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Different Multilayer electrospun nanofibrous membranes were synthesized to study the effects of potentially important steps and parameters including hot-pressing and chitosan / poly (vinyl alcohol) (CTS/PVA) solutions on the membranes performance in Cu(II) ions removal. Due to the importance of continuous adsorption process, in this research, dynamic removal from synthetic wastewater of Cu(II) ions was discussed. Five ENAMs were prepared using different chitosan / poly (vinyl alcohol) (CTS/PVA) solutions. A bead free, smooth ENAM with uniform interconnected porous structure was obtained using 3 % wt./vol CTS solution with CTS/PVA weight ratio of 50/50. The average fiber diameter, porosity and average pore diameter of the as synthesized ENAM were 99.1 nm, 50.6 %, and 0.189 μm, respectively. Multilayer ENAMs were prepared with steady state pure water fluxes of 237–706 LMH/bar as well as time dependent rejections up to 98.6 %. Considering the slight (3 %) reduction of multilayer ENAMs rejection, as well as their high (80 %) flux recovery ratio (FRR) after two regeneration cycles, it can be concluded that the membranes are definitely reusable.

The heart rate-based lactate minimum test is a highly reproducible exercise test. However, the relation between lactate minimum determined by this test and maximal lactate steady state in running and cycling is still unclear. Twelve endurance-trained men performed this test in running and cycling. Exercise intensity at maximal lactate steady state was determined by performing several constant heart rate endurance tests for both exercise modes. Heart rate, power output, lactate concentration, oxygen uptake and rating of perceived exertion at lactate minimum, maximal lactate steady state and maximal performance were analysed. All parameters were significantly higher at maximal lactate steady state compared to lactate minimum for running and cycling. Significant correlations (p

Steady state emission spectra and excited state lifetimes were measured for 1440 distinct heteroleptic [Ir(C^N)2(N^N)]+ complexes prepared via combinatorial parallelized synthesis; 72% of the complexes were found to be luminescent, and the emission maxima of the library spanned the visible spectrum (652-459 nm). Spectral profiles ranged from broad structureless bands to narrow emissions exhibiting vibrational substructure. Measured excited state lifetimes ranged between ∼0.1-14 μs. Automated emission spectral fitting with successive Gaussian functions revealed four distinct measured classes of excited states; in addition to well understood metal-ligand to ligand-charge transfer (3MLLCT) and ligand-centered (3LC) excited states, our classification also identified photophysical characteristics of less explored mixed 3MLLCT/3LC states. Electronic structure features obtained from DFT calculations performed on a large subset of these Ir(III) chromophores offered clear insights into the excited state properties and allowed the prediction of structure/luminescence relationships in this class of commonly used photocatalysts. Models with high prediction accuracy (R2 = 0.89) for emission color were developed on the basis of experimental data. Furthermore, different degrees of nuclear reorganization in the excited state were shown to significantly impact emission energy and excited state lifetimes.

1-(Pyridin-2-yl-hydrazonomethyl)-naphthalen-2-ol (PNOH) is a naphthalene-based fluorescence dual chemo-sensor for Al3+ and Zn2+. The probe (PNOH) is spectroscopically characterised and the chemo-sensing mechanism has been demonstrated through 1H NMR, absorption and both steady state and time resolved fluorescence study. The ‘turn-on’ luminescence property of PNOH is used for the selective detection of trace amount of Al3+and Zn2+via chelation enhanced fluorescence (CHEF) through complex formation. The 1:1 stoichiometry of each sensor-metal complex is observed from Job’s plot based on UV-Vis titration. Most promising advantage of the probe (PNOH) is its application in the one-pot detection of Al3+ (λem- 460 nm) and Zn2+ (λem- 510 nm) exciting at same wavelength (λex- 420 nm) while high intense emission appears at two different wavelengths. Limit of detection (LOD) of PNOH towards Al3+ & Zn2+ are found to be 60 nM & 365 nM respectively. Real water sample analysis has also been demonstrated by using the probe PNOH.

Nature has gifted us many organic molecules which have remarkable influence in our daily life. Amongst many organic molecules, heterocyclic organic molecules have gained potential applications in the advanced field of biomedicine, pharmaceutical, electronics and many more. In the present work fluorescence quenching of biologically active fluorescent probe 8EMOHCC by aniline in different solvents have been studied at room temperature. To understand the molecular behaviour in different media, solvents of different refractive index and dielectric constant have been used. Spectroscopic measurement techniques such as UV/Vis spectroscopy and time related single photon counting are employed to characterise the molecule at room temperature. The fluorescence quenching study shows linear dependence of SV-plot in solvents of different dielectric constants. It reveals that quenching reactions are dynamic in nature. Various parameters of quenching have been determined and identified the type of quenching involved in the quenching reaction. Further, kq is found to be greater than $$ {k}_q^{\prime } $$ in ACN, methanol, propanol and dioxane. Activation energy of quenching (Ea) is found to be greater than energy of diffusion (Ed) in ACN, methanol, propanol, THF solvents and Ed > Ea in dioxane, indicating that quenching reaction is not solely controlled by material diffusion but also activation process.

Herein, we report an intramolecular radical cyclization reaction towards the synthesis of pyrrolidinone derivatives via metal-free photoredox catalysis under irradiation from blue LEDs. Some of the remarkable features of this protocol include synthetic efficiency, green reaction profile, easy isolation of products and short reaction time. The photophysical properties of synthesized compounds were investigated via steady state and time-resolved fluorescence spectroscopy in the solid state. Results showed the promising opportunity for their spectral tuning together with large Stokes-shifted and highly active fluorescence emission, thus indicating that these molecular scaffolds can be effective probes for the biological applications and development of opto-electronic devices.

Changes in the structural dynamics of double stranded (ds)DNA upon ligand binding have been linked to the mechanism of allostery without conformational change, but direct experimental evidence remains elusive. To address this, a combination of steady state infrared (IR) absorption spectroscopy and ultrafast temperature jump IR absorption measurements has been used to quantify the extent of fast (∼100 ns) fluctuations in (ds)DNA·Hoechst 33258 complexes at a range of temperatures. Exploiting the direct link between vibrational band intensities and base stacking shows that the absolute magnitude of the change in absorbance caused by fast structural fluctuations following the temperature jump is only weakly dependent on the starting temperature of the sample. The observed fast dynamics are some two orders of magnitude faster than strand separation and associated with all points along the 10-base pair duplex d(GCATATATCC). Binding the Hoechst 33258 ligand causes a small but consistent reduction in the extent of these fast fluctuations of base pairs located outside of the ligand binding region. These observations point to a ligand-induced reduction in the flexibility of the dsDNA near the binding site, consistent with an estimated allosteric propagation length of 15 Å, about 5 base pairs, which agrees well with both molecular simulation and coarse-grained statistical mechanics models of allostery leading to cooperative ligand binding.

