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Biomedical subjects

R Fries

Publications and source records attributed to R Fries.

At least 37 records · Page 2Linked to original sources

Structural analysis and transcript processing of the bovine proteolipid protein (PLP) gene.

In this study we present the complete genomic structure of the bovine PLP gene and its assignment to the long arm of the X-chromosome (BTXq2.1). We determined a total of 18,767 bp of the bovine PLP gene and compared it to the human heterolog. A very high similarity was detected between the non-coding regions, interrupted primarily by several transposable elements. A deletion of 13 bp in the vicinity to the translation start signal in the promoter of the bovine PLP gene was found. Functional studies of the 3' region showed the use of several polyadenylation signals. Three main transcripts were detected in adult cattle in the range of 3200, 2400, and 1600 nucleotides using Northern blot analysis. An additional shorter transcript was detected in the cerebrum of calves.

Alternative Splicing↗

[Procedures in abattoirs and meat inspection. State of the discussion on the implementation of alternative systems].

Results of field studies which were performed in EU member states and which dealt with meat inspection procedures in fattening pigs were reviewed with respect to their contribution to alternative systems of meat inspection. The following was concluded: Inspection ante mortem (monitoring of circumstances at the farm of origin): Data which should be given to the notice of the authorities have not yet been satisfactorily defined and their informative value still is not yet clear The predictive character of information from the farm of origin regarding the health status of the animals (results of meat inspection) is not yet sound enough The technique of ante mortem inspection should be reconsidered also. Meat inspection (monitoring and assessment of lesions): In all studies the compared techniques (visual and official procedure) did not entirely find all lesions, which were supposed to be on the carcasses and organs. This was true in different percentages. The impact on consumer's health from the lesions monitored has to be discussed more thoroughly. The information presently available is not yet sound enough to generate a new practicable system of meat inspection. Further pilot studies are necessary.

Abattoirs↗

[Reconstructive surgery of the aortic root].

Surgical treatment of proximal aortic disease traditionally consists of composite replacement of valve and aorta. Recent reconstructive procedures on the aortic root allow for treatment of aortic dilatation and concomitant aortic valve regurgitation without the associated disadvantages of mechanical heart valves. From 10/95 to 09/99 we treated 84 patients for regurgitation of the aortic valve and dilatation of the aortic root. Valve preserving replacement of the root consisted of root remodeling (n = 68) or reimplantation of the aortic valve (n = 16). Operative mortality in valve-preserving surgery was not elevated compared to overall results of proximal aortic replacement (3.6% vs 5.6%); this applied to elective procedures (1.8% vs 2.3%) as well as emergency operations (9.3% vs 16.3%). Initial aortic valve function was adequate in all cases. Actuarial freedom from regurgitation grade II or higher was 98% after root remodeling and 92% after valve reimplantation. Freedom from reoperation at two years was 96% in remodeling and 100% in valve reimplantation. Hemodynamic function of the reconstructed valve was investigated in 17 patients under conditions of rest and exercise. These were compared to 9 patients with a mechanical composite valve. Patients with reconstructed valves had almost physiologic gradients during rest and exercise. These gradients were thus significantly lower than the increased gradients of patients after composite replacement. Application of reconstructive procedures to the aortic root allows for restoration of aortic valve function in the majority of patients. Disadvantages of heart valve prostheses can be avoided, and the hemodynamic performance of the reconstructed valve appears almost physiologic.

Adolescent↗

The dynamics of chromosome evolution in birds and mammals.

Comparative mapping, which compares the location of homologous genes in different species, is a powerful tool for studying genome evolution. Comparative maps suggest that rates of chromosomal change in mammals can vary from one to ten rearrangements per million years. On the basis of these rates we would expect 84 to 600 conserved segments in a chicken comparison with human or mouse. Here we build comparative maps between these species and estimate that numbers of conserved segments are in the lower part of this range. We conclude that the organization of the human genome is closer to that of the chicken than the mouse and by adding comparative mapping results from a range of vertebrates, we identify three possible phases of chromosome evolution. The relative stability of genomes such as those of the chicken and human will enable the reconstruction of maps of ancestral vertebrates.

Animals↗

The cattle interleukin-13 gene: genomic organization, chromosomal location, and evolution of the promoter.

