Ultrasonic pretreatment of corn slurry for saccharification: A comparison of batch and continuous systems

The effects of ultrasound on corn slurry saccharification yield and particle size distribution was studied in both batch and continuous-flow ultrasonic systems operating at a frequency of 20 kHz. Ground corn slurry (28% w/v) was prepared and sonicated in batches at various amplitudes (192–320 μm_{peak-to-peak (p–p)}) for 20 or 40 s using a catenoidal horn. Continuous flow experiments were conducted by pumping corn slurry at various flow rates (10–28 l/min) through an ultrasonic reactor at constant amplitude of 12 μm_p–p. The reactor was equipped with a donut shaped horn. After ultrasonic treatment, commercial alpha- and gluco-amylases (STARGEN^TM 001) were added to the samples, and liquefaction and saccharification proceeded for 3 h. The sonicated samples were found to yield 2–3 times more reducing sugars than unsonicated controls. Although the continuous flow treatments released less reducing sugar compared to the batch systems, the continuous flow process was more energy efficient. The reduction of particle size due to sonication was approximately proportional to the dissipated ultrasonic energy regardless of the type of system used. Scanning electron microscopy (SEM) images were also used to observe the disruption of corn particles after sonication. Overall, the study suggests that both batch and continuous ultrasonication enhanced saccharification yields and reduced the particle size of corn slurry. However, due to the large volume involve in full scale processes, an ultrasonic continuous system is recommended.     

Metabolite assignment of ultrafiltered synovial fluid extracted from knee joints of reactive arthritis patients using high resolution NMR spectroscopy

Currently, there are no reliable biomarkers available that can aid early differential diagnosis of reactive arthritis (ReA) from other inflammatory joint diseases. Metabolic profiling of synovial fluid (SF)—obtained from joints affected in ReA—holds great promise in this regard and will further aid monitoring treatment and improving our understanding about disease mechanism. As a first step in this direction, we report here the metabolite specific assignment of ¹H and ¹³C resonances detected in the NMR spectra of SF samples extracted from human patients with established ReA. The metabolite characterization has been carried out on both normal and ultrafiltered (deproteinized) SF samples of eight ReA patients (n = 8) using high-resolution (800 MHz) ¹H and ¹H─¹³C NMR spectroscopy methods such as one-dimensional ¹H CPMG and two-dimensional J-resolved¹H NMR and homonuclear ¹H─¹H TOCSY and heteronuclear¹H─¹³C HSQC correlation spectra. Compared with normal SF samples, several distinctive ¹H NMR signals were identified and assigned to metabolites in the ¹H NMR spectra of ultrafiltered SF samples. Overall, we assigned 53 metabolites in normal filtered SF and 64 metabolites in filtered pooled SF sample compared with nonfiltered SF samples for which only 48 metabolites (including lipid/membrane metabolites as well) have been identified. The established NMR characterization of SF metabolites will serve to guide future metabolomics studies aiming to identify/evaluate the SF-based metabolic biomarkers of diagnostic/prognostic potential or seeking biochemical insights into disease mechanisms in a clinical perspective.     

Effect of Cold Atmospheric Plasma Jet and Gamma Radiation Treatments on Gingivobuccal Squamous Cell Carcinoma and Breast Adenocarcinoma Cells

Surgery, chemotherapy and radiotherapy, the conventional treatment modalities of cancer though successful are limited by presence of residual tumor cells, toxic side-effects and treatment resistance, thus raising the need for investigating other novel approaches. Here, we have used a cold atmospheric plasma (CAP) jet and assessed the in vitro efficacy in gingivobuccal squamous cell carcinoma (GB-SCC) – ITOC-03, breast adenocarcinoma—MCF7 and HEK293 cells. Cells lines were subjected to varying doses of ionizing radiation (0, 2, 4, 6, 8 Gy) and CAP jet treatment (0, 60, 180, 240, 300 s). CAP jet treatment showed time dependent increase in H₂O₂ and NO₂^? concentration. Cell viability assay showed potent effect of CAP jet on all three cell lines in comparison to radiation treatment, while helium gas treatment showed minimal inhibitory effect. Irradiated, CAP jet and helium gas treated cells showed loss of nucleic acid features, 788 cm^?1 and 1340 cm^?1 in Raman spectra, indicating DNA damage. Principal Component Analysis (PCA) showed distinct classification of CAP-treated and control cells, while Principal Component – Linear Discriminant Analysis (PC-LDA) based classification of Raman spectra showed ITOC-03 and HEK293 cells to be sensitive to CAP jet and radiation treatment in comparison to MCF7 cells. Collectively, cell viability assay and Raman spectroscopy have shown potent effect of CAP jet in GB-SCC and breast adenocarcinoma cells.

