[Association and linkage studies for the analysis of candidate genes].
Explore the source record for details and available documents.
Alleles↗
Biomedical subjects
A Ziegler
Publications and source records attributed to A Ziegler.
Influence of salmeterol and benzalkonium chloride on G-protein-mediated exocytotic responses of rat peritoneal mast cells.
The long-acting beta(2)-adrenoceptor agonist salmeterol and the invert soap benzalkonium chloride share physicochemically important structures, namely a polar head group and a long aliphatic chain. Low concentrations of benzalkonium chloride have been shown to inhibit exocytotic responses in rat peritoneal mast cells by selectively interacting with heterotrimeric G-proteins of the G(i)-type. The present study investigates whether salmeterol inhibits, independently of beta-adrenoceptors, exocytotic responses of rat peritoneal mast cells induced by the direct agonists at G-proteins mastoparan or guanosine 5'-O-(3-thiotriphosphate) (++GTP gamma S++). Exocytosis was studied by secretion assays ([3H]5-hydroxytryptamine ([3H]5-HT)-release) using intact, streptolysin O-permeabilised or metabolically inhibited (antimycin, deoxyglucose) rat peritoneal mast cells. Both amphiphilics, salmeterol, and benzalkonium chloride, dose-dependently exerted biphasic effects on mastoparan-induced [3H]5-HT release in intact mast cells. In contrast to benzalkonium chloride, the dose-response curves for secretostatic and celltoxic effects of salmeterol markedly overlapped. Similar to benzalkonium chloride, salmeterol in non-cytotoxic concentrations (1-25 microg/ml) dose-dependently inhibited exocytosis induced by mastoparan (intact cells) or ++GTP gamma S (permeabilised cells). These findings indicate a direct, adrenoceptor-independent affection of G proteins by salmeterol in mast cells.
Adrenergic beta-Agonists↗
2D-spatial/2D-spectral spectroscopic imaging of intracerebral gliomas in rat brain.
1H-MR spectroscopy in vivo is often hampered by poor spectral resolution. Spectral overlap can be avoided with two-dimensional spectroscopic techniques. Correlation peak imaging has been implemented to measure unambiguously the distribution of several metabolites in a rat brain glioma model. Acquisition-weighted spectroscopic imaging reduced the experimental time and provided excellent spatial localization. The choice of an appropriate spectral acquisition window granted good sensitivity. Spectroscopic images presenting a full two-dimensional spectrum in every image pixel were acquired in seven rats at 7 Tesla in 195 min, with a nominal voxel volume of 75 microl. Among other metabolites, the distribution of hypotaurine, phosphoethanolamine, alanine, and even glucose could be visualized both in the C6-glioma and in the unaffected brain.
Animals↗
Determination of morphine 3-esters in rabbit plasma by high-performance liquid chromatography with ultraviolet detection.
A sensitive high-performance liquid chromatographic (HPLC) method for the quantitation of the morphine 3-esters 1[3-(2, 2-dimethylvaleroyl)-morphine (A), 3-(2-phenylbenzoyl)-morphine (B) and 3-(2,2-diphenylpropionyl)-morphine (C)] in rabbit plasma is described. Sample preparation was based on reversed-phase solid-phase extraction. The compounds were separated on C(18) reversed-phase analytical columns and then determined by ultraviolet detection. The recovery from plasma was 78.7 +/- 7.4%, 69.1 +/- 6.9% and 75 +/- 7.2% (mean +/- SD) for A, B, and C, respectively. The present method enabled the detection limit of 0.2, 0.2 and 0.1 ng and quantification limit of 20, 10 and 10 ng/ml for A, B and C, respectively. The developed method was used for determination of the plasmakinetics of these morphine 3-esters in rabbits.
Animals↗
Short treatments of normotensive and hypertensive rats by angiotensin II and nitric oxide inhibitor induce an increase of noradrenaline sensitivity in isolated vena portae preparations.
