Pushing the Detection Limit of Static Wideline NMR Spectroscopy Using Ultrafast Frequency-Swept Pulses.

We report a simple design strategy for wideband uniform-rate smooth truncation (WURST) pulses that enables ultrafast frequency sweeps to maximize the sensitivity of Carr-Purcell-Meiboom-Gill (CPMG) acquisition in static wideline nuclear magnetic resonance (NMR). Three compelling examples showcase the advantage of ultrafast frequency sweeps over currently employed WURST-CPMG protocols, demonstrating the potential of investigating materials that are typically inaccessible to static wideline NMR techniques, e.g., paramagnetic solids with short homogeneous transverse relaxation times.

Structure Determination and Refinement of Paramagnetic Materials by Solid-State NMR.

Paramagnetism in solid-state materials has long been considered an additional challenge for structural investigations by using solid-state nuclear magnetic resonance spectroscopy (ssNMR). The strong interactions between unpaired electrons and the surrounding atomic nuclei, on the one hand, are complex to describe, and on the other hand can cause fast decaying signals and extremely broad resonances. However, significant progress has been made over the recent years in developing both theoretical models to understand and calculate the frequency shifts due to paramagnetism and also more sophisticated experimental protocols for obtaining high-resolution ssNMR spectra. While the field is continuously moving forward, to date, the combination of state-of-the-art numerical and experimental techniques enables us to obtain high-quality data for a variety of systems. This involves the determination of several ssNMR parameters that represent different contributions to the frequency shift in paramagnetic solids. These contributions encode structural information on the studied material on various length scales, ranging from crystal morphologies, to the mid- and long-range order, down to the local atomic bonding environment. In this perspective, the different ssNMR parameters characteristic for paramagnetic materials are discussed with a focus on their interpretation in terms of structure. This includes a summary of studies that have explored the information content of these ssNMR parameters, mostly to complement experimental data from other methods, e.g., X-ray diffraction. The presented overview aims to demonstrate how far ssNMR has hitherto been able to determine and refine the structures of materials and to discuss where it currently falls short of its full potential. We attempt to highlight how much further ssNMR can be pushed to determine and refine structure to deliver a comprehensive structural characterization of paramagnetic materials comparable to what is to date achieved by the combined effort of electron microscopy, diffraction, and spectroscopy.

Antibiotic resistance genes in the subgingival microbiome and implications for periodontitis therapy.

BACKGROUND: Antibiotic resistance is emerging as a global public threat. However, it remains poorly investigated in the context of periodontal therapy. The aim of the study was to investigate the complete diversity of antibiotic resistance genes in a German population. METHODS: Thirty-nine volunteers with periodontitis contributed to the present study with one to four periodontal pockets for a total of 124 subgingival samples. Samples were analyzed using shotgun metagenomics. RESULTS: A total of 19 antibiotic resistance genes from six antibiotic classes were detected in subgingival biofilm. Two thirds of the volunteers (n = 26/39) showed antibiotic resistance genes for at least one of the antibiotic classes used for periodontal treatment in dental practice or research: beta-lactam, lincosamide, macrolide, nitroimidazole, and tetracycline. Macrolide was the most abundant class detected (21/39 patients). CONCLUSIONS: Findings from our study suggest a high prevalence of antibiotic resistance genes in periodontal pockets from German volunteers. We recommend the development and broader use of molecular diagnostic tests for antibiotic resistance in dental practice to ensure treatment success and to minimize antibiotic resistance.

Solid-State Nuclear Magnetic Resonance Techniques for the Structural Characterization of Geminal Alane-Phosphane Frustrated Lewis Pairs and Secondary Adducts.

The first comprehensive solid-state nuclear magnetic resonance (NMR) characterization of geminal alane-phosphane frustrated Lewis pairs (Al/P FLPs) is reported. Their relevant NMR parameters (isotropic chemical shifts, direct and indirect 27 Al-31 P spin-spin coupling constants, and 27 Al nuclear electric quadrupole coupling tensor components) have been determined by numerical analysis of the experimental NMR line shapes and compared with values computed from the known crystal structures by using density functional theory (DFT) methods. Our work demonstrates that the 31 P NMR chemical shifts for the studied Al/P FLPs are very sensitive to slight structural inequivalences. The 27 Al NMR central transition signals are spread out over a broad frequency range (>200 kHz), owing to the presence of strong nuclear electric quadrupolar interactions that can be well-reproduced by the static 27 Al wideband uniform rate smooth truncation (WURST) Carr-Purcell-Meiboom-Gill (WCPMG) NMR experiment. 27 Al chemical shifts and quadrupole tensor components offer a facile and clear distinction between three- and four-coordinate aluminum environments. For measuring internuclear Al⋅⋅⋅P distances a new resonance-echo saturation-pulse double-resonance (RESPDOR) experiment was developed by using efficient saturation via frequency-swept WURST pulses. The successful implementation of this widely applicable technique indicates that internuclear Al⋅⋅⋅P distances in these compounds can be measured within a precision of ±0.1 Å.

