Generation of a whole chromosome painting probe from monochromosomal hybrid cells by the alu-polymerase chain reaction

Fluorescent in situ hybridization (FISH) has become a widespread technique applicable in basic science and diagnostics. Chromosome painting represents a special application of FISH that has found increasing use in identification of complex chromosome rearrangements. Here we present a version of the Alu-PCR method modified to generate a whole chromosome painting probe (WCP) for human chromosome 19 using monochromosomal cell hybrids. In setting up conditions for this method, we established a cheap and fast approach to generation of WCPs for other human chromosomes that could be particularly useful for unambiguous identification of complex chromosomal rearrangements associated with cancer.

Purification and functional analysis of the recombinant protein isolated from E. coli by employing three different methods of bacterial lysis

In this paper, the purification of the human recombinant protein expressed in E. coli using the GSTGene Fusion System, by applying various methods of bacterial lysis: sonication, freeze/thaw and beadbeating, is presented. The study was an attempt to compare the properties of the proteins obtained by the sonication method, recommended by manufacturers but inaccessible for many researchers, with those obtained using two other readily available lysis methods. The data show that all purified proteins were soluble and intact with the highest protein yield being obtained via the freeze/thaw method. The results of functional analysis indicate that the proteins purified using the sonication and freeze/thaw methods of lysis exhibited similar DNA binding affinity, while the protein purified by beadbeating was also functional but with a lower binding affinity. The conclusion of this study is that all three lysis methods could be successfully employed for protein purification

6th Golden Helix Pharmacogenomics Day: pharmacogenomics and individualized therapy

Abstract The Golden Helix Pharmacogenomics Days are international scientific meetings aiming to educate healthcare professionals and biomedical scientists about pharmacogenomics and personalized medicine. In this meeting report, we provide an overview of the scientific lectures and the topics discussed during the 6th Golden Helix Pharmacogenomics Day that was held in Belgrade, Serbia last June 5, 2012. The scientific program included lectures by the local and international speakers from Europe and the United States.</p

Association of gene variants in TLR4 and IL-6 genes with Perthes disease

Uvod Pertesova bolest je idiopatska avaskularna osteonekroza proksimalne epifize femura koja se javlja kod dece. Etiologija ove bolesti je nepoznata. Tokom razvoja Pertesove bolesti zastupljen je proces zapaljenja, za koji je pokazano da utiče na remodelovanje koštanog tkiva. Cilj rada S obzirom na to da genetički faktori koji utiču na proces zapaljenja dosad nisu ispitivani kod Pertesove bolesti, cilj ovog istraživanja je bio da se utvrdi povezanost učestalosti varijanti u genima koji učestvuju u inflamatornom odgovoru, TLR4 (engl. toll-like receptor 4) i IL-6 (interleukin 6), i ove bolesti. Metode rada Ispitano je 37 dece s Pertesovom bolešću i 50 zdravih osoba. Metodom PCR-RFLP određeni su polimorfizmi medijatora zapaljenja TLR4 (Asp299Gly, Thr399Ile) i IL-6 (G-174C, G-597A). Rezultati Pokazano je da su polimorfizmi IL-6 G-174C i G-597A u našem ispitivanju bili u potpunoj gametskoj neravnoteži vezanosti. U kontrolnoj grupi je bilo statistički značajno više nosilaca heterozigotnog genotipa IL-6 G-174C/G-597A u poređenju sa grupom ispitanika s Pertesovom bolešću (p=0,047; OR=2,49; 95% CI=1,00-6,21). Takođe, grupa bolesnika nije bila u Hardi- Vajnbergovoj ravnoteži za polimorfizme IL-6 G-174C/G-597A. Nije primećena statistički značajna razlika u raspodeli genotipova za polimorfizme analizirane u TLR4 genu. Raspodela genotipova među grupama bolesnika formiranih na osnovu uzrasta kada se bolest pojavila nije pokazala statistički značajnu povezanost s analiziranim polimorfizmima. Zaključak Naše istraživanje je pokazalo da su nosioci heterozigotnog genotipa za IL-6 G-174C/G-597A polimorfizme bili značajno češći u kontrolnoj grupi nego u grupi dece obolele od Pertesove bolesti. Na osnovu toga zaključili smo da je kod dece koja su nosioci heterozigotnog genotipa za ove polimorfizme manja verovatnoća za razvoj Pertesove bolesti nego kod nosilaca oba homozigotna genotipa.Introduction Perthes disease is idiopathic avascular osteonecrosis of the hip in children, with unknown etiology. Inflammation is present during development of Perthes disease and it is known that this process influences bone remodeling. Objective Since genetic studies related to inflammation have not been performed in Perthes disease so far, the aim of this study was to analyze the association of frequencies of genetic variants of immune response genes, toll-like receptor 4 (TLR4) and interleukin-6 (IL-6), with this disease. Methods The study cohort consisted of 37 patients with Perthes disease and 50 healthy controls. Polymorphisms of well described inflammatory mediators: TLR4 (Asp299Gly, Thr399Ile) and IL-6 (G-174C, G- 597A) were determined by polymerase chain reaction restriction fragment length polymorphism method. Results IL-6 G-174C and G-597A polymorphisms were in complete linkage disequilibrium. A statistically significant increase of heterozygote subjects for IL-6 G-174C/G-597A was found in controls in comparison to Perthes patient group (p=0.047, OR=2.49, 95% CI=1.00-6.21). Also, the patient group for IL-6 G-174C/G- 597A polymorphisms was not in Hardy-Weinberg equilibrium. No statistically significant differences were found between patient and control groups for TLR4 analyzed polymorphisms. A stratified analysis by the age at disease onset also did not reveal any significant difference for all analyzed polymorphisms. Conclusion Our study revealed that heterozygote subjects for the IL-6 G-174C/G-597A polymorphisms were significantly overrepresented in the control group than in the Perthes patient group. Consequently, we concluded that children who are heterozygous for these polymorphisms have a lower chance of developing Perthes disease than carriers of both homozygote genotypes