Carbon dioxide release rate increased from 0.37 per hour to 0.69 per hour when batch cultures of the ascomycete Aspergillus fumigatus AR04 were compared at 30 and 40° C. This rate increase indicates an approximation to unlimited growth in a stirred vessel., Summary To set a benchmark in fungal growth rate, a differential analysis of prototrophic Aspergillus fumigatus AR04 with three ascomycetes applied in > 103 t year‐1 scale was performed, i.e. Ashbya gosspyii (riboflavin), Aspergillus niger (citric acid) and Aspergillus oryzae (food‐processing). While radial colony growth decreased 0.5‐fold when A. gossypii was cultivated at 40°C instead of 28°C, A. fumigatus AR04 responded with 1.7‐fold faster hyphal growth. A. niger and A. oryzae formed colonies at 40°C, but not at 43°C. Moreover, all A. fumigatus strains tested grew even at 49°C. In chemostat experiments, A. fumigatus AR04 reached steady state at a dilution rate of 0.7 h‐1 at 40°C, 120% more than reported for A. gossypii at 28°C. To study mycelial growth rates under unlimited conditions, carbon dioxide increase rates were calculated from concentrations detected online in the exhaust of batch fermentations for 3 h only. All rates calculated suggest that A. fumigatus AR04 approximates Arrhenius’ rule when comparing short cultivations at 30°C with those at 40°C. Linearization of the exponential phase and comparison of the slopes revealed an increase to 192% by the 10°C up‐shift.

The photoreactivity of triphenylamine in homogeneous media has been investigated by means of laser flash photolysis spectroscopy and preparative experiments. The goal of this study consists in the evaluation of the photophysical and photochemical behavior of transient N-phenyl-4a,4b-dihydrocarbazole (DHC0) under polar and non-polar solvents which was not performed yet. The kinetic parameters of such a key intermediate including lifetime (τd, τR and τox) and rate constants (kd, kR and kox) have been measured and the intermediacy of DHC0 in the co-oxidation of nucleophilic substrates such as sulfides and phosphines was investigated by a combined time-resolved and steady-state approach. Two distinctly oxidizing intermediates, a hydroperoxy radical and a hydroperoxide derivative were formed from transient DHC0 in the presence of molecular oxygen. Radical I oxidizes triarylphosphines, whereas intermediate II converts sulfides into the corresponding sulfoxides.

In the present work, we investigated the anti-tyrosinase activity, mechanism and antioxidant activity of proanthocyanidins (PAs) isolated from Pinus thunbergii needles. The results showed that PAs mainly consisted of catechin/epicatechin and had a strong inhibitory effect on the monophenolase and diphenolase activity of tyrosinase. PAs could reduce the steady state activity of tyrosinase monophenolase while prolonging the reaction delay time in a dose-dependent manner. The inhibitory effect of PAs on tyrosinase diphenolase was reversible, with the IC50 value of 37.64 μg/mL. The inhibition kinetic analysis by Lineweaver-Burk plots showed that PAs were a mixed-type inhibitor of tyrosinase diphenolase, with inhibition constants KI and KIS of 29.92 μg/mL and 128.27 μg/mL, respectively. Further, determination of the metal chelating ability, scanning study, fluorescence quenching, and DOPA (dihydroxyphenylalanine) oxidation were used to study the potential inhibition mechanism of PAs on tyrosinase. The results confirmed that PAs could chelate with copper ions in the active site of tyrosinase to inhibit tyrosinase activity. Moreover, ABTS [2,2′-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid)] and FRAP (ferric reducing antioxidant power) assay showed good antioxidant capacity of PAs. This study revealed the possibility of Pinus thunbergii needle PAs as a new tyrosinase inhibitor in the fields of food, medicine, and cosmetics industries.

Catalysis is an essential function in living systems and provides a way to control complex reaction networks. In natural out-of-equilibrium chemical reaction networks (CRNs) driven by the consumption of chemical fuels, enzymes provide catalytic control over pathway kinetics, giving rise to complex functions. Catalytic regulation of man-made fuel-driven systems is far less common and mostly deals with enzyme catalysis instead of synthetic catalysts. Here, we show via simulations, illustrated by literature examples, how any catalyst can be incorporated in a non-equilibrium CRN and what their effect is on the behavior of the system. Alteration of the catalysts' concentrations in batch and flow gives rise to responses in maximum conversion, lifetime (i.e. product half-lives and t90 – time to recover 90% of the reactant) and steady states. In situ up or downregulation of catalysts' levels temporarily changes the product steady state, whereas feedback elements can give unusual concentration profiles as a function of time and self-regulation in a CRN. We show that simulations can be highly effective in predicting CRN behavior. In the future, shifting the focus from enzyme catalysis towards small molecule and metal catalysis in out-of-equilibrium systems can provide us with new reaction networks and enhance their application potential in synthetic materials, overall advancing the design of man-made responsive and interactive systems., We show, via simulations, how catalytic control over individual paths in a fuel-driven non-equilibrium chemical reaction network in batch or flow gives rise to responses in maximum conversion, lifetime and steady states.

DNA-stabilized silver clusters (Ag(N)-DNAs) exhibit diverse sequence-programmed fluorescence, making these tunable nanoclusters promising sensors and bioimaging probes. Recent advances in the understanding of Ag(N)-DNA structures and optical properties have largely relied on detailed characterization of single species isolated by chromatography. Because most Ag(N)-DNAs are unstable under chromatography, such studies do not fully capture the diversity of these clusters. As an alternate method, we use high-throughput synthesis and spectroscopy to measure steady state Stokes shifts of hundreds of Ag(N)-DNAs. Steady state Stokes shift is of interest because its magnitude is determined by energy relaxation processes which may be sensitive to specific cluster geometry, attachment to the DNA template, and structural engagement of solvent molecules. We identify 305 Ag(N)-DNA samples with single-peaked emission and excitation spectra, a characteristic of pure solutions and single emitters, which thus likely contain a dominant emissive Ag(N)-DNA species. Steady state Stokes shifts of these samples vary widely, are in agreement with values reported for purified clusters, and are several times larger than for typical organic dyes. We then examine how DNA sequence selects Ag(N)-DNA excitation energies and Stokes shifts, comment on possible mechanisms for energy relaxation processes in Ag(N)-DNAs, and discuss how differences in Ag(N)-DNA structure and DNA conformation may result in the wide distribution of optical properties observed here. These results may aid computational studies seeking to understand the fluorescence process in Ag(N)-DNAs and the relations of this process to Ag(N)-DNA structure.

Degenerated strains of Clostridium acetobutylicum lack the ability to produce solvents and to sporulate, allowing the continuous production of hydrogen and organic acids. A degenerated strain of Clostridium acetobutylicum was obtained through successive batch cultures. Its kinetic characterization showed a similar specific growth rate than the wild type (0.25 h−1), a higher butyric acid production of 6.8 g·L−1 and no solvents production. A steady state was reached in a continuous culture at a dilution rate of 0.1 h−1, with a constant hydrogen production of 507 mL·h−1, corresponding to a volumetric rate of 6.10 L·L−1 d−1, and a yield of 2.39 mol of H2 per mole of glucose which represents 60% of the theoretical maximum yield. These results suggest that the degeneration is an interesting alternative for hydrogen production with this strain, obtaining a high hydrogen production in a continuous culture with cells in a permanent acidogenic state.