Frequency-Swept Ultra-Wideline Magic-Angle Spinning NMR Spectroscopy.

Recent years have witnessed the development of solid-state NMR techniques that allow the direct investigation of extremely wide inhomogeneously broadened resonance lines. To date, this typically involves the application of frequency sweeps as offered by wideband uniform rate smooth truncation (WURST) pulses. While the effects of such advanced irradiation schemes on static samples are well understood, the interference between the varying carrier frequency and the time-dependent evolution of the spin system under magic-angle spinning (MAS) conditions is more complex. Herein, we introduce the well-known WURST-Carr-Purcell-Meiboom-Gill (WCPMG) pulse sequence for spinning samples. Using numerical spin-density matrix analysis, an ideal design based on fast frequency sweeps and high truncation of the incorporated WURST pulses is presented that enables uniform excitation/refocusing under MAS conditions with low-to-moderate radio-frequency power requirements. This permits the acquisition of ultra-wideline MAS NMR lines exceeding 500 kHz with chemical shift resolution in a single transmitter step.

Sensitivity-enhanced multiple-quantum MAS NMR for half-integer spin quadrupolar nuclei using WURST-amplitude shaped pulses.

Two-dimensional multiple-quantum MAS (MQMAS) NMR spectroscopy is one of the most widely used solid-state NMR techniques for resolving multiple overlapping central-transition lineshapes for half-integer spin quadrupolar nuclei. In particular when relying on nutation-driven MQ coherence transfers, this technique suffers from low sensitivity that can only be improved by increasing the rf-amplitude of the involved radio-frequency (rf) pulses, which are therefore typically operated at the rf-limit. In such situations, frequently encountered for the three-pulse z-filtered and split-t1 shifted-echo MQMAS NMR sequences, we introduce the advantages of rf-pulses with smoothly truncated amplitude profiles, which we have termed WURST-Amplitude Shaped Pulses (WASPs). When considering the NMR spectrometer hardware, we demonstrate that WASPs feature more suitable properties in comparison to conventional rectangular pulses, enabling a substantial reduction of voltage reflections and transient effects under identical rf-conditions. By employing extensive numerical simulations and experimental validation, we further show that WASPs intrinsically possess a higher potential for nutation-based 3Q excitation involving spin-3/2 and 3Q and 5Q excitation for spin-5/2 quadrupolar nuclei, specifically when large nutation frequencies are available. The concept of smoothly truncating rf-amplitudes is also extended to Fast Amplitude Modulation (FAM) pulses, normally incorporated for rotor-driven 1Q conversion. We additionally evaluate the potential of employing WASPs with peak rf-amplitudes beyond the rf-limit for conventional rectangular rf-pulses.

Minimizing Lineshape Distortions in Static Ultra-wideline Nuclear Magnetic Resonance of Half-Integer Spin Quadrupolar Nuclei.

Chirped excitation using frequency-swept wideline uniform rate smooth truncation (WURST) pulses in combination with Carr-Purcell-Meiboom-Gill acquisition (WCPMG) is currently the state-of-the-art method for the direct observation of the central transition (CT) in static ultra-wideline nuclear magnetic resonance (NMR) of half-integer spin quadrupolar nuclei. However, CT lineshape distortions and an inefficient, large number of transmitter steps in frequency-stepped acquisition are two major drawbacks. Here, we identify three main sources for lineshape distortions occurring in WCPMG NMR spectra of the CT: (I) distortions due to inaccurate setting of the radio frequency field strength, (II) chirped-excitation artifacts, and (III) distortions due to non-selective irradiation. A new and efficient approach for the acquisition minimizing these distortions is presented using low sweep rates (R ≤ 5 kHz/µs) and sweep widths (Δ ≤ 600 kHz). We further demonstrate that such an acquisition strategy also minimizes the number of transmitter steps in ultra-wideline NMR. This is achieved from numerical simulations and theoretical analysis of the orientational dependence for the quadrupolar-perturbed Zeeman states and their transition frequencies. The theoretically derived strategies are validated experimentally, allowing us to set up guidelines for the optimum recording of wideline and ultra-wideline WCPMG NMR spectra.