The aim of the study was to examine the influence of a short-term treatment of conscious Wistar Kyoto rats (WKY) and stroke-prone spontaneously hypertensive rats (SHRSP) by angiotensin II (ANG II) and by ANG II in combination with either l -NAME, HOE 140 or minoxidil on the mean arterial blood pressure (MABP) and the noradrenaline sensitivity in isolated portal vein preparations. MABP was significantly increased by ANG II treatment and ANG II plus l -NAME. However, it was slightly affected by ANG II in association with HOE 140, and significantly lowered by ANG II plus minoxidil. In control animals noradrenaline increased the frequency and the tone of contractile force. While ANG II enhanced the contractile response to noradrenaline, neither in combination with l -NAME, HOE 140 nor minoxidil prevented such an increase in the response to noradrenaline. In the presence of ergotamine, the contractile response to noradrenaline was completely blocked not only in control animals, but also in animals treated with ANG II alone or in combination with minoxidil. However, ergotamine (3 microm) failed to block completely the contractile response to noradrenaline in vessels from animals treated by ANG II in combination with l -NAME or HOE 140. These data suggest that ANG II causes an increase of noradrenaline sensitivity in the isolated portal vein of rat. l -NAME and HOE 140 unmask a contractile response to noradrenaline in the presence of ergotamine which seems to be mediated not only by alpha-adrenoceptors, but may be compensated by an endothelial relaxation.
Angiotensin II↗
Expression of functional recombinant antibody molecules in insect cell expression systems.
Recombinant single-chain variable-fragment molecules (scFv) were constructed from a cell line expressing a monoclonal antibody against African cassava mosaic virus (ACMV) and expressed in Escherichia coli. DNA sequences that encoded the scFv were manipulated to allow scFv expression in insect cell lines. A recombinant baculovirus containing the scFv cDNA was constructed and large amounts of scFv were produced in each of three insect cell lines infected with the baculovirus. However, the scFv were not secreted into the medium by any of the cell lines despite the scFv having been linked to a honeybee melittin leader sequence. The same scFv cDNA construct was introduced into Drosophila DS2 cells and a stable recombinant cell line was obtained that produced scFv that was secreted into the medium. Culture medium containing the scFv was used directly in enzyme-linked immunosorbent assay (ELISA) tests to detect ACMV in plant tissues. Another construct that encoded the Ckappa domain of human IgG was fused to the C-terminus of the scFv that was produced and expressed in Drosophila cells. This scFv derivative also accumulated in the medium and was more active in ELISA than scFv lacking the Ckappa domain.
Animals↗
Extensive gene duplications and a large inversion characterize the human leukocyte receptor cluster.
The chromosomal region 19q13.4 harbors the human leukocyte receptor cluster (LRC) which has been demonstrated to contain 19 genes encoding leukocyte-expressed receptors of the immunoglobulin (Ig) superfamily. A spotted PAC library was used to construct a contig of 65 overlapping clones spanning the complete LRC. Within the 900 kb covered by the contig, we identified one cluster containing killer cell inhibitory receptor genes and two clusters containing Ig-like transcript (ILT) genes. Of these, the second ILT cluster, located at the centromeric end of the LRC, was previously unknown. Detailed analysis of the ILT receptor genes in this cluster revealed one novel (ILT11) and six already known ILT genes. The two ILT clusters are transcribed in opposite directions and are separated by about 200 kb, which contains two leukocyte-associated inhibitory receptor (LAIR) genes. The data suggest that the two ILT clusters, each including one LAIR locus, arose from a single ancestral ILT/LAIR cluster by inverse duplication of a c large genomic fragment. Furthermore, the NK cell-expressed NKp46 gene was localized 20 kb telomeric of FCAR; and 14 novel genes mapping within the LRC were identified by cDNA selection. Together with the gene for the ribosomal protein S9 (RPS9), which had previously been assigned to 19q13.4, the total number of LRC genes is now 44. Of these, 29 belong to the Ig superfamily.
Animals↗
Hemolymph ion composition and volume changes in the supralittoral isopod Ligia pallasii Brandt, during molt.
We analyzed ion composition and volume of the hemolymph of Ligia pallasii in four different stages of the molt cycle using capillary electrophoresis and 3H-inulin. The main ions in the hemolymph were Na+, K+, Mg2+ , Ca2+, and Cl-. The Ca2+ concentration increased significantly during the molt by 47% from intermolt to intramolt and by 37% from intermolt to postmolt, probably due to resorption of Ca2+ from the cuticle and sternal CaCO3 deposits. The K+ concentration increased significantly by 20% during molt. The hemolymph volume normalized to the dry mass of the animals decreased by 36% from intermolt to late premolt. This was due to a reduction in the hemolymph volume and to an increase in dry mass of the animals during premolt. A sudden increase in the hemolymph volume occurring between late premolt and intramolt served to expand the cuticle. Since the Na+, K+, Mg2+, and Cl- concentrations did not change significantly from late premolt to intramolt, the increase in hemolymph volume suggests an uptake of seawater rather than freshwater.
