Two Photon Spectroscopy Can Serve as a Marker of Protein Denaturation Pathway

Rhodamine group of molecules are widely used dyes for imaging of biological molecules. Application of these dyes however includes a limitation that these molecules absorb in the visible range of the spectrum, which does not fall in the ‘biologically transparent window’ (BTW). Two photon absorption (TPA) process could come up with an alternate solution to this as these dyes could be excited in the near infrared (NIR) window to extract similar information. To validate this we have investigated TPA cross section (TPACS, σ₂) of two rhodamine dyes, namely Rhodamine 6G (R6G), Rhodamine B (RhB), site selectively bound with a model protein, bovine serum albumin (BSA), by exciting at 800 nm. Two photon spectroscopy and imaging confirms the binding of the dye to the protein. The decreases in TPACS with increasing temperature at a fixed BSA concentration excellently follows the temperature induced structural transition of BSA as the protein transforms from a molten globule to unfolded conformation beyond 60 °C, which has previously been established through circular dichroism (CD) measurements. The thus established resemblance in TPACS and CD measurement trends thus strongly affirms the suitability of TPA process in protein imaging and as an alternative marker to tracking its conformational transformations using NIR radiation.     

Banchromene and other secondary metabolites from the endophytic fungus Fusarium sp. obtained from Piper guineense inhibit the motility of phytopathogenic Plasmopara viticola zoospores

A new chromene, (S)-banchromene (1), together with seven known compounds, ergosterol, beauvericin (2), fusaproliferin (3), radicinin (4), poly(3-hydroxybutyric acid) (PHB, 5), N-methylpyrrolidone and an inseparable mixture of isochromene derivatives 6a, 6b, were isolated from a culture of Fusarium sp. strain CAMKT24b1, an endophytic fungus from the leaves and twigs of Piper guineense (Piperaceae). The structures of these metabolites were elucidated on the basis of their spectroscopic data; the absolute configuration of 1 was determined by ab initio-calculation of the optical rotation. In tests with the zoospores of the grapevine downy mildew pathogen Plasmopara viticola, compounds 1–4 showed moderate to high levels of motility-impairing activity at concentrations as low as 2.5 μg/mL. Compound 2 was the most active, exhibiting both motility-halting and lytic activities. Furthermore, compounds 2 and 3 displayed significant cytotoxic activity against brine shrimp larvae (Artemia salina) at 10 μg/mL. This is the first report on motility inhibitory and lytic activities of metabolites from an endophytic Fusarium species against the zoospores of the downy mildew pathogen P. viticola.     

Structural characteristics and connection mechanism of gold-catalyzed bridging silicon nanowires

Lateral, single-crystalline silicon nanowires were synthesized using chemical vapor deposition catalyzed by gold nanoparticles deposited on one of the vertical {1 1 1} sidewalls of trenches etched in Si(0 1 1) substrates. Upon encountering the opposing sidewalls of the trenches, the lateral nanowires formed a mechanically strong connection. The bridging connection at the opposing sidewall was observed using high-resolution transmission electron microscopy (TEM) to be epitaxial and unstrained silicon-to-silicon. Using energy-dispersive X-ray spectroscopy in TEM, gold could not be detected at the interface region where the nanowires formed a connection with the opposing sidewall silicon deposit but was detected on the surface adjacent to the impingement region. We postulate that a silicon-to-silicon connection is formed as the gold–silicon liquid eutectic is forced out of the region between the growing nanowire and the opposing sidewall.     

Hollow Reticular Shaped Highly Ordered Rice Husk Carbon for the Simultaneous Determination of Dopamine and Uric Acid

We have prepared activated porous carbon material through simple pyrolysis method from rice husks (rhAC) for sensitive detection of dopamine (DA) and uric acid (UA). The prepared rhAC material was thoroughly characterized using various spectroscopic, microscopic, and electroanalytical techniques. Analysis of the voltammetric data showed that the analytes followed a first order reaction kinetics while following a 2e^?/2H⁺ transfer process. We have discussed the possible oxidation mechanism for the analytes based on the results from our experimental analysis. The rhAC_GCE sensor was tested for interference, reproducibility, and stability. The sensor was also tested to evaluate its applicability in real life.     

Effects of Temperature on The UV-B Sensitivity of Toxic Cyanobacteria Microcystis aeruginosa CS558 and Anabaena circinalis CS537

Rising global temperatures have been suggested to favor cyanobacteria over eukaryotic algae, but UV-B fluxes are also predicted to remain high and may interact with temperature to affect algal growth. To understand the interactive effects of temperature and UV-B radiation, cultures of Microcystis aeruginosa and Anabaena circinalis were grown at either 25 or 30°C and then exposed to an acute irradiance of UV-B (1.4 W m⁻²). Both species showed differences in growth rates at both temperature regimes. The growth rates of M. aeruginosa (0.41 ± 0.02 day⁻¹) and A. circinalis (0.38 ± 0.01 day⁻¹) were higher at 25 and 30°C, respectively. Rates of damage (k) and repair (r) were calculated from the kinetics of change in effective quantum yield, F_v'/F_m'. Analysis of the estimates of r and k shows that M. aeruginosa exhibited relatively high values for both parameters, compared to A. circinalis, at both growth temperatures. In both species, repair rates were higher at 30°C than at 25°C but in A. circinalis damage was also greater at the higher temperature. In contrast, M. aeruginosa showed a lower damage rate at the higher temperature. For both species, the ratio of r:k was higher at the higher temperature. However, the percent inhibition of effective quantum yield by UV-B was greater in A. circinalis than in M. aeruginosa as the r:k was lower A. circinalis. Therefore, it could be concluded that temperature may influence growth and bloom formation of cyanobacteria and that different species may respond differently to UV-B and temperature interactions.     

