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Biomedical subjects

G Brockmann

Publications and source records attributed to G Brockmann.

15 recordsLinked to original sources

Three-dimensional recording of the human face with a 3D laser scanner.

Three-dimensional recording of the surface of the human body or of certain anatomical areas has gained an ever increasing importance in recent years. When recording living surfaces, such as the human face, not only has a varying degree of surface complexity to be accounted for, but also a variety of other factors, such as motion artefacts. It is of importance to establish standards for the recording procedure, which will optimise results and allow for better comparison and validation. In the study presented here, the faces of five male test persons were scanned in different experimental settings using non-contact 3D digitisers, type Minolta Vivid 910). Among others, the influence of the number of scanners used, the angle of recording, the head position of the test person, the impact of the examiner and of examination time on accuracy and precision of the virtual face models generated from the scanner data with specialised software were investigated. Computed data derived from the virtual models were compared to corresponding reference measurements carried out manually between defined landmarks on the test persons' faces. We describe experimental conditions that were of benefit in optimising the quality of scanner recording and the reliability of three-dimensional surface imaging. However, almost 50% of distances between landmarks derived from the virtual models deviated more than 2mm from the reference of manual measurements on the volunteers' faces.

Adult↗

Accuracy and precision of the three-dimensional assessment of the facial surface using a 3-D laser scanner.

Three-dimensional (3-D) recording of the surface of the human body or anatomical areas has gained importance in many medical specialties. Thus, it is important to determine scanner precision and accuracy in defined medical applications and to establish standards for the recording procedure. Here we evaluated the precision and accuracy of 3-D assessment of the facial area with the Minolta Vivid 910 3D Laser Scanner. We also investigated the influence of factors related to the recording procedure and the processing of scanner data on final results. These factors include lighting, alignment of scanner and object, the examiner, and the software used to convert measurements into virtual images. To assess scanner accuracy, we compared scanner data to those obtained by manual measurements on a dummy. Less than 7% of all results with the scanner method were outside a range of error of 2 mm when compared to corresponding reference measurements. Accuracy, thus, proved to be good enough to satisfy requirements for numerous clinical applications. Moreover, the experiments completed with the dummy yielded valuable information for optimizing recording parameters for best results. Thus, under defined conditions, precision and accuracy of surface models of the human face recorded with the Minolta Vivid 910 3D Scanner presumably can also be enhanced. Future studies will involve verification of our findings using test persons. The current findings indicate that the Minolta Vivid 910 3D Scanner might be used with benefit in medicine when recording the 3-D surface structures of the face.

Algorithms↗

Detection of novel single-nucleotide polymorphisms (SNPs) in the CYP21 gene and association analysis of two SNPs for CYP21 and ESR2 with litter size in a commercial sow population.

Altogether 129 F1 sows from a commercial sow farm with at least four litters were genotyped for the oestrogen receptor 2 gene (ESR2) and cytochrome P450 hydroxylase 21 gene (CYP21) and investigated for associations on the litter-size parameters: total number born and number born alive. Five novel polymorphisms were found in the 3'-untranslated region for the CYP21 gene. Genotype and allele frequencies for the CYP21 (position 3462G > A) single-nucleotide polymorphism (SNP) were 0.434 (GG), 0.504 (AG), 0.062 (AA) and 0.69 (G):0.31 (A), respectively. No association was found between this polymorphism and litter-size parameters. For the ESR2 gene, the SNP in exon 5 associated with an amino acid substitution MET (allele A) > VAL (allele G) was investigated. Only two genotypes were found leading to allele frequencies of 0.34 (A):0.66 (G). Only number born alive piglets were significantly increased for the AG genotype (p = 0.034) with 11.64 piglets per sow and litter in comparison with the GG genotype, leading to only 10.96 piglets per sow and litter. From these data, it can be concluded that the investigated SNP of the ESR2 gene is associated with the number of liveborn piglets in the commercial population considered, and hence could be useful in selection for litter size. Therefore, this gene should be investigated in additional populations.

Animals↗

Analysis of properdin (BF) genotypes associated with litter size in a commercial pig cross population.