Cobalt (Co) although, an essential element playing a key role in several physiological and biological processes, can transform into a potential toxin when present in higher amounts or in certain chemical forms. The increasing usage of transition metals in industries, agriculture, medicines and technology has led to an alarming rise in cobalt poisoning cases. Exposure to cobalt can cause several health hazards including neurodegenerative disorders like Alzheimer’s disease, Parkinson’s disease, etc. Cobalt may interact with several proteins in the cellular milieu, generating reactive oxygen species (ROS) and thereby triggering apoptosis of the neural cells via DNA damage. The most common method of removal of such metals from the body is by the use of metal chelators. In this study, we report a facile strategy for the clearance of Co (II) from the physiological milieu using citrate functionalized Mn3O4 NPs (C-Mn3O4 NPs). The chelation ability of C-Mn3O4 NPs has been evaluated using steady state and pico-second resolved optical spectroscopy. Our studies reveal that C-Mn3O4 NPs form a stable complex with cobalt ions, thereby reducing the ability of Co2+ to produce reactive oxygen species (ROS) via Fenton like reaction and hence function as a novel nanotherapeutic for the detoxification of cobalt in the human system.

Carmen Diez-Simon and et al used Maestro software to check the comparison of volatile trapping techniques for the comprehensive analysis of food flavourings by gas chromatography-mass spectrometry. According to RT-qPCR transcript levels of several antioxidant enzymes genes (ascorbate peroxidase, glytathione reductase and glutathione peroxidase) were significantly higher in 'Joker' compared to 'Oitol'. Novel software based methods such as molecular modeling;structure-based drug design, structure-based virtual screening, ligand interaction and molecular dynamics are considered to be powerful tool for investigation of pharmacokinetics and pharmacodyanamic properties of drug, and structural activity relationship between ligand and its target [12]. Raveendra Melanvanki and et al used Maestro software to check the investigation of interaction between boronic acids and sugar: effect of structural change of sugars on binding affinity using steady state and time resolved fluorecencespectroscopy and molecular docking.

The rate of Methanol synthesis over a Cu/ZnO/Al2O3 (60:30:10) catalyst has been measured using CO2/H2 (10:90) and CO/CO2/H2 (10:10:80) streams at 433, 443, 453, 463 and 473 K. Using the CO2/H2 stream, it requires 12 × 103 s to achieve steady-state performance; this time reduces to 5.4 × 103 s on increasing the temperature to 463 K. Using the CO/CO2/H2 stream, steady State performance is not achieved even after 14.4 × 103 s at 433 K but is achieved after 9 × 103 s at 463 K. Significant deviations from steady state behavior (~40% of steady state) are observed only at 453 K and only using the CO2/H2 feed when gas chromatography (GC) is the analysis system. When the reactor output is connected directly into a flame ionization detector (FID), oscillation is observed at all temperatures studied using a CO2/H2 stream. Injection of CO into the CO2/H2 stream, which is synthesizing methanol at 473 K, produces a sharply spiked increase in the rate of methanol synthesis followed by an oscillatory relaxation to steady state behavior. At 433 and 443 K, the injection of CO into the CO2/H2 stream again produces the sharply spiked increase in the rater of methanol synthesis, which returns to the baseline value without oscillations.

Background Inflammation, both acute and chronic, is associated with reductions in the synthesis of retinol-binding protein (RBP) and the concentration of retinol in plasma. Consequently, it is currently recommended that the retinol isotope dilution (RID) method not be used to estimate vitamin A total body stores (TBS) in subjects with inflammation. Objectives To apply compartmental analysis to study the effects of inflammation on hepatic apo-RBP input, plasma retinol pool size, and RID-predicted TBS in theoretical subjects with known steady state values for these parameters. Methods We selected 6 previously generated hypothetical subjects who ingested a dose of stable isotope-labeled vitamin A (day 0). Starting with each subject's published steady state model for retinol tracer kinetics, we developed a parallel model for unlabeled retinol and RBP that incorporated links between these entities and tied liver retinol secretion to RBP availability. Then we perturbed the steady state model by initiating chronic or acute inflammation on day 0 or acute inflammation on day 3 or 9 and simulating results for RBP, plasma retinol, and TBS. Results Chronic inflammation led to substantial reductions in RID-predicted TBS for at least 2 weeks after tracer administration. In contrast, acute inflammation induced on day 0 or 3 resulted in less dramatic impacts on TBS (37% or 20% reduction, respectively, from steady state levels, with levels rebounding by 14 days). When inflammation was induced 9 days after administration of the tracer, the effects on predicted TBS were minimal. Overall, for acute inflammation initiated at 0, 3, or 9 days, accurate predictions of TBS were obtained by 2 weeks. Conclusions Compartmental analysis can be applied in the novel way described here to study the influence of perturbations such as inflammation on predictions of vitamin A status using RID. Such an approach has potential value for studying other perturbations and different nutrients.

Sickle cell disease (SCD) is a chronic inflammatory disease associated with increased levels of multiple cytokines as interferon gamma (IFN-γ) during vaso-occlusive (VOC) crisis and steady state. Therefore, we aimed to explore theimmunological role of serum IFN-γ, interleukin-10 (IL-10) and CD4+ T lymphocytes in children with SCD at steady state and VOC. This study was conducted on 30 children with SCD (15 at steady state and 15 in VOC) and 30 healthy children as controls. Detailed history, complete physical, clinical examination and laboratory investigations as: complete blood count, serum ferritin, IFN-γ, IL-10 and CD4+ T lymphocytes for all patients were recorded. Results: Significant increase in serum IFN-γ in patients with SCD during VOC compared with patients at steady state and controls (P

Research on the interface between solid electrolytes and electrode materials or catholyte is important to effectively and safely use their high energy densities. However, compared to interfaces wit...

In this study, we investigated the production conditions of Quercus coccifera hydrochar, which is an inexpensive and easy available adsorbent, for the adsorption of Basic Red 18 (BR18) azo dye. The hydrochar was produced in the eco-friendly subcritical water medium (SWM). The effects of the pH (2–10), adsorbent size (45–106 μm), adsorbent dose (0.5–1.5 g/L), dye concentration (40–455 mg/L), and contact time (5–120 min) were studied via optimization experiments. The optimum conditions were pH 10, particle size of 45 μm, particle amount of 1.5 g/L, dye concentration of 455 mg/L, and 60 min. The removal efficiency increased sharply for the first 5 min; after that the removal efficiency reached a steady state at 60 min, with a maximum removal of 88.7%. The kinetic studies for the adsorption of BR18 dye in aqueous solution using hydrochar showed pseudo-second-order kinetics. The Langmuir and Freundlich isotherm models were used to explain the relationship between adsorbent and adsorbate, and Freundlich isotherm was the most suitable model because of its high regression coefficient (R2) value. The intraparticle diffusion model was used to determine the adsorption mechanism of BR18 onto Q. coccifera acorn hydrochar. Desorption studies were also carried out using different types of acid and different molarities.

