Protein-tyrosine Kinase Pyk2 Mediates Endothelin-induced p38 MAPK Activation in Glomerular Mesangial Cells

Endothelin-1 (ET-1), a member of a family of 21 amino acid peptides possessing vasoconstrictor properties, is known to stimulate mesangial cell proliferation. In this study, ET-1 (100 nm) induced a rapid activation of p21(ras) in human glomerular mesangial cells (HMC). Inhibition of Src family tyrosine kinase activation with [4-Amino-5-(4-methylphenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine] or chelation of intracellular free calcium with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid acetoxymethyl ester significantly decreased ET-1dependent p21(ras) activation and suggested the involvement of the cytoplasmic proline-rich tyrosine kinase Pyk2. We have observed that Pyk2 was expressed in HMC and was tyrosine-phosphorylated within 5 min of ET-1 treatment. ET-1-induced activation of Pyk2 was further confirmed using phospho-specific anti-Pyk2 antibodies. Surprisingly, Src kinase activity was required upstream of ET-1-induced autophosphorylation of Pyk2. To determine whether Pyk2 autophosphorylation mediated ET-1-dependent p21(ras) activation, adenovirus-mediated transfer was employed to express a dominant-negative form of Pyk2 (CRNK). CRNK expression inhibited ET-1-induced endogenous Pyk2 autophosphorylation, but did not abolish ET-1-mediated increases in GTP-bound p21(ras) levels. ET-1-induced activation of the p38 MAPK (but not ERK) pathway was inhibited in HMC and in rat glomerular mesangial cells expressing the dominant-negative form of Pyk2. These findings suggest that the engagement of Pyk2 is important for ET-1-mediated p38 MAPK activation and hence the biological effect of this peptide in mesangial cells

Priprema i karakterizacija C60/C70 + Ni polikristalnih tankih slojeva naraslih na raznim podlogama

C60/C70 + Ni films were obtained by thermal evaporation method in a wide range of Ni concentration (from 1.5 to 10 wt. %). The polycrystalline structure was detected in layers with Ni concentration of 1.5 wt. %. For this Ni concentration, growth of grains in columnar form was observed by SEM. TEM examination showed existence of crystalline grains of a size of a few micrometers and 10 - 200 nm thick. The electron and X-ray diffraction exhibited Bragg distances of approx. 0.87, 0.83, 0.50 and 0.32 nm, and in electron diffraction 1.0, 0.76 and 0.65 nm interplanar distances were found. In Raman spectra, typical fullerenes and two other bands placed at 580 and 1100 cm-1 were observed. The intensity of the latter bands depends on Ni concentration. For Ni concentration higher then 1.5 wt. %, the degradation of fullerene structure was observed by HRTEM, electron and X-ray diffraction.Tanki slojevi C60/C70 + Ni dobiveni su toplinskim isparavanjem s koncentracijom Ni od 1.5 do 10 tež. %. Polikristalna struktura je opažena u slojevima s 1.5 tež. % Ni. Pomoću SEM je u takvim slojevima opažen stupčast rast zrna. Pomoću TEM ustanovljena su zrna duljine nekoliko µm i debljine 10-200 nm. Ispitivanja su načinjena također elektronskom i rendgenskom difrakcijom. U Ramanovim spektrima opažene su tipične vrpce fulerena pri 580 i 1100 cm−1 . Za koncentracije Ni iznad 1.5 tež. % opažena je degradacija strukture fulerena

Priprema i karakterizacija C60/C70 + Ni polikristalnih tankih slojeva naraslih na raznim podlogama