Properdin (BF) was investigated as a candidate gene influencing litter size in a commercial pig cross population. The BF gene was chosen because of its integral role in influencing uterine epithelium growth and because several quantitative trait loci (QTL) with impact on reproductive traits have been detected near the centromere of porcine chromosome 7. A total of 123 F2 (Large White x Landrace) x Leicoma sows were genotyped using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. The sows were divided into two extreme performance groups, one with a high litter size (n = 61, > or = 14.3 piglets per litter) and the other with a low litter size (n = 62, < or = 11.3 piglets per litter). Although genotype and allele frequencies were uneven with 2.4% (AA), 16.3% (AB), 81.3% (BB) and 0.11 (A): 0.89 (B), the allele A was the unfavourable one, leading to less offspring. With regard to the level of significance at p < 0.05, the total number of born (TNB) and number of born alive (NBA) piglets were associated with BF genotypes. The genotype AA led to 10.55 TNB and 10.00 NBA, whereas the genotype BB led to 13.19 TNB and 12.11 NBA. The genotype AB was intermediate. In future, a systematic mating test is necessary in order to obtain more balanced genotype frequencies. Furthermore, it should be taken into consideration that the investigated polymorphism is located in an intronic region and the causative mutation is not clear yet.

Animals↗

Different isoforms of the soluble leptin receptor in non-pregnant and pregnant mice.

Leptin circulates in murine serum in a free and a bound form. As shown in humans, a soluble leptin receptor (sOB-R), which modulates the effects of its ligand, circulates in murine blood. The aim of our study was to determine abundance and biochemical nature of this protein. For the quantification of sOB-R we developed a ligand-immunofunctional assay (LIFA) which is based on both, leptin binding and immunological recognition. The use of this LIFA revealed that during late gestation sera of pregnant mice had a approximately 290-fold higher level of sOB-R than non-pregnant animals. As investigated by size exclusion chromatography these mice sera demonstrated a co-elution of their leptin binding activity with leptin immunoreactivity and levels of sOB-R measured by LIFA. Therefore, it has to be concluded that sOB-R represents the major leptin binding activity in murine circulation. The molecular analysis of sOB-R by Western blot and by cross-linking with 125I-leptin in sera of pregnant and non-pregnant mice demonstrated two different isoforms of sOB-R, which were capable of leptin binding. The sOB-R in serum migrated at a molecular weight of 150kDa in pregnant and only of 120kDa in non-pregnant animals. Deglycosylation of these isoforms led to sOB-R molecules which were found at the same molecular weight in SDS-PAGE. This finding indicates that both isoforms differ only in the degree of their glycosylation. In conclusion, the non-pregnant and the pregnant states are accompanied by differently glycosylated isoforms of sOB-R whose physiological relevance remains to be determined.

Animals↗

A mammary gland EST showing linkage disequilibrium to a milk production QTL on bovine Chromosome 14.

As part of a genome scan, ESTs derived from mammary gland tissue of a lactating cow were used as candidate genes for quantitative trait loci (QTL), affecting milk production traits. Resource families were genotyped with 247 microsatellite markers and 4 polymorphic ESTs. It was shown by linkage analysis that one of these ESTs, KIEL_E8, mapped to the centromeric region of bovine Chromosome (Chr) 14. Regression analysis revealed the presence of a QTL, with significant effect on milk production, in this chromosome region, and analysis of variance showed no significant interaction of marker genotype and family. The estimated significant differences between homozygous marker genotypes were 140 kg milk, -5.02 kg fat yield, and 2.58 kg protein yield for the first 100 days of lactation. Thus, there was strong evidence for a complete or nearly complete linkage disequilibrium between KIEL_E8 and the QTL. To identify the biological function of KIEL_E8, we extended the sequence for 869 bp by 5'-RACE. A 560-bp fragment of this shows a 90.9% similarity to a gene encoding a cysteine- and histidine-rich cytoplasmic protein in mouse. Although such a protein may have a regulatory function for lactation and a linkage disequilibrium between the EST marker and the QTL has been observed, it remains to be elucidated whether they are identical or not. Nevertheless, KIEL_E8 will be an efficient marker to perform marker-assisted selection in the Holstein-Friesian population.

Alleles↗

[Dealing with asthma for teachers--needs and possibilities].

UNLABELLED: Teachers and schools are confronted with an increasing number of children with asthma. Teachers have to care for 2 to 4 children or adolescents with asthma in an average class; but they feel that they are not adequately informed to cope with such problems. METHODS: In 1999 and 2000, we conducted an inquiry in schools in the region of Osnabrueck. This investigation used approximately the same tools as a previous inquiry in Vienna. Results were similar and demonstrated deficits in the knowledge about asthma and in the ability to manage children with asthma exacerbations. At the same time, we offered a three-hour-programme in 40 schools and nurseries, to be given in the rooms of the schools. We always tried to reach all teachers of one school. Topics were: What is asthma? Self grading of severity; asthma and sports; emergency management; avoidance of triggers. RESULTS: There were 648 participants. There was much interest in the seminars, and we could not always admit all applicans. Theoretical and practical contents taught were, as judged by the participants, helpful in the average school settings. The participants felt that they were now better prepared and more competent in dealing with bronchial asthma in their pupils. CONCLUSION: We conclude that such seminars are helpful for teachers. The program can be conducted with moderate financial and man power resources. One may aim at including such educational units into the regular curriculum of teachers, at least as far as the most common chronic childhood and adolescent diseases are concerned.

