Rapid Quantification of Pharmaceuticals via 1H Solid-State NMR Spectroscopy.

The physicochemical properties of active pharmaceutical ingredients (APIs) can depend on their solid-state forms. Therefore, characterization of API forms is crucial for upholding the performance of pharmaceutical products. Solid-state nuclear magnetic resonance (SSNMR) spectroscopy is a powerful technique for API quantification due to its selectivity. However, quantitative SSNMR experiments can be time consuming, sometimes requiring days to perform. Sensitivity can be considerably improved using 1H SSNMR spectroscopy. Nonetheless, quantification via 1H can be a challenging task due to low spectral resolution. Here, we offer a novel 1H SSNMR method for rapid API quantification, termed CRAMPS-MAR. The technique is based on combined rotation and multiple-pulse spectroscopy (CRAMPS) and mixture analysis using references (MAR). CRAMPS-MAR can provide high 1H spectral resolution with standard equipment, and data analysis can be accomplished with ease, even for structurally complex APIs. Using several API species as model systems, we show that CRAMPS-MAR can provide a lower quantitation limit than standard approaches such as fast MAS with peak integration. Furthermore, CRAMPS-MAR was found to be robust for cases that are inapproachable by conventional ultra-fast (i.e., 100 kHz) MAS methods even when state-of-the-art SSNMR equipment was employed. Our results demonstrate CRAMPS-MAR as an alternative quantification technique that can generate new opportunities for analytical research.

Hydrocarboxyl Radical as a Product of α-Alanine Ultraviolet Photolysis.

UV photodissociation of α-alanine was studied by parahydrogen matrix isolation infrared spectroscopy. The temporal behavior of Fourier transform infrared spectra revealed that UV irradiation at 213 nm yielded the HOCO radical as a direct photoproduct from the S2 excited state. The concentration of HOCO quickly approached a steady state due to secondary photodissociation of HOCO to produce CO2 + H or CO + OH. On the other hand, no photoproducts were detected by S1 excitation at 266 nm. Irradiation of fully deuterated α-alanine at 213 nm yielded ∼2 times more cis-DOCO radicals than the lower energy isomer trans-DOCO, indicating that the conformation of the hydroxyl group is fairly well-preserved upon photodissociation of α-alanine. The present study suggests that HOCO may be a good tracer species in the search for amino acids in interstellar space.

π-Complexes of Diborynes with Main Group Atoms.

We present herein an in-depth study of complexes in which a molecule containing a boron-boron triple bond is bound to tellurate cations. The analysis allows the description of these salts as true π complexes between the B-B triple bond and the tellurium center. These complexes thus extend the well-known Dewar-Chatt-Duncanson model of bonding to compounds made up solely of p block elements. Structural, spectroscopic and computational evidence is offered to argue that a set of recently reported heterocycles consisting of phenyltellurium cations complexed to diborynes bear all the hallmarks of π-complexes in the π-complex/metallacycle continuum envisioned by Joseph Chatt. Described as such, these compounds are unique in representing the extreme of a metal-free continuum with conventional unsaturated three-membered rings (cyclopropenes, azirenes, borirenes) occupying the opposite end.

Exploring Host-Guest Interactions in the α-Zn3(HCOO)6 Metal-Organic Framework.

Metal-organic frameworks (MOFs) are promising gas adsorbents. Knowledge of the behavior of gas molecules adsorbed inside MOFs is crucial for advancing MOFs as gas capture materials. However, their behavior is not always well understood. In this work, carbon dioxide (CO2) adsorption in the microporous α-Zn3(HCOO)6 MOF was investigated. The behavior of the CO2 molecules inside the MOF was comprehensively studied by a combination of single-crystal X-ray diffraction (SCXRD) and multinuclear solid-state magnetic resonance spectroscopy. The locations of CO2 molecules adsorbed inside the channels of the framework were accurately determined using SCXRD, and the framework hydrogens from the formate linkers were found to act as adsorption sites. 67Zn solid-state NMR (SSNMR) results suggest that CO2 adsorption does not significantly affect the metal center environment. Variable-temperature 13C SSNMR experiments were performed to quantitatively examine guest dynamics. The results indicate that CO2 molecules adsorbed inside the MOF channel undergo two types of anisotropic motions: a localized rotation (or wobbling) upon the adsorption site and a twofold hopping between adjacent sites located along the MOF channel. Interestingly, 13C SSNMR spectroscopy targeting adsorbed CO2 reveals negative thermal expansion (NTE) of the framework as the temperature rose past ca. 293 K. A comparative study shows that carbon monoxide (CO) adsorption does not induce framework shrinkage at high temperatures, suggesting that the NTE effect is guest-specific.

