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2-Chloro-1-(1-ethoxyvinyl)pyridinium triflate and several other bench-stable N -(1-alkoxyvinyl) 2-halopyridinium triflates have been developed as reagents for the synthesis of valuable 2-aminopyridine scaffolds via unusually mild S N Ar substitutions with amine nucleophiles. Advantages of this approach include an operationally simple mix-and-stir procedure at room temperature or mild heat and ambient atmosphere and without the need for transition metal catalysts, coupling reagents, or high-boiling solvents. The stable N -(1-ethoxyvinyl) moiety serves as a dual S N Ar-activating group and pyridine N -protecting group that can be cleaved under thermal, acidic, or oxidative conditions. Preliminary results of other nucleophilic substitutions using oxygen-, sulfur-, and carbon-based nucleophiles are also demonstrated.

Two electrons in two orbitals give rise to four states. When the orbitals are weakly coupled as in the case for the d xy orbitals of quadruple bond species, two of the states are diradical in character with electrons residing in separate orbitals and two of the states are zwitterionic with electrons paired in one orbital or the other. By measuring one-and two-photon spectra, the one-electron (ΔW) and two-electron (K) energies may be calculated, which are the determinants of the state energies of the four-state model for the two-electron bond. The K energy is thus especially sensitive to the size of the orbital as K is dependent on the distance between electrons. To this end, one- and two-photon spectra of Mo 2 X 4 (PMe 3 ) 4 are sensitive to secondary bonding interactions of the δ-orbital manifold with the halide orbitals, as reflected in decreasing K energies along the series Cl > Br > I. Additionally, the calculated one-electron energies have been verified with the spectroelectrochemical preparation of the Mo 2 X 4 (PMe 3 ) 4 + complexes, where the δ bond is a one-electron bond, and K is thus absent. The δ → δ* transition shifts over 10,000 cm -1 upon oxidation of Mo 2 X 4 (PMe 3 ) 4 to Mo 2 X 4 (PMe 3 ) 4 + , establishing that transitions within the two-electron δ bond are heavily governed by the two-electron exchange energy.

A fundamental structure for the measurement process is developed. Measurement is defined as the determination of the number of units of a particular quantity or property presented by the measured object or sample. Measurement is distinguished from qualitative observation and data interpretation. The limited sources of measurement numbers (scalar, digital, and counting readouts) reveal that all measurements except manual counting require a measurement device. Measurement systems consist of devices which convert one form of data encoding to another. They begin with the sensor and proceed through intermediate conversion devices until the readout device produces numbers. There are three classes of conversion devices: input (sensor), intermediate, and readout. The characteristics of the conversion devices in a system determine the overall system characteristics. Counting measurements have several unique characteristics. In automated counting, conversion devices are used in event detection, discrimination, and boundary setting. Variance in counting arises from event detection, boundary setting, and random event occurrence. Entire measurement systems or conversion device subsystems may operate by null comparison. Conversion devices in null comparison systems have a different arrangement. The time relationship of multiple or successive measurements affects the information content of data sets. Time-correlation of acquired data is critical in all stimulus-response measurement systems such as chromatography and time-of-flight mass spectrometry and in many subsystems such as lock-in amplifiers and box-car integrators., Competing Interests: The author declares no competing financial interest., (© 2022 The Author. Published by American Chemical Society.)

Modern biomarker and translational research as well as personalized health care studies rely heavily on powerful omics' technologies, including metabolomics and lipidomics. However, to translate metabolomics and lipidomics discoveries into a high-throughput clinical setting, standardization is of utmost importance. Here, we compared and benchmarked a quantitative lipidomics platform. The employed Lipidyzer platform is based on lipid class separation by means of differential mobility spectrometry with subsequent multiple reaction monitoring. Quantitation is achieved by the use of 54 deuterated internal standards and an automated informatics approach. We investigated the platform performance across nine laboratories using NIST SRM 1950-Metabolites in Frozen Human Plasma, and three NIST Candidate Reference Materials 8231-Frozen Human Plasma Suite for Metabolomics (high triglyceride, diabetic, and African-American plasma). In addition, we comparatively analyzed 59 plasma samples from individuals with familial hypercholesterolemia from a clinical cohort study. We provide evidence that the more practical methyl-tert-butyl ether extraction outperforms the classic Bligh and Dyer approach and compare our results with two previously published ring trials. In summary, we present standardized lipidomics protocols, allowing for the highly reproducible analysis of several hundred human plasma lipids, and present detailed molecular information for potentially disease relevant and ethnicity-related materials.