Adolescent↗

Structure of the growth hormone-encoding gene and its promoter in mice.

The isolation and nucleotide sequence determination of the 5' flanking region of the mouse growth hormone (mGH)-encoding gene (mGH) is described. The mGH gene consists of five exons and four introns, as is observed in other mammalian species. The second intron in mGH is much smaller than its rat counterpart, thus being similar in size to human, bovine and porcine GH. The transcription start point was determined to be a C residue 62 bp upstream from the start codon, ATG. Analysis of 1767 bp of the 5' flanking region, with respect to putative regulatory elements, revealed a TATA box, two binding sites for growth hormone factor (GHF1), a GC box (SP1), a thyroid-response element (TRE) and a silencer (SiL) sequence motif. As expected, the mGH promoter shows a higher degree of homology with rat, as compared to the other mammalian species like pig, cattle and human, where an overall homology exists only at the proximal promoter region.

Amino Acid Sequence↗

A 5' splice-region mutation and a dinucleotide deletion in the lysosomal acid lipase gene in two patients with cholesteryl ester storage disease.

Cholesteryl ester storage disease (CESD) results from inherited deficiencies of the lysosomal hydrolase, acid lipase (LAL; E.C. 3.1.1.13). To establish the molecular defects in LAL deficiency, two unrelated probands with severely reduced LAL activity were examined. DNA amplification by reverse-transcription polymerase chain reaction and subsequent sequence analysis of LAL cDNA identified two mutant alleles. Patient 1, presenting with hepatosplenomegaly, mildly elevated liver function tests, and hyperlipidemia, was homozygous for a deletion of nucleotides 823 to 894 of the LAL cDNA. This 72-bp deletion maintained the reading frame and resulted in a loss of 24 amino acids from the LAL protein. Analysis of genomic DNA revealed that the 72 bp corresponded to an exon of the LAL gene. A single G to A point mutation at the last exon position was observed in the genomic DNA of patient 1, indicating a splicing defect with consecutive exon skipping underlying the 72-bp deletion. Patient 2 was a compound heterozygote for the 72-bp deletion and a dinucleotide deletion at positions 967 and 968. This deletion resulted in a shifted reading frame carboxyterminal of codon 296, and 43 random amino acids followed the frame shift. A premature stop at codon 339 truncated the mutant LAL protein by 34 amino acids. Allele-specific hybridization confirmed that patient 1 was homozygous for the 72-bp deletion mutation, and that patient 2 was a compound heterozygote for the 72-bp deletion and the 2-bp deletion.

Adult↗

DNA fingerprinting of trait-selected mouse lines and linkage analysis in reference families.

The first aim of the study was the molecular genetic characterization of long-term trait-selected (growth, adaptability and fertility) lines of mice using multilocus DNA fingerprinting with the simple tandem repetitive oligonucleotide probes (GAA)6 and (GACA)4. Secondly polymorphic markers were screened for association with growth performance based on DNA fingerprint analysis in reference families. Pooled DNA samples of ten unrelated mice of trait-selected mouse lines (over 40 generations) and the unselected control lines were analyzed. Resulting differences in band patterns were reanalyzed comparing individual fingerprints of the animals included in the pooled DNA samples of the different lines. Between the mouse lines about 30% of analyzed fingerprint bands were polymorphic. Individuals of long-term selected mouse lines show only a few individual-specific bands. Most polymorphic bands observed in DNA mixes appear in all animals included in the DNA mix of the corresponding line. Line-specific DNA fingerprint bands were analyzed for their inheritance and linkage with growth performance in reference families using animals with extreme growth performance of the first backcross after crossing of growth-selected with unselected mice. Scanning the distribution of line-specific bands in the reference panel few bands were identified which are associated with growth performance. They appear as useful markers for growth selection. Nevertheless most line-specific bands result from genetic drift rather than from selection.

Adaptation, Physiological↗

Quantification of Y chromosome bearing spermatozoa of cattle using in situ hybridization.

An easy assay for quantification of Y chromosome-bearing sperm (Y-sperm) is needed, especially to monitor sperm separation techniques. In the present study a tritiated bovine male-specific DNA fragment was tested for identification of Y-sperm by in situ hybridization. A protocol for in situ hybridization to bovine sperm was developed and used to study the proportion of Y-sperm of 12 bulls. The usefulness of the method in optimization of sperm separation procedures is illustrated through analysis of fractions of sperm separated by Percoll density gradient centrifugation.

Animals↗

Expression and localization of growth hormone-releasing hormone messenger ribonucleic acid in rat placenta: in vitro secretion and regulation of its peptide product.