C60/C70 + Ni films were obtained by thermal evaporation method in a wide range of Ni concentration (from 1.5 to 10 wt. %). The polycrystalline structure was detected in layers with Ni concentration of 1.5 wt. %. For this Ni concentration, growth of grains in columnar form was observed by SEM. TEM examination showed existence of crystalline grains of a size of a few micrometers and 10 - 200 nm thick. The electron and X-ray diffraction exhibited Bragg distances of approx. 0.87, 0.83, 0.50 and 0.32 nm, and in electron diffraction 1.0, 0.76 and 0.65 nm interplanar distances were found. In Raman spectra, typical fullerenes and two other bands placed at 580 and 1100 cm-1 were observed. The intensity of the latter bands depends on Ni concentration. For Ni concentration higher then 1.5 wt. %, the degradation of fullerene structure was observed by HRTEM, electron and X-ray diffraction.Tanki slojevi C60/C70 + Ni dobiveni su toplinskim isparavanjem s koncentracijom Ni od 1.5 do 10 tež. %. Polikristalna struktura je opažena u slojevima s 1.5 tež. % Ni. Pomoću SEM je u takvim slojevima opažen stupčast rast zrna. Pomoću TEM ustanovljena su zrna duljine nekoliko µm i debljine 10-200 nm. Ispitivanja su načinjena također elektronskom i rendgenskom difrakcijom. U Ramanovim spektrima opažene su tipične vrpce fulerena pri 580 i 1100 cm−1 . Za koncentracije Ni iznad 1.5 tež. % opažena je degradacija strukture fulerena

The use of mathematical models describing the spread of Covid-19 in strategic state security management

Purpose: The security system of each country should be capable of identifying and eliminating potential threats. However, such events' variety and unpredictability make it impossible to be ready for each scenario, should one materialize. Situations posing a national security threat require that effective actions be undertaken, including risk prevention and mitigation of its effects. Decisions must be taken responsibly and competently. However, they also must be taken quickly, as time is a key factor in any emergency. Extreme caution must be exercised, as citizens' lives and health may depend on the strategic level decisions. Therefore, any instruments to support the decisions, enabling the best choices to be made, are very much desirable. Knowledge of the scale of the phenomenon and its evolution is particularly useful and important here. It is easier to fight an opponent knowing their strengths and potential. Design/Methodology/Approach: To know required, mathematical models may be constructed describing the phenomenon in question and its development over time. This paper presents and compares mathematical models describing the SARS-Cov-2 virus's evolution in four EU countries (Poland, Germany, Great Britain, and Italy). Findings: The ability to describe the evolution of the pandemic in mathematical terms and rely on the results of such modeling in strategic state security management. The results obtained were compared with the actions taken in the countries in question. Practical Implications: Furthermore, the importance of reliable information in making crucial decisions on the national level and achieving the appropriate degree of citizen security was emphasized.peer-reviewe

Electronically Excited States of Vitamin B12: Benchmark Calculations Including Time-Dependent Density Functional Theory and Correlated Ab Initio Methods

Time-dependent density functional theory (TD-DFT) and correlated ab initio methods have been applied to the electronically excited states of vitamin B12 (cyanocobalamin or CNCbl). Different experimental techniques have been used to probe the excited states of CNCbl, revealing many issues that remain poorly understood from an electronic structure point of view. Due to its efficient scaling with size, TD-DFT emerges as one of the most practical tools that can be used to predict the electronic properties of these fairly complex molecules. However, the description of excited states is strongly dependent on the type of functional used in the calculations. In the present contribution, the choice of a proper functional for vitamin B12 was evaluated in terms of its agreement with both experimental results and correlated ab initio calculations. Three different functionals, i.e. B3LYP, BP86, and LC-BLYP, were tested. In addition, the effect of relative contributions of DFT and HF to the exchange-correlation functional was investigated as a function of the range-separation parameter, {\mu}. The issues related to the underestimation of charge transfer (CT) excitation energies by TD-DFT was validated by Tozer's L diagnostic, which measures the spatial overlap between occupied and virtual orbitals involved in the particular excitation. The nature of low-lying excited states was also analyzed based on a comparison of TD-DFT and ab initio results. Based on an extensive comparision against experimental results and ab initio benchmark calculations, the BP86 functional was found to be the most appropriate in describing the electronic properties of CNCbl. Finally, an analysis of electronic transitions and a new re-assignment of some excitations are discussed.Comment: Accepted by the Journal of Chemistry

Serum soluble Fas ligand (sFasL) in patients with primary squamous cell carcinoma of the esophagus.

