Validated stability-indicating densitometric thin-layer chromatography: application to stress degradation studies of minocycline

A simple, stability-indicating high-performance thin-layer liquid chromatographic (HPTLC) method for analysis of minocycline was developed and validated. The densitometric analysis was carried out at 345 nm using methanol-acetonitrile-isopropyl alcohol-water (5:4:0.5:0.5, v/v/v/v) as mobile phase.The method employed TLC aluminium plates pre-coated with silica gel 60F-254 as the stationary phase. To achieve good result, plates were sprayed with a 10% (w/v) solution of disodium ethylene diaminetetraacetic acid (EDTA), the pH of which was adjusted to 9.0. Compact spots of minocycline were found at R_f = 0.30 ± 0.02. For proposed procedure, linearity (r = 0.9997), limit of detection (3.7 ng spot⁻¹), recovery (99.23-100.16%), and precision (% R.S.D. ≤ 0.364) was found to be satisfactory. The drug undergoes acidic and basic degradation, oxidation and photodegradation. All the peaks of degradation products were well resolved from the pure drug with significantly different R_f values. The acidic and alkaline degradation kinetics of minocycline, evaluated using this method, is found to be of first order.     

Iron Chelation in Local Infection

Iron is an essential element in multiple biochemical pathways in humans and pathogens. As part of the innate immune response in local infection, iron availability is restricted locally in order to reduce overproduction of reactive oxygen species by the host and to attenuate bacterial growth. This physiological regulation represents the rationale for the therapeutic use of iron chelators to support induced iron deprivation and to treat infections. In this review paper we discuss the importance of iron regulation through examples of local infection and the potential of iron chelation in treating infection.     

Outer-Membrane Protease (OmpT) Based E. coli Sensing with Anionic Polythiophene and Unlabeled Peptide Substrate

E. coli and Salmonella are two of the most common bacterial pathogens involved in foodborne and waterborne related deaths. Hence, it is critical to develop rapid and sensitive detection strategies for near-outbreak applications. Reported is a simple and specific assay to detect as low as 1 CFU mL⁻¹ of E. coli in water within 6 hours by targeting the bacteria's surface protease activity. The assay relies on polythiophene acetic acid (PTAA) as an optical reporter and a short unlabeled peptide (LL37_FRRV) previously optimized as a substrate for OmpT, an outer-membrane protease on E. coli. LL37_FRRV interacts with PTAA to enhance its fluorescence while also inducing the formation of a helical PTAA-LL37_FRRV construct, as confirmed by circular dichroism. However, in the presence of E. coli LL37_FRRV is cleaved and can no longer affect the conformations and optical properties of PTAA. This ability to distinguish between an intact and cleaved peptide was investigated in detail using LL37_FRRV sequence variants.     

A novel apparatus for flame spread study

One of the challenges in the experimental study of flame spread is that, even if the flame spreads at a steady rate, the propagating flame creates an unsteady phenomenon with respect to the laboratory frame of reference. As a result, only a few studies have been done where the detailed flame structure has been experimentally measured along with the spread rates. In this work, we demonstrate the feasibility of a new flame spread apparatus that moves the fuel in the opposite direction of the flame so as to keep the leading edge of the flame stationary with respect to the laboratory. A thermocouple, fixed to the laboratory frame of reference, in front of the leading edge of the flame, senses the presence of the flame and a PID controller keeps the set point temperature constant by moving the sample holder, driven by a stepper motor, in the opposite direction at the velocity of the spread. Unlike conventional studies, this apparatus, called the flame stabilizer, produces real time spread rate with a time resolution of 0.3 s. In this paper, instantaneous flame spread rate and the visible flame structure are compared between a downward spreading flame and the corresponding stabilized flame for spread over ashless filter paper. The results indicate that the difference between the two configurations are within experimental uncertainties and the stabilized flame can represent a spreading flame adequately, including variability of flame spread rate and the flame geometry, for further observations.     

