Size and shape influence of luminescent orthovanadate nanoparticles on their accumulation in nuclear compartments of rat hepatocytes

In this paper the process of nonfunctionalized negatively charged orthovanadate nanoparticle accumulation and redistribution in cells dependent on their shape and size was investigated. Aqueous colloidal solutions of nReVO₄:Eu^3 + (Re = Gd, Y, La) luminescent nanocrystals of different sizes and shapes have been synthesized. The average sizes of spherical particles were 2, 20, and 300 nm, of spindle-like particles – 22 × 6.3 nm, and of rod-like particles – 57 × 4.4 nm. Luminescence of nReVO₄:Eu^3 + nanocrystals was effectively excited by UV and visible irradiation. By means of luminescence microscopy and luminescence microspectroscopy, it has been revealed that spherical nanocrystals with an average diameter of 2 nm tend to accumulate mainly in the rat hepatocyte nuclei in situ and also in the isolated nuclei of these cells. An additional experiment has shown that nanoparticles reveal tropism to nuclear structural components. The penetration into nuclei does not require any modifications of the surface of nanoparticle and is governed by the shape and size of nanoparticle and also is determined by the cellular type.     

Phenomena at three-phase electroslag remelting

The electroslag remelting (ESR) process is widely used to produce high-quality ingots and billets for high-alloyed steels and alloys.Both the single-phase and three-phase alternating current diagram with bifilar and monofilar connection are in use for heavy ingot manufacturing.The numerical simulation of the three-phase bifilar circuit for the 120 t three-phase bifilar six-electrode ESR furnace at different variants of electric connection was presented and discussed.At the bifilar diagram of power supply,the geometrical location of electrodes in a mould holds critical importance for performances:the close location of bifilar pair electrodes provides the highest heat productivity,but the equidistant location of electrodes gives a much more uniform heat distribution.The monofilar mulit-electrode diagram of three-phase connection without phase shift shows the most uniform distribution of potential and heat generation as well as a favorable magnetic field that makes this kind the most promising for providing a high quality of heavy ingots.     

Physicochemical comparison of electroslag remelting with consumable electrode and electroslag refining with liquid metal

The paper is devoted to comparison of electroslag remelting (ESR) with consumable electrode and electroslag refining with liquid metal (ESR LM) processes. The possibility of rearrangement of the heat contributions coming from the consumable electrode and current supplying mould (non-consumable electrode, CSM) makes the core of the ESR process organisation in the CSM. The usage of liquid metal instead of consumable electrodes allows to reduce liquid bath temperature and volume in order to provide low segregation ingot. The formal assessment of physico-chemical conditions and experimental measurements of desulphurisation have shown the same level of refining ability of both processes. Replacing the classic ESR by the ESR LM is a prospective way to produce high-quality ingots from sophisticated and hard-to-deform materials, whereas manufacturing of the consumable electrodes is technically problematic and costly.     

Encapsulation of the Dinuclear Trithiolato-Bridged Arene Ruthenium Complex Diruthenium-1 in an Apoferritin Nanocage: Structure and Cytotoxicity

The effects of encapsulating the cytotoxic dinuclear trithiolato-bridged arene ruthenium complex [(η⁶-p-MeC₆H₄iPr)₂Ru₂(μ₂-S-p-C₆H₄tBu)₃]Cl (DiRu-1) within the apoferritin (AFt) nanocage were investigated. The DiRu-1-AFt nanocarrier was characterized by UV/Vis spectroscopy, ICP-MS, CD and X-ray crystallography. In contrast to previously reported Au- and Pt-based drug-loaded AFt carriers, we found no evidence of direct interactions between DiRu-1 and AFt. DiRu-1-AFt is cytotoxic toward immortalized murine BALB/c-3T3 fibroblasts transformed with SV40 virus (SVT2) and human epidermoid carcinoma A431 malignant cells, and exhibits moderate selectivity for these cancer cells over normal BALB/c-3T3 cells. DiRu-1-AFt triggers the production of reactive oxygen species, depolarization of mitochondrial membrane potential, and induces cell death via p53-mediated apoptosis. Comparison between our data and previous results suggests that the presence of specific interactions between a metal-based drug and AFt within the protein cage is not essential for drug encapsulation.     

Immunohistochemical analysis of S6K1 and S6K2 expression in human breast tumors

AIM: To express recombinant S6K2 in baculovirus expression system; to purify large quantities of recombinant S6K2 for biochemical studies; to generate and characterise specific MABs against recombinant S6K2; to study the patterns S6K1 and S6K2 expression and subcellular localization in normal, benign and malignant breast tissues. METHODS: Recombinant baculovirus, expressing wild type S6K2 was generated using Bac-to-Bac system (Invitrogen); recombinant S6K was purified from infected Sf9 cells using affinity purification approach; monoclonal antibodies against recombinant S6K2 were generated; the specificity of generated MABs towards recombinant and endogenous S6K2 were examined by ELISA, Western blotting, immunoprecipitation and immuhohistochemical staining; immunohistochemical detection of S6K1 and S6K2 in human breast tissues was performed using specific monoclonal antibodies towards S6K1 and S6K2. RESULTS: Large amounts of enzymatically active S6K2 were purified using baculovirus expression system; highly purified preparations of S6K2 were used to generate and characterize anti-S6K2 MABs; elevated levels of S6K1 and S6K2 were found in breast tumors when compared to normal breast tissues; S6K2 is frequently localized in the nuclei of adenocarcinoma tissues, but rarely in fibroadenoma or "normal" breast tissues. CONCLUSION: Production of recombinant S6K2 in large amount and generation of specific monoclonal antibodies towards S6K2 has provided us with excellent tools to study the function and regulation of this important signalling molecule in normal and cancer cells. Immunnohistochemical analysis of S6K1 and S6K2 expression in normal and malignant breast clearly indicates that both kinases are overexpressed in breast tumors, when compared to "normal" tissues. The retention of S6K2 in the nuclei of malignant cells may be caused by disregulation of nucleocytoplasmic shuttling and could subsequently affect cell growth and proliferation.     