Animals↗
Significant association between a silent polymorphism in the neuromedin B gene and body weight in German children and adolescents.
Neuromedin B has been shown to exert an inhibiting effect on food consumption in rats. The corresponding gene NMB maps to chromosome 15q22.3-q23, a region expected to contain a gene for the Bardet-Biedl syndrome type 4 (BBS4). Based on its map position and the putative function of the encoded peptide, NMB can be considered as a candidate gene both for BBS4 and the development of human obesity. To examine its involvement in these phenotypes, we determined the genomic structure of human NMB, and performed a mutation screen in its coding region. In genomic DNA of six BBS4 patients and in a large population sample, two sequence variants were detected: a g.253C-->A transversion creating a P73T substitution and a g.401G-->A silent mutation changing the stop codon TGA into stop codon TAA. A case-control study with 92 extremely obese patients and 94 underweight students revealed a significant association between the g.401G-->A polymorphism and body weight (adjustedp = 0.03), which was confirmed in a validation sample consisting of 95 extremely obese patients, and 95 normal weight and 48 underweight individuals (Mann-Whitney p = 0.02). These results suggest a contribution of NMB or a gene in its close vicinity to genetic weight control in humans.
Adolescent↗
Lisch corneal dystrophy is genetically distinct from Meesmann corneal dystrophy and maps to xp22.3.
PURPOSE: There is an ongoing discussion whether Lisch corneal dystrophy (band-shaped and whorled microcystic dystrophy of the corneal epithelium) represents a disorder that is different from Meesmann corneal dystrophy. The purpose of this study was to evaluate at the molecular level if Lisch and Meesmann corneal dystrophies are genetically distinct. METHODS: We examined at the slit lamp a total of 48 members of a family with an aggregation of Lisch corneal dystrophy. Genomic DNA was extracted from leukocytes of the peripheral blood of seven affected and six unaffected members of this family. Mutational hotspots in the cornea-specific keratin genes K3 and K12 were scanned for mutations by single-strand conformation analysis. To test for linkage to the keratin K3 or K12 loci or for X-chromosomal inheritance, six (K3) and four (K12) microsatellite markers each flanking the keratin loci as well as 22 microsatellite markers covering the X-chromosome were typed. Linkage was analyzed using the MLINK and FASTMAP procedures. RESULTS: A total of 19 trait carriers were identified in six generations of the family. No hereditary transmission from father to son was observed. Linkage was excluded for the keratin K3 and K12 genes. Furthermore, single-strand conformation analysis detected no mutations in these genes. Multipoint linkage analysis revealed linkage with a maximum likelihood of the odds (LOD) score of 2.93 at Xp22.3. Linkage was excluded for Xp22.2 to Xqter. CONCLUSIONS: Lisch corneal dystrophy is genetically different from Meesmann corneal dystrophy. Evidence was found for linkage of the gene for Lisch corneal dystrophy to Xp22.3.
Adolescent↗
Optimizing efficacy of antisense oligodeoxynucleotides targeting inhibitors of apoptosis.
Explore the source record for details and available documents.
Apoptosis↗
Polymorphisms in olfactory receptor genes: a cautionary note.
The hundreds of human olfactory receptor (OR) genes are organized into clusters occurring on nearly every chromosome. Although their sequences are not always closely related, they share stretches of considerable similarity, both at the amino acid and nucleotide levels. We demonstrate here that an HLA complex-linked OR sequence, FAT11, for which recently a number of alleles have been claimed within the Hutterites, contains sequences derived from two closely related, linked OR genes, hs6M1-12 and hs6M1-16. Instead of indicating a difference between alleles of a given locus, two of the polymorphisms described for FAT11 (at amino acids 48 and 220 of the deduced protein sequence, respectively) may in fact reflect distinct sequences of hs6M1-12 and a further, closely related HLA-linked OR locus, hs6M1-13P. As a consequence, recombination rates in Hutterites in the region telomeric of HLA-G may have to be reconsidered.
