A fast and tuneable auxin-inducible degron for depletion of target proteins in budding yeast

The auxin-inducible degron (AID) is a useful technique to rapidly deplete proteins of interest in nonplant eukaryotes. Depletion is achieved by addition of the plant hormone auxin to the cell culture, which allows the auxin-binding receptor, TIR1, to target the AID-tagged protein for degradation by the proteasome. Fast depletion of the target protein requires good expression of TIR1 protein, but as we show here, high levels of TIR1 may cause uncontrolled depletion of the target protein in the absence of auxin. To enable conditional expression of TIR1 to a high level when required, we regulated the expression of TIR1 using the β-estradiol expression system. This is a fast-acting gene induction system that does not cause secondary effects on yeast cell metabolism. We demonstrate that combining the AID and β-estradiol systems results in a tightly controlled and fast auxin-induced depletion of nuclear target proteins. Moreover, we show that depletion rate can be tuned by modulating the duration of β-estradiol preincubation. We conclude that TIR1 protein is a rate-limiting factor for target protein depletion in yeast, and we provide new tools that allow tightly controlled, tuneable, and efficient depletion of essential proteins whereas minimising secondary effects.     

Electrochemical Immuno-Vascular Solid State Point of Care Testing （POCT）

In this work, it is proposed a POCT and innovative method of immunoassay for the detection of C-reactive protein and IgG, using Amperometry coupled to solid state kit, connected to a micro-flow system with comparable sensitivity to a high sensitivity CRP ELISA （hsCRP） and IgG ELISA, with 1-3 min turnaround time to result. Samples of CRP （0 to 250 ng·mL-1） and IgG or diluted spiked human serum are injected through a solid state polymeric kit, micro-flow sensor channels of a SWCT SPE nano-modified biosensor. Preparing two kits immuno-assays, in the same micro-column, built on oxirane groups of polymeric bead surface, with biological support to CRP and IgG biomarkers recognition, in a real time scheme, at the end of analyte injection the initial rate of change in current intensity I/A was proportional to CRP respectively IgG concentration, with low detection limit （LOD） of 0.1 ng·mL-1. It was revealed that CRP/IgG concentrations in serum that might be expected in both normal and pathological conditions can be detected in a real-time-efficient, multi-immunoassay with solid state detection kit technology with determined CRP/lgG concentrations in close agreement with those determined using a commercially available high sensitivity EL1SA.     

Regional Drought Modes in Iran Using the SPI: The Effect of Time Scale and Spatial Resolution

In the present paper, regional drought modes in Iran are identified applying the Principal Component Analysis (PCA) and Varimax rotation to the Standardized Precipitation Index (SPI) computed on different time scales. Data used include gridded monthly precipitation covering the period 1951–2007 retrieved from the Global Precipitation Climatology Centre (GPCC) archive with different spatial resolutions (2.5, 1 and 0.5° resolution). The objective of the study is twofold: (i) Investigate the stability of drought spatial modes as a function of the SPI time scales used for monitoring the different kinds of drought, (ii) Evaluate the impact of the spatial resolution of gridded data on drought regionalization. For the coarse spatial resolution of 2.5°, results show four drought modes of distinct variability, which remain quite stable when the SPI time scale is varied from 1- to 24-month. Differently, for higher spatial resolutions drought modes appear more sensitive to the index time scale and become less spatially homogeneous as the time scale is increased. Moreover, the number of identified modes (sub-regions) may reduce to three or two, but in all cases the most well defined sub-region appears to be the southern one. This suggests that both the spatial resolution of precipitation data and the time scale may affect drought regionalization, i.e. the number of drought modes and their spatial homogeneity.     

The Ketogenic Diet Reduces the Harmful Effects of Stress on Gut Mitochondrial Biogenesis in a Rat Model of Irritable Bowel Syndrome

Functional alterations in irritable bowel syndrome have been associated with defects in bioenergetics and the mitochondrial network. Effects of high fat, adequate-protein, low carbohydrate ketogenic diet (KD) involve oxidative stress, inflammation, mitochondrial function, and biogenesis. The aim was to evaluate the KD efficacy in reducing the effects of stress on gut mitochondria. Newborn Wistar rats were exposed to maternal deprivation to induce IBS in adulthood. Intestinal inflammation (COX-2 and TRL-4); cellular redox status (SOD 1, SOD 2, PrxIII, mtDNA oxidatively modified purines); mitochondrial biogenesis (PPAR-γ, PGC-1α, COX-4, mtDNA content); and autophagy (Beclin-1, LC3 II) were evaluated in the colon of exposed rats fed with KD (IBD-KD) or standard diet (IBS-Std), and in unexposed controls (Ctrl). IBS-Std rats showed dysfunctional mitochondrial biogenesis (PPAR-γ, PGC-1α, COX-4, and mtDNA contents lower than in Ctrl) associated with inflammation and increased oxidative stress (higher levels of COX-2 and TLR-4, SOD 1, SOD 2, PrxIII, and oxidatively modified purines than in Ctrl). Loss of autophagy efficacy appeared from reduced levels of Beclin-1 and LC3 II. Feeding of animals with KD elicited compensatory mechanisms able to reduce inflammation, oxidative stress, restore mitochondrial function, and baseline autophagy, possibly via the upregulation of the PPAR-γ/PGC-1α axis.     

High temperature melt viscosity and fragile to strong transition in Zr–Cu–Ni–Al–Nb(Ti) and Cu47Ti34Zr11Ni8 bulk metallic glasses

The molten viscous behavior of Vitreloy 106, 106a, 105 and 101 bulk metallic glass-forming alloys was investigated at temperatures above T_liq. Couette concentric cylinder viscometry was used to determine the isothermal and continuous cooling melt viscosities. Isothermal three point beam bending measurements of the viscosity in the vicinity of T_g revealed a kinetically stronger liquid at low temperatures than at high temperatures, near T_liq. This behavior is revealed to be the result of a fragile to strong transition in the undercooled liquid, possibly due to polyamorphism.     

Recycling of Screen Glass Into New Traditional Ceramic Materials

The cleaned cathode ray tube (CRT) glass comes from the dismantling plant of TV and PC color kinescopes and has been used to completely replace both feldspathic and inert components of a traditional ceramic body. Panel glass has been added (up to 20 wt%) to a mixture of two commercial ball clays in order to obtain laboratory tiles sintered at 1210°C. Densification has been studied according to ISO rules, while sinterability has been estimated by optical dilatometry. The samples obtained showed main properties similar to commercial ceramic floor and/or wall tiles.     

Graphene sublattice symmetry and isospin determined by circular dichroism in angle-resolved photoemission spectroscopy

The Dirac-like electronic structure of graphene originates from the equivalence of the two basis atoms in the honeycomb lattice. We show that the characteristic parameters of the initial state wave function (sublattice symmetry and isospin) can be determined using angle-resolved photoemission spectroscopy (ARPES) with circularly polarized synchrotron radiation. At a photon energy of hν = 52 eV, transition matrix element effects can be neglected allowing us to determine sublattice symmetry and isospin with high accuracy using a simple theoretical model.