Regulation by FurC in Anabaena Links the Oxidative Stress Response to Photosynthetic Metabolism.

The FUR (Ferric Uptake Regulator) family in Anabaena sp. PCC 7120 consists of three paralogs named FurA (Fur), FurB (Zur) and FurC (PerR). furC seems to be an essential gene in the filamentous nitrogen-fixing strain Anabaena sp. PCC 7120, suggesting that it plays a fundamental role in this organism. In order to better understand the functions of FurC in Anabaena, the phenotype of a derivative strain that overexpresses this regulator (EB2770FurC) has been characterized. The furC-overexpressing variant presented alterations in growth rate, morphology and ultrastructure, as well as higher sensitivity to peroxide than Anabaena sp. PCC 7120. Interestingly, the overexpression of furC led to reduced photosynthetic O2 evolution, increased respiratory activity, and had a significant influence in the composition and efficiency of both photosystems. Comparative transcriptional analyses, together with electrophoretic mobility shift assays allowed the identification of different genes directly controlled by FurC, and involved in processes not previously related to PerR proteins, such as the cell division gene ftsZ and the major thylakoid membrane protease ftsH. The rise in the transcription of ftsH in EB2770FurC cells correlated with reduced levels of the D1 protein, which is involved in the PSII repair cycle. Deregulation of the oxidative stress response in EB2770FurC cells led to the identification of novel FurC targets involved in the response to H2O2 through different mechanisms. These results, together with the effect of furC overexpression on the composition, stability and efficiency of the photosynthetic machinery of Anabaena, disclose novel links between PerR proteins, cell division and photosynthesis in filamentous cyanobacteria.

Local Temperature Increments and Induced Cell Death in Intracellular Magnetic Hyperthermia.

The generation of temperature gradients on nanoparticles heated externally by a magnetic field is crucially important in magnetic hyperthermia therapy. But the intrinsic low heating power of magnetic nanoparticles, at the conditions allowed for human use, is a limitation that restricts the general implementation of the technique. A promising alternative is local intracellular hyperthermia, whereby cell death (by apoptosis, necroptosis, or other mechanisms) is attained by small amounts of heat generated at thermosensitive intracellular sites. However, the few experiments conducted on the temperature determination of magnetic nanoparticles have found temperature increments that are much higher than the theoretical predictions, thus supporting the local hyperthermia hypothesis. Reliable intracellular temperature measurements are needed to get an accurate picture and resolve the discrepancy. In this paper, we report the real-time variation of the local temperature on γ-Fe2O3 magnetic nanoheaters using a Sm3+/Eu3+ ratiometric luminescent thermometer located on its surface during exposure to an external alternating magnetic field. We measure maximum temperature increments of 8 °C on the surface of the nanoheaters without any appreciable temperature increase on the cell membrane. Even with magnetic fields whose frequency and intensity are still well within health safety limits, these local temperature increments are sufficient to produce a small but noticeable cell death, which is enhanced considerably as the magnetic field intensity is increased to the maximum level tolerated for human use, consequently demonstrating the feasibility of local hyperthermia.

High-Repetition-Rate Femtosecond Laser Processing of Acrylic Intra-Ocular Lenses.

The study of laser processing of acrylic intra-ocular lenses (IOL) by using femtosecond laser pulses delivered at high-repetition rate is presented in this work. An ultra-compact air-cooled femtosecond diode laser (HighQ2-SHG, Spectra-Physics) delivering 250 fs laser pulses at the fixed wavelength of 520 nm with a repetition rate of 63 MHz was used to process the samples. Laser inscription of linear periodic patterns on the surface and inside the acrylic substrates was studied as a function of the processing parameters as well as the optical absorption characteristics of the sample. Scanning Electron Microscopy (SEM) Energy Dispersive X-ray Spectroscopy (EDX), and micro-Raman Spectroscopy were used to evaluate the compositional and microstructural changes induced by the laser radiation in the processed areas. Diffractive characterization was used to assess 1st-order efficiency and the refractive index change.

A reagentless enzymatic fluorescent biosensor for glucose based on upconverting glasses, as excitation source, and chemically modified glucose oxidase.

Upon near-infrared excitation Tm(3+)+Yb(3+) doped fluorohafnate glasses present upconversion properties and emit visible light. This property permits to use these glasses (UCG) as excitation sources for fluorescent optical biosensors. Taking this into account, in this work a fluorescent biosensor for glucose determination is designed and evaluated. The biosensor combines the UCG and the fluorescence of the enzyme glucose oxidase chemically modified with a fluorescein derivative (GOx-FS), whose intensity is modified during the enzymatic reaction with glucose. Optical parameters have been optimized and a mathematical model describing the behavior of the analytical signal is suggested. Working in FIA mode, the biosensor responds to glucose concentrations up to, at least, 15mM with a limit of detection of 1.9mM. The biosensor has a minimum lifetime of 9 days and has been applied to glucose determination in drinks. The applicability of the sensor was tested by glucose determination in two fruit juices.

Joining time-resolved thermometry and magnetic-induced heating in a single nanoparticle unveils intriguing thermal properties.

Whereas efficient and sensitive nanoheaters and nanothermometers are demanding tools in modern bio- and nanomedicine, joining both features in a single nanoparticle still remains a real challenge, despite the recent progress achieved, most of it within the last year. Here we demonstrate a successful realization of this challenge. The heating is magnetically induced, the temperature readout is optical, and the ratiometric thermometric probes are dual-emissive Eu(3+)/Tb(3+) lanthanide complexes. The low thermometer heat capacitance (0.021·K(-1)) and heater/thermometer resistance (1 K·W(-1)), the high temperature sensitivity (5.8%·K(-1) at 296 K) and uncertainty (0.5 K), the physiological working temperature range (295-315 K), the readout reproducibility (>99.5%), and the fast time response (0.250 s) make the heater/thermometer nanoplatform proposed here unique. Cells were incubated with the nanoparticles, and fluorescence microscopy permits the mapping of the intracellular local temperature using the pixel-by-pixel ratio of the Eu(3+)/Tb(3+) intensities. Time-resolved thermometry under an ac magnetic field evidences the failure of using macroscopic thermal parameters to describe heat diffusion at the nanoscale.

Mutants of Anabaena sp. PCC 7120 lacking alr1690 and alpha-furA antisense RNA show a pleiotropic phenotype and altered photosynthetic machinery.

Fur proteins are global regulators present in all prokaryotes. In Anabaena sp. PCC 7120 FurA controls iron uptake and modulates an important set of genes related primarily to photosynthesis, nitrogen metabolism and oxidative stress defense. Expression of furA is tuned by the cis-acting antisense alpha-furA RNA that is co-transcribed with the outer-membrane protein Alr1690. Disruption of the alpha-furA-alr1690 message produces the iron-deficient JAH3 mutant that lacks Alr1690 and shows enhanced expression of FurA. JAH3 cells present severe structural disorders related to the number, organization and density of photosynthetic membranes. Quantitative analysis of the fluorescence induction shows that the mutation affects the J-I and I-P phases and causes important alterations in the photosynthetic apparatus, leading to lower photosynthetic performance indexes. These results reveal that expression of the alpha-furA-alr1690 message is required for maintenance of a proper thylakoid arrangement, efficient regulation of iron uptake and optimal yield of the photosynthetic machinery.