Insulin-Like Growth Factor-2 Is Induced Following 5-Aminolevulinic Acid-Mediated Photodynamic Therapy in SW620 Human Colon Cancer Cell Line

The IGF system is a family of polypeptide growth factors, which plays a significant role in the development and growth of many cells. Dysregulation of insulin-like growth factors and their pathway components has been connected with essential tumor properties, such as tumor cell proliferation, antiapoptotic properties, invasive behavior and chemotherapy resistance. However, the effects of photodynamic therapy (PDT), one of the cancer treatment methods for the regulation of the IGF signaling pathway, are still unclear. The aim of this study was to investigate the expression of IGF-2 after 5-aminolevulinic acid (5-ALA)-mediated-PDT in SW620 human colorectal cancer cells with evaluation of cell proliferation and apoptosis and to determine the effects of PDT on the IGF-2 receptor (IGF-2R), IGF-2 binding protein-1 (IGF-2BP-1) and the proapoptotic protein, BAX. Cells were treated with 5-aminolevulinic acid and its methyl ester. Changes of the expression and concentration of IGF-2 before and after treatment were assayed by immunocytochemistry, Western blot and ELISA. We found that IGF-2 was significantly overexpressed in the SW620 cell line, while its receptor and binding protein-1 were not significantly changed. Within this study, we would like to suggest that IGF-2 contributes to the effects of PDT and that its expression will influence post-PDT efficacy.     

In Search of Concerted Strategies for Competitive and Resilient Regions

The European space-economy represents a complex system with a great internal heterogeneity, intensive socioeconomic interactions and differential growth trajectories among countries and regions. The present study aims to investigate the connectivity between spatial competitiveness and resilience in Europe and seeks to design an operational framework for concerted strategies of competitive and resilient regions. To assess the linkage between resilience and competitiveness, we have developed a new measure, viz. the Resilience and Competitiveness Index (RACI) as a function of two constituent sub-indices: Resilience and Competitiveness. This approach is tested on the basis of detailed data on European regions. The empirical results from 268 EU NUTS2 regions offer a solid anchor point for the proposed operational framework for concerted development strategies of competitive and resilient regions. Our research distinguishes and proposes several systematic types of concerted regional strategies according to the performance of a region measured by Resilience and Competiveness sub-indices. A key result of the study is the design of an operational constellation for strategic regional policy evaluation, with a major added value for policy- and decision-making purposes. The use of official data from Eurostat and of standard indicators in our research assures continuity and consistency with the official Regional Competitiveness Index (RCI) classification and measurement, so that policy makers are able to compare the performance of their regions over time and to develop proper concerted strategies accordingly. The clear evidence of a connectivity between regional competitiveness and resilience may help to develop a governance approach that balances competitiveness (mainly represented by productive assets) with resilience (mainly represented by sustainability and ecological awareness) and thus to deal with the complexity in socioeconomic systems.

     

Acoustics and rheological studies of aqueous ethylene glycol blend copper oxide nanofluids

Experimental investigations have been performed to synthesize copper oxide nanoparticles by conventional chemical precipitation method and nanofluids were prepared by two-step method using CuO nanoparticles in different proportions of ethylene glycol–water mixtures (EG–water). Powder X-ray diffraction (PXRD), energy dispersive X-ray (EDX), scanning electron microscope (SEM), particle size, and zeta potential analysis have been studied to characterize both solid and fluid samples for their sizes, shapes, stability, and arrangement. Besides, acoustics and rheological properties such as ultrasonic velocity, density, and viscosity have been measured for all fluid samples at three different temperatures. Interpretations of all these parameters have been made on the basis of particle stability and dispersion capacity of nanoparticles in different proportions of base fluids. The variation of dynamic viscosity with shear rate shows the nanofluids to be behaved like non-Newtonian fluids at very less shear rate but shows Newtonian behavior as the shear rate increases.     