     

Stabilization of Langmuir monolayer of hydrophobic thiocholesterol molecules

The self-assembled monolayer of the thiocholesterol (TCh) exhibits interesting properties that can be used for various technological applications. TCh is predominantly a hydrophobic molecule, and it does not spread at the air–water interface to form a stable Langmuir monolayer. We have stabilized the TCh molecules in the cholesterol (Ch) monolayer. We find the mixed monolayer to be stable upto 0.75 mole fraction of TCh in Ch. The mixed monolayer shows an initial and a final collapse. On compressing the monolayer beyond the initial collapse, the TCh molecules squeeze out irreversibly from the mixed monolayer phase. The calculation of excess area per molecule for the TCh and Ch mixed monolayer system indicates an attractive interaction between the component molecules. Interestingly, the elasticity of the Ch monolayer reduces to less than half, and the monolayer becomes more fluidic due to the presence of even very minute quantity (5%) of TCh.     

Magnesium indium oxide (MgIn2O4) spinel thin films: chemical spray pyrolysis (CSP) growth and materials characterizations

MgIn(2)O(4), which has an inverse spinel structure, has been adopted as the transparent material in optoelectronic device fabrication due to its high optical transparency and electrical conductivity. Such a technologically important material was prepared by the spray pyrolysis technique. Precursors prepared for the cationic ratio Mg/In=0.5 were thermally sprayed onto glass substrates at 400 and 450 degrees C. We report herein the preparation and characterization of the films by X-ray diffraction (XRD), energy-dispersive absorption X-ray spectroscopy (EDAX), and atomic force microscopy (AFM). The XRD results showed the single phase formation of the material that revealed the presence of Mg(2+) and In(3+) in the inverse spinel-related structure. The FTIR and EDAX results further confirmed that the nanocrystalline films were mainly composed of magnesium, indium, and oxygen, in agreement with XRD analysis. We surmised from the AFM micrographs that the atoms have enough diffusion activation energy to occupy the correct site in the crystal lattice. For the 423-nm-thick magnesium indium oxide films grown at 400 degrees C, the electrical conductivity was 5.63x10(-6) Scm(-1) and the average optical transmittance was 63% in the visible range (400-700 nm). Similar MgIn(2)O(4) films deposited at 450 degrees C have a conductivity value of 1.5x10(-5) Scm(-1) and an average transmittance of 75%. Hall coefficient observations showed n-type electrical conductivity and high electron carrier concentration of 2.7x10(19) cm(-3).     

Pi and sigma-phenylethynyl radicals and their isomers o-, m-, and p-ethynylphenyl: structures, energetics, and electron affinities

Molecular structures, energetics, vibrational frequencies, and electron affinities are predicted for the phenylethynyl radical and its isomers. Electron affinities are computed using density functional theory, -namely, the BHLYP, BLYP, B3LYP, BP86, BPW91, and B3PW91 functionals-, employing the double-zeta plus polarization DZP++ basis set; this level of theory is known to perform well for the computation of electron affinities. Furthermore, ab initio computations employing perturbation theory, coupled cluster with single and double excitations [CCSD], and the inclusion of perturbative triples [CCSD(T)] are performed to determine the relative energies of the isomers. These higher level computations are performed with the correlation consistent family of basis sets cc-pVXZ (X = D, T, Q, 5). Three electronic states are probed for the phenylethynyl radical. In C2v symmetry, the out-of-plane (2B1) radical is predicted to lie about 10 kcal/mol below the in-plane (2B2) radical by DFT methods, which becomes 9.4 kcal/mol with the consideration of the CCSD(T) method. The energy difference between the lowest pi and sigma electronic states of the phenylethynyl radical is also about 10 kcal/mol according to DFT; however, CCSD(T) with the cc-pVQZ basis set shows this energy separation to be just 1.8 kcal/mol. The theoretical electron affinities of the phenylethynyl radical are predicted to be 3.00 eV (B3LYP/DZP++) and 3.03 eV (CCSD(T)/DZP++//MP2/DZP++). The adiabatic electron affinities (EAad) of the three isomers of phenylethynyl, that is, the ortho-, meta-, and para-ethynylphenyl, are predicted to be 1.45, 1.40, and 1.43 eV, respectively. Hence, the phenylethynyl radical binds an electron far more effectively than the three other radicals studied. Thermochemical predictions, such as the bond dissociation energies of the aromatic and ethynyl C-H bonds and the proton affinities of the phenylethynyl and ethynylphenyl anions, are also reported.     