Cleaving Carboxyls: Understanding Thermally Triggered Hierarchical Pores in the Metal-Organic Framework MIL-121.

Carboxylic acid linker ligands are known to form strong metal-carboxylate bonds to afford many different variations of permanently microporous metal-organic frameworks (MOFs). A controlled approach to decarboxylation of the ligands in carboxylate-based MOFs could result in structural modifications, offering scope to improve existing properties or to unlock entirely new properties. In this work, we demonstrate that the microporous MOF MIL-121 is transformed to a hierarchically porous MOF via thermally triggered decarboxylation of its linker. Decarboxylation and the introduction of hierarchical porosity increases the surface area of this material from 13 to 908 m2/g and enhances gas adsorption uptake for industrially relevant gases (i.e., CO2, C2H2, C2H4, and CH4). For example, CO2 uptake in hierarchically porous MIL-121 is improved 8.5 times over MIL-121, reaching 215.7 cm3/g at 195 K and 1 bar; CH4 uptake is 132.3 cm3/g at 298 K and 80 bar in hierarchically porous MIL-121 versus zero in unmodified MIL-121. The approach taken was validated using a related aluminum-based MOF, ISOMIL-53. However, many specifics of the decarboxylation procedure in MOFs have yet to be unraveled and demand prompt examination. Decarboxylation, the formation of heterogeneous hierarchical pores, gas uptakes, and host-guest interactions are comprehensively investigated using variable-temperature multinuclear solid-state NMR spectroscopy, X-ray diffraction, electron microscopy, and gas adsorption; we propose a mechanism for how decarboxylation proceeds and which local structural features are involved. Understanding the complex relationship among the molecular-level MOF structure, thermal stability, and the decarboxylation process is essential to fine-tune MOF porosity, thus offering a systematic approach to the design of hierarchically porous, custom-built MOFs suited for targeted applications.

Solid-State NMR Spectroscopy: A Powerful Technique to Directly Study Small Gas Molecules Adsorbed in Metal-Organic Frameworks.

Metal-organic frameworks (MOFs) have shown great potential in gas separation and storage, and the design of MOFs for these purposes is an on-going field of research. Solid-state nuclear magnetic resonance (SSNMR) spectroscopy is a valuable technique for characterizing these functional materials. It can provide a wide range of structural and motional insights that are complementary to and/or difficult to access with alternative methods. In this Concept article, the recent advances made in SSNMR investigations of small gas molecules (i.e., carbon dioxide, carbon monoxide, hydrogen gas and light hydrocarbons) adsorbed in MOFs are discussed. These studies demonstrate the breadth of information that can be obtained by SSNMR spectroscopy, such as the number and location of guest adsorption sites, host-guest binding strengths and guest mobility. The knowledge acquired from these experiments yields a powerful tool for progress in MOF development.

CO Guest Interactions in SDB-Based Metal-Organic Frameworks: A Solid-State Nuclear Magnetic Resonance Investigation.

Metal-organic frameworks (MOFs) are promising materials for greener carbon monoxide (CO) capture and separation processes. SDB-based (SDB = 4,4'-sulfonyldibenzoate) MOFs are particularly attractive due to their remarkable gas adsorption capacity under humid conditions. However, to the best of our knowledge, their CO adsorption abilities have yet to be investigated. In this report, CO-loaded PbSDB and CdSDB were characterized using variable-temperature (VT) 13C solid-state nuclear magnetic resonance (SSNMR) spectroscopy. These MOFs readily captured CO, with the adsorbed CO exhibiting dynamics as indicated by the temperature-dependent changes in the SSNMR spectra. Spectral simulations revealed that the CO simultaneously undergoes a localized wobbling about the adsorption site and a nonlocalized hopping between adjacent adsorption sites. The wobbling and hopping angles were also found to be temperature-dependent. From the appearance of the VT spectra and the extracted motional data, the CO adsorption mechanism was concluded to be analogous to that of CO2. To gain a better understanding on the gas adsorption properties of these MOFs and their CO capture abilities, we subsequently compared the motional data to those reported for CO2 in SDB-based MOFs and CO in MOF-74, respectively. A significant contrast in adsorption strength was observed in both cases because of the different physical properties of the guests (i.e., CO vs CO2) and the MOF frameworks (i.e., SDB-based MOFs vs MOFs with open metal sites). Our results demonstrate that SSNMR spectroscopy can be employed to probe variations in binding behavior.