Amino Acid Substitution↗
Structurally diverse forms of HLA-B27 molecules are displayed in vivo in a cell type-dependent manner.
The formation of a trimeric complex, composed of heavy chain (HC), beta(2)-microglobulin (beta(2)m) and antigenic peptide, is generally believed to be a prerequisite for the expression of HLA class I molecules at the cell surface in vivo. Therefore, a possible role in immunological processes for HC/beta(2)m complexes devoid of peptide has not been seriously considered. Using a novel HLA-B*2705-transgenic rat model and monoclonal antibodies that distinguish between structurally different forms of HLA-B27 molecules, we demonstrate here that class I molecules which appear to lack antigenic peptides are expressed in abundance on a variety of cell types in lymphoid organs. These results imply a role for structurally diverse, possibly empty, MHC molecules in physiological T cell selection which has so far not been sufficiently appreciated.
Amino Acid Sequence↗
In vivo 129Xe NMR in rat brain during intra-arterial injection of hyperpolarized 129Xe dissolved in a lipid emulsion.
Hyperpolarized 129Xe was dissolved in a lipid emulsion and administered to anaesthetized rats by manual injections into the carotid (approximately 1-1.5 mL in a maximum time of 30 s). During injection, 129Xe NMR brain spectra at 2.35 T were recorded over 51 s, with a repetition time of 253 ms. Two peaks assigned to dissolved 129Xe were observed (the larger at 194 +/- 1 ppm assigned to intravascular xenon and the smaller at 199 +/- 1 ppm to xenon dissolved in the brain tissue). Their kinetics revealed a rapid intensity increase, followed by a plateau (approximately 15 s duration) and then a decrease over 5 s. This behaviour was attributed to combined influences of the T1 relaxation of the tracer, of radiofrequency sampling, and of the tracer perfusion rate in rat brain. Similar kinetics were observed in experiments carried out on a simple micro-vessel phantom. An identical experimental set-up was used to acquire a series of 2D projection 129Xe images on the phantom and the rat brain.
Animals↗
A point mutation in PTPRC is associated with the development of multiple sclerosis.
Multiple sclerosis (MS) is the most common demyelinating disease of the central nervous system. It is widely accepted that a dysregulated immune response against brain resident antigens is central to its yet unknown pathogenesis. Although there is evidence that the development of MS has a genetic component, specific genetic factors are largely unknown. Here we investigated the role of a point mutation in the gene (PTPRC) encoding protein-tyrosine phosphatase, receptor-type C (also known as CD45) in the heterozygous state in the development of MS. The nucleotide transition in exon 4 of the gene locus interferes with mRNA splicing and results in altered expression of CD45 isoforms on immune cells. In three of four independent case-control studies, we demonstrated an association of the mutation with MS. We found the PTPRC mutation to be linked to and associated with the disease in three MS nuclear families. In one additional family, we found the same variant CD45 phenotype, with an as-yet-unknown origin, among the members affected with MS. Our findings suggest an association of the mutation in PTPRC with the development of MS in some families.
Base Sequence↗
Epidemic obesity: are genetic factors involved via increased rates of assortative mating?
OBJECTIVE: Prevalence rates of obesity have been increasing in several countries over the past two decades. Mainly secular changes in energy intake and expenditure have been invoked to underly the increasing rates; genetic factors have not been considered because of the very recency of this phenomenon. We hypothesize that genetic factors might very well be involved via an increased rate of assortative mating between obese individuals. We speculate that the recent upsurge in social stigmatization of obese individuals underlies the hypothesized increase in assortative mating. DESIGN: To accumulate evidence for our hypothesis we analysed deduced rates of assortative mating among parents of extremely obese children and adolescents, who belonged to two different large study groups (n = 201 and n = 270). For this purpose we calculated parental body mass indices (BMIs) based on (a) measured current heights and weights, (b) self-reported current heights and weights, and finally (c) measured current heights and recalled weights at ages 20 and 30, respectively. BMI centiles were determined which in turn were attributed to the respective BMI decile. Deduced rates of assortative mating were evaluated in bivariate histograms of the paternal and maternal BMI deciles. RESULTS: High rates of assortative mating were observed as deduced from the bivariate histograms, which revealed a fairly consistent pattern. Thus, in the first study group almost 35% of the parental pairs had a BMI in the tenth decile; over 50% of the mothers and fathers had a BMI in this top decile. Recalled parental BMIs at ages 20 and 30 also clustered in the upper decile. These results were basically replicated in the second study group. In addition, parental loading on the tenth decile was shown to be higher for the subgroup of children and adolescents who had a BMI equal to or exceeding the highest BMI of the population-based age and gender matched control group. CONCLUSIONS: Our results indicate that assortative mating is common among parents of extremely obese children and adolescents, ascertained between 1995 and 1997. In addition, the parental loading on the tenth decile is most prominent for the most obese children. Whereas we are unable to document an increased rate of assortative mating, we interpret our results as being consistent with the hypothesis that an increased rate of assortative mating has contributed to the recent rise in obesity rates in several countries. Thus, assortative mating warrants further studies to assess its impact on obesity prevalence rates through both genetic and non-genetic mechanisms. Our results suggest that assortative mating might especially increase the rates for extreme obesity.