Tks5 Regulates Synaptic Podosome Formation and Stabilization of the Postsynaptic Machinery at the Neuromuscular Junction

Currently, the etiology of many neuromuscular disorders remains unknown. Many of them are characterized by aberrations in the maturation of the neuromuscular junction (NMJ) postsynaptic machinery. Unfortunately, the molecular factors involved in this process are still largely unknown, which poses a great challenge for identifying potential therapeutic targets. Here, we identified Tks5 as a novel interactor of αdystrobrevin-1, which is a crucial component of the NMJ postsynaptic machinery. Tks5 has been previously shown in cancer cells to be an important regulator of actin-rich structures known as invadosomes. However, a role of this scaffold protein at a synapse has never been studied. We show that Tks5 is crucial for remodeling of the NMJ postsynaptic machinery by regulating the organization of structures similar to the invadosomes, known as synaptic podosomes. Additionally, it is involved in the maintenance of the integrity of acetylcholine receptor (AChR) clusters and regulation of their turnover. Lastly, our data indicate that these Tks5 functions may be mediated by its involvement in recruitment of actin filaments to the postsynaptic machinery. Collectively, we show for the first time that the Tks5 protein is involved in regulation of the postsynaptic machinery.     

The age of cows as a factor shaping the antioxidant level during a nutritional experiment with fish oil and linseed supplementation for increasing the antioxidant value of milk

BACKGROUND: So far, in research studies, the age of cows has not been considered as a factor that may influence the changes in the content of milk ingredients with antioxidant properties modified by the feed supplementation. The aim of this study was to determine the influence of supplementation on the content of ingredients having antioxidant properties and to determine the influence of the age of cows taking part in the experiment on these changes. The experiment was conducted using 20 Polish Holstein Friesian cows, 10 primiparous and 10 multiparous. The combined supplementation of fish oil and linseed constituted the experimental factor. RESULTS: The milk of primiparous cows after 21 days of supplementation was characterised by a higher content of C18:1 trans ‐11, C18:2 cis ‐9, trans ‐11, α‐retinol, α‐tocopherol and β‐lactoglobulin compared to the milk of multiparous cows, in which a higher level of lactoferrin, C20:5 and β‐carotene was recorded. In both groups an increase in the total antioxidant status was noted (a higher level in the milk of primiparous cows). CONCLUSIONS: Modification of the diet of cows with fish oil and linseed significantly influenced antioxidant properties of their milk; however, the response of multiparous and primaparous cows was noticeably different to the supplement introduced. Copyright © 2012 Society of Chemical Industry     

Development of Real-time PCR Assays for the Detection of the <Emphasis Type="Italic">pin</Emphasis> II Terminator (t<Emphasis Type="Italic">pin</Emphasis>II) Used in GM Constructs and Its Donor Organism,Potato (<Emphasis Type="Italic">Solanum tuberosum</Emphasis>)

Two singleplex TaqMan methods were developed for the detection of potato targets: one for the detection of the tpinII terminator, which is an emerging terminator used in GM constructs, and one for the detection of the endogenous StLS gene of potato. Performance criteria such as specificity and sensitivity were successfully tested for the two methods, taking into account the recommendations of international guidelines. The presence of the StLS target was checked in 16 potato cultivars. The StLS target is present at low copy number and can be used for quantitation purposes, as demonstrated on transgenic potatoes in this paper. The StLS target is an excellent candidate to replace the presently recommended endogenous target based on the UGP gene, which shows several disadvantages due to its high copy number and lack of specificity. The research also indicates that DNA can easily be extracted from different parts of potato tubers with a classical cetyltrimethylammonium bromide method.     

The role of electrical current mode in calcium carbonate deposition on conductive carbon nanofiber–epoxy coating

Calcium carbonate is one of the most common scaling minerals. In this paper we have used different electrical current modes (direct current [DC], pulsed DC, and alternating current [AC]) to control the amount, morphology, and distribution of calcium carbonate deposit on electroconductive epoxy/carbon nanofiber (CNF) coating. The effect of different current modes on surface scaling was visualized using scanning electron microscopy. It has been shown that both AC and DC anodic polarization limited scale deposition on epoxy/CNF coated surfaces, although the mechanisms of scale inhibition during AC and DC polarization were different. DC polarization of the coating at +2 V resulted in the smallest scale buildup without leading to coating degradation, while DC polarization at potentials as high as +5 V caused the coating to degrade. Interestingly, application of pulsed DC with high pulse frequency (50 Hz) inhibited the degradation. The type of current applied affected also the morphology of the precipitate at the cathode. The results presented in this work show, for the first time, how different modes of electrical current applied to electroconductive composite coatings can be used to control the morphology and distribution of calcium carbonate scale, and how the organic coating degradation at high polarization potentials can be avoided.     