We reported and evaluated a microflow, single-shot, short gradient SWATH MS method intended to accelerate the discovery and verification of protein biomarkers in preclassified clinical specimens. The method uses a 15 min gradient microflow-LC peptide separation, an optimized SWATH MS window configuration, and OpenSWATH software for data analysis. We applied the method to a cohort containing 204 FFPE tissue samples from 58 prostate cancer patients and 10 benign prostatic hyperplasia patients. Altogether we identified 27,975 proteotypic peptides and 4037 SwissProt proteins from these 204 samples. Compared to a reference SWATH method with a 2 h gradient, we found 3800 proteins were quantified by the two methods on two different instruments with relatively high consistency ( r = 0.77). The accelerated method consumed only 17% instrument time, while quantifying 80% of proteins compared to the 2 h gradient SWATH. Although the missing value rate increased by 20%, batch effects reduced by 21%. 75 deregulated proteins measured by the accelerated method were selected for further validation. A shortlist of 134 selected peptide precursors from the 75 proteins were analyzed using MRM-HR, and the results exhibited high quantitative consistency with the 15 min SWATH method ( r = 0.89) in the same sample set. We further verified the applicability of these 75 proteins in separating benign and malignant tissues (AUC = 0.99) in an independent prostate cancer cohort ( n = 154). Altogether, the results showed that the 15 min gradient microflow SWATH accelerated large-scale data acquisition by 6 times, reduced batch effect by 21%, introduced 20% more missing values, and exhibited comparable ability to separate disease groups.

Conductive fabrics have received significant attention because of their widespread applications from smart textiles to energy storage devices. Conductive colloidal materials are preferred as a coating on the fabric to achieve desirable electronic conductivity; however, obtaining a uniform coverage with a simple and effective route is a challenge. Herein, we report exfoliated graphene nanoplatelets (GNPs) in low boiling point solvents and their subsequent coating onto a polyamide fabric surface. Few-layered (average <7 layers) GNPs were obtained by optimizing solubility parameters of solvent mixtures and sonication time. Raman spectroscopy showed that the I D / I G ratio changed from 0.33 to 0.38 in the GNP solution before and after the sonication, confirming an insignificant increase in defects on the basal plane of graphene after sonication treatment. Uniform coating of GNPs was obtained by optimizing concentration and sonication times. Scanning electron microscopy showed a uniform coverage of GNPs, and the surface resistivity of the polyamide fabric decreased from infinity to ∼40 kΩ after 4 h of coating. X-ray diffraction analysis confirmed the minimal effect on the fabric crystallinity during processing. This interface engineering approach is simple and scalable, and it is applicable for the coating of different polymeric fabrics with a great promise in electronic textiles.

G protein-coupled receptors can exist as dimers and higher-order oligomers in biological membranes. The specific oligomeric assembly of these receptors is believed to play a major role in their function, and the disruption of native oligomers has been implicated in specific human pathologies. Computational predictions and biochemical analyses suggest that two molecules of rhodopsin (Rho) associate through the interactions involving its fifth transmembrane helix (TM5). Interestingly, there are several pathogenic loss-of-function mutations within TM5 that face the lipid bilayer in a manner that could potentially influence the dimerization of Rho. Though several of these mutations are known to induce misfolding, the pathogenic defects associated with V209M and F220C Rho remain unclear. In this work, we utilized a variety of biochemical and biophysical approaches to elucidate the effects of these mutations on the dimerization, folding, trafficking, and function of Rho in relation to other pathogenic TM5 variants. Chemical cross-linking, bioluminescence energy transfer, and pulsed-interleaved excitation fluorescence cross-correlation spectroscopy experiments revealed that each of these mutants exhibits a wild type-like propensity to self-associate within the plasma membrane. However, V209M and F220C each exhibit subtle defects in cellular trafficking. Together, our results suggest that the RP pathology associated with the expression of the V209M and F220C mutants could arise from defects in folding and cellular trafficking rather than the disruption of dimerization, as has been previously proposed.

The blood-brain barrier (BBB) is a formidable obstacle for brain delivery of therapeutic antibodies. However, antibodies against the transferrin receptor (TfR), enriched in brain endothelial cells, have been developed as delivery carriers of therapeutic cargoes into the brain via a receptor-mediated transcytosis pathway. In vitro and in vivo studies demonstrated that either a low-affinity or monovalent binding of these antibodies to the TfR improves their release on the abluminal side of the BBB and target engagement in brain parenchyma. However, these studies have been performed with mouse-selective TfR antibodies that recognize different TfR epitopes and have varied binding characteristics. In this study, we evaluated serum pharmacokinetics and brain and CSF exposure of the rat TfR-binding antibody OX26 affinity variants, having K D s of 5 nM, 76 nM, 108 nM, and 174 nM, all binding the same epitope in bivalent format. Pharmacodynamic responses were tested in the Hargreaves chronic pain model after conjugation of OX26 affinity variants with the analgesic and antiepileptic peptide, galanin. OX26 variants with affinities of 76 nM and 108 nM showed enhanced brain and cerebrospinal fluid (CSF) exposure and higher potency in the Hargreaves model, compared to a 5 nM affinity variant; lowering affinity to 174 nM resulted in prolonged serum pharmacokinetics, but reduced brain and CSF exposure. The study demonstrates that binding affinity optimization of TfR-binding antibodies could improve their brain and CSF exposure even in the absence of monovalent TfR engagement.

