Chromatin association of the SMC5/6 complex is dependent on binding of its NSE3 subunit to DNA

SMC5/6 is a highly conserved protein complex related to cohesin and condensin, which are the key components of higher-order chromatin structures. The SMC5/6 complex is essential for proliferation in yeast and is involved in replication fork stability and processing. However, the precise mechanism of action of SMC5/6 is not known. Here we present evidence that the NSE1/NSE3/NSE4 sub-complex of SMC5/6 binds to double-stranded DNA without any preference for DNA-replication/recombination intermediates. Mutations of key basic residues within the NSE1/NSE3/NSE4 DNA-binding surface reduce binding to DNA in vitro. Their introduction into the Schizosaccharomyces pombe genome results in cell death or hypersensitivity to DNA damaging agents. Chromatin immunoprecipitation analysis of the hypomorphic nse3 DNA-binding mutant shows a reduced association of fission yeast SMC5/6 with chromatin. Based on our results, we propose a model for loading of the SMC5/6 complex onto the chromatin

Mechanistic aspects of alloxan diabetogenic activity: a key role of keto-enol inversion of dialuric acid on ionization

The inversion of the keto-enol stability order of dialuric acid on ionization was calculated and verified experimentally. The radical cations in both forms were characterized. The spectrum of the keto form was observed upon direct ionization of dialuric acid under matrix conditions, whereas the enol form was formed upon a sequential electron-proton-proton attachment to alloxan under acidic aqueous condition. Facilitation of the one-electron oxidation of dialuric acid upon its enolization can result in a more effective formation of superoxide radical anion in the process of its auto-oxidation. This process is discussed in reference to the alloxan diabetogenic action. Both neutral keto and enol forms are energetically close, and under favorable conditions, the auto-oxidation of dialuric acid could involve participation of the enol form

A role of the Nse4 kleisin and Nse1/Nse3 KITE subunits in the ATPase cycle of SMC5/6

The SMC (Structural Maintenance of Chromosomes) complexes are composed of SMC dimers, kleisin and kleisin-interacting (HAWK or KITE) subunits. Mutual interactions of these subunits constitute the basal architecture of the SMC complexes. In addition, binding of ATP molecules to the SMC subunits and their hydrolysis drive dynamics of these complexes. Here, we developed new systems to follow the interactions between SMC5/6 subunits and the relative stability of the complex. First, we show that the N-terminal domain of the Nse4 kleisin molecule binds to the SMC6 neck and bridges it to the SMC5 head. Second, binding of the Nse1 and Nse3 KITE proteins to the Nse4 linker increased stability of the ATP-free SMC5/6 complex. In contrast, binding of ATP to SMC5/6 containing KITE subunits significantly decreased its stability. Elongation of the Nse4 linker partially suppressed instability of the ATP-bound complex, suggesting that the binding of the KITE proteins to the Nse4 linker constrains its limited size. Our data suggest that the KITE proteins may shape the Nse4 linker to fit the ATP-free complex optimally and to facilitate opening of the complex upon ATP binding. This mechanism suggests an important role of the KITE subunits in the dynamics of the SMC5/6 complexes

Biochemical changes in the patient’s plasma after red blood cell transfusion

Introduction. The study aimed at in vivo assessment of the impact of administered red blood cells (RBCs) concentrates on the plasma levels of K+, lactate, pH, Na+, Ca++ and glucose, depending on the volume and age of administered products. Biochemical changes occurring during the storage of these products were studied in vitro simultaneously. Materials and methods. Arterial blood samples were collected in vivo from patients before and after RBCs transfusion and plasma levels of biochemical parameters were determined. A group of 80 RBCs samples was analyzed simultaneously, with the samples being equally distributed throughout the recommended storage time of 1–35 days. Results. The age of RBCs concentrate results in increased potassium and lactate levels and decreased pH, sodium and glucose levels in the RBCs samples. The concentrations were in the following ranges: potassium 4.0–40.5 mmol/L; lactate 4.1–28.0 mmol/L; pH 7.0–6.65; sodium 137–116 mmol/L; glucose 29.0–14.0 mmol/L. A prospectively selected group of 46 patients were administered a total of 354 RBCs units. The mean age of RBCs concentrates was 16.18 days. The number of administered RBCs units ranged from 2 to 38, a mean of 7.7 RBCs units/patient. The administration of 1 RBCs unit was associated with a mean in vivo increase of the potassium level by 0.07 mmol/L and lactate level by 0.13 mmol/L. Conclusion. The administration of larger amounts of RBCs concentrates may lead to an increase in the patient’s plasma levels of potassium and lactate. This increase is proportional to the age and volume of RBCs