Cortical Brain Stimulation with Endovascular Electrodes.

Electrical stimulation of neural tissue and recording of neural activity are the bases of emerging prostheses and treatments for spinal cord injury, stroke, sensory deficits, and drug-resistant neurological disorders. Safety and efficacy are key aspects for the clinical acceptance of therapeutic neural stimulators. The cortical vasculature has been shown to be a safe site for implantation of electrodes for chronically recording neural activity, requiring no craniotomy to access high-bandwidth, intracranial EEG. This work presents the first characterization of endovascular cortical stimulation measured using cortical subdural surface recordings. Visual stimulation was used to verify electrode viability and cortical activation was compared with electrically evoked activity. Due to direct activation of the neural tissue, the latency of responses to electrical stimulation was shorter than for that of visual stimulation. We also found that the center of neural activation was different for visual and electrical stimulation indicating an ability of the stentrode to provide localized activation of neural tissue.

Dynamic Disorder and Electronic Structures of Electron-Precise Dianionic Diboranes: Insights from Solid-State Multinuclear Magnetic Resonance Spectroscopy.

The J(11B,11B) coupling constants of various salts of the electron-precise hexacyanodiborane(6) dianion, [B2(CN)6]2-, were obtained using 11B double-quantum-filtered (DQF) J-resolved solid-state nuclear magnetic resonance (SSNMR) spectroscopy. Our results show that the magnitude of the DQF J splitting is influenced by both the crystallographic symmetry of the system and the presence of dynamics. The splittings are amplified by a factor of 3 as compared to the corresponding theoretical J coupling constants for cases where (1) there is an absence of dynamics but the boron pairs are crystallographically equivalent or (2) the boron pairs are crystallographically inequivalent but are rendered magnetically equivalent on the time scale of the experiment due to dynamic disorder, which was identified by 11B and 13C SSNMR experiments. Consequently, molecular motions need to be taken into consideration when interpreting the results of DQF J-resolved experiments, and conversely, these experiments may be used to identify dynamic disorder. Variable-temperature NMR data support the notion of three different motional processes with correlation times ranging from 102 to 106 s-1 over the temperature range of 248-306 K. When molecular motion and crystallographic symmetry are both accounted for, the J(11B,11B) coupling constants for various [B2(CN)6]2- salts were measured to range from 29.4 to 35.8 Hz, and their electronic origins were determined using natural localized molecular orbital and natural bond orbital analyses. The coupling constants were found to strongly correlate to the hybridization states of the boron orbitals that form the B-B bonds and to the strength of the B-B bonds. This study provides a novel tool to study dynamics in ordered and disordered solids and provides new perspectives on electron-precise dianionic diboranes featuring two-center-two-electron bonds in the context of related compounds featuring multiply and singly bonded boron spin pairs.

Effect of acid blue 80, an anthracenedione dye, on rainbow trout liver, gill and gut cells in vitro.

Acid Blue 80 (AB80) is a dark blue colorant that like other synthetic dyes can get into the environment. Cultures of rainbow trout cell lines were dosed with AB80 either directly, which involved mixing AB80 stock solution into the medium over cells, or indirectly, which involved replacing the medium in cultures with medium that had AB80. A dose-dependent decline in cell viability was found in cultures with or without fetal bovine serum (FBS) after direct dosing. However, for FBS cultures, indirect dosing caused no loss of viability over 24h and in the long term was detrimental to RTgill-W1 but not RTL-W1 cultures. After 6 days at 50mg/L cytotoxicity was evident and by 9 days RTgill-W1 cell number had declined. Yet AB at 1mg/L elicited no changes over 9 days in any cell line. AB80 appears to have the potential to be toxic at only very high concentrations.

