1 in the photosystem II complex, at least under normal laboratory conditions. The D1' protein was found to have a degradation rate similar to that of the D1 protein under low- or high-light conditions. Another mutant containing the activated psbA1 gene together with the psbA2 and psbA3 genes produced both the D1 and D1' proteins.

Amino Acid Sequence↗

PubMed1997

Hormonal status of short children born small for gestational age.

The present study was undertaken to evaluate the hormonal status in a subgroup of prepubertal children born small for gestational age (SGA) who lacked postnatal catch-up growth. In this subgroup, a reduced rate of growth hormone (GH) secretion was found, compared with reference groups of healthy children born appropriate for gestational age, of either normal or short stature at the time of investigation. In addition, an abnormal pattern of GH secretion was observed in short children born SGA, which was most pronounced in the younger children, and involved an increased frequency of GH peaks of low amplitude, combined with increased baseline secretion. Levels of insulin-like growth factor I (IGF-I) and IGF-binding protein-3 were also reduced in short children born SGA, compared with the reference groups. These findings may explain, in part, the lack of postnatal catch-up growth in short children born SGA.

Adolescent↗

PubMed1997

Influence of gender on the correlation between plasma growth hormone profiles and urinary growth hormone excretion.

A lot of interest has been directed towards the measurement of urinary growth hormone (GH) excretion instead of plasma GH profiles or provocation tests. We investigated the factors influencing the relationship between 24- and 3-hour plasma GH profiles and urinary GH excretion in a cohort of 113 pediatric patients with growth disorders and healthy volunteers. Plasma and urinary GH were measured by polyclonal immunoassays differing in cross-reactivity with 20 kD GH (100 versus 46%), but not with 22-kD, dimer, deamidated and pituitary GH. In the 24-hour urine samples, only urinary GH excretion expressed as nanograms per 24 h correlated with plasma GH parameters, whereas the correlations for short-term samples were strongest if urinary GH excretion was expressed as nanograms per gram creatinine (r = 0.70-79, p < 0.00005-0.0001). In short-term samples urinary GH excretion depends on urinary volume and should thus be expressed in nanograms per gram creatinine, whereas 24-hour samples correlate best when urinary GH is expressed as nanograms per period. We found a significant sex difference (p < 0.02) in the correlation between 24-hour plasma GH profiles and urinary GH excretion with strong correlations in the female group (r = 0.63-0.78, p < 0.00005-0.0002) and a lack of correlation in the male group. The sex difference in the correlations between serum and urinary GH may reflect sex differences in GH profiles and metabolism, with urinary GH better reflecting the basal and slowly clearing portion of plasma GH than spontaneous GH peaks. The difference in cross-reactivities of molecular GH forms in polyclonal assays may have an impact on the correlation between plasma and urinary GH. Thus, the diagnostic value of urinary GH measurement as compared to serum GH profiles needs to be further evaluated.

Adolescent↗

PubMed1997

Increased proportion of circulating non-22-kilodalton growth hormone isoforms in short children: a possible mechanism for growth failure.

Current knowledge about the interaction between GH and its receptor suggests that the molecular heterogeneity of circulating GH may have important implications for growth. The aim of this study was to investigate the proportion of circulating non-22-kDa GH isoforms in prepubertal children with short stature (height less than -2 SD score) of different etiologies. We have also evaluated the relationships among the ratio of non-22-kDa GH isoforms, auxology, and spontaneous GH secretion. The study groups consisted of 17 girls with Turner's syndrome (TS), aged 3-13 yr, 25 children born small for gestational age (SGA) without postnatal catch-up growth, aged 3-13 yr; and 24 children with idiopathic short stature (ISS), aged 4-15 yr. The results were compared with those from 23 prepubertal healthy children of normal stature (height +/- 2 SD score), aged 4-13 yr. Serum non-22-kDa GH levels, expressed as a percentage of the total GH concentration, were determined by the 22-kDa GH exclusion assay, which is based on immunomagnetic extraction of monomeric and dimeric 22-kDa GH from serum and quantitation of non-22-kDa GH using a polyclonal antibody-based GH assay. All samples were selected from spontaneous GH peaks in 24-h GH profiles. The median proportion of non-22-kDa GH isoforms was increased in children born SGA (9.8%; P = 0.05) and girls with TS (9.9%; P = 0.01), but not in the group of children with ISS (8.9%), compared with that in normal children (8.1%). Individually, increased proportions of non-22-kDa GH isoforms, with values more than 2 SD above the mean for the normal group, were observed in 5 girls with TS, 5 children born SGA, and 4 children with ISS. In children born SGA, the proportion of non-22-kDa GH isoforms was directly correlated with different estimates of spontaneous GH secretion [mean 24-h GH concentration (r = 0.41; P = 0.04), area under the curve over baseline (r = 0.41; P = 0.04), and GH peak area (r = 0.61; P = 0.003)], whereas it was inversely correlated with height SD score (r = -0.42; P = 0.04). In conclusion, an increased proportion of circulating non-22-kDa GH isoforms was observed at spontaneous GH peaks in some non-GH-deficient short children. Our results suggest that the ratio of non-22-kDa GH isoforms in the circulation may have important implications for normal and abnormal growth.