Highly enantioselective organocatalytic direct aldol reaction in an aqueous medium

We have demonstrated that small organic molecules 1 and 2 catalyzed the direct aldol reaction of both acyclic and cyclic ketones with different aldehydes in an excess of water/brine. Excellent enantioselectivities up to >99% and diastereoselectivities up to 99% with very good yields were obtained by using much lower catalyst loadings (0.5 mol %).     

Electrochemically triggered Michael addition on the self-assembly of 4-thiouracil: generation of surface-confined redox mediator and electrocatalysis

Generation of a surface-confined redox mediator (RM) by an electrochemically triggered Michael addition reaction and the electrocatalytic properties of the mediator are described. Electrogenerated o-quinone undergoes Michael addition reaction with the self-assembled monolayer (SAM) of 4-thiouracil (4-TU) on a gold (Au) electrode and yields a surface-confined RM, 1-(3,4-dihydroxyphenyl)-4-mercapto-1H-pyrimidin-2-one (DPTU). The Michael addition reaction depends on the electrolysis potential and time, solution pH, and concentration of catechol (CA) used in the reaction. The redox mediator, DPTU, exhibits reversible redox response, characterstic of a surface-confined species at approximately 0.22 V in neutral pH. The anodic peak potential of DPTU shifts by 58+/-2 mV while changing the solution pH by one unit, suggesting that protons and electrons taking part in the redox reaction are in the ratio of 1:1. The apparent rate constant (ksapp) for the heterogeneous electron-transfer reaction of the RM was determined to be 114+/-5 s-1. The surface coverage (Gamma) of DPTU on the electrode surface was 8.2+/-0.1x10(-12) mol/cm2. DPTU shows excellent electrocatalytic activity toward oxidation of reduced nicotinamide adenine dinucleotide (NADH) with activation overpotential, which is approximately 600 mV lower than that observed at the unmodified Au electrode. The dipositive cations in the supporting electrolyte solution amplify the electrocatalytic activity of DPTU. A 2.5-fold enhancement in the catalytic current was observed in the presence of Ca2+ or Ba2+ ions. The sensitivity of the electrode toward NADH in the presence and absence of Ca2+ ions was 0.094+/-0.011 and 0.04+/-0.0071 nA cm-2 nM-1, respectively. A linear increase in the catalytic current was obtained up to the concentration of 0.8 mM, and the electrode can detect amperometrically as low as 25 nM of NADH in neutral pH.     

Synthesis and anti-tumor activity of β-C-glycoside analogs of the immunostimulant KRN7000

A ring-closing metathesis approach was employed for the synthesis of a β-C-glycoside analog of the immunostimulant KRN7000. The protected C-glycosyl amino acid derivative 18 was converted to amino-olefin 20, and osmylation served to install the diol unit as a mixture of separable syn and anti isomers. Deprotection to the hydroxy-amine 21 was followed by N-acylation and debenzylation to deliver the target compound 5.     

Could diastereoselectivity in the presence of O-2 chiral nonparticipating groups be an indicator of glycopyranosyl oxacarbenium ions in glycosylation reactions?

Although long postulated, the existence of glycopyranosyl oxacarbenium ions as intermediates or transition states (TS) in chemical glycosylation reactions has not been convincingly demonstrated experimentally. It is anticipated that elucidation of such reactive species will greatly assist synthetic chemists to control the α/β stereoselectivity by rational means. Previous density functional theory (DFT) calculations from our group found that the torsion potential about C-2-O-2 in protected glycopyranosyl donors changed from a conventional 3-fold rotor to a 2-fold rotor with a strong syn (CH-2-C-2-O-2-CPg) preference once the donor was ionized to its oxacarbenium ion. This suggested to us that if CPg of the protecting group was a chiral carbon, then diastereoselectivity might be observed in glycosylation reactions that proceed through oxacarbenium ions. The hypothesis to test is as follows: if a nonparticipating O-2 racemic chiral protecting group exhibits diastereoselectivity in glycosylation reactions, then the reaction probably proceeds through an oxacarbenium ion intermediate or TS. We present data for O-2 ether-protected d-glucopyranosyl donors where the racemic protecting group 1-methyl 1'-methylcyclopropylmethyl (MCPM) provides the chirality. MCPM proves to be more activating than the O-2-benzyl ether, and in cases where the donor is otherwise deactivated, several examples of moderate diastereoselectivity are found. These results can be interpreted to indicate that a continuum of reactivity exists where some glycosyl donors form oxacarbenium ions in glycosylation reactions but more reactive donors do not. The strongly activating properties of the cyclopropylmethyl ether functionality and the ability to induce diastereoselectivity with chiral derivatives strongly suggest widespread applications.