Drug-Selected Human Lung Cancer Stem Cells: Cytokine Network, Tumorigenic and Metastatic Properties

Background: Cancer stem cells (CSCs) are thought to be responsible for tumor regeneration after chemotherapy, although direct confirmation of this remains forthcoming. We therefore investigated whether drug treatment could enrich and maintain CSCs and whether the high tumorogenic and metastatic abilities of CSCs were based on their marked ability to produce growth and angiogenic factors and express their cognate receptors to stimulate tumor cell proliferation and stroma formation. Methodology/Findings: Treatment of lung tumor cells with doxorubicin, cisplatin, or etoposide resulted in the selection of drug surviving cells (DSCs). These cells expressed CD133, CD117, SSEA-3, TRA1-81, Oct-4, and nuclear β-catenin and lost expression of the differentiation markers cytokeratins 8/18 (CK 8/18). DSCs were able to grow as tumor spheres, maintain self-renewal capacity, and differentiate. Differentiated progenitors lost expression of CD133, gained CK 8/18 and acquired drug sensitivity. In the presence of drugs, differentiation of DSCs was abrogated allowing propagation of cells with CSC-like characteristics. Lung DSCs demonstrated high tumorogenic and metastatic potential following inoculation into SCID mice, which supported their classification as CSCs. Luminex analysis of human and murine cytokines in sonicated lysates of parental- and CSC-derived tumors revealed that CSC-derived tumors contained two- to three-fold higher levels of human angiogenic and growth factors (VEGF, bFGF, IL-6, IL-8, HGF, PDGF-BB, G-CSF, and SCGF-β). CSCs also showed elevated levels of expression of human VEGFR2, FGFR2, CXCR1, 2 and 4 receptors. Moreover, human CSCs growing in SCID mice stimulated murine stroma to produce elevated levels of angiogenic and growth factors. Conlusions/Significance: These findings suggest that chemotherapy can lead to propagation of CSCs and prevention of their differentiation. The high tumorigenic and metastatic potentials of CSCs are associated with efficient cytokine network production that may represent a target for increased efficacy of cancer therapy. © 2008 Levina et al

The Feasibility of Using Agarophyte Processing Waste

When agarophytes are processed, 16-89% algal waste (AW) is formed. Due to the high content of minerals in this waste and the presence of proteins that are resistant to the action of proteolytic enzymes, such AW is mainly used in the production of feed for farm animals. The significant content of polysaccharides in dry AW indicates that it could be used as a raw material for the production of dietary fiber (DF). Due to the difference in the chemical composition of AW depending on the type of red algae, different approaches for its deproteinization have been used. Thus, a 3% alkali solution should be used for the deproteinization of algal waste from the red algae Gracilaria, a 0.5% alkali solution for GelidiumAW, and a 1% sodium carbonate solution for A. plicataAW.The duration of the deproteinization process is 30 min at a temperature of 97±2 ∘C for all types of AW. In this study, functional and technological solutions of DF from AW were developed. The results showed that their water binding capacity was 6-22 g of water per 1 g of the preparation, the fat binding capacity was 1.6-3.3 g of fat per 1 g of the preparation, and the swelling capacity was 46-312% depending on the type of red algae.The obtained DF was used in the production of minced fish food products of the ‘fish sticks’ type, based on minced fish and consisting of cod and pink salmon. The study of the chemical composition and calorie content of the developed food product showed the possibility of its use in dietary nutrition. Keywords: red algae, algal waste, deproteinization, dietary fibe

The Feasibility of Using Agarophyte Processing Waste

When agarophytes are processed, 16-89% algal waste (AW) is formed. Due to the high content of minerals in this waste and the presence of proteins that are resistant to the action of proteolytic enzymes, such AW is mainly used in the production of feed for farm animals. The significant content of polysaccharides in dry AW indicates that it could be used as a raw material for the production of dietary fiber (DF). Due to the difference in the chemical composition of AW depending on the type of red algae, different approaches for its deproteinization have been used. Thus, a 3% alkali solution should be used for the deproteinization of algal waste from the red algae Gracilaria, a 0.5% alkali solution for GelidiumAW, and a 1% sodium carbonate solution for A. plicataAW.The duration of the deproteinization process is 30 min at a temperature of 97±2 ∘C for all types of AW. In this study, functional and technological solutions of DF from AW were developed. The results showed that their water binding capacity was 6-22 g of water per 1 g of the preparation, the fat binding capacity was 1.6-3.3 g of fat per 1 g of the preparation, and the swelling capacity was 46-312% depending on the type of red algae.The obtained DF was used in the production of minced fish food products of the ‘fish sticks' type, based on minced fish and consisting of cod and pink salmon. The study of the chemical composition and calorie content of the developed food product showed the possibility of its use in dietary nutrition. Keywords: red algae, algal waste, deproteinization, dietary fibe

