HAX-1 overexpression, splicing and cellular localization in tumors

<p>Abstract</p> <p>Background</p> <p>HAX-1 has been described as a protein potentially involved in carcinogenesis and especially metastasis. Its involvement in regulation of apoptosis and cell migration along with some data indicating its overexpression in cancer cell lines and tumors suggests that HAX-1 may play a role in neoplastic transformation. Here we present the first systematic analysis of HAX-1 expression in several solid tumors.</p> <p>Methods</p> <p>Using quantitative RT-PCR, we have determined the mRNA levels of <it>HAX1 </it>splice variant I in several solid tumors. We have also analyzed by semiquantitative and quantitative RT-PCR the expression of five <it>HAX-1 </it>splice variants in breast cancer samples and in normal tissue from the same individuals. Quantitative PCR was also employed to analyze the effect of estrogen on <it>HAX1 </it>expression in breast cancer cell line. Immunohistochemical analysis of HAX-1 was performed on normal and breast cancer samples.</p> <p>Results</p> <p>The results reveal statistically important <it>HAX1 </it>up-regulation in breast cancer, lung cancer and melanoma, along with some minor variations in the splicing pattern. HAX-1 up-regulation in breast cancer samples was confirmed by immunohistochemical analysis, which also revealed an intriguing HAX-1 localization in the nuclei of the tumor cells, associated with strong ER status.</p> <p>Conclusion</p> <p>HAX-1 elevated levels in cancer tissues point to its involvement in neoplastic transformation, especially in breast cancer. The connection between HAX-1 nuclear location and ER status in breast cancer samples remains to be clarified.</p

Tests of multiple molecular markers for the identification of Great Spotted and Syrian Woodpeckers and their hybrids

Great Spotted and Syrian Woodpeckers (Dendrocopos major and D. syriacus) are known to hybridize in nature; however, the extent of this phenomenon is not known due to difficulties in hybrid detection based on plumage analyses. Here, we tested five markers (one mitochondrial and four nuclear) and a set of six microsatellite loci for the identification of these two Woodpeckers and their hybrids. Sequencing of DNA from 26 individuals of both Woodpeckers from different parts of their ranges: one allopatric (D. major; Norway) and two sympatric (Poland and Bulgaria) showed that both species can be clearly separated based on all sequence markers. The highest number of fixed nucleotide sites were found in the mtDNA control region and intron 5 of the transforming growth factor. Analyses of microsatellite data distinguished the two species, but all loci showed a large number of common alleles and their utility in identifying hybrids is therefore doubtful. According to the DNA sequence analyses, 2 out of 18 specimens within the sympatric range in Poland were identified as possible hybrids, most probably paternal backcrosses. Moreover, both hybrids are from synantropic populations (settled in cities), whereas none of the D. major sampled in forests and in its allopatric range (Norway) showed signs of an intermixed genotype. Further research on hybridization and introgression in woodpeckers is undoubtedly needed and could be useful for understanding ecological and ethological interactions among these species, particularly for D. syriacus, which is relatively rare in Europe

Cardiopoietic cell therapy for advanced ischemic heart failure: results at 39 weeks of the prospective, randomized, double blind, sham-controlled CHART-1 clinical trial

Cardiopoietic cells, produced through cardiogenic conditioning of patients' mesenchymal stem cells, have shown preliminary efficacy. The Congestive Heart Failure Cardiopoietic Regenerative Therapy (CHART-1) trial aimed to validate cardiopoiesis-based biotherapy in a larger heart failure cohort

Discrimination of amino acids by electronic tongue

Obecnie potrzebne są metody szybkiego, dokładnego, a przede wszystkim taniego oznaczania aminokwasów w próbkach biologicznych. Do takich technik należy potencjometryczny elektroniczny język, który pozwala na rozróżnianie próbek ciekłych. W pracy przedstawiono zastosowanie potencjometrycznego elektronicznego języka do klasyfikowania 5 aminokwasów. Sprawdzono możliwość rozpoznania poszczególnych analitów w roztworach o różnych pH, a następnie wykorzystując metodę częściowych najmniejszych kwadratów (PLS) rozróżniono poszczególne aminokwasy.In the last few years medical discoveries showed that presence of amino acids can be associated with many diseases. Usually, these compounds are determined using HPLC method (high performance liquid chromatography), however, due to its noxiousness, new methods are investigated. Application of electronic tongue system can be an alternative method for fast, accurate, affordable determination of amino acids in biological samples. This device is used for analysis and classification of complex liquid samples. Its main advantage is the ability to distinguish between samples containing various components on-line, without prior sample preparation and modification. In this paper there are presented preliminary studies on the recognition of 5 amino acids (glutamic acid, alanine, serine, arginine, aspartic acid) with use of a sensor array composed of ion-selective electrodes of various selectivity. Chemical images of the samples were obtained by measuring the electrode signals in solutions of amino acids for various pH ranging from 2.5 to 11.5. The possibility of identification of individual analytes in solutions of different pH was examined with use of the Partial Least Squares Analysis

