Search PubMed⌕ Search

Biomedical subjects

A Kilian

Publications and source records attributed to A Kilian.

32 records · Page 2Linked to original sources
PubMed1995

Comparative mapping of the barley genome with male and female recombination-derived, doubled haploid populations.

Male (anther culture) and female (Hordeum bulbosum) derived, doubled haploid populations were used to map the barley genome and thus determine the different recombination rates occurring during meiosis in the F1 hybrid donor plants. The anther culture-derived (male recombination) population showed an 18% overall increase in recombination rate. This increased recombination rate was observed for every chromosome and most of the chromosome arms. Examination of linkage distances between individual markers revealed eight segments with significantly higher recombination in the anther culture-derived population, and one in the Hordeum bulbosum-derived population. Very strong distortions of single locus segregations were observed in the anther culture-derived population, but map distances were not affected significantly by these distortions. There were 1.047 and 0.912 recombinations per chromosome in the anther culture and Hordeum bulbosum-derived doubled haploid populations, respectively.

Artifacts↗
PubMed1995

Barley telomeres shorten during differentiation but grow in callus culture.

Eukaryotic chromosomes terminate with long stretches of short, guanine-rich repeats. These repeats are added de novo by a specialized enzyme, telomerase. In humans telomeres shorten during differentiation, presumably due to the absence of telomerase activity in somatic cells. This phenomenon forms the basis for several models of telomere role in cellular senescence. Barley (Hordeum vulgare L.) telomeres consist of thousands of TTTAGGG repeats, closely resembling other higher eukaryotes. In vivo differentiation and aging resulted in reduction of terminal restriction fragment length paralleled by a decrease of telomere repeat number. Dedifferentiation in callus culture resulted in an increase of the terminal restriction fragment length and in the number of telomere repeats. Long-term callus cultures had very long telomeres. Absolute telomere lengths were genotype dependent, but the relative changes due to differentiation, dedifferentiation, and long-term callus culture were consistent among genotypes. A model is presented to describe the potential role of the telomere length in regulation of a cell's mitotic activity and senescence.

Journal Article↗
PubMed1995

Rice-barley synteny and its application to saturation mapping of the barley Rpg1 region.

In order to facilitate the map-based cloning of the barley stem rust resistance gene Rpg1, we have demonstrated a high degree of synteny at a micro level between the telomeric region of barley chromosome 1P and rice chromosome 6. We have also developed and applied a simple and efficient method for selecting useful probes from large insert genomic YAC and cosmid clones. The gene order within the most terminal 6.5 cM of barley chromosome 1P was compared with the most terminal 2.7 cM of rice chromosome 6. Nine rice probes, previously mapped in rice or isolated from YAC or cosmid clones from this region, were mapped in barley. All, except one, were in synteny with the rice gene order. The exception, probe Y617R, was duplicated in barley. One copy was located on a different chromosome and the other in a non-syntenic position on barley chromosome 1P. The barley probes from this region could not be mapped to rice, but two of them were inferred to be in a syntenic location based on their position on a rice YAC. This work demonstrates the utility of applying the results of genetic and physical mapping of the small genome cereal rice to map-based cloning of interesting genes from large genome relatives.

Chromosome Mapping↗
PubMed1995

Plasmodium falciparum and Plasmodium berghei: effect of magnesium on the development of parasitemia.

The in vitro growth of Plasmodium falciparum was reduced by 35 and 43% through high concentrations (5 mmole/liter) of magnesium in RPMI medium and magnesium-free medium, respectively, after 48 hr, whereas no significant inhibition could be observed under these conditions after 24 hr cultivation in the respective medium. Levels of magnesium between 0.5 and 3 mmole/liter showed no inhibitory effect on the in vitro growth of P. falciparum even after long-term exposure for 7 days. The 50 and 90% chloroquine inhibitory concentrations of the chloroquine-resistant strain K1 after 24 hr were reduced to some extent in the presence of magnesium at 5 mmole/liter, but less than in the presence of verapamil at 10 mumole/liter, which showed intrinsic activities at this concentration and which completely reversed resistance. However, high physiologic magnesium plasma levels were associated with a significantly longer survival time of NMRI mice infected with P. berghei strain ANKA, compared to normal physiological plasma magnesium levels. It is concluded that in the case of clinically symptomatic magnesium deficiency, supplementation of magnesium will not aggravate concomitant plasmodial infections and therefore should not be withheld.

Analysis of Variance↗
PubMed1995

Variation of nitrate reductase genes in selected grass species.

