Reproductive Performance of Native Pulawska and High Productivity Polish Landrace Sows in the Context of Stress During the Period of Early Pregnancy.

Experiments revealed that it is necessary to get rid of genotypes RYR1C/T and RYR1T/T because of problems with meat quality and reproduction of gilts and sows. This phenomenon, however, is of individual nature and is characterized by high variability. The aim of the study was to analyse the influence of stressogenic factors in native Pulawska and high productivity Polish Landrace breed sows during the period of low pregnancy on reproductive performance and maternal behaviour. Sows in the 2nd reproduction cycle were assigned into groups of five animals as follows: control (C) of RYR1 C/C genotype and experimental (E) of RYR1 C/T genotype. In all, 30 Pulawska breed sows and 30 PL breed sows were analysed. During the early stage of pregnancy (days 42-84), experimental groups were subjected to the action of stressogenic factors: elevated temperature, noisy and rough treatment of sows by service personnel and immobilization stress. The effect of stressogenic factors was referred to the reproductive performance of sows. Pulawska and PL breed sows from the experimental groups were subjected to the evaluation of maternal traits on the basis of perinatal behaviour determined with the help of an ethogram. Analysis of the results of reproduction and the behaviour of the sows confirmed the higher resistance to stressogenic factors of the Pulawska breed. The performed analysis of reproduction and behaviour confirmed that heterozygotic genotypes in the RYR1 locus exhibited less advantageous reproduction, which corroborates the hypothesis about a negative impact of the T allele on swine productivity.

Four-dimensional Fano toric complete intersections.

We find at least 527 new four-dimensional Fano manifolds, each of which is a complete intersection in a smooth toric Fano manifold.

Ensembl 2006.

The Ensembl (http://www.ensembl.org/) project provides a comprehensive and integrated source of annotation of large genome sequences. Over the last year the number of genomes available from the Ensembl site has increased from 4 to 19, with the addition of the mammalian genomes of Rhesus macaque and Opossum, the chordate genome of Ciona intestinalis and the import and integration of the yeast genome. The year has also seen extensive improvements to both data analysis and presentation, with the introduction of a redesigned website, the addition of RNA gene and regulatory annotation and substantial improvements to the integration of human genome variation data.

Ensembl 2005.

The Ensembl (http://www.ensembl.org/) project provides a comprehensive and integrated source of annotation of large genome sequences. Over the last year the number of genomes available from the Ensembl site has increased by 7 to 16, with the addition of the six vertebrate genomes of chimpanzee, dog, cow, chicken, tetraodon and frog and the insect genome of honeybee. The majority have been annotated automatically using the Ensembl gene build system, showing its flexibility to reliably annotate a wide variety of genomes. With the increased number of vertebrate genomes, the comparative analysis provided to users has been greatly improved, with new website interfaces allowing annotation of different genomes to be directly compared. The Ensembl software system is being increasingly widely reused in different projects showing the benefits of a completely open approach to software development and distribution.

Ensembl 2004.

The Ensembl (http://www.ensembl.org/) database project provides a bioinformatics framework to organize biology around the sequences of large genomes. It is a comprehensive and integrated source of annotation of large genome sequences, available via interactive website, web services or flat files. As well as being one of the leading sources of genome annotation, Ensembl is an open source software engineering project to develop a portable system able to handle very large genomes and associated requirements. The facilities of the system range from sequence analysis to data storage and visualization and installations exist around the world both in companies and at academic sites. With a total of nine genome sequences available from Ensembl and more genomes to follow, recent developments have focused mainly on closer integration between genomes and external data.

Ensembl 2002: accommodating comparative genomics.

The Ensembl (http://www.ensembl.org/) database project provides a bioinformatics framework to organise biology around the sequences of large genomes. It is a comprehensive source of stable automatic annotation of human, mouse and other genome sequences, available as either an interactive web site or as flat files. Ensembl also integrates manually annotated gene structures from external sources where available. As well as being one of the leading sources of genome annotation, Ensembl is an open source software engineering project to develop a portable system able to handle very large genomes and associated requirements. These range from sequence analysis to data storage and visualisation and installations exist around the world in both companies and at academic sites. With both human and mouse genome sequences available and more vertebrate sequences to follow, many of the recent developments in Ensembl have focusing on developing automatic comparative genome analysis and visualisation.

The Ensembl genome database project.

The Ensembl (http://www.ensembl.org/) database project provides a bioinformatics framework to organise biology around the sequences of large genomes. It is a comprehensive source of stable automatic annotation of the human genome sequence, with confirmed gene predictions that have been integrated with external data sources, and is available as either an interactive web site or as flat files. It is also an open source software engineering project to develop a portable system able to handle very large genomes and associated requirements from sequence analysis to data storage and visualisation. The Ensembl site is one of the leading sources of human genome sequence annotation and provided much of the analysis for publication by the international human genome project of the draft genome. The Ensembl system is being installed around the world in both companies and academic sites on machines ranging from supercomputers to laptops.