The effect of dichloroacetate on health- and lifespan in C. elegans.

Aging is associated with increased vulnerability to chronic, degenerative diseases and death. Strategies for promoting healthspan without necessarily affecting lifespan or aging rate have gained much interest. The mitochondrial free radical theory of aging suggests that mitochondria and, in particular, age-dependent mitochondrial decline play a central role in aging, making compounds that affect mitochondrial function a possible strategy for the modulation of healthspan and possibly the aging rate. Here we tested such a "metabolic tuning" approach in nematodes using the mitochondrial modulator dichloroacetate (DCA). We explored DCA as a proof-of-principle compound to alter mitochondrial parameters in wild-type animals and tested whether this approach is suitable for reducing reactive oxygen species (ROS) production and for improving organismal health- and lifespan. In parallel, we addressed the potential problem of operator bias by running both unblinded and blinded lifespan studies. We found that DCA treatment (1) increased ATP levels without elevating oxidative protein damage and (2) reduced ROS production in adult C. elegans. DCA treatment also significantly prolonged nematode health- and lifespan, but did not strongly impact mortality doubling time. Operator blinding resulted in considerably smaller lifespan-extending effects of DCA. Our data illustrate the promise of a "metabolic tuning" intervention strategy, emphasize the importance of mitochondria in nematode aging and highlight operator bias as a potential confounder in lifespan studies.

Prospective, randomized, controlled trial comparing lightweight versus heavyweight mesh in chronic pain incidence after TEP repair of bilateral inguinal hernia.

BACKGROUND: This prospective, clinical, randomized, controlled study was performed to define the incidence of chronic pain after total extraperitoneal (TEP) repair with a light-weighted mesh compared with heavyweight mesh in patients with bilateral inguinal hernias. METHODS: Consecutive patients with bilateral inguinal hernias were recruited for TEP inguinal hernia repair under general anesthesia. Heavyweight mesh was randomly assigned to one side of the groin and lightweight mesh to the other. Patients were followed up regularly for up to 1 year by an independent surgeon who was unaware of the mesh assignment. The postoperative pain score by means of a visual analogue scale (VAS) and other data were recorded. RESULTS: Fifty bilateral TEP hernia repairs were performed between September 2007 and February 2009. Six patients (12%) complained of chronic pain 3 months after the operation. A higher average pain score was observed for the side of hernia repaired by heavyweight mesh compared with lightweight mesh, but the difference was not statistically significant. More patients complained about foreign body sensation on the side repaired with heavyweight mesh (24%) compared with the side with lightweight mesh (8%; P < 0.05). There was no recurrence or need for reintervention for either type of mesh. CONCLUSIONS: Lightweight polypropylene mesh may be preferable to heavyweight mesh for TEP inguinal hernia repair because it provides less postoperative foreign body sensation; however, there was no significant difference in the incidence of chronic pain.

A CMOS retinal neurostimulator capable of focussed, simultaneous stimulation.

Restoring vision to the blind by way of medical device technology has been an objective of several research teams for a number of years. It is known that spots of light-phosphenes-can be elicited by way of electrical stimulation of surviving retinal neurons. Beyond this our understanding of prosthetic vision remains rudimentary. We have designed and manufactured an integrated circuit neurostimulator with substantial versatility, able to provide focussed, simultaneous stimulation using current sources and sinks, steering the current to the intended site of stimulation. The ASIC utilizes high-voltage CMOS transistors in key circuits, to manage voltage compliance issues (due to an unknown or changing electrode/tissue interface impedance) given the relatively high stimulation thresholds necessary to elicit physiological excitation of retinal neurons. In addition, a unique multiplexing system comprised of electrodes arranged in a hexagonal mosaic is used, wherein each electrode can be addressed to be a stimulating electrode and all adjacent electrodes serve as the return path. This allows for simultaneous stimulation to be delivered while appropriately managing cross-talk between the stimulating electrodes. Test results indicate highly linear current sources and sinks (differential nonlinearity error of 0.13 least significant bits -2.6 microA), with the ASIC clearly able to provide focussed stimulation using electrodes immersed in a saline solution.

Focal activation of the feline retina via a suprachoroidal electrode array.