In order to study the variation of nitrate reductase (NR) genes among grass species, gene number, intron size and number, and the heme-hinge fragment sequence of 25 grass species were compared. Genomic DNA cut with six restriction enzymes and hybridized with the barley NAD(P)H and NADH NR gene probes revealed a single NAD(P)H NR gene copy and two or more NADH NR gene copies per haploid genome in most of the species examined. Major exceptions were Hordeum vulgare, H. vulgare ssp. spontaneum, and Avena strigosa, which appeared to have a single NADH NR gene copy. The NADH NR gene intron number and lengths were examined by polymerase chain reaction amplification. Introns I and III appeared to be absent in at least one of the NADH NR genes in the grass species, while intron II varied from 0.8 to 2.4 kilobases in length. The NADH NR gene heme-hinge regions were amplified and sequenced. The estimated average overall nucleotide substitution rate in the sequenced region was 7.8 x 10(-10) substitutions/site per year. The synonymous substitution rate was 2.11 x 10(-9) substitutions/synonymous site per year and the nonsynonymous substitution rate was 4.10 x 10(-10) substitutions/nonsynonymous site per year. Phylogenetic analyses showed that all of the wild Hordeum species examined clustered in a group separate from H. vulgare and H. vulgare ssp. spontaneum.

Base Sequence↗
PubMed1993

Anther culture and Hordeum bulbosum-derived barley doubled haploids: mutations and methylation.

Anther culture and Hordeum bulbosum-derived doubled haploid (DH) lines of barley (Hordeum vulgare L.) were analyzed for RFLP and RAPD polymorphisms. Polymorphisms were not detected in the anther culture- or H. bulbosum-derived DH lines among 273 RFLP and 89 polymerase chain reaction (PCR)-amplified DNA fragments assayed. It was calculated that base substitution or small deletion/insertion mutations had not been induced among 401,640 bp screened. Large deletion/insertion mutations were not observed among 33 Mb screened. Polymorphisms were observed when DNA was digested with the methylation-sensitive restriction enzymes HpaII and MspI: these RFLPs originated primarily from the anther culture-derived doubled haploids. The data indicate that heritable DNA methylation changes had occurred during DH production, particularly with the anther culture method.

Base Sequence↗
PubMed1992

Cloning and mapping of telomere-associated sequences from Hordeum vulgare L.

We present a novel approach to the efficient cloning of telomere-associated sequences and demonstrate its application to the cloning and mapping of these sequences from barley. The method is a modification of the Vectorette PCR technique and allows specific amplification and cloning of subtelomeric sequences. Telomere-associated sequences isolated from barley include hypervariable, repetitive sequences. Polymorphisms detected by subtelomeric markers behaved as Mendelian factors and were mapped to the most distal positions of barley chromosomes 1, 2, and 4.

Base Sequence↗
PubMed1981

[The effect of AR-L 115 BS on contractility dynamics of the isolated, metabolically supported dog heart (author's transl)].

In this study the contractility dynamics of the left ventricle are analysed independent of respiration, pre- and afterload, heart rate and perfusion pressure. These parameters alter varyingly following the administration of 2-[(2-methoxy-4-methylsufinyl)phenyl]-1H-imidazo[4,5-b]pyridine (AR-L 115 BS) so that the study plan selected must take these factors into account. The model employed consisted of a metabolically supported isolated pulsating dog's heart. The dose of AR-L 115 Bs used was 0.125 mg/kg body weight and it was infused over a period of 30 min. Altogether the study demonstrates that AR-L 115 BS has a marked positive inotropic effect on the isolated heart even under the conditions of infusion. The coronary resistance diminishes and the cardiac output increases.

Animals↗
PubMedDate not supplied

The hTERTalpha splice variant is a dominant negative inhibitor of telomerase activity.

The telomerase catalytic subunit (hTERT) is an essential component of the holoenzyme complex that adds telomeric repeats to the ends of human chromosomes. Maintenance of telomeres by telomerase or another mechanism is required for cell immortalization, and loss of telomeric DNA has been proposed as a trigger for cellular senescence. Available evidence suggests that regulation of telomerase activity primarily depends on transcriptional control of hTERT. However, several human tissues as well as some normal cell strains have been shown to express low levels of hTERT mRNA even though they lack telomerase activity. We have previously identified six splice variants of hTERT, including a "deletion" variant (hTERTalpha) that is missing conserved residues from the catalytic core of the protein. Several of the deletion variants have been detected in normal and developing human tissues. We now show that hTERTalpha inhibits endogenous telomerase activity, which results in telomere shortening and chromosome end-to-end fusions. Telomerase inhibition induced a senescence-like state in HT1080 cells and apoptosis in a jejunal fibroblast cell line. These results suggest a possible role for hTERT splice variants in the regulation of telomerase activity.

Alternative Splicing↗
PubMedDate not supplied

RFLP markers linked to the durable stem rust resistance gene Rpg1 in barley.

The gene, Rpg1, conferring stable resistance in barley to the wheat stem rust pathogen (Puccinia graminis f. sp. tritici) was mapped using two doubled haploid populations. Rpg1 mapped to the extreme subteleomeric region of barley chromosome 1P 0.3 and 1.1 cM proximal from the molecular markers ABG704 and plastocyanin (Plc), respectively, and 2.2 cM distal from MWG036B. The closest marker, ABG704, was sequenced and PCR-based markers were developed.

Base Sequence↗