Cancer and genomics.

Identification of the genes that cause oncogenesis is a central aim of cancer research. We searched the proteins predicted from the draft human genome sequence for paralogues of known tumour suppressor genes, but no novel genes were identified. We then assessed whether it was possible to search directly for oncogenic sequence changes in cancer cells by comparing cancer genome sequences against the draft genome. Apparently chimaeric transcripts (from oncogenic fusion genes generated by chromosomal translocations, the ends of which mapped to different genomic locations) were detected to the same degree in both normal and neoplastic tissues, indicating a significant level of false positives. Our experiment underscores the limited amount and variable quality of DNA sequence from cancer cells that is currently available.

Integration of cytogenetic landmarks into the draft sequence of the human genome.

We have placed 7,600 cytogenetically defined landmarks on the draft sequence of the human genome to help with the characterization of genes altered by gross chromosomal aberrations that cause human disease. The landmarks are large-insert clones mapped to chromosome bands by fluorescence in situ hybridization. Each clone contains a sequence tag that is positioned on the genomic sequence. This genome-wide set of sequence-anchored clones allows structural and functional analyses of the genome. This resource represents the first comprehensive integration of cytogenetic, radiation hybrid, linkage and sequence maps of the human genome; provides an independent validation of the sequence map and framework for contig order and orientation; surveys the genome for large-scale duplications, which are likely to require special attention during sequence assembly; and allows a stringent assessment of sequence differences between the dark and light bands of chromosomes. It also provides insight into large-scale chromatin structure and the evolution of chromosomes and gene families and will accelerate our understanding of the molecular bases of human disease and cancer.

Malignancy: Case report: Myelodysplasia Following Treatment of Chronic Lymphocytic Leukaemia with Fludarabine.

Recently there has been an increased awareness of a possible link between the use of purine nucleoside analogues and myelodysplasia. We report the case of a patient who developed myelodysplasia with complex cytogenetic changes after receiving fludarabine. We review the literature, discuss the possible links between myelodysplasia and nucleoside analogues and putative mechanisms for secondary neoplasia.

Investigation of clonal involvement of myeloid cells in Philadelphia-positive and high hyperdiploid acute lymphoblastic leukemia.

Acute lymphoblastic leukemia (ALL) with a high hyperdiploid clone has a good prognosis for both childhood and adult patients while patients with Philadelphia-positive (Ph) ALL do badly at all age groups. It has been suggested that different responses to treatment might be related to the cell of origin of the leukemia with 'stem cell' cases responding less well than those arising in a lymphoid committed progenitor cell. The clonal involvement of different cell lineages in 12 patients with acute lymphoblastic leukemia has been examined by applying fluorescence in situ hybridization (FISH) to detect chromosomal abnormalities in bone marrow cells previously identified by morphology and/or immunology. The karyotype of the malignant clone was either high hyperdiploid or Philadelphia translocation (Ph) positive with a breakpoint in the minor breakpoint cluster region of the BCRgene (m-BCR) or in the major breakpoint cluster region of the BCR gene (M-BCR). The high hyperdiploid clone, in each case, was found in cells of the B-lymphoid (CD19+) lineage but not in T cells (CD3+) or in cells of the myeloid (CD13+) or erythroid (glycophorin A+) lineages, indicative of a lymphoid committed progenitor cell. Heterogeneity of lineage involvement was found in Ph+ ALL: the m-BCR Ph+ clone was found in lymphoid/blast cells but not in neutrophils or eosinophils. In contrast both M-BCR Ph+ clones were detected in myeloid as well as lymphoid lineages, indicative of a stem cell origin.

Myelodysplasia Following Treatment of Chronic Lymphocytic Leukaemia with Fludarabine.

Recently there has been an increased awareness of a possible link between the use of purine nucleoside analogues and myelodysplasia. We report the case of a patient who developed myelodysplasia with complex cytogenetic changes after receiving fludarabine. We review the literature, discuss the possible links between myelodysplasia and nucleoside analogues and putative mechanisms for secondary neoplasia.

Increased sensitivity of minimal residual disease detection by interphase FISH in acute lymphoblastic leukemia with hyperdiploidy.

Interphase fluorescent in situ hybridization (FISH) for the detection of minimal residual disease detection (MRD) in patients with acute lymphoblastic leukemia (ALL) and a high hyperdiploid clone (>50 chromosomes) at diagnosis is presented. By simultaneous targetting of three chromosomes gained, a sensitivity of 10(4) was achieved. Control values (mean + s.d. x 2 of false positive cells in normal peripheral blood) using probes singly, in pairs or as a triplet were 1.4-2.0%, 0.01-0.02% and zero (in 10(4) cells), respectively. A serial dilution experiment mixing control blood lymphocytes with bone marrow from a patient with a >85% cells with >50 chromosomes and probing it with single, dual or triple probes, revealed the clone down to dilutions of 10(-1), 10(-3) and 10(-4), respectively. FISH confirmed chromosomal gains at diagnosis (13 cases) and in relapse (three cases). Positive remission samples (0.01 to 0.06% cells with chromosome gains) were more frequent during the first 7 months (7/12) from diagnosis than later (3/11). Serial studies showed a decline in clone size with time. Conversion from a negative to a positive sample heralded relapse in one case. FISH confirmed an isolated central nervous system relapse and detected a hyperdiploid clone in a chromosomally normal relapse. This technique could be applied whenever three cytogenetic/genetic changes are found.