Effects of branching characteristics and copolymer composition distribution on non-isothermal crystallization kinetics of metallocene LLDPEs

The effects of branch content (BC) and copolymer composition distribution (CCD) on the non-isothermal crystallization kinetics of metallocene m-LLDPEs were studied using modified Avrami analysis, modulated differential scanning calorimetry (MDSC), and Crystaf. Several m-LLDPEs and an m-HDPE - all having comparable M_w and PDI - were experimented. In addition, a ZN-LLDPE was used for comparison purposes. The branch content, unlike the used cooling rates (2-6 °C/min), significantly affected the crystallization behavior. Crystallization peak temperature, , decreased linearly with increasing BC. All the m-LLDPEs showed primary and secondary crystallizations. The secondary crystallization showed to be more pronounced at high BC. The primary crystallization Avrami parameter n for m-HDPE ranged between 3.72 and 4.50, indicating spherulitic crystal growth whereas that for the m-LLDPEs, varied from 2.02 to 5.70. The ZN-LLDPE (having broader composition distribution) offered higher values of and than the m-LLDPEs with similar BC, M_w, and PDI. It, unlike the m-LLDPEs and m-HDPE, fairly agreed with the crystallization kinetic model proposed by Liu et al. The lamella thickness of the m-LLDPEs, L, calculated as per Gibbs-Thomson equation, showed to be in the range 2-16 nm, depending on BC and it decreased approximately following the relationship: L (nm) = 15.0 e^{(−0.0498BC)}.     

Interaction of Fusarium solani lectin with monosaccharides and oligosaccharides: a fluorometric study

The intrinsic fluorescence intensity of Fusarium solani lectin was quenched upon binding to mono- and oligosaccharides without any change in the emission maximum and it was used to determine association constants for several sugars and glycans. The lectin interacted very poorly with monosaccharides but well with disaccharides (T-antigen and LacNAc) with a distinction between beta1-->4 and beta1-->3 linkages. Among the monosaccharides, the interaction was observed only with Gal/GalNAc derivatives and not with Glc/Man derivatives. Thermodynamic studies revealed that the binding of the lectin with all the saccharides is enthalpically driven and exothermic in nature. Asialo-triantennary N-glycan and asialo-biantennary N-glycan showed higher affinity than monovalent LacNAc with significant increase in binding enthalpy, pointing towards the importance of multivalency in the lectin-ligand interactions. Time-resolved fluorescence measurement indicated the lectin has two lifetimes for tryptophan and the shorter lifetime is affected on ligand binding.     

Interaction of Sulfadiazine with Model Water Soluble Proteins: A Combined Fluorescence Spectroscopic and Molecular Modeling Approach

The binding behavior of antibacterial drug sulfadiazine (SDZ) with water soluble globular proteins like bovine as well as human serum albumin (BSA and HSA, respectively) and lysozyme (LYS) was monitored by fluorescence titration and molecular docking calculations. The experimental data reveal that the quenching of the intrinsic protein fluorescence in presence of SDZ is due to the strong interaction in the drug binding site of the respective proteins. The Stern-Volmer plot shows positive deviation at higher quencher concentration for all the proteins and was explained in terms of a sphere of action model. The calculated fluorophore-quencher distances vary within 4?~?11 Å in different cases. Fluorescence experiments at different temperature indicate thermodynamically favorable binding of SDZ with the proteins with apparently strong association constant (~10⁴–10⁵ M^?1) and negative free energy of interaction within the range of ?26.0?~??36.8 kJ mol^?1. The experimental findings are in good agreement with the respective parameters obtained from best energy ranked molecular docking calculation results of SDZ with all the three proteins.     

Extending Petri net to reduce control strategies of railway interlocking system

In our previous articles we gave step by step refinement process towards the development of safety properties of moving block interlocking system (MBRIS). The refinement process started from abstraction to fuzzy based safety properties using Z and then fuzzy multi agent specification language. However, one dimensional control of train passing through a switch and level crossing were not discussed. This paper reduces the existing two dimensional controls along the switch and level crossing to one dimensional for shifting it to a train only. For example, in the existing model the train movement along components switches and level crossings depends on both the train and components control. Whereas, in one dimensional control train is the only authority to control a switch and level crossing required for its desired operation. For this reduction, concurrent and mobile agent concepts are required. Therefore, we integrate mobile agent concepts with Petri nets to develop the mobile Petri net (MPN) a new class of PNs. This supports both mobility and concurrency. Further, we prove that the collection of different MPNs in a connected network is a PN. This proof allowed us to use the properties of PN to verify the system. Finally, we use MPN to model the safety properties of MBRIS along the switch and level crossing. This provides one dimensional control to a train along a switch and level crossing which increases the safety of the railway interlocking system. Moreover, we use reachability graph (RG) to verify the switch and level crossing models.     

Generation of higher degree chaos by controlling harmonics of the modulating signal in EDFRL

Erbium doped fiber ring lasers (EDFRL) are being used to generate optical chaos for secure communication by modulating the cavity loss/pump power or exploiting nonlinearities. The security level in chaotic communication depends on degree of chaos quantified by the Lyapunov exponent and its variability which is determined by the number of tuneable system parameters which were limited to five main parameters, i.e. modulation index, modulation frequency, pump power, cavity gain and loss. In this study we have increased the number of tuneable parameters using square, triangular and sum of harmonics waveforms. We have analysed the effect on degree of chaos of phase and duty cycle of square modulating signal with gradual addition of harmonics. For the given cavity parameters, the Lyapunov exponents can be increased by more than fifteen times using square wave modulating signal and a duty cycle of 60%. The electrical parameters identified make generation of new chaotic sequences more flexible in a field deployed EDFRL chaotic system.