To simultaneously quantify and profile the complex mixture of short-, median-, and long-chain CPs (SCCPs, MCCPs, and LCCPs) in Australian sewage sludge, we applied and further validated a recently developed novel instrumental technique, using quadrupole time-of-flight high resolution mass spectrometry running in the negative atmospheric pressure chemical ionization mode (APCI-qTOF-HRMS). Without using an analytical column the cleaned extracts were directly injected into the qTOF-HRMS followed by quantification of the CPs by a mathematical algorithm. The recoveries of the four SCCP, MCCP and LCCP-spiked sewage sludge samples ranged from 86 to 123%. This APCI-qTOF-HRMS method is a fast and promising technique for routinely measuring SCCPs, MCCPs, and LCCPs in sewage sludge. Australian sewage sludge was dominated by MCCPs with concentrations ranging from 542 to 3645 ng/g dry weight (dw). Lower SCCPs concentrations (<57-1421 ng/g dw) were detected in the Australian sewage sludge, which were comparable with the LCCPs concentrations (116-960 ng/g dw). This is the first time that CPs were reported in Australian sewage sludge. The results of this study gives a first impression on the distribution of the SCCPs, MCCPs, and LCCPs in Australia wastewater treatment plants (WWTPs).

Fluorescent deoxynucleosides possessing the modified bases 6-(2-benzo[b]furyl)- and 6-(2-furyl)pyrrolocytosine (BFpC and FpC) have been synthesized along with the quencher nucleosides possessing 6-{4-[(4-dimethylamino)azo]phenyl}pyrrolocytosine (DABCYLpC) and 6-(p-nitrophenyl)pyrrolocytosine (p-NO2-PhpC) nucleobase analogs. Standard treatment of BFpC, FpC, DABCYLpC, and p-NO2-PhpC with dimethoxytrityl chloride (DMT-Cl) led to the unusual substitution on the C7 of the pyrrolocytosine skeleton. The desired 5'-O-DMT-protected nucleoside analogs were synthesized from suitably protected 5'-O-DMT cytidines. Subsequent phosphitylation smoothly afforded BFpC-, FpC-, DABCYLpC-, and p-NO2-PhpC-derived monomers suitable for standard oligonucleotide synthesis.

A prochiral bridged compound of C2v symmetry, the norbornadiene (NBD), oriented in a chiral liquid crystal composed of various mixtures of poly-γ-benzyl-l-glutamate (PBLG) and poly-ε-carboxy-l-lysine (PCBLL), two chiral homopolypeptides, is investigated using natural abundance deuterium 2D-NMR (NAD 2D-NMR) spectroscopy. In such chiral oriented solvents, enantiotopic directions are spectrally nonequivalent, and two distinct (2)H quadrupolar doublets associated with enantioisotopomeric pairs of NBD are detected. As the two homopolypeptides have the same absolute configuration but distinct chemical functions in their side chains, the variation of residual quadrupolar couplings (RQC's) allows the determination of the relative solute-fiber affinities toward the two polypeptides in these lyotropic bipolymeric systems. Besides the experimental measurement of RQC's and the determination of their signs at each inequivalent (2)H site, the elements of the second-rank order tensor, Sαβ, are calculated by assuming a modeled structure. The variations of RQC's and diagonalized order parameters, Sα'α', are followed versus the relative proportion of two polypeptides in the chiral oriented mixture. The influence of the solute mass fraction in the two-homopolypeptide oriented samples is also examined as well as the case of homogeneous and uniform achiral mesophases "PBG-PCBL" made of two pairs of mirror-image homopolypeptides (PBLG/PBDG and PCBLL/PCBDL). In the latter, the solute ordering is modulated by the proportion of each type of homopolypeptide (chemical nature and absolute configuration), leading to eliminate the enantiodiscrimination mechanisms on the average. In the frame of a model, new insights on the solute-homopolypeptide fiber interactions are discussed.

Pesticide exposure threatens many freshwater and estuarine ecosystems around the world. This study examined the temporal and spatial trends of pesticide concentrations in a waterway within an agriculturally developed dry-tropics catchment using a combination of grab and passive sampling methods over a continuous two-year monitoring program. A total of 43 pesticide residues were detected with 7 pesticides exceeding ecologically relevant water quality guidelines/trigger values during the study period and 4 (ametryn, atrazine, diuron, and metolachlor) of these exceeding guidelines for several months. The presence and concentration of the pesticides in the stream coincided with seasonal variability in rainfall, harvest timing/cropping cycle, and management changes. The sampling approach used demonstrates that the application of these complementary sampling techniques (both grab and passive sampling methods) was effective in establishing pesticide usage patterns in upstream locations where application data are unavailable.