Interleukin (IL)-13 plays a key role in the T-helper 2 type immune response. In domestic animal species no sequence information is available for the IL13 gene. Here, we report the isolation and sequencing of the cattle IL13 gene. Since a heterologous cDNA probe failed to hybridize to cattle genomic DNA the gene was identified by a positional sequencing approach. In human and mouse both cytokines are located within a region of about 30 kilobases (kb). Therefore the region upstream of the cattle IL4 gene has been sequenced by a shotgun approach. The cattle IL13 gene was found to be located about 20 kb upstream of IL4 in a head-to-tail orientation. The exon-intron structure and the promoter region is well conserved across species. The alignment of the IL13 sequences from different species allowed the identification of highly conserved amino acid positions and transcriptional elements. Two conserved amino acid positions (glu(31) and pro(94)) are most probably homologous to the interleukin-4 signalling residues. One of the potential binding sites for transcription factors is located at position -197 to -189 and is closely related to the P elements of the IL4 promoter. Nevertheless, the overall identity of the promoter sequences of IL13 and IL4 is low. Therefore we conclude that both cytokines use diverse regulatory elements. There is evidence that IL-13, but not IL-14, is the key mediator in allergic asthma. Furthermore IL-13 is involved in resistance to some infections. Therefore IL-13 is of interest for disease association studies in domestic animals.

Amino Acid Sequence↗

Molecular cloning and chromosomal assignment of the porcine 54 and 56 kDa vacuolar H(+)-ATPase subunit gene (V-ATPase).

Vacuolar proton-translocating ATPases (V-ATPase) are multisubunit enzyme complexes located in the membranes of eukaryotic cells regulating cytoplasmic pH. So far, nothing is known about the genomic organization and chromosomal location of the various subunit genes in higher eukaryotes. Here we describe the isolation and analysis of a cDNA coding for the 54- and 56-kDa porcine V-ATPase subunit alpha and beta isoforms. We have determined the genomic structure of the V-ATPase subunit gene spanning at least 62 kb on Chromosome (Chr) 4q14-q16. It consists of 14 exons with sizes ranging from 54 bp to 346 bp, with a non-coding first exon and an alternatively spliced seventh exon leading to two isoforms. The 5' end of the V-ATPase cDNA was isolated by RACE-PCR. The V-ATPase alpha isoform mRNA, lacking the seventh exon, has an open reading frame of 1395 nucleotides encoding a hydrophilic protein of 465 amino acids with a calculated molecular mass of 54.2 kDa and a pI of 7.8, whereas the beta isoform has a length of 1449 nucleotides encoding a protein of 483 amino acids with a calculated molecular mass of 55.8 kDa. Amino acid and DNA sequence comparison revealed that the porcine V-ATPase subunit exhibits a significant homology to the VMA13 subunit of Saccharomyces cerevisiae V-ATPase complex and V-ATPase subunit of Caenorhabditis elegans.

Amino Acid Sequence↗

High-resolution, human-bovine comparative mapping based on a closed YAC contig spanning the bovine mh locus.

A closed YAC contig spanning the mh locus was assembled by STS content mapping with seven microsatellite markers, eight genes or EST, and nine STS corresponding to YAC ends. The contig comprises 27 YACs, has an average depth of 4.3 YACs, and spans an estimated 1.2 Mb. A linkage map was constructed based on five of the microsatellite markers anchored to the contig and shown to span 7 cM, yielding a ratio of 160 kb/1 cM for the corresponding chromosome region. Comparative mapping data indicate that the constructed contig spans an evolutionary breakpoint connecting two chromosome segments that are syntenic but not adjacent in the human. Consolidation of human gene order by means of whole genome radiation hybrids and its comparison with the bovine order as inferred from the contig confirm conservation of gene order within segments.

Animals↗

Construction and characterization of a porcine P1-derived artificial chromosome (PAC) library covering 3.2 genome equivalents and cytogenetical assignment of six type I and type II loci.

A porcine P1-derived artificial chromosome (PAC) library of a male German Landrace pig was constructed in pCYPAC2. In total 90,240 clones were generated and individually transferred into microtiter plates. An average insert size of 119.1 kb was determined by analyzing 150 randomly selected PAC clones by pulsed field electrophoresis, yielding approximately 3.2 genome equivalents. The stability of nine clones was followed through 110 generations showing no reduction of the insert size. The probability of identifying a specific chromosomal region within the library was tested by screening for the presence of seven type I and five type II loci. The analysis showed that most loci (10/12) were present in the library at least twice. To determine the percentage of chimerism, six clones were analyzed by fluorescence in situ hybridization (FISH) on metaphase chromosomes. We assign one type I locus (Triadin) and three type II loci (SW855, S0300, SW1129).