US energy conservation and efficiency policies: Challenges and opportunities

Expanding energy conservation and efficiency in every sector nationwide is one of the most cost-effective instruments for reducing US energy imports, the trade deficit and energy’s environmental impacts. For these reasons, energy conservation and efficiency have been essential elements of US energy policy since the oil embargos and price spikes of the 1970s. The Energy Independence and Security Act of 2007 (EISA) is the latest federal legislation to expand and strengthen US energy conservation and efficiency policies, programs, and practices. Specifically, EISA and its recent predecessor, the Energy Policy Act of 2005 (EPAct05), contain almost 200 titles with new provisions for energy conservation and efficiency aimed at improvements in vehicle fuel economy. These provisions include efficiency of appliances and lighting; energy savings in residential, commercial, and government buildings; the efficiency of industrial manufacturing plants; and the efficiency of electric power delivery and end-use. These actions have begun to contribute to new federal, state, and local policies, programs, and practices across the US, and expectations are high for increases in the level of energy savings. This paper summarizes the history of US energy conservation and efficiency policies, outlines EISA’s and EPAct05’s key provisions, and considers prospects for the future.     

The Impact of Hypergravity and Vibration on Gene and Protein Expression of Thyroid Cells

Experiments in space either on orbital missions on-board the ISS, or in suborbital missions using sounding rockets, like TEXUS as well as parabolic flight campaigns are still the gold standard to achieve real microgravity conditions in the field of gravitational biology and medicine. However, during launch, and in flight, hypergravity and vibrations occur which might interfere with the effects of microgravity. It is therefore important to know these effects and discriminate them from the microgravity effects. This can be achieved by ground-based facilities like centrifuges or vibration platforms. Recently, we have conducted several experiments with different thyroid cancer cell lines. This study, as part of the ESA-CORA-GBF 2010-203 project, focused on the influence of vibration and hypergravity on benign human thyroid follicular epithelial cells (Nthy-ori 3-1 cell line). Gene and in part protein expression regulation under both conditions were analyzed for VCAN, ITGA10, ITGB1, OPN, ADAM19, ANXA1, TNFA, ABL2, ACTB, PFN2, TLN1, EZR, RDX, MSN, CTGF, PRKCA, and PRKAA1 using quantitative real-time PCR and Western Blot. We found that hypergravity and vibration affected genes and proteins involved in the extracellular matrix, the cytoskeleton, apoptosis, cell growth and signaling. Vibration always led to a down-regulation, whereas hypergravity resulted in a more heterogeneous expression pattern. Overall we conclude that both conditions can influence gene regulation and production of various genes and proteins. As a consequence, it is important to perform control experiments on hypergravity and vibration facilities in parallel to flight experiments.     

A Realtime and Continuous Assessment of Cortisol in ISF Using Electrochemical Impedance Spectroscopy

This study describes the functioning of a novel sensor to measure cortisol concentration in the interstitial fluid (ISF) of a human subject. ISF is extracted by means of vacuum pressure from micropores created on the stratum corneum layer of the skin. The pores are produced by focusing a near infrared laser on a layer of black dye material attached to the skin. The pores are viable for approximately three days after skin poration. Cortisol measurements are based on electrochemical impedance (EIS) technique. Gold microelectrode arrays functionalized with Dithiobis (succinimidyl propionate) self-assembled monolayer (SAM) have been used to fabricate an ultrasensitive, disposable, electrochemical cortisol immunosensor. The biosensor was successfully used for in-vitro measurement of cortisol in ISF. Tests in a laboratory setup show that the sensor exhibits a linear response to cortisol concentrations in the range 1 pm to 100 nM. A small pilot clinical study showed that in-vitro immunosensor readings, when compared with commercial evaluation using enzyme-linked immunoassay (ELISA) method, correlated well with cortisol levels in saliva and ISF. Further, circadian rhythm could be established between the subject's ISF and the saliva samples collected over 24 hours time-period. Cortisol levels in ISF were found reliably higher than in saliva. This Research establishes the feasibility of using impedance based biosensor architecture for a disposable, wearable cortisol detector. The projected commercial in-vivo real-time cortisol sensor device, besides being minimally invasive, will allow continuous ISF harvesting and cortisol monitoring over 24 hours even when the subject is asleep. Forthcoming, this sensor could be interfaced to a wireless health monitoring system that could transfer sensor data over existing wide-area networks such as the internet and a cellular phone network to enable real-time remote monitoring of subjects.