Phenotypic cell-based screening is a powerful approach to small-molecule discovery, but a major challenge of this strategy lies in determining the intracellular target and mechanism of action (MoA) for validated hits. Here, we show that the small-molecule BRD0476, a novel suppressor of pancreatic β-cell apoptosis, inhibits interferon-gamma (IFN-γ)-induced Janus kinase 2 (JAK2) and signal transducer and activation of transcription 1 (STAT1) signaling to promote β-cell survival. However, unlike common JAK-STAT pathway inhibitors, BRD0476 inhibits JAK-STAT signaling without suppressing the kinase activity of any JAK. Rather, we identified the deubiquitinase ubiquitin-specific peptidase 9X (USP9X) as an intracellular target, using a quantitative proteomic analysis in rat β cells. RNAi-mediated and CRISPR/Cas9 knockdown mimicked the effects of BRD0476, and reverse chemical genetics using a known inhibitor of USP9X blocked JAK-STAT signaling without suppressing JAK activity. Site-directed mutagenesis of a putative ubiquitination site on JAK2 mitigated BRD0476 activity, suggesting a competition between phosphorylation and ubiquitination to explain small-molecule MoA. These results demonstrate that phenotypic screening, followed by comprehensive MoA efforts, can provide novel mechanistic insights into ostensibly well-understood cell signaling pathways. Furthermore, these results uncover USP9X as a potential target for regulating JAK2 activity in cellular inflammation., Competing Interests: Notes The authors declare no competing financial interest.

A series of thiazoloquin(az)olinones were synthesized and found to have potent inhibitory activity against CD38. Several of these compounds were also shown to have good pharmacokinetic properties and demonstrated the ability to elevate NAD levels in plasma, liver, and muscle tissue. In particular, compound 78c was given to diet induced obese (DIO) C57Bl6 mice, elevating NAD > 5-fold in liver and >1.2-fold in muscle versus control animals at a 2 h time point. The compounds described herein possess the most potent CD38 inhibitory activity of any small molecules described in the literature to date. The inhibitors should allow for a more detailed assessment of how NAD elevation via CD38 inhibition affects physiology in NAD deficient states.

The structural and conformational elucidation of flexible bioactive molecules in solution is currently a crucial goal for the scientific community, but it is rarely achievable by available techniques. The anti-inflammatory drug diflunisal is presented here as a model case for supporting the efficiency of NMR spectroscopy combined with the use of weakly ordering media as a promising methodology for the conformational investigation of small bioactive molecules. Starting from NMR anisotropic data (40 independent dipolar couplings), a quite accurate description of its torsional distribution around the inter-ring C-C bond was found, characterized by a pair of two couples of conformers. According to the relative configuration of the carboxylic group and the fluorine atom in the ortho position to the inter-ring C-C bond, the more stable couple of conformers are defined as "trans" type conformers (F opposite to the carboxylic group) whereas the less stable couple are "cis" type conformers (F and carboxylic group on the same side). In order to study the influence of fluorine nuclei on the structure and conformational distribution, the same analytical strategy has been applied to investigate the phenylsalicylic acid, its nonfluorinated analogue.

Investigation of fluorescence quenching of probes, such as ATTO dyes, is becoming an increasingly important topic owing to the use of these dyes in super-resolution microscopies and in single-molecule studies. Photoinduced electron transfer is their most important nonradiative pathway. Because of the increasing frequency of the use of ATTO and related dyes to investigate biological systems, studies are presented for inter- and intramolecular quenching of ATTO 590 with tryptophan. In order to examine intramolecular quenching, an ATTO 590-tryptophan conjugate was synthesized. It was determined that tryptophan is efficiently quenching ATTO 590 fluorescence by excited-state charge transfer and two charge transfer complexes are forming. In addition, it was discovered that an exciplex (whose lifetime is 5.6 ns) can be formed between tryptophan and ATTO 590, and it is suggested that the possibility of such exciplex formation should be taken into account when protein fluorescence is monitored in a system tagged with ATTO dyes.

Four novel heterometallic RE-Na-organic frameworks, [(RE)Na3(PZTC)2(H2O)4]·2H2O (RE = Yb (1), Ho (2), Er (3), Y (4); PZTC = pyrazine-2,3,5-tricarboxylate), were synthesized via solvothermal reactions and characterized by IR, elemental analysis, and single-crystal X-ray diffraction. The results show that the four complexes are isostructural. In the frameworks, the trinuclear Na cluster and RE ion acting as nodes are bridged by the multifunctional PZTC ligand to give a 3-D framework. Codoping in the frameworks was realized due to their isostructural characteristics. The codoped complexes were calcinated at 800 °C to give rise to the corresponding oxides. Investigation of their photophysical properties shows that the upconversion luminescence (UCL) of the Ho system is green while that of the Er system is red upon excitation at 980 nm. With regard to the luminescence color and intensity, the Er system is preferable to the Ho system for application in bioimaging. Both the red and the green UCL of the Ho(3+) and Er(3+) systems involve a two-photon process. In addition, the UCL mechanism is given. The UCL comparison of Na-doped oxides with non-Na-doped oxides indicates that doping Na can greatly enhance the UCL of the Er system.