Antiviral activity of temporin-1CEb analogues against gingival infection with herpes simplex virus type 1

IntroductionOral herpes infections caused by herpes simplex virus type 1 (HSV-1) are one of the most common in the human population. Recently, they have been classified as an increasing problem in immunocompromised patients and those suffering from chronic inflammation of the oral mucosa and gums. Treatment mainly involves nucleoside analogues, such as acyclovir and its derivatives, which reduce virus replication and shedding. As drug-resistant strains of herpes emerge rapidly, there is a need for the development of novel anti-herpes agents. The aim of the study was to design an antiviral peptide, based on natural compounds, non-toxic to the host, and efficient against drug-resistant HSV-1. Here, we designed a lysine-rich derivative of amphibian temporin-1CEb conjugated to peptides penetrating the host cell membrane and examined their activity against HSV-1 infection of oral mucosa.MethodsWe assessed the antiviral efficiency of the tested compound in simple 2D cell models (VeroE6 and TIGKs cells) and a 3D organotypic model of human gingiva (OTG) using titration assay, qPCR, and confocal imaging. To identify the molecular mechanism of antiviral activity, we applied the Azure A metachromatic test, and attachment assays techniques. Toxicity of the conjugates was examined using XTT and LDH assays.ResultsOur results showed that temporin-1CEb analogues significantly reduce viral replication in oral mucosa. The mechanism of peptide analogues is based on the interaction with heparan sulfate, leading to the reduce attachment of HSV-1 to the cell membrane. Moreover, temporin-1CEb conjugates effectively penetrate the gingival tissue being effective against acyclovir-resistant strains. Collectively, we showed that temporin-1CEb can be regarded as a novel, naturally derived antiviral compound for HSV-1 treatment

Oxidized polyethylene wax as a potential carbon source for PHA production

© 2016 The Authors. Published by MDPI. This is an open access article available under a Creative Commons licence. The published version can be accessed at the following link on the publisher’s website: https://doi.org/10.3390/ma9050367We report on the ability of bacteria to produce biodegradable polyhydroxyalkanoates (PHA) using oxidized polyethylene wax (O-PEW) as a novel carbon source. The O-PEW was obtained in a process that used air or oxygen as an oxidizing agent. R. eutropha H16 was grown for 48 h in either tryptone soya broth (TSB) or basal salts medium (BSM) supplemented with O-PEW and monitored by viable counting. Study revealed that biomass and PHA production was higher in TSB supplemented with O-PEW compared with TSB only. The biopolymers obtained were preliminary characterized by nuclear magnetic resonance (NMR), gel permeation chromatography (GPC), differential scanning calorimetry (DSC), and thermogravimetric analysis (TGA). The detailed structural evaluation at the molecular level was performed by electrospray ionization tandem mass spectrometry (ESI-MS/MS). The study revealed that, when TSB was supplemented with O-PEW, bacteria produced PHA which contained 3-hydroxybutyrate and up to 3 mol % of 3-hydroxyvalerate and 3-hydroxyhexanoate co-monomeric units. The ESI-MS/MS enabled the PHA characterization when the content of 3-hydroxybutyrate was high and the appearance of other PHA repeating units was very low

Magnetic quantum ratchet effect in (Cd,Mn)Te- and CdTe-based quantum well structures with a lateral asymmetric superlattice

We report on the observation of magnetic quantum ratchet effect in (Cd, Mn) Te-and CdTe-based quantum well structures with an asymmetric lateral dual grating gate superlattice subjected to an external magnetic field applied normal to the quantum well plane. A dc electric current excited by cw terahertz laser radiation shows 1/B oscillations with an amplitude much larger as compared to the photocurrent at zero magnetic field. We show that the photocurrent is caused by the combined action of a spatially periodic in-plane potential and the spatially modulated radiation due to the near-field effects of light diffraction. Magnitude and direction of the photocurrent are determined by the degree of the lateral asymmetry controlled by the variation of voltages applied to the individual gates. The observed magneto-oscillations with enhanced photocurrent amplitude result from Landau quantization and, for (Cd, Mn) Te at low temperatures, from the exchange enhanced Zeeman splitting in diluted magnetic heterostructures. Theoretical analysis, considering the magnetic quantum ratchet effect in the framework of semiclassical approach, describes quite well the experimental results