The placenta is the source of many hypothalamic peptides. We now report that GH-releasing hormone (GHRH) mRNA was detectable in rat placenta. On Northern blot hybridization analysis, the size of the placental GHRH transcript was the same with the putative GHRH precursor mRNA. On in situ hybridization, the cells expressing the GHRH mRNA had the morphological characteristics of cytotrophoblasts. In addition, immunoreactive (IR) postranslational products of GHRH were present in rat placental extracts and in the effluent of in vitro perifused placentae. On gel filtration chromatography, the bulk of IR-GHRH present in placental extracts and perifusion effluent had the same size as the authentic hypothalamic GHRH (5.2K). A higher mol wt form of IR-GHRH of about 10K was also present and may represent the pro-GHRH predicted from the sequence of the GHRH cDNA. The mean basal release of IR-GHRH in the perifusion effluent from full thickness rat placental fragments was 337.3 +/- 38.5 (+/- SE; n = 48) pg/10 min fraction.g tissue. Depolarization by 56 mM KCl increased the concentration of the secreted immunoreactive peptide to 632.2 +/- 50.5. A 10-min exposure to 8-bromo-cAMP caused an immediate, monophasic, and dose-dependent increase in IR-GHRH secretion, which lasted between 20-30 min. The ensuing response to a KCl pulse was similar in size and pattern to that in the control channels. In contrast, a 10-min pulse of phorbol 12-myristate 13-acetate (a protein kinase-C-irreversible activator) induced a gradual, prolonged, and dose-dependent increase in basal GHRH secretion which lasted for at least 4 h. Additionally, phorbol 12-myristate 13-acetate enhanced KCl-induced GHRH secretion. In conclusion, our data suggest that the GHRH gene is expressed in the rat placenta. The placental GHRH transcript and its peptide products appear to have the same size as their hypothalamic counterparts, while the site of its placental GHRH synthesis is the cytotrophoblast. Finally, the secretion of placental GHRH seems to be regulated by both the adenyl cyclase and the protein kinase-C pathways.

Animals↗

Effect of hypertonic saline infusion on the level of immunoreactive dynorphin in extracted human plasma.

Dynorphin-A and its related peptides are derived from prodynorphin, one of the three known endogenous opioid precursors. The prodynorphin gene is expressed in the vasopressinergic magnocellular neurons of the hypothalamus, while its peptide products are present in the vasopressin (AVP) neurosecretory vesicles of the neurohypophysis. The concentration of immunoreactive (IR) dynorphin is orders of magnitude higher in the neurohypophysis than in any other tissue, suggesting that perhaps the prodynorphin-derived peptides are secreted from the hypothalamic-neurohypophyseal unit into the general circulation. Experiments in rats have shown that osmotic stimuli increase both AVP and prodynorphin in the hypothalamus. To determine whether human hypothalamic prodynorphin is also under osmotic regulation, we measured plasma IR-dynorphin, plasma IR-AVP, and serum sodium immediately before and during the infusion of normal or hypertonic saline in normal human volunteers. Because of the unusual susceptibility of the prodynorphin-derived peptides to cleavage by endopeptidases, we also developed an appropriate plasma dynorphin extraction technique. We found that the IR-dynorphin present in human plasma was composed of 6K- and 4K-sized peptides and that no larger than 6K or smaller than 4K dynorphins were present. The infusion of normal saline did not have any significant effect on plasma IR-dynorphin, while 3% hypertonic saline increased its plasma levels. Thus, the mean IR-dynorphin level in the plasma of the volunteers infused with normal saline was 40.3 +/- 6.4 fmol/mL (mean +/- SE; n = 6) at zero time; after 30 min of infusion, plasma IR-dynorphin was 36.0 +/- 6.3, after 60 min it was 29.9 +/- 5, after 90 min it was 36.0 +/- 4.7, after 120 min it was 36.8 +/- 3.2, and after 150 min it was 36.0 +/- 6.1. The plasma IR-dynorphin level in the volunteers infused with hypertonic saline was 31.7 +/- 3.5 fmol/mL (mean +/- SE; n = 10) at zero time. After 30 min of infusion it increased to 37.4 +/- 3.8, after 60 min to 46.4 +/- 7.7, after 90 min to 56.2 +/- 9.1, after 120 min to 53.6 +/- 8.7, and after 150 min to 99.0 +/- 14.2. The increase in plasma IR-dynorphin with time was significant (P less than 0.0001) and correlated positively with serum sodium and plasma AVP. The physiological role of the prodynorphin-derived peptides of the hypothalamic-neurohypophyseal unit is not yet known.(ABSTRACT TRUNCATED AT 400 WORDS)

Chromatography, Affinity↗