Regulation of biofilm formation by marT in Salmonella Typhimurium

Molecular Biology Reports - In this study, we aimed at identifying the regulatory role of marT gene, known as the regulator of misL, on 15 different biofilm-related genes in S. Typhimurium 14028...     

An Augmented Lagrangian Based Compressed Sensing Reconstruction for Non-Cartesian Magnetic Resonance Imaging without Gridding and Regridding at Every Iteration

Background

Non-Cartesian trajectories are used in a variety of fast imaging applications, due to the incoherent image domain artifacts they create when undersampled. While the gridding technique is commonly utilized for reconstruction, the incoherent artifacts may be further removed using compressed sensing (CS). CS reconstruction is typically done using conjugate-gradient (CG) type algorithms, which require gridding and regridding to be performed at every iteration. This leads to a large computational overhead that hinders its applicability.

Methods

We sought to develop an alternative method for CS reconstruction that only requires two gridding and one regridding operation in total, irrespective of the number of iterations. This proposed technique is evaluated on phantom images and whole-heart coronary MRI acquired using 3D radial trajectories, and compared to conventional CS reconstruction using CG algorithms in terms of quantitative vessel sharpness, vessel length, computation time, and convergence rate.

Results

Both CS reconstructions result in similar vessel length (P = 0.30) and vessel sharpness (P = 0.62). The per-iteration complexity of the proposed technique is approximately 3-fold lower than the conventional CS reconstruction (17.55 vs. 52.48 seconds in C++). Furthermore, for in-vivo datasets, the convergence rate of the proposed technique is faster (60±13 vs. 455±320 iterations) leading to a ∼23-fold reduction in reconstruction time.

Conclusions

The proposed reconstruction provides images of similar quality to the conventional CS technique in terms of removing artifacts, but at a much lower computational complexity.     

Prime-Boost Vaccination with SA-4-1BBL Costimulatory Molecule and Survivin Eradicates Lung Carcinoma in CD8+ T and NK Cell Dependent Manner

Subunit vaccines containing universal tumor associated antigens (TAAs) present an attractive treatment modality for cancer primarily due to their safety and potential to generate long-term immunological responses that can safeguard against recurrences. However, TAA-based subunit vaccines require potent adjuvants for therapeutic efficacy. Using a novel form of the 4-1BBL costimulatory molecule, SA-4-1BBL, as the adjuvant of choice, we previously demonstrated that a single vaccination with survivin (SVN) as a bona fide self TAA was effective in eradicating weakly immunogenic 3LL tumors in >70% of C57BL/6 mice. The present study was designed to i) assess the therapeutic efficacy of a prime-boost vaccination and ii) investigate the mechanistic basis of vaccine efficacy. Our data shows that a prime-boost vaccination strategy was effective in eradicating 3LL lung carcinoma in 100% of mice. The vaccine efficacy was correlated with increased percentages of CD8⁺ T cells expressing IFN-γ as well as potent killing responses of both CD8⁺ T and NK cells in the absence of detectable antibodies to ssDNA as a sign of autoimmunity. Antibody depletion of CD8⁺ T cells one day before vaccination completely abrogated therapeutic efficacy, whereas depletion of CD4⁺ T cells had no effect. Importantly, NK cell depletion had a moderate (∼50% reduction), but significant (p<0.05) effect on vaccine efficacy. Taken together, these results shed light on the mechanistic basis of the SA-4-1BBL/SVN subunit vaccine formulation in a lung carcinoma model and demonstrate the robust therapeutic efficacy of the prime-boost immunization strategy with important clinical implications.     

Contribution of Gamma amino butyric acid (GABA) to salt stress responses of Nicotiana sylvestris CMSII mutant and wild type plants

Plants accumulate high levels of Gamma amino butyric acid (GABA) in response to different environmental stresses and GABA metabolism has different functions such as osmotic and pH regulation, bypass of tricarboxylic acid cycle, and C:N balance. The cytoplasmic male sterile (CMS) II mutant of Nicotiana sylvestris has a deletion in the mitochondrial gene nad7 which encodes the NAD7 subunit of complex I which causes increased leaf respiration, impaired photosynthesis, slower growth and increased amino acid levels. In this study we aimed to elucidate the role of GABA and GABA metabolism in different genotypes of the same plant system under salt stress (100mM NaCl) in short (24h) and long (7, 14 and 21 days) terms. We have investigated the differences in leaf fresh and dry weights, relative water content, photosynthetic efficiency (F(v)/F(m)), glutamate dehydrogenase (GDH, EC 1.4.1.4) and glutamate decarboxylase (GAD, EC 4.1.1.15) enzyme activities, GABA content and GAD gene expression profiles. GDH activity showed variations in CMSII and wild type (WT) plants in the first 24h. GAD gene expression profiles were in good agreement with the GAD enzyme activity levels in CMSII and WT plants after 24h. In long-term salinity, GAD activities increased in WT but, decreased in CMSII. GABA accumulation in WT and CMSII plants in short and long term was induced by salt stress. Variations in GDH and GAD activities in relation to GABA levels were discussed and GABA metabolism has been proposed to be involved in better performance of CMSII plants under long term salinity.     