Important requirements for exogenous dyes or contrast agents in magnetic resonance imaging (MRI) include an effective concentration of paramagnetic or superparamagnetic ions at the target to be imaged. We report the concise synthesis and characterization of several new enantiopure bifunctional derivatives of (α(1)R,α(4)R,α(7)R,α(10)R)-α(1),α(4),α(7),α(10)-tetramethyl-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTMA) (and their 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) analogues as controls) that can be covalently attached to a contrast agent delivery system using either click or peptide coupling chemistry. Gd complexes of these derivatives can be attached to delivery systems while maintaining optimal water residence time for increased molecular imaging sensitivity. Long chain biotin (LC-biotin) derivatives of the Eu(III) and Gd(III) chelates associated with avidin are used to demonstrate higher efficiencies. Variable-temperature relaxometry, (17)O NMR, and nuclear magnetic resonance dispersion (NMRD) spectroscopy used on the complexes and biotin-avidin adducts measure the influence of water residence time and rotational correlation time on constrained and unconstrained systems. The Gd(III)-DOTMA derivative has a shorter water residence time than the Gd(III)-DOTA derivative. Compared to the constrained Gd(III)-DOTA derivatives, the rotationally constrained Gd(III)-DOTMA derivative has ∼40% higher relaxivity at 37 °C, which could increase its sensitivity as an MRI agent as well as reduce the dose of the targeting agent.

A novel, potent, and orally bioavailable inhibitor of hepatitis C RNA replication targeting NS4B, compound 4t (PTC725), has been identified through chemical optimization of the 6-(indol-2-yl)pyridine-3-sulfonamide 2 to improve DMPK and safety properties. The focus of the SAR investigations has been to identify the optimal combination of substituents at the indole N-1, C-5, and C-6 positions and the sulfonamide group to limit the potential for in vivo oxidative metabolism and to achieve an acceptable pharmacokinetic profile. Compound 4t has excellent potency against the HCV 1b replicon, with an EC50 = 2 nM and a selectivity index of >5000 with respect to cellular GAPDH. Compound 4t has an overall favorable pharmacokinetic profile with oral bioavailability values of 62%, 78%, and 18% in rats, dogs, and monkeys, respectively, as well as favorable tissue distribution properties with a liver to plasma exposure ratio of 25 in rats.

Apple (Malus domestica Borkh.) fruit volatile production is regulated by a variety of factors including low oxygen storage conditions. This study examined the impact of low pO2 controlled atmospheres on 'Scarlett Spur Red Delicious' apple volatile production and disorder development. Accumulation of apple volatile compounds was characterized during long-term cold storage at 0.5 °C in air or low pO2 (0.3, 0.8, or 1.5 kPa) with 1 kPa CO2. Volatile accumulation differed quantitatively with pO2 as acetaldehyde, ethanol, and ethyl ester accumulation increased with decreased pO2 during the first weeks in storage. Differences in volatile accumulation among atmospheres were evident through 6 months. The rate of ethanol accumulation increased with decreased pO2 and could potentially be used to monitor low O2 stress. Incidence of low oxygen disorders after 9 months was highest in fruit held at the lowest pO2. The sesquiterpene α-farnesene was not detected throughout the storage period.

Toxicity of commercial nanoparticles of titania, silica, and zinc oxides is being investigated in this in vitro study. Particles of these compositions are found in many food items, and thus this study is directed toward particle behavior in simulated digestion media and their interaction with intestinal epithelial cell line C2BBe1, a clone of Caco-2 cells, originally isolated from a human colon cancer. Even though the primary particle size of all three particles was below 50 nm, the particles appeared as aggregates in culture media with a negatively charged surface. In the presence of pepsin (pH 2), the charge on the titania became positive, and silica was almost neutral and aggregated extensively, whereas ZnO dissolved. For silica and titania, treatment with simulated intestinal digestive solution led to a strongly negatively charged surface and particle sizes approaching values similar to those in media. On the basis of infrared spectroscopy, we concluded that the surface of silica and titania was covered with bile salts/proteins after this treatment. Transmission electron microscopy indicated that the C2BBe1 cells internalized all three particles. Toxicity assays included investigation of necrosis, apoptosis, membrane damage, and mitochondrial activity. Titania and SiO₂ particles suspended in media at loading levels of 10 μg/cm² exhibited no toxicity. With ZnO at the same loading level, mild toxicity was observed based only on the LDH assay and decrease of mitochondrial activity and not necrosis or apoptosis. Titania particles exposed to the simulated digestion media exhibited mild toxicity based on decrease of mitochondrial activity, likely due to transport of toxic bile salts via adsorption on the particle surface.