This work aimed to analyze the dynamics of methane production and the conditions leading to a transient-inhibited steady-state phase in anaerobic digestion of physically pretreated slaughterhouse wastewater. Mathematical analysis provided complementary information for applications at the real scale. The pretreatments consisted of autoclaving (120 °C, 1.2 atm, 60 min), which generated the semisolid, and homogenization (30000 rpm, 1 minute). The semisolid was the substrate for a biochemical methane production test, which was carried out at room temperature with a substrate to inoculum ratio of 3:1. The methane production kinetics were complex and followed a stepped curve with four stages: 1) an exponential phase, 2) a transient methanogenesis inhibition phase, 3) a recovery phase and 4) a stationary phase. FTIR analysis showed that both the wastewater and semisolid had high contents of aliphatic and nitrogen compounds, while anaerobic digestion led to reductions of amines I and II, other amides, and aliphatic structures and the production of several amino acids. Mathematical models employed to predict cumulative methane production (Gompertz, Richards, Luedeking-Piret, and Modified first-rate) showed a good fit and low percentage error, although none of them accurately forecasted the transient inhibited steady-state (2nd phase).

Molecular dynamics (MD) simulations are conducted to systematically benchmark the effects of molecular weight, chain number, and cooling rate on the glass transition temperature (Tg) and coefficient of thermal expansion (CTE) of poly(ethylene oxide) (PEO). Hyperbolic regression as an objective identified method is used to extract Tg and CTE. The results show that for a cooling rate higher than 5 × 1013 K/min, Tg and CTE are both strongly affected by rapid quenching. For a cooling rate lower than 5 × 1013 K/min, Tg and CTE in the high-temperature domain still slightly depend on the cooling rate. Eventually, to eliminate the finite size effect of the model, a threshold molecular weight of 11,240 g/mol should be satisfied in the system. In addition, the chain number must be more than 10, at least for an oligomer system (50 monomers). This study comprehensively reveal the critical parameters of poly(ethylene oxide) (PEO) in the MD simulation that causes the numerical effect on polymer simulation and the variation of thermodynamics property. The threshold value of total molecular weight of 11,240 g/mol must be satisfied, which leads the polymer that exhibits its steady state bulk properties. For the oligomer system (Mw

Protein arginine methyltransferases (PRMTs) are of great interest for the development of therapeutics due to their involvement in a number of malignancies, such as lung and colon cancer. PRMT5 catalyzes the formation of symmetrical dimethylarginine of a wide variety of substrates and is responsible for the majority of this mark within cells. To gain insight into the mechanism of PRMT5 inhibition, we co-expressed the human PRMT5:MEP50 complex (hPRMT5:MEP50) in insect cells for a detailed mechanistic study. In this report, we carry out steady state, product, and dead-end inhibitor studies that show hPRMT5:MEP50 uses a rapid equilibrium random order mechanism with EAP and EBQ dead-end complexes. We also provide evidence of ternary complex formation in solution using hydrogen/deuterium exchange mass spectrometry. Isotope exchange and intact protein mass spectrometry further rule out ping-pong as a potential enzyme mechanism, and finally, we show that PRMT5 exhibits a pre-steady state burst that corresponds to an initial slow turnover with all four active sites of the hetero-octamer being catalytically active.

The dynamics of leaf photosynthesis in fluctuating light affects carbon gain by plants. Mesophyll conductance (gm) limits CO2 assimilation rate (A) under the steady state, but the extent of this limitation under non-steady-state conditions is unknown. In the present study, we aimed to characterize the dynamics of gm and the limitations to A imposed by gas diffusional and biochemical processes under fluctuating light. The induction responses of A, stomatal conductance (gs), gm, and the maximum rate of RuBP carboxylation (Vcmax) or electron transport (J) were investigated in Arabidopsis (Arabidopsis thaliana (L.)) and tobacco (Nicotiana tabacum L.). We first characterized gm induction after a change from darkness to light. Each limitation to A imposed by gm, gs and Vcmax or J was significant during induction, indicating that gas diffusional and biochemical processes limit photosynthesis. Initially, gs imposed the greatest limitation to A, showing the slowest response under high light after long and short periods of darkness, assuming RuBP-carboxylation limitation. However, if RuBP-regeneration limitation was assumed, then J imposed the greatest limitation. gm did not vary much following short interruptions to light. The limitation to A imposed by gm was the smallest of all the limitations for most of the induction phase. This suggests that altering induction kinetics of mesophyll conductance would have little impact on A following a change in light. To enhance the carbon gain by plants under naturally dynamic light environments, attention should therefore be focused on faster stomatal opening or activation of electron transport.

Bioaerosols are emitted during the biological treatment of water, soil, and air pollutants. The elimination of these pollutants has become a priority due to their detrimental effects on human health. Advanced oxidation technologies have been used to control bioaerosol emissions specially to improve indoor air quality. This investigation was focused on evaluating the biofiltration of ethyl acetate vapors in terms of removal efficiency and bioaerosol emission. Also, a continuous photocatalytic process to inactivate bioaerosols emitted from the biofilter was assessed as a post-treatment. The photocatalysis was developed with ZnO and TiO2 immobilized onto Poraver glass beads. Flow cytometry (FC) coupled with fluorochromes was used to characterize and quantify bioaerosol emissions in terms of live, dead, and injured cells. Ethyl acetate removal efficiencies were maintained in a steady state with values of 100% under 60-g m−3 h−1 inlet load (IL). Biomass concentration in the biofilter reached values up to 228 mgbiomass gperlite−1 at day 56 of operation, but the spontaneous occurrence of predatory mites diminished biomass concentration by 33%. Bioaerosols emitted during the steady-state operation of the biofilter were composed mainly by bacteria (~ 94%) and in a less extent of fungal spores (0.29–6%). The most efficient photocatalytic system comprised TiO2/Poraver with 78% inactivation of bioaerosols during the first 2 h of the process, whereas the ZnO/Poraver system showed null activity (~ 0%) of inactivation. FC results show that the main mechanism of inactivation of TiO2/Poraver was cell death.