Phenolic composition, antioxidant capacity and antibacterial activity of selected Irish Brassica vegetables

Vegetables belonging to the Brassicaceae family are rich in polyphenols, flavonoids and glucosinolates, and their hydrolysis products, which may have antibacterial, antioxidant and anticancer properties. In the present study, phenolic composition, antibacterial activity and antioxidant capacity of selected Brassica vegetables, including York cabbage, Brussels sprouts, broccoli and white cabbage were evaluated after extraction with aqueous methanol. Results obtained showed that York cabbage extract had the highest total phenolic content, which was 33.5, followed by 23.6, 20.4 and 18.4 mg GAE/g of dried weight (dw) of the extracts for broccoli, Brussels sprouts and white cabbage, respectively. All the vegetable extracts had high flavonoid contents in the order of 21.7, 17.5, 15.4 and 8.75 mg QE/g of extract (dw) for York cabbage, broccoli, Brussels sprouts and white cabbage, respectively. HPLC-DAD analysis showed that different vegetables contain a mixture of distinct groups of phenolic compounds. All the extracts studied showed a rapid and concentration dependent antioxidant capacity in diverse antioxidant systems. The antibacterial activity was determined against Gram-positive and Gram-negative bacteria. York cabbage extract exhibited significantly higher antibacterial activity against Listeria monocytogenes (100%) and Salmonella abony (94.3%), being the most susceptible at a concentration of 2.8%, whereas broccoli, Brussels sprouts and white cabbage had moderate to weak activity against all the test organisms. Good correlation (r2 0.97) was found between total phenolic content obtained by spectrophotometric analysis and the sum of the individual polyphenols monitored by HPLC-DAD.     

Solvent-free sonochemical one-pot three-component synthesis of 2H-indazolo[2,1-b]phthalazine-1,6,11-triones and 1H-pyrazolo[1,2-b]phthalazine-5,10-diones

A rapid and efficient one-pot three-component protocol for the synthesis of 2H-indazolo[2,1-b]phthalazine-1,6,11-triones 4 and 1H-pyrazolo[1,2-b]phthalazine-5,10-diones 6 has been developed by domino coupling of phthalhydrazide, 1,3-diketones, and aldehydes under solvent-free conditions at 80 °C as well as under solvent-free ultrasound irradiation at room temperature promoted by (S)-camphorsulfonic acid.     

Structure and binding of the H4 histone tail and the effects of lysine 16 acetylation

The H4 histone tail plays a critical role in chromatin folding and regulation--it mediates strong interactions with the acidic patch of proximal nucleosomes and its acetylation at lysine 16 (K16) leads to partial unfolding of chromatin. The molecular mechanism associated with the H4 tail/acidic patch interactions and its modulation via K16 acetylation remains unknown. Here we employ a combination of molecular dynamics simulations, molecular docking calculations, and free energy computations to investigate the structure of the H4 tail in solution, the binding of the H4 tail with the acidic patch, and the effects of K16 acetylation. The H4 tail exhibits a disordered configuration except in the region Ala15-Lys20, where it exhibits a strong propensity for an α-helical structure. This α-helical region is found to dock very favorably into the acidic patch groove of a nucleosome with a binding free energy of approximately -7 kcal mol(-1). We have identified the specific interactions that stabilize this binding as well as the associated energetics. The acetylation of K16 is found to reduce the α-helix forming propensity of the H4 tail and K16's accessibility for mediating external interactions. More importantly, K16 acetylation destabilizes the binding of the H4 tail at the acidic patch by mitigating specific salt bridges and longer-ranged electrostatic interactions mediated by K16. Our study thus provides new microscopic insights into the compaction of chromatin and its regulation via posttranslational modifications of histone tails, which could be of interest to chromatin biology, cancer, epigenetics, and drug design.     