Strategies for carrying out the reaction of 4,6-dichloropyrimidine-5-carboxaldehyde with various hydrazines to generate 1-substituted 4-chloropyrazolo[3,4-d]pyrimidines in a selective and high-yielding manner are presented. For aromatic hydrazines, the reaction is performed in the absence of an external base, which promotes exclusive hydrazone formation. The hydrazones subsequently cyclize at an elevated temperature to form the desired pyrazolo[3,4-d]pyrimidine products. For aliphatic hydrazines, the reaction sequence proceeds as a single step in the presence of an external base.

The conformation and interconversion dynamics of two derivatives of the 18-membered hexathia metacyclophane 1 and 2 were studied by (1)H NMR spectroscopy in isotropic solvents and by (2)H NMR in chiral liquid crystalline (CLC) solutions, as well as by molecular structure computations. For the analysis of the dynamic effects, we made use of the concepts of "average symmetry" and "isodynamic groups", introduced by Altmann (Altmann, Proc. R. Soc.1967, 184, A298). Compound 1, which is unsubstituted in the inner aromatic site, has, according to the NMR and molecular force field calculations, a boat shaped ground conformation with C(2) symmetry. It is highly flexible and in the NMR spectrum exhibits two successive dynamic processes. There is a low temperature (170-210 K, E(a) = 10.5 kcal/mol) alternate "wing flipping", which corresponds to interchange between pairs of enantiomers and results, in the fast exchange limit, in an average prochiral molecule with C(2v) symmetry. This process is followed, at higher temperatures (290-320 K, E(a) = 28.5 kcal/mol), by an umbrella flipping type inversion with an average structure of D(2h) symmetry. This second process involves averaging of effective enantiotopic into homotopic sites and can only be studied in chiral solvents. The origin of the chiral discrimination and of their stepwise averaging is discussed. Compound 2, which is substituted with methoxy groups at the inner sites of the benzene rings, is much less flexible and exhibits dynamic effects in the NMR spectrum only at temperatures above 370 K. We were able to study the kinetic parameters of this process in isotropic solvents (E(a) = 21.4 kcal/mol). As for 1, the detailed mechanism of this process can in principle be established using dynamic NMR in CLC; however, experimental limitation precluded us from doing so. Possible alternatives and their effect on the 1D and 2D exchange spectra in CLC are discussed in a concluding section.

SET and MYND domain-containing protein 2 (SMYD2) is a protein lysine methyltransferase that catalyzes the transfer of methyl groups from S-adenosylmethionine (AdoMet) to acceptor lysine residues on histones and other proteins. To understand the kinetic mechanism and the function of individual domains, human SMYD2 was overexpressed, purified, and characterized. Substrate specificity and product analysis studies established SMYD2 as a monomethyltransferase that prefers nonmethylated p53 peptide substrate. Steady-state kinetic and product inhibition studies showed that SMYD2 operates via a rapid equilibrium random Bi Bi mechanism at a rate of 0.048 ± 0.001 s(-1), with K(M)s for AdoMet and the p53 peptide of 0.031 ± 0.01 μM and 0.68 ± 0.22 μM, respectively. Metal analyses revealed that SMYD2 contains three tightly bound zinc ions that are important for maintaining the structural integrity and catalytic activity of SMYD2. Catalytic activity was also shown to be dependent on the GxG motif in the S-sequence of the split SET domain, as a G18A/G20A double mutant and a sequence deletion within the conserved motif impaired AdoMet binding and significantly decreased enzymatic activity. The functional importance of other SMYD2 domains including the MYND domain, the cysteine-rich post-SET domain, and the C-terminal domain (CTD), were also investigated. Taken together, these results demonstrated the functional importance of distinct domains in the SMYD family of proteins and further advanced our understanding of the catalytic mechanism of this family., (© 2011 American Chemical Society)

The conformational dynamics and orientational behavior of two model cyclic molecules, cis-decalin (cis-dec) and tetrahydrofurane (THF), dissolved in weakly ordering, polypeptidic chiral liquid crystals (CLCs) are theoretically discussed and experimentally investigated using deuterium and carbon-13 NMR spectroscopies. The analysis of enantiomeric and enantiotopic discriminations in these compounds is shown to depend on the rate of conformational exchange regime, slow or fast. The slow exchange regime is illustrated through the case of cis-dec at low temperature (243 K). We show that the deuterium NMR spectra in this regime can be qualitatively and quantitatively interpreted by restricting the conformational pathway of cis-dec to two enantiomeric conformers of C(2)-symmetry. The orientational order parameters of these interconverting enantiomers are calculated by matching the (2)H quadrupolar splittings with calculated conformer structures. The fast exchange regime is investigated through the examples of cis-dec at high temperature (356 K) and THF at room temperature (300 K). The (2)H NMR spectra above the coalescence temperature are analyzed by introducing the concept of "average molecular structure". This fictitious structure allows easily identifying NMR equivalences of solutes dissolved in CLC. However, it cannot be applied to determine consistent orientational order parameters. This study emphasizes that enantiotopic discriminations observed for flexible molecules in the fast exchange regime can be quantitatively interpreted only by considering the orientational order of each conformer.