Background Individuals with sickle cell disease (SCD) are susceptible to infective conditions that predispose them to hemolysis and anemia. Folic acid is recommended as a preventative measure against anemia in SCD patients; however, there is scarce literature on the implications of this practice. Patients and Methods Plasma concentrations of folate were measured in acutely ill pediatric SCD patients presenting with malaria or bacteremia and compared with those of SCD patients in steady state, or acutely ill non-SCD patients with confirmed malaria. Results The proportion of individuals with high (>45.3 nmol/L) folate concentrations was 29.5% (13/44), 18.2% (8/44), 33.3% (6/18), and 0% in the SCD-malaria, SCD steady state, SCD bacteremia, and the non-SCD malaria groups, respectively. The proportion of SCD patients with high folate levels did not vary significantly at steady state and during confirmed malaria (p = 0.216), and during acute bacteremia (p = 0.20). The median (interquartile range) plasma folate levels were 34.50 (24.40-52.00 nmol/L), 33.40 (15.83-60.85 nmol/L), 30.85 (24.68-39.65 nmol/L), and 13.30 (10.03-17.18 nmol/L), respectively, in the SCD malaria, SCD bacteremia, SCD steady state, and the non-SCD malaria sub-groups. The median folate levels of SCD steady state, SCD malaria, and SCD bacteremia sub-groups differed significantly (p < 0.0001) when compared with non-SCD patients, but the levels in the SCD bacteremia and malaria groups were not significantly different from the SCD steady state group. Conclusion Elevated levels of plasma folate were found in a high proportion of pediatric SCD patients. The implications of such elevated folate levels in pediatric SCD patients are unknown but may suggest a need for review of current recommendations for prophylactic doses of folic acid in SCD patients.

Photophysical and diffusion properties of excitons generated in perylene and pyrene nanoaggregates have been investigated using steady state as well as time-resolved absorption and emission spectro...

In this study, the kinetic resolution of (R,S)-2-pentanol via transesterification to achieve S-2-pentanol, a key intermediate required in the synthesis of anti-Alzheimer drugs, was investigated in continuous-flow packed-bed reactors. The effects of residence time, substrate concentration, and operation time of the enzyme were investigated. Under steady state conditions, 50% conversion and enantiomeric excess of the substrate (eeS)>99% were achieved at a residence time of 0.04 min. Productivity of the continuous-flow process (1.341 mmol/min/g)was about 4 times higher than that of the corresponding batch process (0.363 mmol/min/g). In addition, the mathematical modeling of the packed-bed reactor was conductedusing an axial dispersion model. Ping Pong Bi Bi kinetics was used in this model. Design parameters were determined and the developed equations were solved using an algorithm for solving boundary value problems for ordinary differential equations by collocation (bvp4c) using MATLAB. The results, obtained from the model, fitted the experimental data very well.

The chiral ligand N-methylephedrine (NME) was found to catalyse the addition of dimethylzinc to benzaldehyde in an enantiodivergent way, with a monomeric and a homochiral dimeric complex both catalysing the reaction at a steady state and giving opposite product enantiomers. A change in the sign of the enantiomeric product was thus possible by simply varying the catalyst loading or the ligand ee, giving rise to an enantiodivergent non-linear effect. Simulations using a mathematical model confirmed the possibility of such behaviour and showed that this can lead to situations where a reaction gives racemic products, although the system is composed only of highly enantioselective individual catalysts. Furthermore, depending on the dimer's degree of participation in the catalytic conversion, enantiodivergence may or may not be observed experimentally, which raises questions about the possibility of enantiodivergence in other monomer/dimer-catalysed systems. Simulations of the reaction kinetics showed that the observed kinetic constant kobs is highly dependent on user-controlled parameters, such as the catalyst concentration and the ligand ee, and may thus vary in a distinct way from one experimental setup to another. This unusual dependency of kobs allowed us to confirm that a previously observed U-shaped catalyst order vs. catalyst loading-plot is linked to the simultaneous catalytic activity of both monomeric and dimeric complexes., An asymmetric reaction consisting of competing monomeric and dimeric catalysts may explain enantiodivergent non-linear effects.

The isothermal oxidation behavior of Fe-10Al, Fe-15Al, Fe-20Al alloys with and without the addition of 0.1 at% Y was studied at 800 °C under 1 × 105 Pa of flowing pure O2 for 24 h. The oxidation of three Fe-Al alloys can be divided into transient state and steady state oxidation stages. The oxidation of each stage is approximately in accordance with the parabolic law. The addition of 0.1 at% Y changes the oxidation behavior obviously and leads to a significant increase of the weight gain of Fe-10Al and Fe-15Al. The scale grown on Fe-10Al is much thicker and more complicated than that grown on Fe-20Al, which is composed of an exclusive thin layer of Al2O3 protective film. Due to the formation of a large number of nodules, the scales grown on Fe-15Al cannot provide full protection for the alloy. Scale microstructure of the three Fe-Al-0.1Y alloys is similar to their corresponding Fe-Al alloys. However, nodules with very small size still appear on the surface of Fe-20Al-0.1Y alloy. The critical Al concentration to form an exclusive Al2O3 protective layer for Fe-Al binary alloy is on the borderline between 15 at%–20 at%. For Fe-Al-0.1Y alloy, the presence of 20 at% Al is not enough to inhibit the growth of nodules.

Blue-light-induced stomatal opening has been studied by applying a short pulse (~5 to 60 s) of blue light to a background of saturating photosynthetic red photons, but little is known about steady-state stomatal responses. Here we report stomatal responses to blue light at high and low CO2 concentrations. Steady-state stomatal conductance (gs ) of C3 plants increased asymptotically with increasing blue light to a maximum at 20% blue (120 µmol m-2 s-1 ). This response was consistent from 200 to 800 µmol mol-1 atmospheric CO2 (Ca ). In contrast, blue light induced only a transient stomatal opening (~5 min) in C4 species above a Ca of 400 µmol mol-1 . Steady-state gs of C4 plants generally decreased with increasing blue intensity. The net photosynthetic rate of all species decreased above 20% blue because blue photons have lower quantum yield (moles carbon fixed per mole photons absorbed) than red photons. Our findings indicate that photosynthesis, rather than a blue-light signal, plays a dominant role in stomatal regulation in C4 species. Additionally, we found that blue light affected only stomata on the illuminated side of the leaf. Contrary to widely held belief, the blue light-induced stomatal opening minimally enhanced photosynthesis and consistently decreased water use efficiency. This article is protected by copyright. All rights reserved.

This study describes the use of a prototype for the continuous photocatalytic reaction process using Fe/Nb2O5-immobilized catalyst for triclosan and 2.8-dichlorodibenzene-p-dioxin (2.8-DCDD)’s degradation. The experiments were carried out with different parameters and matrices in a steady state. In addition, photolysis and photocatalytic tests were performed. The results indicated that the generation of 2.8-DCDD was observed in matrices with Cl−. The Fe/Nb2O5-immobilized catalysts were efficient in the degradation of triclosan and 2.8-dichlorodibenzene-p-dioxin. However, 2.8-DCDD formation was not observed in the ultra-pure water matrix, which indicated influence of ions. The photocatalysis was more efficient than the photolysis when comparing both matrices and radiation. Even with a radiation oscillation, the solar process showed positive results.