Accurate dispensing of volatile reagents on demand for chemical reactions in EWOD chips

Digital microfluidic chips provide a new platform for manipulating chemicals for multi-step chemical synthesis or assays at the microscale. The organic solvents and reagents needed for these applications are often volatile, sensitive to contamination, and wetting, i.e. have contact angles of <90° even on the highly hydrophobic surfaces (e.g., Teflon? or Cytop?) typically used on digital microfluidic chips. Furthermore, often the applications dictate that the processes are performed in a gas environment, not allowing the use of a filler liquid (e.g., oil). These properties pose challenges for delivering controlled volumes of liquid to the chip. An automated, simple, accurate and reliable method of delivering reagents from sealed, off-chip reservoirs is presented here. This platform overcomes the issues of evaporative losses of volatile solvents, cross-contamination, and flooding of the chip by combining a syringe pump, a simple on-chip liquid detector and a robust interface design. The impedance-based liquid detection requires only minimal added hardware to provide a feedback signal to ensure accurate volumes of volatile solvents are introduced to the chip, independent of time delays between dispensing operations. On-demand dispensing of multiple droplets of acetonitrile, a frequently used but difficult to handle solvent due to its wetting properties and volatility, was demonstrated and used to synthesize the positron emission tomography (PET) probe [(18)F]FDG reliably.     

A systematic approach to magnetic resonance imaging evaluation of epiphyseal lesions

Magnetic Resonance Imaging (MRI) is the preferred modality of choice to image epiphyseal lesions. It provides excellent soft tissue resolution and extent of disease. A wide spectrum of tumor and tumor like lesions can involve the epiphysis. Early and accurate diagnosis as well as appropriate management of epiphyseal lesions is critical as these conditions may lead to disabling complications such as, limb length discrepancy, angular or joint surface deformities and secondary osteoarthritis. In this article, we discuss the role of conventional sequences, such as T1W, fluid sensitive T2W and intravenous (IV) Gadolinium enhanced sequences as well as the additional value of problem solving MRI sequences such as, chemical shift and diffusion weighted imaging. Based on the imaging findings on various MRI sequences and lesion characteristics, a systematic approach directed to the diagnoses of epiphyseal lesions is presented and discussed. MRI features of clinically and biopsy proven examples of the epiphyseal lesions, such as osteomyelitis, intra-osseous abscess, infiltrative malignancy, metastases, transient osteoporosis, subchondral insufficiency fracture, avascular necrosis, osteochondral fracture, osteochondritis dissecans, eosinophilic granuloma and geode are demonstrated. Using this systematic approach, the reader will be able to better characterize epiphyseal lesions with a potential to positively affect patient management.     

Water Vapor Sorption in Cementitious Materials—Measurement,Modeling and Interpretation

The rate and extent of uptake and release of moisture are critical in controlling the behavior of cementitious materials ranging from fluid transport to hygral deformations. While classically determined using an equilibrium (static) salt solution method (Baroghel-Bouny in Cem Concr Res 37:414–437, 2007), advanced capabilities offered by gravimetric dynamic vapor sorption (DVS) analyzers, are now permitting acquisition of sorption spectra at microgram ( $\upmu \hbox {g}$ ) resolution on the order of a few weeks. This work highlights new multicycle determinations of adsorption/desorption isotherms, acquired using a custom-built DVS analyzer for well-hydrated alite and ordinary portland cement pastes over a range of water-to-solid ratios ( $w/s$ , mass basis). Special focus is paid to describe measurement aspects relevant to acquiring reliable spectra, and their interpretation. Sorption isotherms are used to assess transport properties, and sorption hysteresis and its irreversibility following first drying. Based on an optimization-based criterion, the Young-Nelson model is selected to simulate sorption evolutions, including the effects of hysteresis. Sensitivity analyses carried out using this model are used to understand the role of parameters, including porosity and $w/s$ , on the hysteresis that develops from the first to subsequent sorption cycles.