The longitudinal relaxation time of ortho-hydrogen (the spin isomer directly observable by NMR) has been measured in various organic solvents as a function of temperature. Experimental data are perfectly interpreted by postulating two mechanisms, namely intramolecular dipolar interaction and spin-rotation, with activation energies specific to these two mechanisms and to the solvent in which hydrogen is dissolved. This permits a clear separation of the two contributions at any temperature. Contrary to the self-diffusion coefficients at a given temperature, the rotational correlation times extracted from the dipolar relaxation contribution do not exhibit any definite trend with respect to solvent viscosity. Likewise, the spin-rotation correlation time obeys Hubbard's relation only in the case of hydrogen dissolved in acetone-d6, yielding in that case a spin-rotation constant in agreement with literature data. Concerning para-hydrogen, which is NMR-silent, the only feasible approach is to dissolve para-enriched hydrogen in these solvents and to follow the back-conversion of the para-isomer into the ortho-isomer. Experimentally, this conversion has been observed to be exponential, with a time constant assumed to be the relaxation time of the singlet state (the spin state of the para-isomer). A theory, based on intermolecular dipolar interactions, has been worked out for explaining the very large values of these relaxation times which appear to be solvent-dependent.

[reaction: see text] 1-Benzyl-3,4-unsaturated-4-piperidinyl benzyldimethylsilane has been prepared and observed to readily undergo palladium-catalyzed cross-coupling reactions with a variety of aryl iodides and aryl bromides to generate 3,4-unsaturated 4-arylpiperidines, often at ambient temperature.

This study deals with a spin system constituted of three nonequivalent protons, two of them originating from para-hydrogen (p-H(2)) after a hydrogenation reaction carried out in the earth magnetic field. It is shown that three singlet states are created provided indirect (J) couplings exist between the three spins, implying hyperpolarization transfer toward the third spin. Upon insertion of the sample in the NMR (Nuclear Magnetic Resonance) high field magnet, the following events occur: (i) the longitudinal two-spin orders which are parts of the singlet states survive; (ii) the other two terms (of these singlet states) tend to be destroyed by magnetic field gradients but at the same time are partly converted into differences of longitudinal polarizations. Nuclear spin relaxation is studied by appropriate NMR measurements when evolution takes place in the high field magnet or in the earth field. In the former case, relaxation is classical although complicated by numerous relaxation rates associated with both longitudinal two-spin orders and longitudinal polarizations. In the latter case, an equilibration between the singlet states first occur, their disappearance being thereafter driven by relaxation rates which remain very small because of the absence of any dipolar contribution. Thus, even in the case of a three-spin system, long-lived states exist; this unexpected property could be very useful for many applications.

The scope of the triphenylsilyl perrhennate (O3ReOSiPh3, 1) catalyzed 1,3-isomerization of allylic alcohols has been thoroughly explored. It was found to be effective for a wide variety of secondary and tertiary allylic alcohol substrates bearing aryl, alkyl, and cyano substituents. Two general reaction types were found which gave high levels of product selectivity: those driven by formation of an extended conjugated system and those driven by selective silylation of a particular isomer. The efficiency of chirality transfer with various substrates was investigated, and conditions were found in which secondary and tertiary allylic alcohols could be formed with high levels of enantioselectivity. Consideration of selectivity trends with respect to the nature of the substituents around the allylic system revealed that this is a reliable and predictable method for allylic alcohol synthesis.

The excited-state properties of the layered La[Ag(CN)(2)](3) and La[Au(CN)(2)](3) systems have been examined and compared with mixed-metal systems of varying metal ratios such as La[Ag(0.78)Au(0.22)(CN)(2)](3), La[Ag(0.55)Au(0.45)(CN)(2)](3), La[Ag(0.33)Au(0.67)(CN)(2)](3), and La[Ag(0.19)Au(0.81)(CN)(2)](3). We have found that these mixed-metal systems luminesce quite strongly at room temperature at an energy that is tunable and depends on the Au:Ag stoichiometric ratio. The emission energy of the mixed-metal samples lies between those of the pure Au and Ag systems. This provides evidence that the excited states responsible for this emission are delocalized over the Ag and Au centers. The strong luminescence of the mixed-metal systems at ambient temperatures is in stark contrast to the weak luminescence behavior of pure La[Au(CN)(2)](3) and La[Ag(CN)(2)](3) samples, which makes the mixed-metal systems more viable than the pure systems for practical applications.