Adolescent↗
Rates of psychiatric disorders in a clinical study group of adolescents with extreme obesity and in obese adolescents ascertained via a population based study.
OBJECTIVE: To compare rates of DSM-IV psychiatric disorders between (1) a clinical study group of extremely obese adolescents and young adults, (2) gender-matched population-based obese controls and (3) a population-based control group of the same age range. DESIGN: Rates of psychiatric disorders were assessed in (1) the clinical study group of obese adolescents and (2) the population based sample of obese adolescents, and compared to (3) a large population-based control group using a standardized psychiatric interview. SUBJECTS: (1) Clinical study group: 30 female and 17 male extremely obese adolescents and young adults (age range: 15-21 y; mean BMI:42.4 kg/m2). (2) Thirty females and 17 males with the highest BMI (age range 15-21 y; mean BMI: 29.8 kg/m2) of a population-based control group encompassing 1655 (805 males) adolescents and young adults. (3) The population based control group excluding the 30 females and 17 males with the highest BMI (n = 1608; 788 males). MEASUREMENTS: Munich-Composite International Diagnostic Interview (M-CIDI) allowing for DSM-IV diagnoses. RESULTS: High rates of mood, anxiety, somatoform and eating disorders were detected in the clinical sample of obese adolescents which exceeded those observed in population controls (all P-values < 0.01). Rates between population-based obese adolescents and young adults and population controls did not differ. In most patients the psychiatric disorders set in after onset of obesity. 57% and 35% of the female and male patients, respectively, reported eating binges with lack of control. However, less than one-half of these patients qualified for a DSM-IV diagnosis of an eating disorder. CONCLUSIONS: Extremely obese adolescents and young adults who seek long-term inpatient treatment have a high lifetime prevalence for affective, anxiety, somatoform and eating disorders. Because the mean BMI of the clinical study group was considerably higher than that of the obese population controls, we were not able to clarify whether the high rate of psychopathology in the study group was related to the extreme obesity or to their treatment-seeking behavior.
Adipose Tissue↗
Leptin suppresses semi-starvation induced hyperactivity in rats: implications for anorexia nervosa.
Semi-starvation induced hyperactivity (SIH) occurs in rodents upon caloric restriction. We hypothesized that SIH is triggered by the decline in leptin secretion associated with food restriction. To test this hypothesis, rats, which had established a stable level of activity, were treated with leptin or vehicle via implanted minipumps concomitantly to initiation of food restriction for 7 days. In a second experiment treatment was initiated after SIH had already set in. In contrast to the vehicle-treated rats, which increased their baseline activity level by 300%, the development of SIH was suppressed by leptin. Furthermore, leptin was able to stop SIH, after it had set in. These results underscore the assumed major role of leptin in the adaptation to semi-starvation. Because SIH has been viewed as a model for anorexia nervosa, we also assessed subjective ratings of motor restlessness in 30 patients with this eating disorder in the emaciated state associated with hypoleptinemia and after increments in leptin secretion brought upon by therapeutically induced weight gain. Hypoleptinemic patients ranked their motor restlessness higher than upon attainment of their maximal leptin level during inpatient treatment. Thus, hypoleptinemia might also contribute to the hyperactivity frequently associated with anorexia nervosa.
Animals↗