Evolution and current development trends of the Russian Federation’s higher education system

The aim of the study is to substantiate evolutionary development stages and current trends in the higher education system in the Russian Federation via general scientific methods of cognition, quantitative and qualitative assessment, methods of comparative and factor analysis, analysis and synthesis, grouping and comparison, the method of logical generalization. As a result, increasing globalization changes the approach to education. The authors came to the conclusion that it is necessary to take into account the impact of external and inter systemic factors on the development of higher education organizations in the Russian Federation.El objetivo del estudio es sustanciar las etapas de desarrollo evolutivo y las tendencias actuales en el sistema de educación superior en la Federación de Rusia a través de métodos científicos generales de cognición, evaluación cuantitativa y cualitativa, métodos de análisis comparativo y factorial, análisis y síntesis, agrupación y comparación, El método de generalización lógica. Como resultado, la creciente globalización cambia el enfoque de la educación. Los autores llegaron a la conclusión de que es necesario tener en cuenta el impacto de los factores externos e intersistémicos en el desarrollo de las organizaciones de educación superior en la Federación de Rusia

Membrane-Disrupting Activity of Cobra Cytotoxins Is Determined by Configuration of the N-Terminal Loop

In aqueous solutions, cobra cytotoxins (CTX), three-finger folded proteins, exhibit conformational equilibrium between conformers with either cis or trans peptide bonds in the N-terminal loop (loop-I). The equilibrium is shifted to the cis form in toxins with a pair of adjacent Pro residues in this loop. It is known that CTX with a single Pro residue in loop-I and a cis peptide bond do not interact with lipid membranes. Thus, if a cis peptide bond is present in loop-I, as in a Pro-Pro containing CTX, this should weaken its lipid interactions and likely cytotoxic activities. To test this, we have isolated seven CTX from Naja naja and N. haje cobra venoms. Antibacterial and cytotoxic activities of these CTX, as well as their capability to induce calcein leakage from phospholipid liposomes, were evaluated. We have found that CTX with a Pro-Pro peptide bond indeed exhibit attenuated membrane-perturbing activity in model membranes and lower cytotoxic/antibacterial activity compared to their counterparts with a single Pro residue in loop-I

Flow cytometry for pediatric platelets

The ability of platelets to carry out their hemostatic function can be impaired in a wide range of inherited and acquired conditions: trauma, surgery, inflammation, pre-term birth, sepsis, hematological malignancies, solid tumors, chemotherapy, autoimmune disorders, and many others. Evaluation of this impairment is vitally important for research and clinical purposes. This problem is particularly pronounced in pediatric patients, where these conditions occur frequently, while blood volume and the choice of blood collection methods could be limited. Here we describe a simple flow cytometry-based screening method of comprehensive whole blood platelet function testing that was validated for a range of pediatric and adult samples (n = 31) in the hematology hospital setting including but not limited to: classic inherited platelet function disorders (Glanzmann’s thrombasthenia; Bernard-Soulier, Wiscott-Aldrich, and Hermasky-Pudlak syndromes, MYH9-dependent thrombocytopenia), healthy and pre-term newborns, acute and chronic immune thrombocytopenia, chronic lympholeukemia, effects of therapy on platelet function, etc. The method output includes levels of forward and side scatter, levels of major adhesion and aggregation glycoproteins Ib and IIb-IIIa, active integrins’ level based on PAC-1 binding, major alpha-granule component P-selectin, dense granule function based on mepacrine uptake and release, and procoagulant activity quantified as a percentage of annexin V-positive platelets. This analysis is performed for both resting and dual-agonist-stimulated platelets. Preanalytical and analytical variables are provided and discussed. Parameter distribution within the healthy donor population for adults (n = 72) and children (n = 17) is analyzed