Animals↗

A 5x genome coverage bovine BAC library: production, characterization, and distribution.

A bovine large-insert DNA library has been constructed in a Bacterial Artificial Chromosome (BAC) vector. The source DNA was derived from lymphocytes of a Jersey male. High-molecular-weight DNA fragments were produced by treatment with EcoRI/EcoRI methylase and cloned into the EcoRI site of pBACe3.6. In total, 157,240 individual BACs have been picked into 384-well plates. Approximately 190 randomly chosen clones have been characterized by Pulsed Field Gel Electrophoresis (PFGE) and have an average insert size of 105 kb, suggesting library coverage representing 5-6 genome equivalents. The frequency of clones without inserts is 4%. The chromosomal location of 51 BACs was studied by FISH; 3 showed more than one signal, indicating a chimerism frequency of roughly 6%. Approximately 50% of the clones in the library contain Simple Repeat Sequences (microsatellites), and 4% of the clones contain centromeric repeats. Insert stability was assessed by restriction digestion of DNA prepared from 20 clones after serial culture for one and three nights. Only one clone showed any evidence of an altered restriction pattern. Clones from 360 x 384-well plates (138,240 colonies) were gridded onto high-density membranes, and PCR superpools were produced from the same set of clones. Both membranes and superpools are available from the RZPD, Berlin (http://www.rzpd.de). PCR 4-D superpools have been prepared from an additional 23,000 clones. The library has been screened for a total of 24 single-copy sequences; positive clones have been obtained in all cases.

Animals↗

Water and air in two poultry processing plants' chilling facilities--a bacteriological survey.

1. Water, aerosols and air in chilling facilities in 2 poultry abattoirs were bacteriologically examined. The different types of samples displayed a variety of bacterial loads and genera. 2. The aerobic plate count in water from spray nozzles was about log10 2 to 3 in both plants. Flavobacterium, Alcaligenes, Acinetobacter and Pseudomonadaceae were predominant in these samples. 3. The microbial load in air of the 2 plants was quite different, consisting mainly of Micrococcaceae and Gram-positive irregular rods. 4. The aerobic plate count in aerosols was extremely high. The flora was the same as was found in the spray nozzles samples, plus Enterobacteriaceae, Micrococcaceae and streptococci, which presumably came from the air in the system. Cross-contamination may well occur in evaporative chilling.

Abattoirs↗

Clinical significance of sleep-related breathing disorders in patients with implantable cardioverter defibrillators.

The prevalence and clinical significance of sleep-related breathing disorders (SRBDs) in patients with cardiac disease and a history of life-threatening ventricular tachyarrhythmias is unclear. Forty consecutive recipients of implantable cardioverter defibrillators (ICDs) with cardiac disease and a documented history of spontaneous, life-threatening, ventricular tachyarrhythmias underwent full night polysomnography. SRBDs were diagnosed if the apnea/hypopnea index was > 10. SRBD were diagnosed in 16 of 40 patients (40%): central sleep apnea (CSA) was present in 9 of these 16 patients (56%), 8 of whom had associated Cheyne-Stoke respiration. Seven of the 16 patients with SRBD (44%) had obstructive sleep apnea (OSA). Patients with and without SRBDs were comparable with respect to left ventricular ejection fraction, NYHA classification, underlying heart disease, ICD indications, and concomitant antiarrhythmic drug and beta-blocker therapy. Patients were followed prospectively for 2 years. ICD-treated ventricular tachyarrhythmias occurred in 10 of 24 patients (42%) without SRBD, in 4 of 9 patients (44%) with CSA, and in 3 of 7 patients (44%) with OSA (NS). The numbers and circadian distributions of episodes recorded during follow-up in patients without SRBD versus with CSA or OSA were not significantly different (14 +/- 25, median = 4 vs 4 +/- 5, median = 2.5 vs 15 +/- 15, median = 7, respectively). The 2-year mortality, which was entirely attributable to nonsudden cardiac events, was highest in patients with CSA (4/9 [44%], vs 0/7 [0%] with OSA, vs 3/24 patients (12.5%) without SRBD; P < 0.05).