Esophageal carcinomas have been shown to express Fas ligand (FasL) and down-regulate Fas to escape from host immune surveillance. Circulating soluble FasL (sFasL) has been suggested to provide protection from Fas-mediated apoptosis. The aim of this study was to assess serum sFasL levels in esophageal cancer. The pretreatment levels of sFasL in the serum of 100 patients with esophageal squamous cell cancer and 41 healthy volunteers were determined by ELISA. Probability of survival was calculated according to the method of Kaplan-Meier. The prognostic influence of high and low level of sFasL was analyzed with the log-rank test. The mean serum level of sFasL in patients with esophageal cancer was significantly higher than that in healthy donors (1.567+/-1.786 vs 0.261+/-0.435,

The Araucaria Project: A study of the classical Cepheid in the eclipsing binary system OGLE LMC562.05.9009 in the Large Magellanic Cloud

We present a detailed study of the classical Cepheid in the double-lined, highly eccentric eclipsing binary system OGLE-LMC562.05.9009. The Cepheid is a fundamental mode pulsator with a period of 2.988 days. The orbital period of the system is 1550 days. Using spectroscopic data from three 4-8-m telescopes and photometry spanning 22 years, we were able to derive the dynamical masses and radii of both stars with exquisite accuracy. Both stars in the system are very similar in mass, radius and color, but the companion is a stable, non-pulsating star. The Cepheid is slightly more massive and bigger (M_1 = 3.70 +/- 0.03M_sun, R_1 = 28.6 +/- 0.2R_sun) than its companion (M_2 = 3.60 +/- 0.03M_sun, R_2 = 26.6 +/- 0.2R_sun). Within the observational uncertainties both stars have the same effective temperature of 6030 +/- 150K. Evolutionary tracks place both stars inside the classical Cepheid instability strip, but it is likely that future improved temperature estimates will move the stable giant companion just beyond the red edge of the instability strip. Within current observational and theoretical uncertainties, both stars fit on a 205 Myr isochrone arguing for their common age. From our model, we determine a value of the projection factor of p = 1.37 +/- 0.07 for the Cepheid in the OGLE-LMC562.05.9009 system. This is the second Cepheid for which we could measure its p-factor with high precision directly from the analysis of an eclipsing binary system, which represents an important contribution towards a better calibration of Baade-Wesselink methods of distance determination for Cepheids.Comment: Accepted to be published in Ap

Phosphorylation of Serine 43 Is Not Required for Inhibition of c-Raf Kinase by the cAMP-dependent Protein Kinase

The activity of the serine/threonine kinase c-Raf (Raf) is inhibited by increased intracellular cAMP. This is believed to require phosphorylation with the cAMP-dependent protein kinase (PKA), although the mechanism by which PKA inhibits Raf is controversial. We investigated the requirement for PKA phosphorylation using Raf mutants expressed in HEK293 or NIH 3T3 cells. Phosphopeptide mapping of (32)P-labeled Raf (WT) or a mutant lacking a putative PKA phosphorylation site (serine to alanine, S43A) confirmed that serine 43 (Ser(43)) was the major cAMP (forskolin)-stimulated phosphorylation site in vivo. Interestingly, the EGF-stimulated Raf kinase activity of the S43A mutant was inhibited by forskolin equivalently to that of the WT Raf. Forskolin also inhibited the activation of an N-terminal deletion mutant Delta5-50 Raf completely lacking this phosphorylation site. Although WT Raf was phosphorylated by PKA, phosphorylation did not inhibit Raf catalytic activity in vitro, nor did forskolin treatment inhibit the activity of an N-terminally truncated Raf protein (Raf 22W) or a full-length Raf protein (Raf-CAAX) expressed in NIH 3T3 cells. In contrast, forskolin inhibited the EGF-dependent activation of a Raf isoform (B-Raf), lacking an analogous phosphorylation site to Ser(43). Thus, these results demonstrate that PKA exerts its inhibitory effects independently of direct Raf phosphorylation and suggests instead that PKA prevents an event required for the EGF-dependent activation of Raf