Leveraging an enzyme/artificial substrate system to enhance cellular persulfides and mitigate neuroinflammation

Persulfides and polysulfides, collectively known as the sulfane sulfur pool along with hydrogen sulfide (H₂S), play a central role in cellular physiology and disease. Exogenously enhancing these species in cells is an emerging therapeutic paradigm for mitigating oxidative stress and inflammation that are associated with several diseases. In this study, we present a unique approach of using the cell''s own enzyme machinery coupled with an array of artificial substrates to enhance the cellular sulfane sulfur pool. We report the synthesis and validation of artificial/unnatural substrates specific for 3-mercaptopyruvate sulfurtransferase (3-MST), an important enzyme that contributes to sulfur trafficking in cells. We demonstrate that these artificial substrates generate persulfides in vitro as well as mediate sulfur transfer to low molecular weight thiols and to cysteine-containing proteins. A nearly 100-fold difference in the rates of H₂S production for the various substrates is observed supporting the tunability of persulfide generation by the 3-MST enzyme/artificial substrate system. Next, we show that the substrate 1a permeates cells and is selectively turned over by 3-MST to generate 3-MST-persulfide, which protects against reactive oxygen species-induced lethality. Lastly, in a mouse model, 1a is found to significantly mitigate neuroinflammation in the brain tissue. Together, the approach that we have developed allows for the on-demand generation of persulfides in vitro and in vivo using a range of shelf-stable, artificial substrates of 3-MST, while opening up possibilities of harnessing these molecules for therapeutic applications.

A persulfide/hydrogen sulfide generation strategy through artificial substrates for 3-mercaptopyruvate sulfurtransferase (3-MST) is reported, which enhances cellular persulfides, attenuates reactive oxygen species (ROS), and alleviates inflammation.     

Integrated quality by design (QbD) and design of experiments (DoE) approach for UFLC determination of telaprevir in rat serum

An ultrafast liquid chromatographic bioanalytical method was developed and validated for the determination of telaprevir in Wistar albino rat serum. Principles of quality by design (QbD) were implemented for enhancing the bioanalytical liquid–liquid extraction of telaprevir from rat serum. A Box–Behnken design was utilized in the studies by selecting extraction time, centrifugation speed, and vortex time as the critical method variables for evaluating their effect on the critical analytical attribute, i.e., %recovery of telaprevir. Chromatographic separation was achieved within a run time of 10?min using a C-18 column and mobile phase comprising of methanol:borate buffer of pH 9 (90:10 v/v) flowing at 1.2?mL/min. Photodiode array detection was performed at 270?nm. Results of validation studies were satisfactory. The method was linear over a concentration of 25–10,000?ng/mL. Limit of detection for the developed method was 10?ng/mL. Further, design of experiments (DoE) used for inter-day accuracy and precision study suggested superior method reliability. This integrated QbD- and DoE-based approach ensured the development of a validated and reliable analytical method for optimum bioanalysis of telaprevir in biological matrix.     

Photoluminescence properties of Gd:ZnO nano phosphor

Journal of Sol-Gel Science and Technology - Gd (0.1, 0.5, 1.0?mol%) doped ZnO nano phosphor, prepared by wet chemical method followed by sintering in air at 700?°C, was...     

A General Strategy Toward pH-Resistant Phenolic Fluorophores for High-Fidelity Sensing and Bioimaging Applications

Aryl alcohol-type or phenolic fluorophores offer diverse opportunities for developing bioimaging agents and fluorescence probes. Due to the inherently acidic hydroxyl functionality, phenolic fluorophores provide pH-dependent emission signals. Therefore, except for developing pH probes, the pH-dependent nature of phenolic fluorophores should be considered in bioimaging applications but has been neglected. Here we show that a simple structural remedy converts conventional phenolic fluorophores into pH-resistant derivatives, which also offer “medium-resistant” emission properties. The structural modification involves a single-step introduction of a hydrogen-bonding acceptor such as morpholine nearby the phenolic hydroxyl group, which also leads to emission bathochromic shift, increased Stokes shift, enhanced photo-stability and stronger emission for several dyes. The strategy greatly expands the current fluorophores’ repertoire for reliable bioimaging applications, as demonstrated here with ratiometric imaging of cells and tissues.     

US Coast Guard air station location with respect to distress calls: A spatial statistics and optimization based methodology

We study the problem of suitably locating US Coast Guard air stations to respond to emergency distress calls. Our goal is to identify robust locations in the presence of uncertainty in distress call locations. Our analysis differs from the literature primarily in the way we model this uncertainty. In our optimization and simulation based methodology, we develop a statistical model and demonstrate our procedure using a real data set of distress calls. In addition to guiding strategic decisions of placement of various stations, our methodology is also able to provide guidance on how the resources should be allocated across stations.     

A categorical invariant for cubic threefolds

We prove a categorical version of the Torelli theorem for cubic threefolds. More precisely, we show that the non-trivial part of a semi-orthogonal decomposition of the derived category of a cubic threefold characterizes its isomorphism class.     

A branch-and-cut method for 0-1 mixed convex programming

We generalize the disjunctive approach of Balas, Ceria, and Cornuéjols [2] and devevlop a branch-and-cut method for solving 0-1 convex programming problems. We show that cuts can be generated by solving a single convex program. We show how to construct regions similar to those of Sherali and Adams [20] and Lovász and Schrijver [12] for the convex case. Finally, we give some preliminary computational results for our method. Received January 16, 1996 / Revised version received April 23, 1999?Published online June 28, 1999     

A disjunctive cutting plane procedure for general mixed-integer linear programs

In this paper we develop a cutting plane algorithm for solving mixed-integer linear programs with general-integer variables. A novel feature of the algorithm is that it generates inequalities at all γ-optimal vertices of the LP-relaxation at each iteration. The cutting planes generated in the procedure are found by considering a natural generalization of the 0-1 disjunction used by Balas, Ceria, and Cornuéjols in the context of solving binary mixed-integer linear programs [3, 4]. Received: January 1999 / Accepted: March 2000?Published online December 15, 2000     

Min-cut clustering

We describe a decomposition framework and a column generation scheme for solving a min-cut clustering problem. The subproblem to generate additional columns is itself an NP-hard mixed integer programming problem. We discuss strong valid inequalities for the subproblem and describe some efficient solution strategies. Computational results on compiler construction problems are reported.This paper is dedicated to Phil Wolfe on the occasion of his 65th birthday.This research was supported by NSF grants DMS-8719128 and DDM-9115768, and by an IBM grant to the Computational Optimization Center, Georgia Institute of Technology.     

Finding an interior point in the optimal face of linear programs

We study the problem of finding a point in the relative interior of the optimal face of a linear program. We prove that in the worst case such a point can be obtained in O(n ³ L) arithmetic operations. This complexity is the same as the complexity for solving a linear program. We also show how to find such a point in practice. We report and discuss computational results obtained for the linear programming problems in the NETLIB test set.Research supported in part by NSF Grant CCR-8810107, CCR-9019469 and a grant from GTE Laboratories.Research supported in part by NSF Grant DDM-8922636 and NSF Coop. Agr. No. CCR-8809615 through Rice University.