Aged↗

Transvenous atrial cardioversion threshold in patients with implantable cardioverter defibrillator: influence of active pectoral can.

Recent studies have shown that transvenous atrial cardioversion is feasible with lead configurations primarily designed for implantable cardioverter defibrillators (ICD). The purpose of this study was to examine the influence of an active pectoral ICD can on the atrial cardioversion threshold (ADFT). Forty consecutive patients received a transvenous single lead system (Endotak DSP 0125, CPI, St. Paul, MN, USA) in combination with a left subpectoral ICD (Ventak Mini, CPI) for treatment of malignant ventricular tachyarrhythmias. Patients were randomized into two groups: 21 received a Hot Can 1743 and 19 patients a Cold Can 1741. Step-down testing of the ventricular defibrillation threshold (VDFT) was performed intraoperatively and evaluation of the ADFT for induced atrial fibrillation (AF) at predischarge. After testing, each patient received a 2-J shock and was asked to quantify discomfort on a numerical scale ranging from 0 to 10. Both groups were comparable with regard to all clinical parameters studied. The mean VDFT in patients with a Hot Can device was significantly lower than in patients with a Cold Can (7.5 +/- 2.3 J vs 9.8 +/- 3.8 J; P < 0.03). The mean ADFT in the Hot Can group tended to be lower than in the group with Cold Cans (3.4 +/- 1.4 J vs 4.5 +/- 2.4 J; P = 0.07), and the proportion of patients in whom atrial cardioversion was accomplished at low energies (< or = 3 J) was higher in patients with active compared with patients with inactive pulse generators (57% vs 26%; P < 0.04). The mean discomfort reported after delivery of a 2-J shock was comparable in both groups (Hot Can 5.2 +/- 1.9; Cold Can: 5.3 +/- 2.1; P = NS). We conclude that the inclusion of an active left subpectoral can in the defibrillation vector of a ventricular ICD seems to reduce the energy requirements for atrial cardioversion without increasing the discomfort caused by low energy shocks.

Atrial Fibrillation↗

[Associations between farm characteristics in fattening pigs and occurrence of lesions at meat inspection].

A revised inspection system for fattening pigs in the EU has been proposed emphasizing the need for a more detailed documentation of the fattening process. It is supposed that the availability of such information enables a discrimination of conspicuous and non-conspicuous lots of fattening pigs delivered to the abattoir allowing for a modified, only visual inspection of non-conspicuous lots. This proposal motivated a study performed in 1996 on about 22,000 pigs, searching, among other things, for first hints on associations between management/farm conditions for fattening pigs and lesions found at meat inspection, which can be used for discrimination. Taking the two lesions 'pneumonia' and 'callous affections of the legs' as examples this contribution demonstrates the use of regression trees to visualize association structures and to develop hypotheses on relations between lesions and farm characteristics. An association of 'callous affections of the legs' with flooring systems is clearly shown in the generated tree. The occurrence of 'pneumonia' seems to be associated with the application of disinfection measures in this data set. However, this result should be interpreted with caution as some problems for data analysis arise because of the low number of farms in this study in relation to the high number and complexity of farm characteristics considered. A further problem is the high variability of daily deliveries within farms that cannot be explained referring to farm characteristics common to all deliveries. As a conclusion data from this study have been useful to develop hypotheses on associations between farm characteristics and lesions at meat inspection. However, there is need for further, more specific studies before such associations can be utilized to discriminate between conspicuous and non-conspicuous lots of fattening pigs.

Animal Husbandry↗

Influence of beta-blockers on the frequency of arrhythmia recurrences in patients with implantable cardioverter-defibrillator: an intraindividual comparison.

We studied retrospectively 60 consecutive recipients of an implantable cardioverter-defibrillator and identified 16 patients who were temporarily on and off beta-blockers (further medication unchanged). An intraindividual analysis revealed that 56% of the patients experienced more arrhythmic episodes during follow-up off beta-blockers compared to 44% while being on beta-blockers. Also, the mean episode frequency during follow-up time on and off beta-blockers was comparable (0.4+/-0.6 vs. 0.5+/-0.5, ns).

Adrenergic beta-Antagonists↗