The nitrate radical (NO3) and dinitrogen pentoxide (N2O5) play an important role in the nocturnal atmosphere chemistry. Observations of NO3 radicals and N2O5 were performed in a semirural ground site at Tai'Zhou in polluted southern China using cavity ring down spectroscopy (CRDS) from 23 May to 15 June 2018. The observed NO3 and N2O5 concentrations were relatively low, with 1 min average value of 4.4 ± 2.2 and 26.0 ± 35.7 pptV, respectively. The N2O5 uptake coefficient was determined to be from 0.027 to 0.107 based on steady state lifetime method. Fast N2O5 hydrolysis was the largest contributor to the loss of NO3 and contributed to substantial nitrate formation, with an average value of 14.83 ± 6.01 µg/m3. Further analysis shows that the N2O5 heterogeneous reactions dominated the nocturnal NOx loss and the nocturnal NOx loss rate is 0.14 ± 0.02 over this region.

Systematic variation of Mo/Al = {0.10, 0.25, 0.35} in Mo/H-ZSM-5 catalysts effects ∼20x change of Mo-to-H+ ratio during methane dehydroaromatization (DHA) reactions at steady state and on deactivating catalysts. Mo and Bronsted-acid site count, respectively enumerated by ethane hydrogenolysis probe reaction and dimethyl ether chemical titration, evolve disparately during DHA catalysis, indicating the two sites reduce in number by distinct mechanisms. The deactivation of Mo/H-ZSM-5 catalytic activity is uniquely attributed to the loss of active, or accessible, Mo sites, evinced by (i) the non-selective nature of deactivation (i.e. that deactivation occurs without modification of residual active sites), (ii) invariance in product distribution from Mo/H+ = 0.10 – 2.1, and (iii) linear correspondence between Mo-catalyzed ethane hydrogenolysis rate and active site count during DHA on deactivating samples. DHA forward rate – the intrinsic kinetic descriptor of catalyst activity – normalizes by Mo content at steady state and on deactivating catalysts with Mo/Al = 0.10 – 0.35 for Mo-based contact times in the range τMo = 0.79 – 44 molMo s mol C -1 , unequivocally establishing Mo aggregates as the sole kinetically-relevant active site on Mo/H-ZSM-5. Bronsted-acid site content over a ∼40x range in τH+ = 1.4 – 54 molH+ s mol C -1 has no discernable correlation with or effect on DHA rate, methane conversion, or product distribution – demonstrating H+ does not catalyze any rate- or selectivity-determining steps in the benzene formation pathway and suggesting that the benefits of zeolitic acid sites are only to disperse Mo during catalyst synthesis and, conceivably, to catalyze equilibrated reaction steps.

We study the following Neumann problem in one-dimension space arising from population genetics: { u t = d u x x + h ( x ) u 2 ( 1 − u ) in ( − 1 , 1 ) × ( 0 , ∞ ) , 0 ≤ u ≤ 1 in ( − 1 , 1 ) × ( 0 , ∞ ) , u ′ ( − 1 , t ) = u ′ ( 1 , t ) = 0 in ( 0 , ∞ ) , where h changes sign in ( − 1 , 1 ) and d is a positive parameter. Lou and Nagylaki (2002) [6] conjectured that if ∫ − 1 1 h ( x ) d x ≥ 0 , then this problem has at most one nontrivial steady state (i.e., a time-independent solution which is not identically equal to zero or one). Nakashima (2018) [15] proved this uniqueness under some additional conditions on h ( x ) . Unexpectedly, in this paper, we discover 3 nontrivial steady states for some h ( x ) satisfying ∫ − 1 1 h ( x ) d x ≥ 0 . Moreover, bi-stable phenomenon occurs in this scenario: one with two layers is stable; two with one layer each are ordered with the smaller one being stable and the larger one being unstable.

A combined steady-state and transient kinetic study on the mechanism of diphenylmethane (DPM) formation in HZSM-5 from HCHO and benzene revealed two kinetically relevant steps—the alkylation of ben...

UDP‐glucuronic acid 4‐epimerase is a short‐chain dehydrogenase/reductase that uses tightly bound NAD+ to catalyze interconversion of UDP‐glucuronic acid and UDP‐galacturonic acid. Here, the authors present a biochemical mechanistic study of the enzyme from Bacillus cereus. Epimerization involves oxidation at the substrate C4 and nonstereoselective reduction in a 4‐keto intermediate. Substrate positioning in a kinetically slow binding step can establish stereo‐electronic constraints to prevent decarboxylation of the intermediate., UDP‐glucuronic acid (UDP‐GlcA) is a central precursor in sugar nucleotide biosynthesis and common substrate for C4‐epimerases and decarboxylases releasing UDP‐galacturonic acid (UDP‐GalA) and UDP‐pentose products, respectively. Despite the different reactions catalyzed, the enzymes are believed to share mechanistic analogy rooted in their joint membership to the short‐chain dehydrogenase/reductase (SDR) protein superfamily: Oxidation at the substrate C4 by enzyme‐bound NAD+ initiates the catalytic pathway. Here, we present mechanistic characterization of the C4‐epimerization of UDP‐GlcA, which in comparison with the corresponding decarboxylation has been largely unexplored. The UDP‐GlcA 4‐epimerase from Bacillus cereus functions as a homodimer and contains one NAD+/subunit (k cat = 0.25 ± 0.01 s−1). The epimerization of UDP‐GlcA proceeds via hydride transfer from and to the substrate’s C4 while retaining the enzyme‐bound cofactor in its oxidized form (≥ 97%) at steady state and without trace of decarboxylation. The k cat for UDP‐GlcA conversion shows a kinetic isotope effect of 2.0 (±0.1) derived from substrate deuteration at C4. The proposed enzymatic mechanism involves a transient UDP‐4‐keto‐hexose‐uronic acid intermediate whose formation is rate‐limiting overall, and is governed by a conformational step before hydride abstraction from UDP‐GlcA. Precise positioning of the substrate in a kinetically slow binding step may be important for the epimerase to establish stereo‐electronic constraints in which decarboxylation of the labile β‐keto acid species is prevented effectively. Mutagenesis and pH studies implicate the conserved Tyr149 as the catalytic base for substrate oxidation and show its involvement in the substrate positioning step. Collectively, this study suggests that based on overall mechanistic analogy, stereo‐electronic control may be a distinguishing feature of catalysis by SDR‐type epimerases and decarboxylases active on UDP‐GlcA.

A steady-state kinetic model for the NH3-SCR and NH3 oxidation by a monolithic Cu-exchanged chabazite (Cu-CHA) catalyst has been developed by a combination of flow reactor experiments and kinetic modeling with an in-house computational code. Arrhenius parameters for NH3-SCR and NH3 oxidation and reaction orders with respect to H2O and O2 are collected using a Cu-CHA catalyst coated onto a honeycomb cordierite substrate. While H2O is shown to affect NH3-SCR and low temperature NH3 oxidation to N2 negatively, it promotes high temperature NH3 oxidation to NO. By a combination of calculations for heat and mass transfer and the parameters of the monolithic catalyst, a kinetic model is proposed. The model is validated with eight new experiments for NH3-SCR and NH3 oxidation. The calculated conversions of NO and NH3 are in good agreement with the experimental values over a wide range of temperatures (150 °C–600 °C), indicating that the model can be used for predicting the catalyst performance of the monolithic Cu-CHA for NH3-SCR.