Molecular Characterisation of Uterine Endometrial Proteins during Early Stages of Pregnancy in Pigs by MALDI TOF/TOF

The molecular mechanism underlying embryonic implantation is vital to understand the correct communications between endometrium and developing conceptus during early stages of pregnancy. This study’s objective was to determine molecular changes in the uterine endometrial proteome during the preimplantation and peri-implantation between 9 days (9D), 12 days (12D), and 16 days (16D) of pregnant Polish Large White (PLW) gilts. 2DE-MALDI-TOF/TOF and ClueGO^TM approaches were employed to analyse the biological networks and molecular changes in porcine endometrial proteome during maternal recognition of pregnancy. A total of sixteen differentially expressed proteins (DEPs) were identified using 2-DE gels and MALDI-TOF/TOF mass spectrometry. Comparison between 9D and 12D of pregnancy identified APOA1, CAPZB, LDHB, CCT5, ANXA4, CFB, TTR upregulated DEPs, and ANXA5, SMS downregulated DEPs. Comparison between 9D and 16D of pregnancy identified HP, APOA1, ACTB, CCT5, ANXA4, CFB upregulated DEPs and ANXA5, SMS, LDHB, ACTR3, HP, ENO3, OAT downregulated DEPs. However, a comparison between 12D and 16D of pregnancy identified HP, ACTB upregulated DEPs, and CRYM, ANXA4, ANXA5, CAPZB, LDHB, ACTR3, CCT5, ENO3, OAT, TTR down-regulated DEPs. Outcomes of this study revealed key proteins and their interactions with metabolic pathways involved in the recognition and establishment of early pregnancy in PLW gilts.     

Radiosensitization of Human Leukemic HL-60 Cells by ATR Kinase Inhibitor (VE-821): Phosphoproteomic Analysis

DNA damaging agents such as ionizing radiation or chemotherapy are frequently used in oncology. DNA damage response (DDR)—triggered by radiation-induced double strand breaks—is orchestrated mainly by three Phosphatidylinositol 3-kinase-related kinases (PIKKs): Ataxia teleangiectasia mutated (ATM), DNA-dependent protein kinase (DNA-PK) and ATM and Rad3-related kinase (ATR). Their activation promotes cell-cycle arrest and facilitates DNA damage repair, resulting in radioresistance. Recently developed specific ATR inhibitor, VE-821 (3-amino-6-(4-(methylsulfonyl)phenyl)-N-phenylpyrazine-2-carboxamide), has been reported to have a significant radio- and chemo-sensitizing effect delimited to cancer cells (largely p53-deficient) without affecting normal cells. In this study, we employed SILAC-based quantitative phosphoproteomics to describe the mechanism of the radiosensitizing effect of VE-821 in human promyelocytic leukemic cells HL-60 (p53-negative). Hydrophilic interaction liquid chromatography (HILIC)-prefractionation with TiO₂-enrichment and nano-liquid chromatography—tandem mass spectrometry (LC-MS/MS) analysis revealed 9834 phosphorylation sites. Proteins with differentially up-/down-regulated phosphorylation were mostly localized in the nucleus and were involved in cellular processes such as DDR, all phases of the cell cycle, and cell division. Moreover, sequence motif analysis revealed significant changes in the activities of kinases involved in these processes. Taken together, our data indicates that ATR kinase has multiple roles in response to DNA damage throughout the cell cycle and that its inhibitor VE-821 is a potent radiosensitizing agent for p53-negative HL-60 cells.