Multiple exposures to unloading decrease bone's responsivity but compound skeletal losses in C57BL/6 mice

A single exposure to mechanical unloading can result in significant bone loss, but the consequences of multiple exposures are largely unknown. Within a 18-wk period, adult C57BL/6 male mice were exposed to 2 wk of hindlimb unloading (HLU) followed by 4 wk of reambulation (RA) once (1x-HLU), twice (2x-HLU), or three times (3x-HLU), or served as ambulatory age-matched controls. In vivo μCT longitudinally tracked changes in trabecular and cortical compartments of the femur. Normally ambulating control mice experienced significant age-related loss in trabecular bone volume fraction throughout the course of the experiment. This loss was compounded by HLU with 2x- and 3x-HLU mice experiencing a 27% and 24% greater reduction in trabecular bone and a 60% and 63% inhibition of age-related trabecular thickening. The recovery of cortical bone was also incomplete during each 4-wk RA period and, at completion of the experiment, cortical area in 3x-HLU mice was 5% smaller than in control and 1x-HLU. When eliminating age as a confounding variable, comparison between individual HLU/RA cycles showed that the magnitude of the response diminished during subsequent exposures. The extent of trabecular thinning in mice unloaded for the first time was 1.6-fold greater than the second time and nearly twofold greater than the third time. Similarly, the increase in trabecular thickness during the first RA cycle was twofold greater than during the second and third RA cycle. Together, our data demonstrate that even though multiple exposures to mechanical unloading are more detrimental than a single unloading period, bone's mechanosensitivity is reduced with consecutive unloading/reambulation cycles.     

The influence of copper on alkaline protease stability toward autolysis and thermal inactivation

Alkaline proteases are one of the most important group of enzymes that are indispensable in a number of different industrial sectors. In this work, the effect of copper ions (Cu²⁺) was investigated for improving the thermostability and hydrolytic performance of Bacillus clausii GMBE 42 alkaline protease at different temperatures (45–65°C). Maximal residual activity was observed in the presence of 5 mM CuCl₂. The enzyme was thermoinactivated according to first‐order kinetics. A stabilization effect caused by copper ions was the result of a decrease in both autolysis and thermoinactivation rates. Thermodynamic analysis of the thermoinactivation process showed that E_a,i, ΔG_i, and ΔH_i values of the enzyme were higher in the presence of copper ions, but there was no measurable change in ΔS_i values. These results show the thermostabilizing potential of copper ions on the enzyme. Lower K_m values and higher k_cat and k_cat/K_m values were obtained in the presence of copper ions, which is an indication of the nonessential activation of the enzyme by copper ions. Thermodynamic analysis of casein hydrolysis showed that in presence of copper ions E_a, ΔG^≠, ΔH^≠, , and values of enzyme were lower, but there was no change in ΔS^≠ values. This is so far the first study that investigates the effect of cations on the basic catalytic and thermodynamic properties of an alkaline serine protease, which may be used to remove protein wastes from various industries such as food and leather processing.     

Synthesis and structural characterization of a novel seven-coordinate cobalt(II) complex: 2,9-Bis(ethanolamine)-1,10-phenanthrolinechlorocobalt(II) chloride

Condensation reaction of 2,9-dicarboxaldehyde-1,10-phenanthroline with 2-aminoethanol followed by NaBH₄ reduction yielded the polydentate Schiff base ligand 2,9-bis(ethanolamine)-1,10-phenanthroline in its reduced form. This ligand was characterized by elemental analysis, LC-MS, IR, UV-Vis and NMR spectroscopy. Reaction of the reduced Schiff base ligand with aqueous solution of cobalt(II) chloride affords 2,9-bis(ethanolamine)-1,10-phenanthrolinechlorocobalt(II) chloride in high yield. Single crystals of the cobalt(II) complex were obtained from the crystallization in ethanol and its structure was elucidated by X-ray structural analysis. The cobalt(II) complex ion was found to be seven-coordinated in a pentagonal bipyramidal geometry, whereby cobalt(II) ion is surrounded by the six donor atoms in the ligand molecule and a chloride ion.     

Is a single nucleotide polymorphism a risk factor for lung cancer in the matrix metalloproteinase-2 promoter?

We aimed to investigate the association of polymorphism frequencies of MMP-2 C1306T and MMP-2 plasma enzyme activity in lung cancer patients. In this study 300 genomic DNA (200 lung cancer patients + 100 no lung cancer) were analyzed. Polymorphisms were determined by using polymerase chain reaction-restriction fragment length polymorphism (RFLP) and electrophoresis. Plasma MMP-2 enzyme activity levels were measured by using ELISA. Sex, asbestos expose and smoking might be risk factors for lung cancer. The frequencies of C1306T genotypes in controls CC 65%, CT 32%, TT 3% and in patients CC 61%, CT 37%, TT2 % were found. It was determined that CC genotype frequency increase significantly in patients and controls. Plasma MMP-2 enzyme activity levels were increased in lung cancer patients according to controls. Finally, we claimed that determining of MMP-2 enzyme level can be used as a marker in lung cancer.     

Effects of surface-active block copolymers with oxyethylene and fluoroalkyl side chains on the antifouling performance of silicone-based films

Block copolymers made from a poly(dimethyl siloxane) (Si) and a poly(meth)acrylate carrying oxyethylene (EG) or fluoroalkyl (AF) side chains were synthesized and incorporated as surface-active components into a silicone matrix to produce cross-linked films with different surface hydrophilicity/phobicity. Near-edge X-ray absorption fine structure (NEXAFS) studies showed that film surfaces containing Si-EG were largely populated by the siloxane, with the oxyethylene chains present only to a minor extent. In contrast, the fluorinated block was selectively segregated to the polymer–air interface in films containing Si-AF as probed by NEXAFS and X-ray photoelectron spectroscopy (XPS) analyses. Such differences in surface composition were reflected in the biological performance of the coatings. While the films with Si-EG showed a higher removal of both Ulva linza sporelings and Balanus amphitrite juveniles than the silicone control, those with Si-AF exhibited excellent antifouling properties, preventing the settlement of cyprids of B. amphitrite.