BACKGROUND The objective of this study was to investigate the serum concentrations of olanzapine in relation to age, sex, and other factors in Chinese patients aged between 10 and 90 years. METHODS Data for 884 olanzapine patients, deposited between 2016 and 2017, were retrieved from the therapeutic drug monitoring database of the Affiliated Brain Hospital of Guangzhou Medical University. The effects of covariates on serum olanzapine concentration, dose-normalized concentration (C/D ratio), and normalized concentration (C/D/weight) were investigated. RESULTS Generally, male patients had lower olanzapine concentration, C/D ratio, and C/D/weight than female patients (P < 0.001). Smoking and drinking reduced olanzapine concentration, C/D ratio, and C/D/weight (P < 0.001). Coadministration with valproate decreased olanzapine concentration, C/D ratio, and C/D/weight by about 16%, 30%, and 40%, respectively (P < 0.001). Patients younger than 60 years had higher olanzapine concentrations (P < 0.05) but lower C/D ratios and C/D/weight (P < 0.001) than patients older than 60 years. Age was correlated with olanzapine concentration (r = -0.082, P < 0.05), C/D ratio (r = 0.196, P < 0.001), and C/D/weight (r = 0.169, P < 0.001). Sample timing after dose and diagnostic factors also contributed to the olanzapine concentrations. Multiple linear regression analysis revealed significant influences of dosage, age, sex, valproate comedication, smoking, postdose interval, and schizophrenia (vs bipolar affective disorders) on serum olanzapine concentrations. CONCLUSIONS The metabolism of olanzapine may be altered by several factors. Patients characterized with a combination of factors may benefit from therapeutic drug monitoring for the adjustment of olanzapine dose to minimize adverse reactions.

Elevation of intracellular Zn2+ following ischemia contributes to cell death by affecting mitochondrial function. Zn2+ is a differential regulator of the mitochondrial enzyme lipoamide dehydrogenase (LADH) at physiological concentrations (Ka = 0.1 µM free zinc), inhibiting lipoamide and accelerating NADH dehydrogenase activities. These differential effects have been attributed to coordination of Zn2+ by LADH active-site cysteines. A detailed kinetic mechanism has now been developed for the diaphorase (NADH-dehydrogenase) reaction catalyzed by pig heart LADH using 2,6-dichlorophenol-indophenol (DCPIP) as a model quinone electron acceptor. Anaerobic stopped-flow experiments show that two-electron reduced LADH is 15-25-fold less active towards DCPIP reduction than four-electron reduced enzyme, or Zn2+-modified reduced LADH (the corresponding values of the rate constants are (6.5 ± 1.5) × 103 M–1·s–1, (9 ± 2) × 104 M–1·s–1, and (1.6 ± 0.5) × 105 M–1·s–1, respectively). Steady-state kinetic studies with different diaphorase substrates show that Zn2+ accelerates reaction rates exclusively for two-electron acceptors (duroquinone, DCPIP), but not for one-electron acceptors (benzoquinone, ubiquinone, ferricyanide). This implies that the two-electron reduced form of LADH, prevalent at low NADH levels, is a poor two-electron donor compared to the four-electron reduced or Zn2+-modified reduced LADH forms. These data suggest that zinc binding to the active-site thiols switches the enzyme from one- to two-electron donor mode. This zinc-activated switch has the potential to alter the ratio of superoxide and H2O2 generated by the LADH oxidase activity.

The effects of itraconazole, a strong CYP3A4 inhibitor, on the steady-state pharmacokinetics of vemurafenib were evaluated in a phase 1, multicenter, open-label, fixed-sequence study. Patients with BRAFV600 mutation-positive metastatic malignancies received oral vemurafenib 960 mg twice daily on days 1 to 20 (period A) and oral vemurafenib 960 mg twice daily with oral itraconazole 200 mg once daily on days 21 to 40 (period B). A mixed-effects analysis of variance model was used to compare log-transformed area under the concentration-time curve during the dosing interval and maximum plasma concentration values for vemurafenib in 8 patients between period B (vemurafenib plus itraconazole) and period A (vemurafenib alone). Multiple doses of itraconazole increased steady-state exposure of vemurafenib by approximately 40%, with geometric least squares mean ratios (period B/period A) of 140% (90% confidence interval, 121-161) for both maximum plasma concentration and area under the concentration-time curve during the dosing interval. There was no apparent increase in incidence or severity of adverse events during coadministration of vemurafenib with itraconazole. In conclusion, coadministration of itraconazole with vemurafenib resulted in a modest increase in exposure of vemurafenib at steady state and was generally well tolerated.

Homogeneous crystal nucleation in polyamide 66 (PA 66) was studied by a combination of atomic-force microscopy (AFM) and fast-scanning chip calorimetry (FSC), with a specific experimental setup all...

Thiopental is an ultra-short-acting barbiturate and has been used commonly in the induction phase of general anesthesia. However, the toxic effect of thiopental is not completely clear. The effect of thiopental on intracellular Ca2+ ([Ca2+]i) levels was investigated in non-excitable cells. Experiments were carried out using a flow-cytometric technique, rat thymic lymphocytes (as non-excitable cells), and appropriate fluorescent probes. Treatment of cells with 300 µM thiopental increased Fluo-3 fluorescence intensity, indicating elevation of [Ca2+]i. This increase was partially attenuated by a chelator of intracellular Zn2+. Thus, thiopental elevated both [Ca2+]i and intracellular Zn2+ ([Zn2+]i) levels. Under intracellular Zn2+-free conditions, 100–300 µM thiopental was still able to induce a statistically significant increase in [Ca2+]i, whereas removal of extracellular Ca2+ greatly reduced the increase in [Ca2+]i induced by this dose of thiopental. Therefore, the thiopental-induced increase in [Ca2+]i was mainly due to an increased influx of Ca2+. Treatment of cells with 300 µM thiopental increased FluoZin-3 fluorescence intensity, indicating the presence of [Zn2+]i, both in the presence and absence of extracellular Zn2+. The thiopental-induced elevation of [Zn2+]i was due to an increase in both influx of Zn2+ and intracellular Zn2+ release. Concanavalin A (10 µg/mL) augmented Fluo-3 fluorescence in the presence of an intracellular Zn2+ chelator. The combination of concanavalin A and 100–300 µM thiopental synergistically increased [Ca2+]i. Results suggest that thiopental increases [Ca2+]i in both quiescent and activated lymphocytes, possibly resulting in modulation of immune system function.