Adolescent↗

PubMed1997

Growth response to growth hormone (GH) treatment relates to serum insulin-like growth factor I (IGF-I) and IGF-binding protein-3 in short children with various GH secretion capacities. Swedish Study Group for Growth Hormone Treatment.

The purpose of the study was to evaluate the relationship between the 1-yr (n = 193) and 2-yr (n = 128) growth response and the individual serum concentrations of insulin-like growth factor I (IGF-I) and IGF-binding protein 3 (IGFBP-3) before and during GH treatment. Our study group of prepubertal short children had from very low to high GH secretory capacity, estimated during an arginine-insulin tolerance test, and the ages ranged from 3-15 yr at the start of treatment. Their serum levels of IGF-I and IGFBP-3 were low before treatment compared to those in an age-related reference group of prepubertal children and increased significantly from the start to 1 month of GH treatment. The mean increase in height SD score was 0.80 SD score after 1 yr of GH treatment and 1.26 SD score after 2 yr, with a wide range. In univariate analyses the highest correlation coefficients to the 2-yr growth response were found to be vs. the following variables from the start of treatment: IGF-I SD score (r = -0.49), log maximum GH concentration (log GHmax) during the arginine-insulin tolerance test (r = -0.47), difference between the height SD score of the individual child and the midparental height SD score (diffSD score; r = -0.45), IGFBP-3 SD score (r = -0.39), age (r = -0.30), short term change in IGFBP-3 SD score (r = 0.37), and IGF-I SD score (r = 0.34). In multivariate stepwise regression analysis, 41% of the variation in the 2-yr growth response could be explained by IGF-I SD score or log GHmax together with age at the start of treatment, weight SD score at 1 yr of age, and diffSD score. When both IGF-I SD score and GHmax were included and when the short term changes in IGF-I SD score were added, 46% and 58% of the variation, respectively, could be explained. The regression algorithms using different combinations of variables and their corresponding prediction intervals are also presented.

Adolescent↗

PubMed1997

Circulating non-22 kDa growth hormone isoforms in healthy children of normal stature: relation to height, body mass and pubertal development.

The proportion of non-22 kDa GH isoforms was evaluated in 93 healthy children (48 boys aged 6.8-18.4 years and 45 girls aged 3.9-18.4 years) of normal stature (height +/- 2 s.d. score) at different stages of puberty. In addition, correlations among the proportion of non-22 kDa GH isoforms, auxology, spontaneous GH secretion and biochemical measurements were investigated. Serum non-22 kDa GH levels, expressed as percentage of total GH concentration in the samples, were determined by the 22 kDa GH exclusion assay, in which monomeric and dimeric 22 kDa GH are removed from serum and the non-22 kDa GH isoforms are quantitated using a polyclonal antibody GH assay. Samples were selected from spontaneous GH peaks in 24-h GH profiles. For boys, the median proportion of non-22 kDa GH isoforms was 8.5% (range 3.2-26.6%) and for girls it was 9.6% (1.8-17.4%), with no influence of age and no sex-related difference in prepubertal (boys, 7.2%; girls, 8.8%) or pubertal children (boys, 9.1%; girls, 9.9%). However, the median proportion of non-22 kDa GH isoforms was significantly higher in pubertal boys (9.1%) than in prepubertal boys (7.2%; P = 0.03). In pubertal boys, height S.D. scores (SDS) were inversely correlated to the proportion of non-22 kDa GH isoforms (r = -0.38; P = 0.02), especially at mid-puberty (r = -0.7; P = 0.01), indicating that the presence of increased amounts of circulating non-22 kDa GH isoforms was associated with less growth. In prepubertal children, positive correlations between non-22 kDa GH and weight SDS (r = 0.46; P = 0.03), weight-for-height SDS (r = 0.51; P = 0.01) and body mass index (r = 0.42; P = 0.04) were observed. No significant correlations were seen with spontaneous GH secretion or measurements of IGF-1, IGF-binding protein-3, insulin and leptin. These findings in normal children indicate that the proportion of circulating non-22 kDa GH isoforms may have physiologic significance for growth and metabolism in different stages of development, and emphasize the importance of evaluating the circulating ratio of 22 kDa and non-22 kDa GH in children with growth disorders.

Adolescent↗

PubMed1997

Growth hormone (GH) assays: influence of standard preparations, GH isoforms, assay characteristics, and GH-binding protein.

The impact of the adoption of the new biosynthetic growth hormone (GH) WHO International Reference Preparation (IRP 88/624), and the recommendation to report results in microgram/L instead of mU/L, is described. Conversion factors were determined by comparing both the linear and nonlinear relations of the GH values. The Pharmacia polyclonal IRMA (p-IRMA) and the DELFIA monoclonal time-resolved immunofluorometric assay (trIFMA) with kit calibrators calibrated either against the pituitary-derived WHO IRP 80/505 or the new 88/624 were evaluated. Conversion factors of 4.17 mU/L = 1 microgram/L for the p-IRMA and 4.31 mU/L = 1 microgram/L for the trIFMA were necessary. Different cross-reactivity patterns for the deaminated and dimer 22-kDa, 20-kDa, and 17-kDa GH isoforms were found. Expected GH recovery was similar when the measured values were adjusted according to the results of the cross-reactivity study.

Adolescent↗