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1

Optimalization and validation of ADSRRS-fingerprinting

100%
Krawczyk B., Lewandowski K.
Biotechnologia
|
2007
|
issue 2
95-113
EN
Macrorestriction analysis of genomic DNA followed by pulsed-field gel electrophoresis (REA-PFGE) has become the 'gold standard' for molecular typing (10). In recent years, some alternative techniques have been successfully applied. However, there is still a need for a new method which may appear to be less complex, cheaper, and have a power of discrimination at least similar to that of REA-PFGE. Recently, we showed the performance and convenience of a novel assay, based on the fingerprinting of bacterial genomes by amplification of DNA fragments surrounding rare restriction sites (ADSRRS-fingerprinting), for its potential usefulness in epidemiological investigations. In this study, ADSRRS-fingerprinting method was optimized at each stage of the procedure. The modified method, named ADSRRS-fingerprinting plus, differs from the original technique in several points. The optimized method considerably shortens the time of experiment, offers good discriminatory power and also demonstrates excellent reproducibility. Next, ADSRRS-fingerprinting plus was validated. These experiments proved that the method is specific, selective and suitable for intended purpose and also confirmed its reliability, reproducibility and simplicity in the interpretation of the results. Optimized and validated procedure of ADSRRS-fingerprinting plus was used in the development of a diagnostic kit (ADSRRS-KIT) for strain differentiation of bacteria below the species level, using a simple set of ready-to-use reagents and standard equipment.
2
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Diblock Copolymer Melt in Spherical Unit Cells of Higher Dimensionalities

81%
Banaszak M., Koper A., Knychała P., Lewandowski K.
Acta Physica Polonica A
|
2012
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vol. 121
|
issue 3
703-710
EN
Using self-consistent field theory in spherical unit cells of various dimensionality, D = 1, 2, 3, and 4, we calculate phase diagram of a diblock, A-b-B, copolymer melt in 4-dimensional space, d = 4. The phase diagram is parameterized by the chain composition, f, and incompatibility between A and B, quantified by the product χ N. We predict 4 stable nanophases: layers, cylinders, 3D spherical cells, and 4D spherical cells. We also calculate order-disorder and order-order transition lines. In the strong segregation limit, that is for large χ N, the order-order transition compositions are determined by the strong segregation theory in its simplest form, for D = 1, 2, 3, and 4.
3

Blood proliferative diseases ? application of the DNA microarrays for transcriptome analysis

71%
Handschuh L., Lewandowski K., Kazmierczak M., Komarnicki M., Figlerowicz M.
Biotechnologia
|
2009
|
issue 3
22-43
EN
Hematopoiesis is a complex process precisely regulated by a wide spectrum of cooperating factors. Dysfunction of hematopoietic cell proliferation, differentiation or maturation usually leads to the malignant transformation. The DNA microarray-based transcriptome analysis helped to revise the traditional classification of hematological disorders, predict their outcome, test potential therapeutic agents and better understand basic mechanisms underlying cancer origin and development. Here, the results of gene expression profiling in myelo- and lymphoproliferative diseases such as leukemia, lymphoma and myelodysplastic syndromes, are presented. Two microarray technologies were applied in this area of research: Affymetrix gene chips and cDNA microarrays. Among them, Lymphochip is a prominent example of a specialized cDNA microarray tool designed to investigate gene expression in the immunological system and hematological diseases. It seems that typical problems connected with microarray results analysis ? small number of patients, loss of reproducibility can be overcome by increasing the number of samples and application of identical protocols, equipment and reagents in different laboratories.
4

Dasatinib treatment can overcome imatinib and nilotinib resistance in CML patient carrying F359I mutation of BCR-ABL oncogene

61%
Baranska M., Lewandowski K., Gniot M., Iwola M., Lewandowska., Komarnicki M.
Journal of Applied Genetics
|
2008
|
vol. 49
|
issue 2
201-203
EN
Point mutations of bcr-abl tyrosine kinase are the most frequent causes of imatinib resistance in chronic myeloid leukaemia (CML) patients. In most CML cases with BCR-ABL mutations leading to imatinib resistance the second generation of tyrosine kinase inhibitors (TKI- e.g. nilotinib or dasatinib) may be effective. Here, we report a case of a CML patient who during imatinib treatment did not obtain clinical and cytogenetic response within 12 months of therapy. The sequencing of BCR-ABL kinase domains was performed and revealed the presence of a F359I point mutation (TTC-to-ATC nucleotide change leading to Phe-to-Ile amino acid substitution). After 1 month of nilotinib therapy a rapid progression of clinical symptoms was observed. In the presence of the F359I point mutation only dasatinib treatment overcame imatinib and nilotinib resistance.
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