Single-cell trisomy in hematologic malignancy. Random change or tip of the iceberg?

Finding a clone in the bone marrow of a patient with a hematologic disorder is important to confirm the neoplastic nature of the disease and may be indicative of prognosis. Since cytogenetic analysis detects only actively dividing clones, the presence of a single abnormal cell among 20 cells analyzed raises doubts about its clonal nature. Fluorescence in situ hybridization (FISH) enables rapid detection of certain chromosomal abnormalities in metaphase and interphase cells, thus enabling detection of minor or inactive clones. Seven patients with hematologic malignancy each having random cell(s) were investigated thus: at diagnosis, with MDS and a cell with +8 (two cases) or +9 (one case) and with AML and a cell with +4 (one case), +7 (one case), or two cells with +9, +22/ +10, +17, +17 (one case). One patient with ALL in remission had one cell with trisomy 4. One patient, a male aged 66 years with refractory anemia with ringed sideroblasts, was found to have a minor trisomy 8 clone in his diagnostic marrow. A follow-up marrow 42 months later showed no trisomy 8 cell among 62 metaphases analyzed, and the percentage of trisomic cells using FISH on interphase cells was within the control range. This patient has survived for more than 42 months requiring no treatment. Single-cell abnormalities in the other six cases proved to be random events. Thus it appears that single-cell abnormalities may not be clonal or at most indicate the presence of a minor clone well below the level of cytogenetic detection. The prognostic significance of such minor clones is at present unclear.

Detection of the BCR-ABL gene by reverse transcription/polymerase chain reaction and fluorescence in situ hybridization in a patient with Philadelphia chromosome negative acute lymphoblastic leukaemia.

The Philadelphia (Ph) chromosome is detected in leukaemia cells in approximately 20% of adults with acute lymphoblastic leukaemia (ALL). When treated with chemotherapy alone, Ph-positive ALL has a poor prognosis, and patients may benefit from bone marrow transplantation in first remission. Here we report a patient with chromosomally normal bone marrow, in all 60 cells analysed, who was found to have the p210-type BCR-ABL chimaeric transcript by RT/PCR. Fluorescence in situ hybridization was labelled cosmid probes for BCR and ABL showed the presence of BCR-ABL juxtaposition on a normal chromosome 22 in leukaemia cell metaphases. We conclude that molecular and cytogenetic methods should be used in conjunction to detect the BCR-ABL gene rearrangement in ALL.

[The dynamics of the drug release from suppositories. Part 5: Liberation of chlorpromazine from suppository bases (author's transl)]. / Dynamik der Arzneimittelabagabe aus Suppositorien. 5. Mitteilung: Liberation von Chlorpromazin aus Zäpfchengrundlagen.

The authors determined the liberation of chlorpromazine hydrochloride from suppositories prepared with the polyethylene glycols PEG 1000, 1500, 2000, 4000, 6000 and their mixtures as well as with the lipophil bases cocoa butter, Witepsol W35 and Witepsol H15. The determinations were made with the apparatus described by Kerckhoffs and Huinziga using a phosphate buffer (pH = 7.4). It was found that the liberation was most rapid from hydrophilic polyethylene glycol bases; and the release rates decreased in the following order: PGE 4000 (75%) + PEG (25%) leads to PEG 4000 (75%) + glycerol (25%) leads to PEG 2000 leads to PEG 1500 (70%( + PEG 6000 (30%) leads to PEG 4000 leads to PEG 6000. The release from lipophil bases was poor.

[The dynamics of drug release from suppositories. Part 6: The effects of lipophil and hydrophilic suppository bases on the release of adeiphenin (author's transl)]. / Dynamik der Arzneimittelabgabe aus Suppositorien. 6. Mitteilung: Einfluss lipophiler und hydrophiler Zäpfchengrundlagen auf die Liberation von Adiphenin.

On studying the release of adiphenin from suppositories, the authors found that it is most rapid from hydrophilic bases. The release rate decreased in the following order: PEG 1500 leads to PEG 2000 leads to PEG 4000 (70%) + Glycerol (30%) leads to PEG 1000 (70%) + Peg 4000 (30%) leads to PEG 4000 leads to PEG 6000. The release is considerably slower from the bases Witepsol W35, Witepsol H15 and cacao butter.