In this study, a correlation of kinetic parameters was established for the decolorization of a methyl orange (MO) azo dye by Fenton oxidation. The experiments were carried out in a batch reactor at room temperature and the operating conditions were optimized using the 24−1 fractional factorial design. Each of the selected factors, i.e. the initial concentration of the MO dye, the catalyst dosage (Fe2+), the initial concentration of H2O2 and the pH of the solution, were varied on two levels. Regressions equations were constructed by relating the parameters of dye oxidation rate in aqueous phase (the initial rate Vi, the rate constant k and the maximum yield by unit time) to four operating conditions. MO dye decolorization profiles as a function of time were satisfactorily adjusted for the majority of the experiments by second-order kinetics on the Levenberg Marquart algorithm using Minitab 17. The validation of the model equations, carried out by the Analysis of Variance (ANOVA), showed that these equations explain about 98% of the variability of the responses and that these simple linear models should cover 92% of the future responses. It has been shown that there is robust relationship between the dynamic behaviour of this process at the initial time and the maximum yield of decolorization at steady state with dye concentration and hydrogen peroxide.

C 3 F 7 CN/CO 2 and C 3 F 7 CN/N 2 gas mixtures are regarded as the most promising substitutes to replace SF 6 in high-voltage devices. In this study, a measurement system for steady state Townsend (SST) method was built and the effective ionization coefficient (α), density normalized critical electric field (E/N) and the coefficient of synergistic effect (C) of the two gas mixtures were obtained. The effective ionization coefficients were measured in the variety of E/N between 150 and 1050 Td (1 Td = 10−21 V-m2). The density normalized critical electric fields ((E/N) lim ) of gas mixtures shows nonlinear increase with the increase of C 3 F 7 CN concentration, from 0 to 100%. It was found that the (E/N) Um of C 3 F 7 CN/N 2 gas mixture is higher than that of C 3 F 7 CN/CO 2 at a C 3 F 7 CN mole ratio lower than 60.5%, and the difference between two gas mixtures becomes very small when the ratio of C 3 F 7 CN is higher than 60.5%. The synergistic effect rises sharply and then decreases slowly with increasing ratio of C 3 F 7 CN. When the mole ratio is 6.9% and 6.6%, the coefficients of synergistic effect in C 3 F 7 CN/CO 2 and C 3 F 7 CN/N 2 gas mixtures reach to the highest point respectively. In the range where the ratio is less than 20.5%, the synergistic effect of gas mixture using N 2 as the buffer gas is stronger. This paper will supply the basic data for C 3 F 7 CN/CO 2 and C 3 F 7 CN/N 2 gas mixtures measured by SST.

Aims Phospholamban (PLB) stoichiometrically regulates the cardiac Ca2+ pump (SERCA2a) in the sarcoplasmic reticulum (SR); but in the nuclear envelope (NE) of cardiomyocytes (CMs), the PLB to SERCA2a molar ratio is higher, which highlights our poor understanding of how SR proteins distribute to their functional subcompartments. By tracking newly made PLB and SERCA2a in CMs, we will elucidate underlying cellular pathways responsible for their unique intracellular distributions. Methods and results Highly specific monoclonal antibodies were used to compare the subcellular distributions of SERCA2a, PLB, and junctin (JCN) in dog heart tissue. The data supported a view that both non-junctional and junctional SR proteins are all prominently enriched in transverse stretches of SR tubules, along the edges of sarcomeres (SR z-tubules). To understand the genesis of these steady state distributions, we analyzed confocal immunofluorescence images of adult rat CMs after acute expression (12-48 h) of the dog ortholog of PLB (dPLB) or dSERCA2a. Newly made dog proteins in rat CMs were detected using dog–specific monoclonal antibodies. By 12-24 h, dSERCA2a had accumulated within the NE in a punctate pattern, presumably reflecting initial sites of biosynthesis. Over the next 24-48 h, higher levels of dSERCA2a immunofluorescence accumulated in transverse/radial SR tubules, aligned along sarcolemmal transverse (T)-tubules, and extending from NE puncta. The patterns of SR tubules carrying dSERCA2a overlapped with those for newly made JCN, suggesting a common Nuclear Envelope to SR along T-tubules or NEST pathway for SR proteins. In contrast to the SERCA2a distribution pattern, dPLB accumulated uniformly in the NE, without visible puncta. With co-expression of dSERCA2a, however, PLB no longer uniformly filled the NE, but instead moved together with SERCA2a to form bright NE puncta, from which the two proteins then trafficked anterogradely. Conclusion Expression of dog SR protein orthologs (dSERCA2a, dPLB, and dJCN) for as little as 48 h reproduces their characteristic steady state distributions. Detailed analyses of the time courses of protein accumulation suggest a possible mechanism by which PLB distributes to both the NE and SR, unlike SERCA2a. SERCA2a moves in SR z-tubules directly from rough ER, along pathways that are in common with those used by junctional SR proteins. A different trafficking route for PLB away the rough ER/NE led to its accumulation in the NE, a process that may account for its enrichment in NE in situ. Association of SERCA2a with PLB from this NE pool enhanced PLB trafficking along the NEST pathway, contributing to steady state stoichiometry and physiologically regulated SERCA2a.

The kinetic theory of enzymes that modify insoluble substrates is still underdeveloped, despite the prevalence of this type of reaction both in vivo and industrial applications. Here, we present a steady-state kinetic approach to investigate inhibition occurring at the solid–liquid interface. We propose to conduct experiments under enzyme excess (E0 ≫ S0), i.e. the opposite limit compared with the conventional Michaelis–Menten framework. This inverse condition is practical for insoluble substrates and elucidates how the inhibitor reduces enzyme activity through binding to the substrate. We claim that this type of inhibition is common for interfacial enzyme reactions because substrate accessibility is low, and we show that it can be analyzed by experiments and rate equations that are analogous to the conventional approach, except that the roles of enzyme and substrate have been swapped. To illustrate the approach, we investigated the major cellulases from Trichoderma reesei (Cel6A and Cel7A) acting on insoluble cellulose. As model inhibitors, we used catalytically inactive variants of Cel6A and Cel7A. We made so-called inverse Michaelis–Menten curves at different concentrations of inhibitors and found that a new rate equation accounted well for the data. In most cases, we found a mixed type of surface-site inhibition mechanism, and this probably reflected that the inhibitor both competed with the enzyme for the productive binding-sites (competitive inhibition) and hampered the processive movement on the surface (uncompetitive inhibition). These results give new insights into the complex interplay of Cel7A and Cel6A on cellulose and the approach may be applicable to other heterogeneous enzyme reactions.

The multi-route reversible chemical scheme involves several chemical species and needs to inspect their behavior, activation energy, and the transition period before attaining equilibrium. The comparison between the transition periods of each reaction route is perceived. However, the behavior of the involved species in a multi-routes of the reaction mechanism is presented graphically. The computational procedure is adopted by using MATLAB.