Dairy products discrimination according to the milk type using an electrochemical multisensor device coupled with chemometric tools

This study shows the potential application of a potentiometric electronic tongue coupled with a lab-made DataLogger device for the classification of dairy products according to the type of milk used in their production, i.e., natural, fermented and UHT milk. The electronic tongue device merged a commercial pH electrode and 15 lipid/polymeric membranes, which were obtained by a drop-by-drop technique. The potentiometric signal profiles gathered from the 16 sensors, during the analysis of the 11 dairy products (with ten replicate samples), together with principal component analysis showed that dairy samples could be naturally grouped according to the three types of milk evaluated. To further investigate and verify this capability, a linear discriminant analysis together with a simulated annealing variable selection algorithm was also applied to the electrochemical data, which were randomly split into two datasets, one used for model training and internal-validation using a repeated K-fold cross-validation procedure (with 64% of the data); and the other for external validation purposes (containing the remaining 36% of the data). The multivariate supervised strategy used allowed establishing a classification model, based on the potentiometric information of four sensor lipid membranes, which enabled achieving a successful discrimination rate of 100% for both internal-and external-validation processes. The demonstrated versatility of the built electronic tongue for discriminating Dairy products according to the type of milk used in their production combined with its simplicity, low-cost and fast time analysis may envisage a possible future application in dairy industry

Antioxidant synergism and antagonism between tocotrienols, quercetin and rutin in model system

The aim of the study was to reveal antioxidant synergism or antagonism between quercetin, rutin and selected tocotrienols in linoleic acid emulsion. The oxidative stress was generated by 2,2′-azobis(2-amidinopropane) dihydrochloride (AAPH) or CuSO4; the increase of the concentration of peroxidation products was monitored using fluorescence probe 2,7-dichlorofluorescein (DCF). The antioxidant activity of tested substances depends on the form of the antioxidant (aglycone, glycoside), its concentration, localization in the emulsion, and the factors generating oxidative stress. The synergistic effect occurred when the effectiveness of individual antioxidant was relatively weak and mainly when the concentration of antioxidants was in the physiologically significant range of 1 μM. We suggest that tocotrienols were regenerated by flavonoids. The synergism benefitted from the proximity of the localization of interacting antioxidants (e.g. the presence of one of the antioxidants at the oil-water interface)

Electrochemical methods in bioanalytics : selected aspects

The cooperation of scientists specializing in different fields has given rise to the integration of previously distinct thematic areas and creation of new multidimensional disciplines as a result. Biochemistry, which has derived from the borderline of chemistry and biology, can be set as a good example. In this short review an insight into electrochemical studies, which are currently carried out in the Department of Microbioanalytics at the Faculty of Chemistry (Warsaw University of Technology), was presented. Three independent scientific pathways introducing electrochemical methods for biochemical and bioanalytical purposes can be distinguished among the ongoing researches. The first one embraces the design of the so-called electronic tongue – a system used for the qualitative and quantitative analysis of liquid samples of complex composition. In this work, potentiometric sensor arrays were applied to develop an electronic tongue system enabling the evaluation of the effectiveness of bitter taste masking of pharmaceuticals. The second scientific pathway involves voltammetric studies of the interactions of biologically active peptides with copper(II) ions. The interest was drawn to clarify and describe the role of β-amyloid and NSFRY copper(II) complexes, relevant to Alzheimer’s disease occurrence and cardiovascular system disorder respectively. Finally, boronic acids and their derivatives, exhibiting the affinity for molecules possessing 1,2 or 1,3-diol group in their structure, were used as selective molecular receptors in the third research project. The studies include the selection of the optimal method and conditions of the immobilization process, providing the most favorable receptor layer structure, and the determination of the performances of constructed electrochemical sensor towards particular bioanalytes