Two reaction strategies are developed to promote the highly selective 1,3-isomerization of a variety of allylic alcohols using O3ReOSiPh3 as a catalyst. The first strategy utilizes substrates whose 1,3-regioisomer contains a conjugated alkene, which relies on thermodynamics to obtain high selectivity. The second strategy employs N,O-bis(trimethylsilyl)acetamide as an additive to selectively and irreversibly remove the product from the reaction equilibrium and works well for the isomerization of tertiary allylic alcohols into primary allylic alcohols containing trisubstituted alkene components. High stereoselectivity is also observed in the 1,3-isomerization of enantioenriched allylic alcohols.

The deuterium/hydrogen (D/H)(i) ratio measurement by quantitative (2)H NMR spectroscopy is a method of choice for the analysis of kinetic isotopic effects associated with enzyme-catalyzed reactions during a biosynthetic pathway. However, the efficiency of the current isotropic (2)H-[(1)H] NMR can be limited by the rather small chemical shift dispersion of deuterium nuclei. In addition, this method does not allow the enantiotopic deuterons in prochiral molecules to be spectrally discriminated, hence precluding the quantification of isotopic fractionation on methylene prostereogenic sites. In this work, we explore another analytical strategy able to circumvent these disadvantages. This approach is based on the use of natural abundance (2)H 2D NMR experiments on solutes embedded in polypeptidic, chiral liquid crystalline solvent. Thus, we show that NMR in these oriented phases is a powerful way to separate deuterium signals on the basis of the quadrupolar interactions, providing a promising alternative to overcrowded (2)H NMR spectra obtained in liquid state. To illustrate our purpose, we have experimentally investigated the case of 1,1'-bis(phenylthio)hexane derived by cleavage from methyl linoleate of safflower. The (2)H NMR results in chiral liquid crystals are presented and discussed. We show, for the first time, that (D/H)(pro-R) and (D/H)(pro-S) can be measured at the same methylene position of a fatty acid chain.

Functionalized vinyl pinacol boronates suitable for Suzuki cross-coupling reactions are synthesized using ruthenium-catalyzed olefin cross-metathesis of 1-propenyl pinacol boronate and various alkenes, including functionalized and 1,1-disubstituted alkenes. The resultant boronate cross products are stereoselectively transformed into predominantly Z-vinyl bromides and E-vinyl iodides. The vinyl bromides may be synthesized in a two-step, one-pot synthesis from a variety of olefins, resulting in a Z-selective formal vinyl bromide cross-metathesis reaction.

[reaction: see text] Natural abundance deuterium 2D NMR spectroscopy in chiral liquid crystal was successfully used to efficiently analyze the enantiomeric composition of organic chiral building blocks involved in the syntheses of natural and synthetic bioactive products. The results reported here emphasize the high potential of this analytical strategy and prove its applicability for routinely determining enantiomeric excesses.

The antiaromaticity of a series of dications of p-substituted diphenylmethylidene fluorenes was explored using three criteria attributed to aromaticity/antiaromaticity. The relative stability of the dications (energetic criterion) was measured via the redox potentials obtained by electrochemical oxidation under very fast sweep rates with microelectrodes. Comparison of redox potentials with those of a model system, p-substituted tetraphenylethylenes, shows relatively small destabilization of the potentially antiaromatic fluorenylidene dication. However, the amount of destabilization is comparable with the limited electrochemical data available for other antiaromatic systems. Nucleus independent chemical shifts (NICS) were calculated for these dications (magnetic criterion) and indicated their antiaromaticity. A good linear relationship between experimental and calculated (B3LYP/6-31G(d)) (1)H and (13)C NMR shifts for the three dications, 3c, 3e, and 3f, for which NMR data has been reported, validated the accuracy of the NICS values. Bond length alternation/elongation (structural criterion) was explored via the harmonic oscillator model of aromaticity (HOMA) using the geometries calculated with density functional theory, but there was insufficient variation to evaluate relative antiaromaticity. In addition, the presence of benzannulation appears to restrict bond length alternation to such an extent that the magnitude of the HOMA index is of little use in evaluating the antiaromaticity of many polycyclic hydrocarbons. Both NICS values and redox potentials for formation of the dication in these systems show a strong linear correlation with sigma(p)(+) values, with the more antiaromatic fluorenylidene dication possessing the more electron-withdrawing substituent. The correlation between NICS values and redox potentials is also good, as might be expected, suggesting a strong relationship between magnetic and energetic characteristics of antiaromaticity. However, magnetic characteristics appear to be a more sensitive probe than energetic characteristics evaluated through redox potentials or structural characteristics evaluated through HOMA calculations.

[reaction: see text] The selective ring opening cross metathesis of 1,5-cyclooctadiene and trisubstituted cycloolefins with acroyl species is described. The ring-opened products contain electronically differentiated olefins suitable for additional metathesis reactions. Trisubstituted cycloolefins open regioselectively, placing the acroyl cap on the less-substituted terminus.