45 research outputs found

    Power efficiency management of photovoltaic energy source based on MPPT algorithm

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    In the paper energy supply system based on photovoltaic (PV) arrays was described. Also models of a single PV cell and a voltage boost converter were described. The boost converter was used for holding an appropriate work state of the PV arrays associated with its maximum power point level in various work conditions associated with irradiance level and the arrays temperature. Finally, comparison of two strategies of voltage level control in PV arrays system was put forward. These strategies were used to attain the maximum power point, and to define the work conditions, in which described control algorithms are the most effective

    Liposome-based DNA carriers may induce cellular stress response and change gene expression pattern in transfected cells

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    <p>Abstract</p> <p>Background</p> <p>During functional studies on the rat stress-inducible <it>Hspa1b </it>(<it>hsp70.1</it>) gene we noticed that some liposome-based DNA carriers, which are used for transfection, induce its promoter activity. This observation concerned commercial liposome formulations (LA), Lipofectin and Lipofectamine 2000. This work was aimed to understand better the mechanism of this phenomenon and its potential biological and practical consequences.</p> <p>Results</p> <p>We found that a reporter gene driven by <it>Hspa1b </it>promoter is activated both in the case of transient transfections and in the stably transfected cells treated with LA. Using several deletion clones containing different fragments of <it>Hspa1b </it>promoter, we found that the regulatory elements responsible for most efficient LA-driven inducibility were located between nucleotides -269 and +85, relative to the transcription start site. Further studies showed that the induction mechanism was independent of the classical HSE-HSF interaction that is responsible for gene activation during heat stress. Using DNA microarrays we also detected significant activation of the endogenous <it>Hspa1b </it>gene in cells treated with Lipofectamine 2000. Several other stress genes were also induced, along with numerous genes involved in cellular metabolism, cell cycle control and pro-apoptotic pathways.</p> <p>Conclusions</p> <p>Our observations suggest that i) some cationic liposomes may not be suitable for functional studies on <it>hsp </it>promoters, ii) lipofection may cause unintended changes in global gene expression in the transfected cells.</p

    Differential expression of HSPA1 and HSPA2 proteins in human tissues; tissue microarray-based immunohistochemical study

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    In the present study we determined the expression pattern of HSPA1 and HSPA2 proteins in various normal human tissues by tissue-microarray based immunohistochemical analysis. Both proteins belong to the HSPA (HSP70) family of heat shock proteins. The HSPA2 is encoded by the gene originally defined as testis-specific, while HSPA1 is encoded by the stress-inducible genes (HSPA1A and HSPA1B). Our study revealed that both proteins are expressed only in some tissues from the 24 ones examined. HSPA2 was detected in adrenal gland, bronchus, cerebellum, cerebrum, colon, esophagus, kidney, skin, small intestine, stomach and testis, but not in adipose tissue, bladder, breast, cardiac muscle, diaphragm, liver, lung, lymph node, pancreas, prostate, skeletal muscle, spleen, thyroid. Expression of HSPA1 was detected in adrenal gland, bladder, breast, bronchus, cardiac muscle, esophagus, kidney, prostate, skin, but not in other tissues examined. Moreover, HSPA2 and HSPA1 proteins were found to be expressed in a cell-type-specific manner. The most pronounced cell-type expression pattern was found for HSPA2 protein. In the case of stratified squamous epithelia of the skin and esophagus, as well as in ciliated pseudostratified columnar epithelium lining respiratory tract, the HSPA2 positive cells were located in the basal layer. In the colon, small intestine and bronchus epithelia HSPA2 was detected in goblet cells. In adrenal gland cortex HSPA2 expression was limited to cells of zona reticularis. The presented results clearly show that certain human tissues constitutively express varying levels of HSPA1 and HSPA2 proteins in a highly differentiated way. Thus, our study can help designing experimental models suitable for cell- and tissue-type-specific functional differences between HSPA2 and HSPA1 proteins in human tissues

    Validation of selected molecular methods for the mutations determination in codons 12 and 13 of K-RAS gene in five Polish oncological research centers

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    Chorzy na raka jelita grubego z przerzutami mogą osiągnąć korzyść z leczenia panitumumabem jedynie, jeśli w guzie nie stwierdzono mutacji w genie K-RAS. W związku z tym konieczne jest zbadanie statusu tego genu w celu wyłonienia chorych, którzy mogą być poddani takiemu leczeniu. Celem pracy było opracowanie standardowej procedury oznaczania statusu genu K-RAS w materiale izolowanym z bloczków parafinowych. Kolejnym celem była walidacja wybranych technik molekularnych oznaczania mutacji w pięciu ośrodkach w Polsce, w których odbywa się leczenie chorych na raka jelita grubego. Ocenie poddano cztery różne techniki oznaczania mutacji: SSCP, DHPLC, RFLP/PCR i bezpośrednie sekwencjonowanie. Stwierdzono, że wszystkie jednostki uczestniczące w procesie walidacji są odpowiednio przygotowane do podjęcia działalności diagnostycznej w zakresie oznaczania statusu genu K-RAS. Przyjęto następujące zalecenia dla laboratoriów diagnostycznych: 1. Materiał do izolacji DNA powinien zawierać przynajmniej 70% utkania nowotworowego; 2. Ujednolicenie procedury izolacji DNA ze skrawków parafinowych wymaga stosowania gotowego zestawu do izolacji DNA; 3. W przypadku braku jednoznacznego wyniku konieczne jest stosowanie dwóch metod oznaczania mutacji, przy czym jedną z nich powinno być sekwencjonowanie bezpośrednie.Metastatic colorectal cancer patients will benefit from treatment with panitumumab only when they don't have mutation in K-RAS gene. Therefore, estimation of mutational status of K-RAS is necessary for the selection of patients, who should be treated with panitumumab. The aim of this study was to evolve a standard method of estimation of K-RAS mutational status in the material isolated from paraffin blocs. The second aim was the validation of selected molecular methods of K-RAS mutation evaluation in five Polish oncological centers where mCRC patients are treated. Four methods were evaluated: SSCP, DHPLC, RFLP/PCR and direct sequencing. We found that all groups in five selected oncological centers, who took part in the validation process, were well prepared for molecular diagnosis of K-RAS mutational status. The following recommendations for diagnostic laboratories were approved: 1. At least 70% of cancer cells should be present in a tissue for DNA isolation; 2. The method of DNA isolation should be standardized, the most appropriate is usage of DNA isolation kits; 3. In case of equivocal results two independent molecular methods should be employed, one of them should be direct sequencing

    Mieczysław Chorąży

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