This paper presents the results of the first investigations into the use of bipolar electrical stimulation of the retina with a suprachoroidal vision prosthesis, and the effects of different electrode configurations on localization of responses on the primary visual cortex. Cats were implanted with electrodes in the suprachoroidal space, and electrically evoked potentials were recorded on the visual cortex. Responses were elicited to bipolar and monopolar stimuli, with each stimulating electrode coupled with either six-return electrodes, two-return electrodes, or a single-return electrode. The average charge threshold to elicit a response with bipolar stimulation and six-return electrodes was 76.47+/-8.76 nC. Bipolar stimulation using six-return electrodes evoked responses half the magnitude of those elicited with a single or two-return electrodes. Monopolar stimulation evoked a greater magnitude, and area of cortical activation than bipolar stimulation. This study showed that suprachoroidal, bipolar stimulation can elicit localized activity in the primary visual cortex, with the extent of localization and magnitude of response dependent on the electrode configuration.

Effect of maternal age, birth weight and infant sex on total nucleated cell (TNC) count and volume of umbilical cord blood (UCB) collected.

This study evaluates the effect of maternal age, birth weight and infant sex on two main UCB parameters for use and long-term cryopreservation: TNC and volume. Data from 1000 UCB units were collected and analyzed in this study. The results indicate that TNC is correlated to infant birth weight and sex but not maternal age at delivery. Volume is only correlated to birth weight but not maternal age and infant sex.

Incomplete cellular depopulation may explain the high failure rate of bovine ureteric grafts.

BACKGROUND: The aim was to assess the results of a decellularized bovine ureter graft (SynerGraft) for complex venous access. METHODS: Bovine ureter conduits were implanted in patients with a failed fistula or access graft in whom native vessels were unsuitable as conduits. Graft histories were obtained from all patients who had undergone this procedure at one institution. Failed grafts were explanted and subjected to histological examination. A sample of fresh bovine ureter was immunostained for galactose (alpha1 --> 3) galactose (alpha-Gal). RESULTS: Nine patients with a median age of 46 (range 25-70) years underwent complex venous access surgery between August 2004 and November 2006 using a SynerGraft. Graft types included loop superficial femoral artery to stump of long saphenous vein (four patients), loop brachial artery to vein (two), brachial artery to axillary vein (two) and left axillary artery to innominate vein (one). Three grafts developed aneurysmal dilatation and two thrombosed. Histological assessment of the explanted bovine ureters revealed acute and chronic transmural inflammation. Immunostaining of fresh bovine ureter suggested residual cells and the xenoantigen alpha-Gal. CONCLUSION: Graft failure with aneurysmal dilatation and thrombosis in complex arteriovenous conduits using bovine ureter may be due to residual xenoantigens.

Efficacy of supra-choroidal, bipolar, electrical stimulation in a vision prosthesis.

The key to successful, clinical application of therapeutic neurostimulators lies primarily with the safety and efficacy of their electrode-tissue interfaces. The authors posit that for electrical stimulation of the visual system, supra-choroidal electrode placement provides a safe, stable and readily-accessible site for implantation and the provision of electrical stimulation. The present paper explores the efficacy of supra-choroidal electrical stimulation of retinal neurons. Based upon recordings made with surface electrodes placed on the primary visual cortex, areas of activation in the cortex were shown to change when different areas on the supra-choroidal space were stimulated. Finally, the threshold to elicit a response from neurons in the visual cortex, was found to be 77.55 +/- 29.85 nC.

Charge recovery during concurrent stimulation for a vision prosthesis.

Parallel or concurrent stimulation in an epiretinal neuroprosthesis is likely necessary in order to deliver sufficient phosphenes for effective vision. Important issues with concurrent stimulation are the effect of current distribution which introduces current leakage or 'cross talk' between adjacent electrodes and charge recovery which determines balanced charge being delivered/recovered at each electrode from the previous phase. In this paper, we present the effect of concurrent stimulation of two hexagonally arranged platinum electrode arrays on charge recovery. Balanced and imbalanced (unequal) currents were delivered to the hexagonal arrays when they were immersed in physiological saline. Both simulation and experimental results revealed that charge was not recovered at individual electrodes, particularly when imbalanced currents were delivered. However, total charge injected to both hexagonal arrays was recovered.